EA046157B1 - COMPOSITIONS AND METHODS FOR SELECTIVE REGULATION OF GENE EXPRESSION - Google Patents

Abstract

) The present invention provides engineered transcription factors for selective activation of SCN1a and their use in the treatment of diseases and disorders such as Dravet syndrome. Also provided are microRNA binding sites and their use for selective expression in parvalbumin-containing neurons.

Description

Link

This patent application claims priority to U.S. Provisional Patent Application No. 62/854,238, filed May 29, 2019; US Provisional Patent Application No. 62/857727, filed June 5, 2019; and U.S. Provisional Patent Application No. 63/008569, filed April 10, 2020, each of which is incorporated herein by reference in its entirety.

List of sequences

This application contains a Sequence Listing that has been submitted electronically in ASCII format and is incorporated herein by reference in its entirety. The ASCII copy in question, created on May 28, 2020, is called 46482-724_601_SL.txt and is 418483 bytes in size.

BACKGROUND OF THE INVENTION

A wide range of human diseases are associated with abnormal gene expression. In some cases, a genetic mutation in a gene causes its dysregulation, suppression, or lack of expression at all, resulting in haploinsufficiency. In some cases, a genetic mutation in a gene causes it to become activated, resulting in overexpression of the gene. There are many challenges when treating genetic disorders or diseases. One approach is gene therapy, which involves the therapeutic delivery of a nucleic acid into a patient's cells. However, various problems associated with gene therapy remain unresolved, such as unwanted immune response caused by gene therapy, off-target effects, limitations in the cloning ability of gene therapy carriers (such as viruses), maintenance of therapeutic effect over a longer period of time, and etc. The central nervous system (CNS) poses many unique challenges to the development of therapies that address the underlying disorder of gene and/or protein expression. Although there are medications that help manage the symptoms of CNS diseases/disorders, many CNS diseases/disorders, such as Dravet syndrome, have no specific treatment or cure. Thus, there is a need for new compositions and methods capable of modulating the expression of any endogenous gene to help reverse the effects of a disease or disorder, in particular, therapies with reduced immunogenicity, reduced off-target effects, increased specificity for the target gene and/or increased therapeutic efficiency.

Summary of the present invention

In one aspect, the present application provides an expression cassette comprising a sequence encoding a non-naturally occurring transcription factor that increases expression of the SCN1A gene in a cell, wherein the non-naturally occurring transcription factor contains a DNA binding domain (DBD) operably linked to at least with two transcription-activating domains (TADs) as follows: TAD1-TAD2-DBD, DBD-TAD3-TAD4, or TAD1TAD2-DBD-TAD3-TAD4. In certain embodiments, TAD1, TAD2, TAD3, and TAD4 are independently selected from the following: VP16, VP64, Viper, CITED2, CITED4, CREB3, or functional fragments thereof. In certain embodiments, TAD1 and TAD2 are the same TAD. In certain embodiments, TAD1 and TAD2 are CITED2 or a functional fragment thereof. In certain embodiments, TAD1 and TAD2 are CITED4 or a functional fragment thereof. In certain embodiments, TAD3 and TAD4 are the same TAD. In certain embodiments, TAD3 and TAD4 are CITED2 or a functional fragment thereof. In certain embodiments, TAD3 and TAD4 are CITED4 or a functional fragment thereof. In certain embodiments, TAD1, TAD2, TAD3, and TAD4 are the same TAD. In certain embodiments, TAD1, TAD2, TAD3, and TAD4 are CITED2 or a functional fragment thereof. In certain embodiments, TAD1, TAD2, TAD3, and TAD4 are CITED4 or a functional fragment thereof. In certain embodiments, there is no linker between at least two TAD domains.

In certain embodiments, there is a linker between at least two TAD domains. In certain embodiments, the linker comprises or consists of GGSGGGSG (SEQ ID NO: 177) or GGSGGGSGGGSGGGSG (SEQ ID NO: 178).

In certain embodiments, the DBD binds to a region of the genome containing 18-27 nucleotides.

In certain embodiments, the DBD has at least 80% sequence identity to its closest human counterpart. In certain embodiments, the DBD has at least 90% sequence identity to its closest human counterpart. In certain embodiments, each of the DBDs and at least two TADs has at least 80% sequence identity to their closest human counterparts. In certain embodiments, each of the DBDs and at least two TADs is characterized by sequence identity, composition

- 1 046157 lying at least 90% in relation to their closest human analogues.

In certain embodiments, the DBD comprises a guide RNA and a nuclease-inactivated Cas protein. In certain embodiments, the nuclease-inactivated Cas protein is a nuclease-inactivated Cas9 protein.

In certain embodiments, the DBD comprises a zinc finger domain. In certain embodiments, the DBD contains six to nine zinc finger domains. In certain embodiments, the DBD contains six zinc fingers. In certain embodiments, the DBD binds to a region of the genome containing 18 nucleotides. In certain embodiments, the DBD contains nine zinc fingers. In certain embodiments, the DBD binds to a 27 nucleotide region of the genome.

In certain embodiments, the DBD contains a sequence having at least 95% sequence identity to any of SEQ ID NOs: 148-151. In certain embodiments, the DBD comprises a sequence comprising any of SEQ ID NOs: 148-151.

In certain embodiments, the DBD is derived from human EGR1 or human EGR3.

In certain embodiments, the DBD contains a sequence having at least 90% identity to any of SEQ ID NOs: 77-98. In certain embodiments, the DBD contains SEQ ID NO: 77-98.

In certain embodiments, the DBD comprises a sequence having at least 90% identity to SEQ ID NO: 92. In certain embodiments, the DBD comprises SEQ ID NO: 92.

In certain embodiments, the non-naturally occurring transcription factor comprises a sequence having at least 90% identity to SEQ ID NO: 130 or 131. In certain embodiments, the non-naturally occurring transcription factor comprises SEQ ID NO: 130 or 131.

In certain embodiments, the expression cassette comprises a nucleotide sequence having at least 90% identity to either SEQ ID NO: 72 or 73. In certain embodiments, the expression cassette comprises a nucleotide sequence in either SEQ ID NO: 72 or 73.

In certain embodiments, the expression cassette further comprises a regulatory element that directs expression of the transcription factor at a higher level in PV-containing neurons than in other cell types. In certain embodiments, the regulatory element comprises any of SEQ ID NOs: 1-4. In certain embodiments, the regulatory element comprises SEQ ID NO: 2 or 3.

In certain embodiments, the expression cassette further comprises a PV-selective microRNA binding site. In certain embodiments, the PV-selective miRNA binding site has at least 90% identity to any of SEQ ID NO: 7, 14, or 15. In certain embodiments, the PV-selective miRNA binding site comprises any of SEQ ID NO: 7, 14 or 15.

In certain embodiments, the expression cassette is part of a viral vector. In certain embodiments, the viral vector is an AAV virus. In certain embodiments, the AAV virus is an AAV9 virus or a scAAV9 virus. In certain embodiments, the viral vector is a lentivirus.

In another aspect, the application provides an expression cassette comprising a sequence encoding a non-naturally occurring transcription factor that increases expression of the SCN1A gene in a cell, wherein the non-naturally occurring transcription factor comprises a DNA binding domain operably linked to a transcription activating domain, wherein the DNA binding domain represents is a zinc finger protein containing the sequence LEPGEKP - [YKCPECGKSFS X HQRTH TGEKP]n - YKCPECGKSFS X HQRTH - TGKKTS (SEQ ID NO: 147), and where there is no HA tag (SEQ ID NO: 303) between the DNA binding domain and the activating domain transcription domain. In certain embodiments, the transcription activating domain comprises a VP16, VPR, or VP64 sequence or a functional fragment thereof. In certain embodiments, the transcription activating domain comprises VP64.

In certain embodiments, the DNA binding domain binds to a genomic region containing 18-27 nucleotides. In certain embodiments, the DNA binding domain is a zinc finger domain comprising SEQ ID NO: 147, where n=6-9. In certain embodiments, the DNA binding domain is

- 2 046157 is a zinc finger domain containing SEQ ID NO: 147, where n=6. In certain embodiments, the DNA binding domain binds to a region of the genome containing 18 nucleotides. In certain embodiments, the DNA binding domain is a zinc finger domain comprising SEQ ID NO: 147, where n=9. In certain embodiments, the DNA binding domain binds to a genomic region containing 27 nucleotides.

In certain embodiments, the DNA binding domain comprises a sequence having at least 95% sequence identity to any of SEQ ID NOs: 148-151. In certain embodiments, the DNA binding domain comprises a sequence comprising any of SEQ ID NOs: 148-151.

In certain embodiments, the DNA binding domain comprises a sequence having at least 90% identity to any of SEQ ID NOs: 77-91. In certain embodiments, the DNA binding domain comprises any of SEQ ID NOs: 77-91.

In certain embodiments, the expression cassette further comprises a regulatory element that directs expression of the transcription factor at a higher level in PV-containing neurons than in other cell types. In certain embodiments, the regulatory element comprises any of SEQ ID NOs: 1-4. In certain embodiments, the regulatory element comprises SEQ ID NO: 2 or 3.

In certain embodiments, the non-naturally occurring transcription factor comprises a sequence having at least 90% identity to SEQ ID NO: 127. In certain embodiments, the non-naturally occurring transcription factor comprises SEQ ID NO: 127.

In certain embodiments, the expression cassette comprises a nucleotide sequence having at least 90% identity to either SEQ ID NO: 93 or 71. In certain embodiments, the expression cassette comprises a nucleotide sequence in either SEQ ID NO: 93 or 71.

In certain embodiments, the expression cassette further comprises a PV-selective microRNA binding site. In certain embodiments, the PV-selective miRNA binding site has at least 90% identity to any of SEQ ID NO: 7, 14, or 15. In certain embodiments, the PV-selective miRNA binding site comprises any of SEQ ID NO: 7, 14 or 15.

In certain embodiments, the expression cassette is part of a viral vector. In certain embodiments, the viral vector is an AAV virus. In certain embodiments, the AAV virus is an AAV9 virus or a scAAV9 virus. In certain embodiments, the viral vector is a lentivirus. In another aspect, the application provides a polynucleotide comprising a PV-selective miRNA binding site comprising a sequence having at least 80% sequence identity to SEQ ID NO: 14 or 15, wherein the miRNA binding site reduces transgene expression in excitatory neurons. In certain embodiments, the PV-selective miRNA binding site comprises SEQ ID NO: 14. In certain embodiments, the PV-selective miRNA binding site comprises SEQ ID NO: 15. In another aspect, the application provides an expression cassette comprising a PV-selective microRNA binding site and promoter and/or enhancer. In certain embodiments, the promoter and/or enhancer is a PV-selective regulatory element that drives expression of the transgene at a higher level in parvalbumin-containing neurons (PVs) than in other cell types. In certain embodiments, the PV-selective regulatory element is operably linked to the transgene. In another aspect, the present application provides an expression cassette comprising a regulatory element operably linked to a transgene and at least one microRNA binding site, wherein the regulatory element controls expression of the transgene at a higher level in parvalbumin-containing (PV) neurons than in other neurons. cell types, and where the microRNA binding site reduces transgene expression in excitatory neurons. In certain embodiments, the expression cassette does not contain SEQ ID NO: 67. In certain embodiments, the microRNA binding site contains at least one binding site for MIR128 (SEQ ID NO: 9). In certain embodiments, the microRNA binding site comprises at least one binding site for MIR221 (SEQ ID NO: 11). In certain embodiments, the microRNA binding site comprises at least one binding site for MIR222 (SEQ ID NO: 13). In certain embodiments, the microRNA binding site comprises at least one binding site for MIR128 (SEQ ID NO: 9) and at least one binding site for MIR221 (SEQ ID NO:

- 3 046157

eleven). In certain embodiments, the microRNA binding site comprises at least one binding site for MIR128 (SEQ ID NO: 9), at least one binding site for MIR221 (SEQ ID NO: 11), and at least one binding site for MIR222 (SEQ ID NO: 13). In certain embodiments, the miRNA binding site comprises a sequence having at least 90% identity to any of SEQ ID NO: 7, 14, or 15. In certain embodiments, the miRNA binding site comprises SEQ ID NO: 7, 14 or 15.

In certain embodiments, the transgene encodes a polypeptide containing a non-naturally occurring transcription factor that increases expression of the SCN1A gene in a cell. In certain embodiments, the transcription factor binds to a region of the genome containing 18-27 nucleotides. In certain embodiments, the transcription factor comprises a DNA binding domain. In certain embodiments, the transcription factor comprises a DNA binding domain and a transcription activating domain. In certain embodiments, the DNA binding domain has at least 80% sequence identity to its closest human counterpart. In certain embodiments, the DNA binding domain has at least 90% sequence identity to its closest human counterpart. In certain embodiments, both the DNA binding domain and the transcription activating domain have at least 80% sequence identity to their closest human counterparts. In certain embodiments, both the DNA binding domain and the transcription activating domain have at least 90% sequence identity to their closest human counterparts.

In certain embodiments, the DNA binding domain comprises a guide RNA and a nuclease-inactivated Cas protein. In certain embodiments, the nuclease-inactivated Cas protein is a nuclease-inactivated Cas9 protein.

In certain embodiments, the DNA binding domain comprises a zinc finger domain. In certain embodiments, the DNA binding domain comprises six to nine zinc finger domains. In certain embodiments, the DNA binding domain contains six zinc fingers. In certain embodiments, the DNA binding domain binds to a region of the genome containing 18 nucleotides. In certain embodiments, the DNA binding domain contains nine zinc fingers. In certain embodiments, the DNA binding domain binds to a genomic region containing 27 nucleotides.

In certain embodiments, the DNA binding domain comprises a sequence having at least 95% sequence identity to any of SEQ ID NOs: 148-151. In certain embodiments, the DNA binding domain comprises a sequence comprising any of SEQ ID NOs: 148-151. In certain embodiments, the DNA binding domain comprises a sequence having at least 90% identity to any of SEQ ID NOs: 92-98. In certain embodiments, the DNA binding domain comprises any of SEQ ID NOs: 92-98.

In certain embodiments, the DNA binding domain is a zinc finger protein containing the sequence LEPGEKP - [YKCPECGKSFS X HQRTH TGEKP]n - YKCPECGKSFS X HQRTH - TGKKTS (SEQ ID NO: 147).

In certain embodiments, the DNA binding domain comprises a sequence having at least 90% identity to any of SEQ ID NOs: 77-91. In certain embodiments, the DNA binding domain comprises any of SEQ ID NOs: 77-91.

In certain embodiments, the DNA binding domain is derived from human EGR1 or human EGR3.

In certain embodiments, the transcription activating domain comprises a VP16, VPR, VP64, CITED2, CITED4, or CREB3 sequence or a functional fragment thereof. In certain embodiments, the transcription activating domain comprises a human CITED2, CITED4, or CREB3 sequence or a functional fragment thereof.

In certain embodiments, the regulatory element comprises a sequence comprising any of SEQ ID NOs: 1-4. In certain embodiments, the regulatory element comprises a sequence comprising SEQ ID NO: 2 or 3.

In certain embodiments, the non-naturally occurring transcription factor comprises a sequence having at least 90% identity to any of SEQ ID NOs: 105, 106, and 127-129. According to certain options

- 4 046157 implementation of a non-naturally occurring transcription factor contains any of SEQ ID NOs: 105, 106 and 127-129.

In certain embodiments, the transgene comprises a nucleotide sequence having at least 90% identity to any of SEQ ID NO: 71, 74, 75, 76, or 184. In certain embodiments, the transgene comprises any of SEQ ID NO: 71, 74, 75, 76 or 184.

In certain embodiments, the expression cassette is part of a viral vector. In certain embodiments, the viral vector is an AAV virus. In certain embodiments, the AAV virus is an AAV9 virus or a scAAV9 virus. In certain embodiments, the viral vector is a lentivirus.

In another aspect, the present application provides a method for selectively expressing a transgene in parvalbumin-containing neurons (PVs) of primates, comprising administering to the primate a viral vector containing the transgene and at least one miRNA binding site, wherein the miRNA binding site reduces expression of the transgene in excitatory neurons.

In certain embodiments, the viral vector further comprises a regulatory element operably linked to the transgene, wherein the regulatory element controls expression of the transgene at a higher level in parvalbumin-containing neurons (PVs) than in other cell types. In certain embodiments, the microRNA binding site comprises at least one binding site for MIR128 (SEQ ID NO: 9). In certain embodiments, the microRNA binding site comprises at least one binding site for MIR221 (SEQ ID NO: 11). In certain embodiments, the microRNA binding site comprises at least one binding site for MIR222 (SEQ ID NO: 13). In certain embodiments, the microRNA binding site comprises at least one binding site for MIR128 (SEQ ID NO: 9) and at least one binding site for MIR221 (SEQ ID NO: 11). In certain embodiments, the microRNA binding site comprises at least one binding site for MIR128 (SEQ ID NO: 9), at least one binding site for MIR221 (SEQ ID NO: 11), and at least one binding site for MIR222 (SEQ ID NO: 13). In certain embodiments, the miRNA binding site comprises a sequence having at least 90% identity to any of SEQ ID NO: 7, 14, or 15. In certain embodiments, the miRNA binding site comprises SEQ ID NO: 7, 14 or 15.

In certain embodiments, the transgene comprises a sequence encoding a non-naturally occurring transcription factor that increases expression of the SCNIA gene in a cell.

In certain embodiments, the transcription factor binds to a region of the genome containing 18-27 nucleotides.

In certain embodiments, the transcription factor comprises a DNA binding domain.

In certain embodiments, the transcription factor comprises a DNA binding domain and a transcription activating domain.

In certain embodiments, the DNA binding domain has at least 80% sequence identity to its closest human counterpart. In certain embodiments, the DNA binding domain has at least 90% sequence identity to its closest human counterpart. In certain embodiments, both the DNA binding domain and the transcription activating domain have at least 80% sequence identity to their closest human counterparts. In certain embodiments, both the DNA binding domain and the transcription activating domain have at least 90% sequence identity to their closest human counterparts.

In certain embodiments, the DNA binding domain comprises a guide RNA and a nuclease-inactivated Cas protein. In certain embodiments, the nuclease-inactivated Cas protein is a nuclease-inactivated Cas9 protein.

In certain embodiments, the DNA binding domain comprises a zinc finger domain. In certain embodiments, the DNA binding domain comprises six to nine zinc finger domains. In certain embodiments, the DNA binding domain contains six zinc fingers. In certain embodiments, the DNA binding domain binds to a region of the genome containing 18 nucleotides. In certain embodiments, the DNA binding domain contains nine zinc fingers. In certain embodiments, the DNA binding domain

- 5 046157 is called with a genomic region containing 27 nucleotides.

In certain embodiments, the DNA binding domain comprises a sequence having at least 95% sequence identity to any of SEQ ID NOs: 148-151. In certain embodiments, the DNA binding domain comprises a sequence having any of SEQ ID NOs: 148-151.

In certain embodiments, the DNA binding domain is derived from human EGR1 or human EGR3.

In certain embodiments, the DNA binding domain comprises a sequence having at least 90% identity to any of SEQ ID NOs: 92-98. In certain embodiments, the DNA binding domain comprises any of SEQ ID NOs: 92-98.

In certain embodiments, the DNA binding domain is a zinc finger protein containing the sequence LEPGEKP - [YKCPECGKSFS X HQRTH TGEKP]n - YKCPECGKSFS X HQRTH - TGKKTS (SEQ ID NO: 147).

In certain embodiments, the DNA binding domain comprises a sequence having at least 90% identity to any of SEQ ID NOs: 77-91. In certain embodiments, the DNA binding domain comprises any of SEQ ID NOs: 77-91.

In certain embodiments, the transcription activating domain comprises a VP16, VPR, VP64, CITED2, CITED4, or CREB3 sequence or a functional fragment thereof. In certain embodiments, the transcription activating domain comprises a human CITED2, CITED4, or CREB3 sequence or a functional fragment thereof.

In certain embodiments, the regulatory element comprises a sequence comprising any of SEQ ID NOs: 1-4. In certain embodiments, the regulatory element comprises a sequence comprising SEQ ID NO: 2 or 3.

In certain embodiments, the non-naturally occurring transcription factor comprises a sequence having at least 90% identity to any of SEQ ID NOs: 105, 106, and 127-129. In certain embodiments, the non-naturally occurring transcription factor comprises any of SEQ ID NOs: 105, 106, and 127-129.

In certain embodiments, the transgene comprises a nucleotide sequence having at least 90% identity to any of SEQ ID NO: 71, 74, 75, 76, or 184. In certain embodiments, the transgene comprises any of SEQ ID NO: 71, 74, 75, 76 or 184.

In certain embodiments, the viral vector is an AAV virus. In certain embodiments, the AAV virus is an AAV9 virus or a scAAV9 virus. In certain embodiments, the viral vector is a lentivirus.

In certain embodiments, the primo is a human. In certain embodiments, the primate is a non-human primate. In certain embodiments, the non-human primate is an old world monkey, an orangutan, a gorilla, a chimpanzee, a marmoset, a cynomolgus monkey, a rhesus monkey, or a pig-tailed monkey.

In another aspect, the present application provides an expression cassette comprising a sequence encoding a non-naturally occurring transcription factor that increases expression of the SCN1A gene in a cell, wherein the non-naturally occurring transcription factor contains a sequence having at least 90% identity to with respect to SEQ ID NO: 128 or 129. In certain embodiments, the non-naturally occurring transcription factor comprises SEQ ID NO: 128 or 129.

In another aspect, the present application provides a method of increasing the expression of SCNIA in a cell by introducing any of the expression cassettes provided herein. In certain embodiments, the cell is a nerve cell. In certain embodiments, the nerve cell is selected from the group consisting of unipolar, bipolar, multipolar, or pseudounipolar neurons. In certain embodiments, the cell is a GABAergic neuron. In certain embodiments, the cell is a PV-containing neuron. In certain embodiments, the cell is not a nerve cell. In certain embodiments, the cell is a glial cell. In certain embodiments, the glial cell is selected from the group consisting of astrocytes, oligodendrocytes, ependymal cells, Schwann cells, and satellite cells. In certain embodiments, the cell is located within a subject. In certain embodiments, the subject is a mammal. In certain embodiments, the subject represents

- 6 046157 a person. In certain embodiments, increasing the expression of SCN1A treats a disease, disorder, or symptom. In certain embodiments, the disorder is a disorder of the central nervous system. In certain embodiments, the disorder is epilepsy associated with SCN1A haploinsufficiency. In certain embodiments, the haploinsufficiency results from the subject being heterozygous for a loss of function mutation in the SCN1A gene. In certain embodiments, the disorder is epilepsy associated with an insertion, deletion, or substitution in the SCN1A gene. In certain embodiments, the disorder is epilepsy associated with a point mutation in the SCN1A gene. In certain embodiments, the disorder is Dravet syndrome. In certain embodiments, the symptom of a central nervous system disorder is neuronal hyperactivity. In certain embodiments, treatment of a central nervous system disorder involves reducing neuronal hyperactivity. In certain embodiments, the symptom of the central nervous system disorder is seizures. In certain embodiments, treatment of a central nervous system disorder involves reducing the frequency of seizures. In certain embodiments, treatment of a central nervous system disorder involves reducing the severity of seizures.

In another aspect, the present application provides a method of increasing the expression of SCNIA in the CNS by introducing any of the expression cassettes provided herein. In certain embodiments, the expression cassette is administered via unilateral intracerebroventricular (ICV) administration. In certain embodiments, the expression cassette is administered via bilateral intracerebroventricular (ICV) administration. In certain embodiments, increased expression of SCN1A occurs in the brain. In certain embodiments, increased expression of SCN1A occurs in the frontal cortex, parietal cortex, temporal cortex, hippocampus, medulla oblongata, and/or occipital cortex. In certain embodiments, increased expression of SCN1A occurs in the spine. In certain embodiments, increased expression of SCN1A occurs in the spinal cord and/or intervertebral ganglion.

Inclusion by link

All publications, patents and patent applications referenced herein are incorporated herein by reference to the same extent as if each individual publication, patent or patent application had been specifically and individually designated for inclusion by reference.

Brief description of graphic materials

The novel features of the present invention are set forth in detail in the accompanying claims. A better understanding of the features and advantages of the present invention will be obtained by reference to the following detailed description, which sets forth illustrative cases in which the principles of the present invention are used, and the accompanying drawings, which show the following.

In fig. Figure 1 shows activation of endogenous SCN1A using engineered transcription factors that bind to different regions on chromosome 2 (referring to GRCh38.p12). Data are presented as fold change in SCN1A expression compared to the control (EGFP-KASH) condition.

In fig. 2A, FIG. 2B and FIG. Figure 2C shows the relative expression of endogenous SCN1A in HEK293 cells using SCN1A-specific transcriptional activators (see Table 1). Data are presented as fold change relative to control conditions and shown on a Logi ₀ scale.

In fig. Figure 3A shows the relative expression of endogenous SCN1A in GABA neurons using an SCN1A-specific transcriptional activator (construct 30). Data are presented as fold change relative to control conditions (CBA-EGFP).

In fig. Figure 3B shows the relative expression of endogenous SCN1A in GABA neurons using SCN1A-specific transcriptional activators (constructs 25 and 16). Data are presented as fold change relative to control conditions (CBA-EGFP) in Log ₁₀ .

In fig. Figure 4 shows the relative expression of endogenous SCN1A and 40 immediate neighboring genes driven by the specific transcription factor SCN1A (construct 30). Data are presented as fold change relative to control conditions (CBA-EGFP-KASH) in Log ₁₀ .

In fig. 5A and FIG. 5B shows expression of the SCN1A-specific transcriptional activator in vivo compared to a control expression cassette that expressed eGFP. In fig. 5A shows the relative expression of the SCN1A gene in mice injected with either control eGFP or construct 4 containing the SCN1A transcriptional activator. In fig. Figure 5B shows the change in SCN1A expression in terms of the average percentage of eGFP. These experiments indicate that transcriptional activation by construct 4 resulted in an approximately 20-30% increase in SCN1A expression.

In fig. 6A, FIG. 6B, FIG. 6C, FIG. 6D, FIG. 6E, FIG. 6F and FIG. Figure 6G shows the effect on hyperthermic seizures in the Scn1a ^tm1Kea knockout mouse model of Dravet syndrome using different SCN1A-specific transcription factors compared to controls. P1 Scn1a +/− (heterozygous; HET) mice were injected with either AAV9-EGFP or an AAV9 vector expressing a SCN1A-specific transcription factor (one of constructs 31–34, 42, and 43). Infused P26-P28 mice were subjected to a hyperthermia-induced seizure test and the core temperature at which they experienced a tonic-clonic seizure was recorded. In fig. 6D shows a direct comparison between design 32, which contains an HA tag located between the DBD and TAD, and design 34, which does not contain an HA tag. In fig. 6E shows a direct comparison between construct 31, which contains the miRNA m1 binding site located between the coding region and the polyA tail, and construct 32, which does not contain the miRNA m1 binding site. In fig. 6H illustrates the effect on hyperthermic seizures in the Scn1a ^RX mutant mouse model of Dravet syndrome using construct 31 (compared to PBS-infused control).

In fig. 7A, FIG. 7B, FIG. 7C and FIG. Figure 7D shows survival in the Scn1a ^tm1Kea knockout mouse model of Dravet syndrome under various conditions. In fig. 7A shows a comparison of wild type (PBS WT) and Scn1a +/- (PBS HET) mice in a survival assay. P1 Scn1a +/− (N=53) and Scn1a +/+ (N=54) mice were infused with PBS. Mice were observed in their home cage daily and the date of any mortality was recorded. There was a significant difference in survival between Scn1a +/- and Scn1a +/+ animals (P<0.0001). In fig. 7B-D shows the effect on survival in a mouse model of Dravet syndrome for mice treated with various SCN1A-specific transcription factors compared to controls. P1 Scn1a +/− mice were infused with either PBS or an AAV9 vector expressing a transcription factor specific for SCN1A (constructs 31 or 33). Mice were observed in their home cage daily and the date of any mortality was recorded. In fig. 7D shows a direct comparison between construct 31, which contains the miRNA m1 binding site located between the coding region and the polyA tail, and construct 33, which does not contain the miRNA m1 binding site. In fig. 7E shows survival in the Scn1a ^RX mutant mouse model of Dravet syndrome using construct 31 (compared to PBS-injected control).

In fig. 8 shows the relative expression of Scn1A mRNA in various brain tissues following intraparenchymal delivery of an AAV9 vector encoding a SCN1A-specific transcription factor (construct 33) administered to two cynomolgus monkeys at a dose of 1.2 x ^{10 12} genome copies/animal, normalized to two untreated control animals. All animals were sacrificed 28 days postinjection, and Scn1A mRNA was quantified in tissue samples using Taqman PCR. Data are presented as normalized target mRNA expression in different brain tissue sections. Similar results were reported with a different primer/probe set derived from the Scn1a gene.

In fig. 9A-F show the expression pattern of EGFP in the dentate gyrus region of the marmoset hippocampus following treatment with AAV9 vectors containing the EGFP transgene under the control of the EF1a promoter, the RE 2 promoter (SEQ ID NO: 2), or the RE 2 promoter (SEQ ID NO: 2) with a binding site. microRNA m1 (SEQ ID NO: 7), located between the EGFP coding region and the polyA site. A representative region of the hippocampal dentate gyrus is shown for each processing vector. The top row shows cell nuclei stained with DAPI, and the bottom row shows GFP-positive areas stained with anti-GFP antibody. In fig. 9A (EF1a treatment), the hylus CA4 region of the hippocampus is circled and the arrows point to the dentate cell granule cell body (DG) layer. Fig. 9B and FIG. 9C are concentrated in the same area. Region CA4, which is a mixture of excitatory and inhibitory interneurons, is highlighted because it was the only region with significant expression in the RE 2 + m1 condition. During EF1a and RE 2 driven transgene expression, GFP expression was more widespread and included other areas of the hippocampus. The DG cell layer is thought to contain mainly excitatory neurons. GFP expression driven by EF1a and RE 2 is visible in the DG cell layer (Fig. 9D and Fig. 9E) but is not present in RE 2 + m1-treated animals (Fig. 9F) (white arrows).

In fig. 10A-L show the expression pattern of EGFP in the dentate gyrus region of the marmoset hippocampus after treatment with AAV9 vectors containing the EGFP transgene under the control of the EF1a promoter, the RE 2 promoter (SEQ ID NO: 2) or the RE 2 promoter (SEQ ID NO: 2) with The microRNA m1 binding site (SEQ ID NO: 7), located between the EGFP coding region and the polyA site, is primarily localized to parvalbumin-positive (PV) cells in RE 2 and RE 2 + m1-treated animals. A representative region of the hippocampal dentate gyrus is shown for each processing vector. The top row shows GFP-positive areas, and the next bottom row shows the same areas stained with the inhibitory interneuron marker for PV. The boxed area in FIG. 10A-F is shown at higher magnification in FIG. 10G-L. GFP expression driven by RE 2 and RE 2 + m1 is mainly colocalized with the inhibitory marker of PV-containing interneurons (white arrows in Figs. 10H and 10K, 10I and 10L), whereas in EG-EF1a GFP expression is not as readily localized to PV-positive cells (white arrows in Fig. 10G and 10J). In addition, GFP-positive cells are characterized by a distinct interneuron morphology of highly branched cells with pi

- 8 046157 ramid cell body in animals treated with RE 2 and RE 2 + ml (arrows in Fig. 10H and 10I), compared with less distinct cell body morphology in animals treated with EF1a (arrows in Fig. 10G).

In fig. 11 shows the VG/diploid genome in tissue samples from the frontal cortex (FC), rostral parietal cortex (Rostral PC), temporal cortex (TC), caudal parietal cortex (Caudal PC), hippocampus (Hip), medulla oblongata (Med), and occipital cortex. (OC) animals treated with AAV9-RE ^gaba eTF ^SC NiA administered at a dose of 4.8E+13 or 8E+13 μg/animal via unilateral intracerebroventricular (ICV) administration (Example 10 and Example 11). Each data point represents the VG/diploid genome for a tissue sample, and the horizontal bars represent the average VG/diploid genome across all tissue samples for each animal.

In fig. Figure 12 shows transcripts/μg RNA in tissue samples from the frontal cortex (FC), rostral parietal cortex (Rostral PC), temporal cortex (TC), caudal parietal cortex (Caudal PC), hippocampus (Hip), medulla oblongata (Med), and occipital cortex. (OC) animals treated with AAV9-RE ^gaba eTF ^SC N ^1A administered at a dose of 4.8E+13 or 8E+13 vector genomes/animal via unilateral intracerebroventricular (ICV) administration (Example 10 and Example 11). Each data point represents the vector genomes/diploid genome for a tissue sample, and the horizontal bars represent the average vector genomes/diploid genome ratio across all tissue samples for each animal. Average transcripts for ARFGAP2 were 1.85E+6/μg RNA and are indicated by the dashed upper limit line. The detection limit is indicated by the dashed lower limit line.

In fig. Figure 13 shows vector biodistribution (vector genomes/diploid genome) and transgene expression (transcripts/μg RNA) in peripheral tissue samples outside the brain. Peripheral tissue samples shown are spinal cord C2/L4 (SC C2/L4), intervertebral ganglion C2/L4 (DRG C2/L4), liver, spleen, heart, kidney, lung, pancreas, and testes/ovaries. The average VCN (vector biodistribution) and transcript (transgene expression) values in the primate brain are shown by the dotted line.

Detailed Description of the Present Invention

Presented herein are engineered transcription factors or eTFs that are not naturally occurring and have been designed to bind to a target site in the genome and modulate the expression of an endogenous gene of interest. Such eTFs can be designed to enhance or suppress the expression (RNA and/or protein expression) of a gene of interest. This document also provides microRNA binding sites that can be incorporated into a viral vector and allow selective expression of the transgene in parvalbumin-containing neurons (PVs).

In one aspect, the present application provides eTFs that are capable of increasing the expression of the sodium voltage channel alpha subunit 1 (SCN1A) gene and increasing the expression of its corresponding protein product Nav1.1, as well as methods of using them to treat diseases or disorders associated with Nav1 deficiency .1, such as Dravet syndrome.

In another aspect, the present application provides microRNA binding sites that reduce the expression of miRNA binding site-containing mRNA in excitatory neurons, thereby leading to selective gene expression in GABAergic neurons or parvalbumin containing neurons (PV), and methods of using them to selectively express a representative gene. gene of interest in PV neurons.

Definitions.

The singular forms used herein are intended to include the plural unless the context clearly indicates otherwise. Moreover, to the extent that the terms including, includes, having, has, with, or variants thereof are used either in the detailed description or in the claims, such terms are intended to be included in the same manner as the term containing.

The term approximately or roughly means within the acceptable range of error for a particular value as determined by one skilled in the art, which will depend in part on how the value is measured or determined, i.e. limitations of the measurement system. For example, approximately may mean within one or more than one standard deviation in accordance with practice in the art. Alternatively, approximately may mean a range of up to 20%, up to 15%, up to 10%, up to 5%, or up to 1% of the target value.

The terms definition, measurement, assessment, evaluation, analysis, and their grammatical equivalents may be used interchangeably herein to refer to any form of measurement and include determining whether an item is present or not (eg, detection). These terms may include both quantitative and/or qualitative definitions. The rating can be relative or absolute.

The term expression refers to the process by which a nucleic acid sequence or polynucleotide is transcribed from a DNA template (eg, into mRNA or other RNA transcript), and/or the process by which the transcribed mRNA is subsequently translated into peptides, polypeptides or proteins. Transcripts and encoded polypeptides can be called together

- 9 046157 to produce a gene product. If the polynucleotide is derived from genomic DNA, expression may involve splicing of the mRNA in the eukaryotic cell.

As used herein, the terms operably linked, functionally linked, or their grammatical equivalents refer to the juxtaposition of genetic elements, eg, promoter, enhancer, polyadenylation sequence, etc., where the elements are in a relationship that allows them to function in the expected manner. For example, a regulatory element, which may include promoter and/or enhancer sequences, is operably linked to a coding region if the regulatory element helps initiate transcription of the coding sequence. There may be intermediate residues between the regulatory element and the coding region as long as this functional relationship is maintained. As used herein, the term vector refers to a macromolecule or association of macromolecules that contains or is associated with a polynucleotide and that can be used to mediate delivery of the polynucleotide into a cell. Examples of vectors include plasmids, viral vectors, liposomes and other gene delivery vehicles. The vector typically contains genetic elements, for example, regulatory elements, operably linked to the gene to facilitate expression of the gene in the target.

As used herein, the terms expression cassette and nucleic acid cassette interchangeably refer to a combination of nucleic acid sequences or elements that are expressed together or operably linked for expression. In some cases, an expression cassette refers to a combination of regulatory elements and the gene or genes to which they are operably linked for expression.

The term AAV is an abbreviation for adeno-associated virus and can be used to refer to the virus itself or a derivative of it. The term covers all serotypes, subtypes, and naturally occurring and recombinant forms, unless otherwise stated. The abbreviation rAAV refers to recombinant adeno-associated virus, also called recombinant AAV vector (or rAAV vector). The term AAV includes AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, AAV12, rh10 and their hybrids, avian AAV, bovine AAV, canine AAV, equine AAV, primate AAV , non-primate AAV and sheep AAV. The genomic sequences of various AAV serotypes, as well as the sequences of native terminal repeats (TRs), Rep proteins and capsid subunits are known in the art. Such sequences can be found in the literature or in public databases such as GenBank. As used herein, the term rAAV vector refers to an AAV vector containing a polynucleotide sequence not of AAV origin (ie, a polynucleotide heterologous to AAV), typically a sequence of interest for the genetic transformation of a cell. In general, a heterologous polynucleotide is flanked by at least one, and usually two, AAV inverted terminal repeat (ITR) sequences. The rAAV vector can be single-stranded (ssAAV) or self-complementary (scAAV). An AAV virus or AAV viral particle refers to a viral particle consisting of at least one AAV capsid protein and an encapsidated rAAV polynucleotide vector. If the particle contains a heterologous polynucleotide (ie, a polynucleotide different from the wild-type AAV genome, such as a transgene to be delivered into a mammalian cell), it is commonly referred to as an rAAV vector particle or simply an rAAV particle. Thus, production of an rAAV particle necessarily involves production of an rAAV vector, since such a vector is contained within the rAAV particle.

As used herein, the terms treat, treatment, therapy, etc. refer to alleviating, delaying or delaying the progression of, preventing, mitigating, reducing the effects or symptoms of, preventing the occurrence of, inhibiting or alleviating the onset of diseases or disorders. The methods of the present disclosure can be used with any mammal. Examples of mammals include, but are not limited to, rats, cats, dogs, horses, cows, sheep, pigs, and, more preferably, humans. The therapeutic effect includes eradication or improvement of the underlying disease being treated. In addition, a therapeutic effect is achieved by eliminating or alleviating one or more physiological symptoms associated with the underlying disorder, such that improvement is observed in the subject although the subject may still be suffering from the underlying disorder. In some cases, for prophylactic purposes, a therapeutic agent can be administered to a subject at risk of developing a particular disease or to a subject reporting one or more physiological symptoms of a disease, even if the diagnosis of that disease has not been made. The methods of the present disclosure can be used with any mammal. In some cases, treatment may result in a reduction or cessation of symptoms (eg, reduction in the frequency, duration, and/or severity of seizures). A prophylactic effect includes delaying or eliminating the onset of a disease or condition, delaying or eliminating the onset of symptoms of a disease or condition, slowing, stopping or reversing the progression of a disease or condition, or any combination thereof.

The term effective amount or therapeutically effective amount refers to that amount of a composition described herein that is sufficient to affect

- 10 046157 intended use, including, without limitation, the treatment of a disease as defined below. The therapeutically effective amount may vary depending on the intended therapeutic use (in vivo) or the subject and the pathological condition being treated, for example, the weight and age of the subject, the severity of the pathological condition, the route of administration, etc., as can be readily determined by one skilled in the art. technology. The term also applies to the dose that will produce a specific response in the target cell. The specific dosage will vary depending on the particular composition chosen, the dosage schedule to be followed, whether it is administered in combination with other compounds, the time of administration, the tissue into which it is administered, and the physical delivery system in which it is administered.

A nucleotide or peptide sequence fragment refers to a sequence that is shorter than the reference or full-length sequence.

Molecule variant refers to allelic variations of such sequences, i.e. to a sequence substantially similar in structure and biological activity to either the entire molecule or a fragment thereof.

A functional fragment of a DNA or protein sequence refers to a fragment that retains biological activity (functional or structural) that is substantially similar to the biological activity of the full-length DNA or protein sequence. The biological activity of a DNA sequence may lie in its ability to influence expression in a manner known to be attributable to the full-length sequence.

The terms subject and individual are used interchangeably herein to refer to a vertebrate animal, preferably a mammal, more preferably a human. The methods described herein may be useful for human therapeutic purposes, veterinary medicine, and/or preclinical studies in animal models of a disease or condition.

The term in vivo refers to an event that occurs in the body of a subject.

The term in vitro refers to an event that occurs outside the subject's body. For example, an in vitro assay includes any assay performed outside the subject. In vitro assays include cell-based assays that use live or dead cells. In vitro assays also include cell-free assays, which do not use intact cells.

In general, sequence identity or sequence homology, which can be used interchangeably, refers to the exact nucleotide-nucleotide or amino acid-amino acid match of two polynucleotide or polypeptide sequences, respectively. Typically, methods for determining sequence identity involve comparing two nucleotide or amino acid sequences and determining their percentage identity. Comparison of sequences, for example for the purpose of identity assessment, can be performed using any suitable alignment algorithm, including, without limitation, the Needleman-Wunsch algorithm (see, for example, the EMBOSS Needle aligner, available at www.ebi.ac.uk /Tools/psa/emboss_needle/, optional with default settings), the BLAST algorithm (see, for example, the BLAST alignment tool available at blast.ncbi.nlm.nih.gov/Blast.cgi, optional with default settings), and Smith-Waterman algorithm (see, for example, the EMBOSS Water equalizer, available at www.ebi.ac.uk/Tools/psa/emboss_water/, optionally with default settings). Optimal alignment can be assessed using any appropriate parameters of the selected algorithm, including default parameters. Percent identity, also called percent homology, between two sequences can be calculated as the number of exact matches between two optimally aligned sequences divided by the length of the reference sequence and multiplied by 100. Percent identity can also be determined, for example, by comparing sequence information using improved BLAST computer program, including version 2.2.9, available from the National Institutes of Health. The BLAST program is based on the alignment method of Karlin and Altschul, Proc. Natl. Acad. Sci. USA 87:2264-2268 (1990) and as described in Altschul, et al., J. Mol. Biol. 215:403-410 (1990); Karlin and Altschul, Proc. Natl. Acad. Sci. USA 90:5873-5877 (1993) and Altschul et al., Nucleic Acids Res. 25:3389–3402 (1997). Briefly, the BLAST program defines identity as the number of identical aligned characters (i.e., nucleotides or amino acids) divided by the total number of characters in the shorter of the two sequences. The program can be used to determine the percentage of identity over the entire length of the sequences being compared. Default parameters are provided to optimize searches using short sequences of queries, such as using the blastp program. The program also allows the SEG filter to be used to mask segments of query sequences, as defined by the SEG program Wootton and Federhen, Computers and Chemistry 17: 149-163 (1993). High sequence identity generally includes sequence identity ranges from approximately 80% to 100% and integers in between. As used herein, the term engineered protein refers to a non-naturally occurring protein, including, but not limited to

- 11 046157 nia, a protein derived from a naturally occurring protein, or when a naturally occurring protein is modified or reprogrammed to have a specific property.

As used herein, the terms synthetic and artificial are used interchangeably to refer to a protein or domain thereof that has low sequence identity (eg, less than 50% sequence identity) to a naturally occurring human protein. For example, the VPR and VP64 domains are synthetic transactivation domains.

As used herein, the term engineered transcription factor or eTF refers to a non-naturally occurring DNA-binding protein or non-naturally occurring transcription modulator that has been modified or reprogrammed to bind to a specific target binding site and/or to include a modified or replaced transcription effector domain. .

As used herein, the term DNA binding domain can be used to refer to one or more DNA binding motifs, such as a zinc finger or a basic helix-loop-helix (bHLH) motif, individually or together as part of a DNA-binding protein.

The terms transcription activation domain, transcription activation domain, transactivation domain, trans-activation domain, and TAD are used interchangeably herein and refer to a protein domain that, together with a DNA binding domain, can activate transcription from a promoter by contacting the transcription machinery (e.g. general transcription factors and/or RNA polymerase) either directly or through other proteins known as coactivators.

The terms transcriptional repressor domain and TRD are used interchangeably herein and refer to a protein domain that, in combination with a DNA-binding domain, can repress transcription from a promoter by contact with transcription machinery (e.g., general transcription factors and/or RNA polymerase) or directly , or through other proteins known as corepressors.

The term GRCh38.p12 refers to Human Genome Reference Consortium Build 38 patch release 12 (GRCh38.p12), having GenBank build accession number GCA_000001405.27 and dated December 21, 2017.

Unless otherwise specified, all terms used herein have the same meaning as those of ordinary skill in the art, and the practice of the present invention will utilize conventional techniques of molecular biology, microbiology, and recombinant DNA technology that are within the scope of knowledge specialists in the present field of technology.

Engineered transcription factors (eTFs) that activate SCN1A

In one aspect, the present application provides eTFs that are capable of increasing the expression of the voltage-regulated sodium channel alpha subunit 1 (SCN1A) gene and increasing the expression of its corresponding protein product Nav1.1. The SCN1A gene belongs to a family of genes that encode subunits used to assemble sodium channels. These channels, which transport positively charged sodium ions into cells, play a key role in the cell's ability to generate and transmit electrical signals. The SCN1A gene encodes one part (alpha subunit) of a sodium channel called Nav1.1. These channels are primarily found in the brain, where they control the flow of sodium ions into cells. Nav1.1 channels are involved in transmitting signals from one nerve cell (or neuron) to another. Several mutations in the SCN1A gene have been found to cause genetic epilepsy with febrile seizures plus (GEFS+), which is a spectrum of seizure disorders of varying severity. These conditions include simple febrile (fever-associated) seizures, which begin in infancy and usually resolve by age 5, and febrile seizures plus (FS+). FS+ includes febrile and other types of seizures, including those not associated with fever (afebrile seizures) that continue into adulthood. The GEFS+ spectrum also includes other conditions, such as Dravet syndrome (also known as severe myoclonic epilepsy of infancy, or SMEI), which cause more severe seizures that last longer and are difficult to control. These recurring seizures (epilepsy) can get worse over time and are often accompanied by decreased brain function. Many other mutations have been linked to familial hemiplegic migraine, a form of migraine headache that runs in families, and at least one mutation has been linked to the effectiveness of some anticonvulsant drugs. Thus, the eTF presented herein, which increases SCN1A expression, can be used to treat a variety of diseases or disorders associated with mutations in the Nav1.1 channel.

Transcription factors (TFs) are proteins that bind to specific sequences in the genome and control gene expression. The engineered transcription factors or eTFs provided herein that activate SCN1A are non-naturally occurring proteins that contain a DNA binding domain (DBD) and

- 12 046157 at least one domain that is a transcription modulator, for example, either a transcription activation domain (TAD) or a transcription repressor domain (TRD). In one embodiment, the eTF that activates SCN1A may comprise a DBD and a TAD (eg, TADDBD or DBD-TAD), wherein the DBD and TAD may be derived from the same protein or from different proteins. In another embodiment, an eTF that activates SCN1A may comprise a DBD and two TADs, wherein the DBD and one TAD are derived from the same protein, the DBD is derived from the first protein and both TADs are derived from the second protein, the DBD and one TAD are derived from the first protein and a second TAD is derived from a second protein, or a DBD is derived from a first protein, one TAD is derived from a second protein, and a second TAD is derived from a third protein (e.g., TAD1-DBD-TAD1, TAD1-DBDTAD2, TAD1-TAD1-DBD, TAD1-TAD2-DBD, DBD-TAD1-TAD1 or DBD-TAD1-TAD2). In another embodiment, an eTF that activates SCN1A may comprise a DBD and three TADs, wherein the DBD and TAD are derived from the same protein, the DBD is derived from the first protein, and the TADs are derived from one or more different proteins, or the DBD and all TADs come from different proteins, for example TAD _X -TAD _X -TAD _X -DBD, TAD _X -TAD _X -DBD-TAD _X , TAD _X -DBD-TAD _X -TAD _X or DBDTAD _X -TAD _X -TAD _X. where each X is independently selected and may be the same or different from one or all other TADs. Examples include, for example, TAD1-TAD1-DBD-TAD1, TAD1-TAD1DBD-TAD2, TAD1-TAD2-DBD-TAD1, TAD1-TAD2-DBD-TAD2, TAD1-TAD2-DBD-TAD3, TAD1-DBDTAD1-TAD1 , TAD1-DBD-TAD2-TAD2, TAD1-DBD-TAD1-TAD2, TAD2-DBD-TAD1-TAD2, TAD1DBD-TAD2-TAD3, TAD1-TAD1-TAD1-DBD, TAD1-TAD2-TAD2-DBD, TAD1-TAD2 -TAD2-DBD, TAD1TAD2-TAD3-DBD, DBD-TAD1-TAD1-TAD1, DBD-TAD1-TAD1-TAD2, DBD-TAD1-TAD2-TAD2 or

DBD-TAD1-TAD2-TAD3, etc. In another embodiment, an eTF that activates SCN1A may comprise a DBD and four TADs, wherein the DBD and TAD are derived from the same protein, the DBD is derived from the first protein, and the TADs are derived from one or more different proteins, or wherein the DBD and all TADs come from different proteins, for example TADX-TADX-TADX-TADX-DBD, TADXTADX-TADX-DBD-TADX, TADX-TADX-DBD-TADX-TADX, TADX-DBD-TADX-TADX-TADX or DBDTAD _X -TAD _X -TAD _X -TAD _X , where each X is independently selected and may be the same or different from one or all other TADs. Examples include, for example, TAD1-TAD1-DBD-TAD1-TAD1,

TAD 1 -TAD 1-DBD-TAD2-TAD2, TAD1-TAD2-DBD-TAD1-TAD3, TAD 1 -TAD 1 -TAD 1-DBD-TAD2, TAD 1-DBD-TAD2-TAD3-TAD4, TAD 1-DBD -TAD2-TAD3-TAD4,

TAD 1-TAD2-DBD-TAD1-TAD2, TAD 1-TAD3-DBD-TAD1-TAD2, TAD 1-TAD2-TAD3-DBD-TAD4, TAD 1 -DBD-TAD 1-TAD2-TAD3, TAD 1 -TAD 1 -TAD 1 -TAD 1 -DBD,

TAD 1-TAD2-DBD-TAD2-TAD1, TAD 1-TAD2-DBD-TAD3-TAD4, TAD 1 -DBD-TAD 1 -TAD 1-TAD2, TAD2-DBD-TAD1-TAD2-TAD3, TAD 1-TAD2- TAD2-TAD3-DBD,

TAD1-TAD2-TAD3-TAD4-DBD, DBD-TAD1-TAD1-TAD1-TAD1, DBD-TAD1-TAD1-TAD2-TAD2, DBDTAD1-TAD2-TAD3-TAD4 or DBD-TAD1-TAD2-TAD3-TAD3, etc. d. In one embodiment, the eTF that activates SCN1A comprises a DBD and two TADs that are located at one end of the DBD (e.g., the N-terminus or the C-terminus), wherein the DBD is derived from a first protein and both TADs are derived from a second protein, or A DBD is derived from a first protein, one TAD is derived from a second protein, and a second TAD is derived from a third protein (e.g., TAD1-TAD1-DBD, TAD1-TAD2-DBD, DBD-TAD1-TAD1, or DBD-TAD1-TAD2). In certain embodiments, the DBD may be a synthetic construct comprising domains from multiple proteins.

In certain embodiments, the DBD and the TAD and/or the two TADs can be directly conjugated, eg, without an intervening amino acid sequence, the DBD and the TAD and/or the two TADs can be conjugated using a peptide linker or combinations thereof. In certain embodiments, the DBD is conjugated to a TAD and/or one TAD is conjugated to a second TAD via a linker containing 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 , 16, 17, 18, 19, 20, 25, 30, 35, 40, 50, 60, 70, 75, 80, 90 or 100 amino acids or 1-5, 1-10, 1-20, 1-30, 1-40, 1-50, 1-75,

1-100, 5-10, 5-20, 5-30, 5-40, 5-50, 5-75, 5-100, 10-20, 10-30, 10-40, 10-50, 10- 75, 10-100, 20-30, 20-40, 2050, 20-75 or 20-100 amino acids. In some cases, a DBD and a TAD and/or two TADs are conjugated through naturally occurring intermediate residues found in the naturally occurring proteins from which the domains are derived. In other embodiments, the DBD and the TAD and/or two TADs are conjugated through a synthetic or exogenous linker sequence. Suitable linkers can be flexible, cleavable, non-cleavable, hydrophilic and/or hydrophobic. In certain embodiments, a DBD and a TAD and/or two TADs can be fused together through a linker containing multiple glycine and/or serine residues. Examples of glycine/serine peptide linkers include [GS]n, [GGGS]n (SEQ ID NO: 179), [GGGGS]n (SEQ ID NO: 180), [GGSG]n (SEQ ID NO: 181), where n is an integer equal to or greater than 1. In certain embodiments, the linker used to conjugate a DBD and a TAD and/or two TADs is GGSGGGSG (SEQ ID NO: 177). In certain embodiments, the linker used to conjugate a DBD and a TAD and/or two TADs is GGSGGGSGGGSGGGSG (SEQ ID NO: 178). In certain embodiments, when a DBD is conjugated to two TADs, the first and second TAD may be conjugated to the DBD with the same or different linkers, or one TAD may be conjugated to the DBD by a linker and

- 13 046157 the other TAD may be conjugated directly to the DBD (eg, without intervening linker sequences), or both TADs may be directly conjugated to the DBD (eg, without intervening linker sequences). In certain embodiments, when a DBD is conjugated to two TADs at the same end (e.g., N-terminus or C-terminus), the linker connecting the two TADs may be the same or different from the linker connecting the TAD to the DBD, or TADs can be conjugated to each other by a linker, but TADs are directly conjugated to a DBD (e.g., without an intervening linker sequence), or TADs can be directly conjugated to each other (e.g., without intervening linker sequences), but TADs are conjugated to a DBD with using a linker. In certain embodiments, eTFs provided herein that activate SCN1A do not contain one or more HA tags (eg, SEQ ID NO: 171) located between the DBD and one or more TADs.

The eTFs presented herein that activate SCN1A have properties distinct from natural transcription factors. In certain embodiments, an eTF that activates SCN1A comprises a DBD derived from a naturally occurring protein that has been modified such that the DBD binds to a different target site than the naturally occurring protein from which it is derived, and an eTF comprising such a modified DBD modulates the expression of a different gene (eg, SCN1A) compared to the native protein from which the DBD is derived (eg, a non-SCN1A gene). In other embodiments, an eTF provided herein that activates SCN1A contains a TAD derived from a naturally occurring protein that has been modified such that the eTF containing such modified TAD modulates the expression of another gene (e.g., SCN1A) relative to with the naturally occurring protein from which the TAD is derived (e.g., a non-SCN1A gene), and/or the eTF containing such a modified TAD modulates SCN1A expression differently (e.g., upregulates or represses) compared to the naturally occurring protein , from which the TAD is derived. In certain embodiments, an eTF provided herein that activates SCN1A comprises a DBD derived from a naturally occurring protein and a TAD derived from a naturally occurring protein (the same or different proteins), wherein both the DBD and the TAD have been modified. In such embodiments, the DBD may bind to a different target site compared to the naturally occurring protein from which it was derived, the eTF containing such a modified DBD and TAD modulates the expression of a different gene (eg, SCN1A) compared to the naturally occurring protein , from which the domains are derived (e.g., gene(s) other than SCN1A), and/or eTF containing such modified DBDs and TADs, modulate SCN1A expression differently (e.g., activate or repress) compared to naturally occurring proteins from which the DBD and TAD domains are derived.

DNA binding domains (DBDs).

eTFs provided herein that activate SCN1A may comprise any suitable DBD that binds to a target site of interest (eg, a target site that results in up-regulation of SCN1A when bound by an eTF provided herein). In certain embodiments, the DBD may be a synthetically engineered DBD. In other embodiments, the DBD may be derived from a naturally occurring protein. DBD families include basic helix-loop-helix (bHLH) (e.g., c-Myc), basic leucine zipper (e.g., C/EBP), helix-turn-helix (e.g., Oct-1), and zinc fingers (e.g. , EGR1 or EGR3). These families exhibit a wide range of DNA binding and gene target specificities. As contemplated herein, any of the known human transcription factor proteins can serve as a protein platform to engineer and/or reprogram the DBD to recognize a specific target site, resulting in modulation of endogenous SCN1A gene expression. In exemplary embodiments, the DBD provided herein comprises a zinc finger domain binding a TALEN domain or a gRNA/Cas complex.

The DBD presented herein can be designed to recognize any target site that leads to activation of SCN1A. In exemplary embodiments, the DBD is designed to recognize a genomic location and enhance expression of an endogenous SCN1A gene associated with an eTF. Binding sites capable of modulating the expression of the endogenous SCNIA gene when bound by the eTF provided herein can be located anywhere in the genome, resulting in modulation of SCN1A gene expression. In various embodiments, the binding site may be located on a chromosome other than SCN1A, on the same chromosome as SCN1A, upstream of the transcription start site (TSS) of the SCN1A gene, downstream of the TSS of the SCN1A gene, proximal to TSS of the SCN1A gene, distal to the TSS of the SCN1A gene, in the coding region of the SCN1A gene, in the intron of the SCN1A gene, downstream of the polyA tail of the SCN1A gene, in the promoter sequence that regulates the SCN1A gene, or in the enhancer sequence that regulates the SCN1A gene .

- 14 046157

A DBD can be designed to bind to a target binding site of any length, as long as it provides specific sequence recognition of the target binding site by the DBD, for example, with minimal or no off-target binding. In certain embodiments, the target binding site may modulate SCN1A expression upon binding to eTF at a level that is at least 2-fold, 5-fold, 10-fold, 20-fold, 50-fold, 75-fold, 100-fold , 150-fold, 200-fold, 250-fold, 500-fold or more compared to all other genes. In certain embodiments, the target binding site may modulate SCNIA expression upon binding to eTF at a level that is at least 2-fold, 5-fold, 10-fold, 20-fold, 50-fold, 75-fold, 100-fold , 150-fold, 200-fold, 250-fold, 500-fold, or more than the 40 nearest neighboring genes (e.g., the 40 genes closest to the chromosome, upstream or downstream of the SCN1A coding sequence ). In certain embodiments, the target binding site may be at least 5 bp, 10 bp, 15 bp, 20 bp, 25 bp, 30 bp, 35 bp .n., 40 bp, 45 bp. or 50 bp or more. The specific length of the binding site is determined by the type of DBD in the eTF. In general, the longer the binding site, the greater the specificity of binding and modulation of gene expression (e.g., longer binding sites have fewer off-target effects). In certain embodiments, an eTF comprising a DBD recognizing a longer target binding site has fewer off-target effects associated with non-specific binding (such as, for example, modulation of off-target or non-SCN1A gene expression) relative to the off-target effects observed with the eTF. having a DBD that binds to a shorter target site. In some cases, the reduction in off-target binding is at least 1.2, 1.3, 1.4, 1.5, 2, 3, 4, 5, 6, 7, 8, 9 or 10 times lower compared to comparable eTF , having a DBD that recognizes a shorter target binding site.

In certain embodiments, the DBD provided herein can be modified to have increased binding affinity such that it binds to the target binding site for a longer period of time so that the TAD conjugated to the DBD can recruit more transcription factors and/or recruit such a transcription factor for a longer period of time to have a greater impact on the expression level of the endogenous SCN1A gene. In certain embodiments, the DBD may be modified to increase its specific binding (or on-target binding) to a desired target site and/or modified to reduce its non-specific or off-target binding.

In various embodiments, the binding between the DBD or eTF and the target binding site can be determined using various methods. In certain embodiments, specific binding between a DBD or eTF and a target binding site can be determined using a mobility shift assay, a DNase protection assay, or any other in vitro method known in the art for protein-DNA binding assays. In other embodiments, the specific binding between the eTF and the target binding site can be determined using a functional assay, for example, by measuring the expression (RNA or protein) of a gene (eg, SCN1A) when the target binding site is bound to the eTF. For example, the target binding site may be located upstream of a reporter gene (such as eGFP) or the SCN1A gene in a vector contained in a cell or integrated into the genome of a cell where the cell expresses the eTF. Alternatively, the eTF expression vector can be introduced into a cell type that naturally contains the SCN1A gene. Higher levels of reporter gene (or SCN1A) expression in the presence of eTF compared to a control (e.g., no eTF or an eTF that recognizes a different target site) indicates that the DBD of the eTF binds to the target site. Suitable in vitro (eg, non-cellular) transcription and translation systems can also be used in a similar manner. In certain embodiments, the eTF that binds to the target site may have at least a 2-fold, 3-fold, 5-fold, 10-fold, 15-fold, 20-fold, 30-fold, 50-fold, 75-fold , 100-fold, 150-fold, or greater expression of a reporter gene or SCN1A compared to a control (eg, without eTF or with an eTF that recognizes a different target site).

In certain embodiments, an eTF disclosed herein that activates SCN1A recognizes a target binding site that is at least 9 bp, 12 bp, 15 bp, 18 bp, 21 bp, 24 bp, 27 bp, 30 bp, 33 bp. or 36 bp; more than 9 bp, 12 bp, 15 bp, 18 bp, 21 bp, 24 bp, 27 bp or 30 bp; or 9-33 bp, 9-30 bp, 9-27 bp, 9-24 bp, 9-21 bp, 9-18 bp, 9 -15 bp, 9-12 bp, 12-33 bp, 12-30 bp, 12-27 bp, 12-24 bp, 12-21 bp, 12-18 bp, 12-15 bp, 15-33 bp, 15-30 bp, 15-27 bp, 15-24 bp. n., 15-21 bp., 15-18 bp., 18-33 bp., 18-30 bp., 18-27 bp., 1824 bp., 18 -21 bp, 21-33 bp, 21-30 bp, 21-27 bp, 21-24 bp, 24-33 bp, 24-30 bp, 24-27 bp, 27-33 bp, 27-30 bp. or 30-33 bp. In exemplary embodiments, the eTF disclosed herein that activates SCN1A recognizes a target binding site that constitutes

- 15 046157

18-27 bp, 18 bp or 27 bp

In certain embodiments, an eTF disclosed herein that activates SCN1A recognizes a target binding site located on chromosome 2. In certain embodiments, an eTF disclosed herein that activates SCN1A recognizes a target binding site located on chromosome 2 within 110 kb, 100 kb, 90 kb, 80 kb, 70 kb, 60 kb, 50 k. bp, 40 kb, 30 kb, 20 kb, 10 kb, 5 kb, 4 kb. b., 3 kb., 2 kb. or 1 kb. upstream or downstream of TCC SCN1A. In certain embodiments, the eTF disclosed herein that activates SCN1A recognizes a target binding site that is located within 110 kb on chromosome 2. upstream of the TSS of SCN1A. In certain embodiments, the eTF disclosed herein that activates SCN1A recognizes a target binding site that is located within 110 kb on chromosome 2. downstream of the TSS SCN1A. In exemplary embodiments, such target binding sites are 18-27 bp, 18 bp. or 27 bp

In certain embodiments, the eTF disclosed herein that activates SCN1A recognizes a target binding site that is located on chromosome 2 within positions 166179652-165989571, within positions 166128050-166127958, within positions 166155414-166140590, within positions 166179652- 1661777272 or within the provisions of 1659990246-165989592 (all with reference to GRCh38.p12). In exemplary embodiments, such target binding sites are 18-27 bp, 18 bp. or 27 bp

In certain embodiments, an eTF disclosed herein that activates SCN1A recognizes a target binding site that is (i) 18-27 bp, 18 bp. or 27 bp, (ii) overlaps with a position on chromosome 2 selected from 166178880, 166177369, 166177362, 166177299, 166177299, 166155393, 166155264, 166149373, 166149176 , 166149165, 166149118, 166148953, 166148565, 166142396, 166142391, 166142344 , 166142239, 166141162, 166140928, 166140590, 165990076, 165989684, 165989571, 166155255, 166155099, 166148843, 166148361, 166142219, 166141090, 165990246, 165990193, 166149168, 166127991, 166128002, 166128037 or 166128025 (all referencing GRCh38.p12) and (iii) is capable of causing at least a 1.2-fold increase in SCN1A expression upon binding to an eTF disclosed herein.

In certain embodiments, an eTF disclosed herein that activates SCN1A (i) binds to a target site containing or consisting of any of SEQ ID NOs: 18, 25, 30, 31, or 35-66, and (ii) is capable of cause at least a 1.2-fold increase in SCN1A expression upon binding to an eTF disclosed herein.

In certain embodiments, an eTF disclosed herein that activates SCN1A recognizes a target binding site that is (i) 18-27 bp, 18 bp. or 27 bp, (ii) overlaps with a position on chromosome 2 selected from 166155255, 166155099, 166148843, 166148361, 166142219, 166141090, 165990246, 165990193, 166149168 , 166127991, 166128002, 166128037 or 166128025 (all referring to GRCh38 .p12), and (iii) is capable of causing at least a 2-fold increase in SCNIA expression upon binding to the eTF disclosed herein.

In certain embodiments, an eTF disclosed herein that activates SCN1A (i) binds to a target site comprising any of SEQ ID NO: 18, 30, 31, 37, 38, 45, 47, 48, 49, 55, 61 , 62 or 64 or consisting of it, and (ii) is capable of causing at least a 2-fold increase in SCNIA expression upon binding to an eTF disclosed herein.

In certain embodiments, an eTF disclosed herein that activates SCN1A recognizes a target binding site that is (i) 18-27 bp, 18 bp. or 27 bp, and (ii) overlaps with a position on chromosome 2 selected from 166149168, 166127991, 166128002, 166128037 or 166128025 (all referring to GRCh38.p12), and (iii) is capable of causing at least 5- fold increase in SCNIA expression upon binding of the eTF disclosed herein.

In certain embodiments, an eTF disclosed herein that activates SCN1A (i) binds to a target site containing or consisting of any of SEQ ID NOs: 18, 30, 31, 37, or 38, and (ii) is capable of inducing at least a 5-fold increase in SCN1A expression upon binding to the eTF disclosed herein.

In certain embodiments, an eTF disclosed herein that activates SCN1A recognizes a target binding site that is (i) 18-27 bp, 18 bp. or 27 bp, (ii) overlaps with a position on chromosome 2 selected from 166128002, 166128037, or 166128025 (all referring to GRCh38.p12), and (iii) is capable of causing at least a 15-fold increase in SCN1A expression when linked to the eTF disclosed herein.

In certain embodiments, an eTF disclosed herein that activates SCN1A (i) binds to a target site containing or consisting of any of SEQ ID NO: 30, 37, or 38, and (ii) is capable of inducing at least 15- fold increase in SCN1A expression upon binding of the eTF disclosed herein.

- 16 046157

In certain embodiments, an eTF disclosed herein that activates SCN1A recognizes a target binding site that is (i) 18-27 bp, 18 bp. or 27 bp, (ii) overlaps with a position on chromosome 2 selected from 166128037 or 166128025 (all referring to GRCh38.p12), and (iii) is capable of causing at least a 20-fold increase in SCN1A expression when bound open in this eTF document.

In certain embodiments, an eTF disclosed herein that activates SCN1A (i) binds to a target site containing or consisting of any of SEQ ID NO: 30 or 38, and (ii) is capable of causing at least a 20-fold increase in expression of SCN1A upon binding of an eTF disclosed herein.

In certain embodiments, an eTF disclosed herein that activates SCN1A recognizes a target binding site that is (i) 18-27 bp, 18 bp. or 27 bp, (ii) overlaps with a position on chromosome 2 at position 166128025 and (iii) is capable of causing at least a 25-fold increase in SCNIA expression when bound by an eTF disclosed herein.

In certain embodiments, an eTF disclosed herein that activates SCN1A (i) binds to a target site containing or consisting of SEQ ID NO: 30, and (ii) is capable of causing at least a 25-fold increase in SCN1A expression when bound by the disclosed in this eTF document.

In certain embodiments, an eTF disclosed herein that activates SCN1A recognizes a target binding site that is (i) 18-27 bp, 18 bp. or 27 bp and (ii) binds to a genomic region that is within at least 1 kb, 750 bp, 500 bp, 400 bp, 300 bp, 200 bp .b., 100 bp. or 50 bp genomic location having the sequence of any of SEQ ID NO: 18, 25, 30, 31 or 35-66. In certain embodiments, the target binding site is capable of causing at least a 1.2-fold, 2-fold, 5-fold, 15-fold, 20-fold, or 25-fold increase in SCN1A expression upon binding to an eTF disclosed herein.

In certain embodiments, an eTF disclosed herein that activates SCN1A recognizes a target binding site that is (i) 18-27 bp, 18 bp. or 27 bp and (ii) binds to a genomic region that at least partially overlaps with a genomic location having the sequence of any of SEQ ID NOs: 18, 25, 30, 31, or 35-66. In certain embodiments, the target binding site is capable of causing at least a 1.2-fold, 2-fold, 5-fold, 15-fold, 20-fold, or 25-fold increase in SCNIA expression upon binding to an eTF disclosed herein.

In certain embodiments, an eTF disclosed herein that activates SCN1A recognizes a target binding site having any of the following sequences: SEQ ID NO: 18, 25, 30, 31, or 35-66. In certain embodiments, the target binding site is capable of causing at least a 1.2-fold, 2-fold, 5-fold, 15-fold, 20-fold, or 25-fold increase in SCN1A expression upon binding to an eTF disclosed herein.

In certain embodiments, an eTF disclosed herein that activates SCN1A results in at least 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8 -x, 9x, 10x, 15x, 20x, 25x, 50x, 100x or more or at least 50%, 60%, 70%, 75%, 80% , 90%, 100%, 125%, 150%, 200%, 250%, 300%, 400%, or 500% or more activation of SCN1A expression (SCN1A RNA and/or Nav1.1 protein) in cell or in vivo vs. with control (eg, without eTF or with eTF that does not recognize the target site). In various embodiments, activation of SCNIA expression can be detected using PCR, Western blotting, or immunoassay methods.

In certain embodiments, an eTF disclosed herein that activates SCN1A binds to a target site that is capable of increasing SCNIA expression by at least 1.2-fold, 1.3-fold, 1.4-fold, 1.5-fold, 1. 6 times, 1.7 times, 1.8 times, 1.9 times, 2 times, 3 times, 4 times, 5 times, 8 times, 10 times, 12 times, 15 times, 18 times, 20 times, 25 times , 30 times, 40 times, 50 times, 75 times, 100 times or more or at least 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 90%, 100 %, 125%, 150%, 200%, 250%, 300%, 400%, or 500% or more relative to the control in the transcription activation assay. An exemplary SCN1A transcriptional activation assay is presented herein in Example 3. Briefly, HEK293 was transfected with a plasmid carrying eTF or an eGFP control reporter construct. 48 hours after transfection, cells are harvested, RNA is isolated and reverse transcribed, and the resulting cDNA samples are analyzed by qPCR (eg, using primers having SEQ ID NOs: 185 and 186) to quantify endogenous SCN1A transcript content. GAPDH can be used as a reference gene to determine the relative expression levels of SCN1A.

In certain embodiments, an eTF disclosed herein that

- 17 046157 activates SCN1A, is characterized by minimal off-target effects, for example, off-target effects associated with non-specific binding, such as, for example, modulation of the expression of a non-target gene or a gene other than SCN1A. In one embodiment, an eTF disclosed herein that activates SCN1A specifically activates SCN1A relative to control by at least 5-fold, 10-fold, 15-fold, 20-fold, 25-fold, 30-fold, 40-fold, or 50-fold more than the expression produced by eTF for one or more non-target genes compared to the control. In an exemplary embodiment, an eTF disclosed herein that activates SCN1A specifically activates transcription of the SCN1A gene relative to control to at least 15 times greater than transcription of the 40 nearest neighboring genes (e.g., the 40 closest genes located to the coding sequence SCN1A on chromosome 2) produced by eTF relative to control, e.g. PLA2R1, ITGB6, RBMS1, TANK, PSMD14, TBR1, SLC4A10, DPP4, FAP, IFIH1, GCA, FIGN, GRB14, COBLL1, SLC38A11, SCN3A, SCN2A, CSRNP3, GALNT3 , TTC21B, SCN9A, SCN7A, B3GALT1, STK39, CERS6, NOSTRIN, SPC25, ABCB11, DHRS9, BBS5, KLHL41, FASTKD1, PPIG, CCDC173, PHOSPHO2, KLHL23, SSB, METTL5, UBR3 and MYO3B (see Table 14). In various embodiments, transcriptional activation of the SCN1A gene can be detected using PCR methods.

In certain embodiments, an eTF disclosed herein that activates SCN1A is capable of reducing seizure frequency in a hyperthermic seizure (HTS) assay in the Scn1a ^tm1kea mouse model of Dravet syndrome. In certain embodiments, an eTF disclosed herein is capable of reducing seizure frequency at 42.6°C in the HTS assay by at least 1.2-fold, 1.3-fold, 1.4-fold, 1.5-fold, 1.6-fold times, 1.7 times, 1.8 times, 1.9 times, 2.0 times or more or by at least 20%, 30%, 40%, 50%, 55%, 60%, 65%, 70 %, 75%, 80%, 85%, 90%, 95%, 100% or more compared to control (eg, treated with PBS or exposed to an AAV vector containing an eGFP coding sequence). In certain embodiments, an eTF disclosed herein is capable of reducing seizure frequency at 42.6°C in an HTS assay such that at least 60%, 62%, 65%, 70%, 75%, 76%, 80% , 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of mice analyzed , no seizures were observed at a temperature of 42.6°C. An exemplary HTS assay is described herein in Example 6. Briefly, litters of pups generated from male Scn1a +/- mice crossed with female C57Bl/6J mice can be injected with an AAV9 vector encoding an eTF that activates SCN1A as provided herein document, or a control vector encoding eGFP via bilateral ICV in P1. Mice can be dosed with ~1.0E10-5.0E12 genome copies/mouse. The HTS assay is performed on heterozygous P26-P28 SCNIA and wild-type SCNIA mice on a mixed 129Stac X C57BL/6 background by increasing the body temperature of the mice (under controlled conditions and with body temperature controlled) by ~0.5°C every 2 minutes until the first a tonic-clonic seizure accompanied by loss of body position, or until the body temperature reaches 43°C. A mouse was considered to be seizure-free if no seizure with loss of body position was detected during the entire experimental period.

In certain embodiments, an eTF disclosed herein that activates SCN1A is capable of increasing the survival of a mouse heterozygous for SCN1A, for example, the Scn1a ^tm1kea mouse strain. In certain embodiments, an eTF disclosed herein is capable of increasing survival of heterozygous SCN1A mice at P100 by at least 1.2-fold, 1.3-fold, 1.4-fold, 1.5-fold, 1.6-fold, 1.7-fold , 1.8 times, 1.9 times, 2.0 times or more, or by at least 20%, 30%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100% or more, as compared to a control (eg, treated with PBS or treated with an AAV vector containing an eGFP coding sequence). In certain embodiments, an eTF disclosed herein is capable of increasing the survival of SCN1A heterozygous mice at P100 such that at least 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% of mice in the assay are still alive at P100. An exemplary survival assay is described herein in Example 7. Briefly, litters of pups generated from male Scn1a +/- mice crossed with female C57Bl/6J mice can be administered the AAV9 vector via bilateral ICV at P1. Mice can be dosed with ~1.0E10-5.0E12 genome copies/mouse. The number of mice surviving to P100 is determined.

In certain embodiments, an eTF provided herein that activates SCN1A may comprise a DBD of a zinc finger protein derived from a zinc finger protein, or the nuclease is an inactivated zinc finger protein. The zinc finger is a small protein structural motif that is characterized by the coordination of one or more zinc ions (Zn ²⁺ ) to stabilize the fold. Zinc finger (Znf) domains are relatively small protein motifs that contain multiple finger-like protrusions that make tandem contacts with a target region of DNA. The modular nature of the zinc finger motif allows it to bind a large number of DNA sequence combinations with high affinity and specificity, and therefore

- 18 046157 is ideal for creating a protein that can target and bind specific DNA sequences. Many engineered zinc finger arrays are based on the zinc finger domain of the mouse transcription factor Zif268. Zif268 contains three distinct zinc finger motifs that together bind a 9-bp sequence. with high affinity. A large number of zinc finger proteins have been identified and have been characterized into different types based on structure, which are further described herein. Any such zinc finger protein is useful in connection with the DBDs described herein.

Various methods are available to construct zinc finger proteins. For example, methods for designing zinc finger proteins to bind to a target DNA sequence of interest have been described, see, for example, Liu Q, et al., Design of polydactyl zinc-finger proteins for uniquely addressing within complex genomes, Proc Natl Acad Sci USA. 94 (11): 5525-30 (1997); Wright D. A. et al., Standardized reagents and protocols for engineering zinc finger nucleases by modular assembly, Nat Protoc. Nat Protoc. 2006; 1(3): 1637-52 and C. A. Gersbach and T Gaj, Synthetic Zinc Finger Proteins: The Advent of Targeted Gene Regulation and Genome Modification Technologies, Am Chem Soc 47: 2309-2318 (2014). In addition, various web-based tools for designing zinc finger proteins to bind to a target DNA sequence of interest are publicly available, see, for example, software tools for designing zinc finger nucleases and OmicX's engineered genome data analysis website, available online at omictools.com/zfns-category; and Scripps' Zinc Finger Tools design website, available online at scripps.edu/barbas/zfdesign/zfdesignhome.php. In addition, various commercially available services are available to design zinc finger proteins to bind to a target DNA sequence of interest, see for example the commercially available services or kits offered by Creative Biolabs (online at creative-biolabs.com/Design-and -Synthesis-of-ArtificialZinc-Finger-Proteins.html), the Zinc Finger Consortium Modular Assembly Kit available from Addgene (online at addgene.org/kits/zfc-modular-assembly/), or the CompoZr Custom ZFN Service from Sigma Aldrich (Online at sigmaaldrich.com/life-science/zinc-finger-nuclease-technology/customzfn.html).

In certain embodiments, eTFs provided herein that activate SCN1A contain a DBD containing one or more zinc fingers, or are derived from a zinc finger protein DBD. In some cases, the DBD contains multiple zinc fingers, with each zinc finger linked to another zinc finger or other domain at either its N-terminus, C-terminus, or both via an amino acid linker. In some cases, the DBD provided herein contains one or more zinc fingers from one or more of the types of zinc fingers described in table. 9. In some cases, the DBD provided herein contains a plurality of zinc finger structures or motifs or a plurality of zinc fingers having one or more of SEQ ID NOs: 152-167 or any combination thereof. In certain embodiments, the DBD comprises X-[ZF-X]n and/or [X-ZF]n-X, wherein the ZF is a zinc finger domain containing any of the motifs listed in Table 1. 9 (for example, any of SEQ ID NO: 136-146), X is an amino acid linker containing 1-50 amino acids, and n is an integer from 1 to 15, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15, wherein each ZF may independently have the same sequence or a different sequence from other ZF sequences in the DBD, and wherein each linker X may independently have the same sequence or sequence different from other X sequences in the DBD. Each zinc finger may be linked to another sequence, a zinc finger, or a domain at its C-terminus, N-terminus, or both. In a DBD, each linker X may be identical in sequence, length and/or property (eg, flexibility or charge) or different in sequence, length and/or property. In some cases, two or more linkers may be identical while other linkers are different. In exemplary embodiments, the linker may be derived from or derived from zinc finger linking sequences found in one or more of the naturally occurring zinc finger proteins presented in Table 1. 9. In other embodiments, suitable linker sequences include, for example, linkers of 5 or more amino acids in length. See also US Patent No. 6479626; 6903185 and 7153949 for exemplary linker sequences of 6 or more amino acids in length, each of which is incorporated herein in its entirety. DBD proteins provided herein may include any combination of suitable linkers between individual zinc fingers of the protein. The DBD proteins described herein may include any combination of suitable linkers between individual zinc fingers of the protein.

In certain embodiments, eTFs provided herein that activate SCN1A comprise a DBD containing one or more classical zinc fingers. The classical zinc finger C2H2 contains two cysteines in one chain and two his residues

- 19 046157 tidine in another chain, coordinated by a zinc ion. The classical zinc finger domain contains two β-sheets and one α-helix, with the α-helix interacting with the DNA molecule and forming the basis for DBD binding to the target site and can be called the recognition helix. In exemplary embodiments, the zinc finger recognition helix contains at least one amino acid substitution at position -1, 2, 3, or 6, thereby altering the binding specificity of the zinc finger domain. In other embodiments, the DBD provided herein contains one or more non-classical zinc fingers, such as C2-H2, C2-CH, and C2-C2.

In another embodiment, an eTF provided herein that activates SCN1A contains a DBD containing a zinc finger motif having the following structure: LEPGEKP - [YKCPECGKSFS X HQRTH TGEKP]n - YKCPECGKSFS X HQRTH - TGKKTS (SEQ ID NO: 147), where n is an integer from 1 to 15, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15, and each X is independently a sequence recognition helix (e.g., recognition helix) capable of binding to 3 bp. target sequence. In exemplary embodiments, n is 3, 6, or 9. In a particularly preferred embodiment, n is 6. In various embodiments, each X may independently have the same amino acid sequence or a different amino acid sequence compared to other X sequences in the DBD. In an exemplary embodiment, each X is a sequence containing 7 amino acids that was designed to interact with 3 bp. target binding site of interest using Scripps' Zinger Finger Design Tool located on the Internet at scripps.edu/barbas/zfdesign/zfdesignhome.php.

Since each zinc finger in a DBD recognizes 3 bp, the number of zinc fingers included in the DBD informs the length of the binding site recognized by the DBD, e.g., a DBD with 1 zinc finger recognizes a target binding site that is 3 bp, a DBD with 2 zinc finger DBD recognizes a 6 bp target binding site, 3 zinc finger DBD recognizes a 9 bp target binding site, 4 zinc finger DBD recognizes a 12 bp target binding site, DBD 5 zinc finger DBD recognizes a 15 bp target binding site, 6 zinc finger DBD recognizes a 18 bp target binding site, 9 zinc finger DBD recognizes a 27 bp target binding site, etc. In general, DBDs that recognize longer target binding sites will exhibit greater binding specificity (e.g., less off-target binding or nonspecific binding).

In other embodiments, an eTF provided herein that activates SCN1A comprises a DBD that is derived from a naturally occurring zinc finger protein by making one or more amino acid substitutions in one or more zinc finger domain recognition helices to alter the binding specificity of the DBD ( for example, changing the target site recognized by the DBD). The DBD presented herein can be derived from any naturally occurring zinc finger protein. In various embodiments, such a DBD may be derived from any species of zinc finger protein, such as mouse, rat, human, etc. In an exemplary embodiment, the DBD provided herein is derived from human zinc finger protein. In certain embodiments, the DBD provided herein is derived from a naturally occurring protein listed in Table. 9. In an exemplary embodiment, the DBD protein provided herein is derived from a human EGR zinc finger protein, such as EGR1, EGR2, EGR3, or EGR4.

In certain embodiments, an eTF provided herein that activates SCN1A contains a DBD that is derived from a naturally occurring protein by modifying the DBD to increase the number of zinc finger domains in the DBD protein by repeating one or more zinc fingers within the DBD found in nature of protein. In certain embodiments, such modifications include duplication, tripling, quadrupling, or further multiplication of zinc fingers within the DBD of a naturally occurring protein. In some cases, one zinc finger is multiplied from the DBD of a human protein, for example, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or more copies of the same zinc finger motif are repeated in the DBD of eTF. In some cases, a set of zinc fingers from the DBD of a naturally occurring protein is multiplied, for example, a set of 3 zinc fingers from the DBD of a naturally occurring protein is duplicated to produce an eTF having a DBD with 6 zinc fingers, repeated three times to produce a DBD of an eTF with 9 zinc fingers or repeated four times to obtain DBD eTF with 12 zinc fingers, etc. In some cases, a set of zinc fingers from the DBD of a naturally occurring protein is partially replicated to form a DBD eTF having more zinc fingers, e.g., a DBD eTF contains four zinc fingers, with the zinc fingers being one copy of the first zinc finger, one copy of the second zinc finger and two copies of the third zinc finger from a naturally occurring protein,

- 20 046157 only four zinc fingers in DBD eTF. Such DBDs are then further modified by making one or more amino acid substitutions in one or more zinc finger domain recognition helices to change the binding specificity of the DBD (eg, changing the target site recognized by the DBD). In exemplary embodiments, the DBD is derived from a naturally occurring human protein, such as a human EGR zinc finger protein, such as EGR1, EGR2, EGR3, or EGR4.

Human EGR1 and EGR3 are characterized by a three-fingered C2H2 zinc finger DBD. The general binding rules for zinc fingers stipulate that all three fingers interact with their cognate DNA sequence of similar geometry, using the same amino acids in the alpha helix of each zinc finger to determine the specificity or sequence recognition of the target binding site. Such binding rules allow the DBD of EGR1 or EGR3 to be modified to create a DBD that recognizes the desired target binding site. In some cases, the 7 amino acid DNA recognition helix in the EGR1 or EGR3 zinc finger motif is modified according to published zinc finger design rules. In certain embodiments, each zinc finger in a three-finger EGR1 or EGR3 DBD is modified, for example, by changing the sequence of one or more recognition helices and/or by increasing the number of zinc fingers in the DBD. In certain embodiments, EGR1 or EGR3 is reprogrammed to recognize a target binding site of at least 9, 12, 15, 18, 21, 24, 27, 30, 33, 36, or more base pairs in the desired target site. In certain embodiments, such a DBD derived from ERG1 or EGR3 contains at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or more zinc fingers. In an exemplary embodiment, the one or more zinc fingers in the DBD contain at least one amino acid substitution at position -1, 2, 3, or 6 of the recognition helix.

In various embodiments, an eTF that activates SCN1A containing a DBD derived from EGR1 or EGR3 has a DNA binding specificity that is different from the binding specificity of naturally occurring EGR1 or EGR3, e.g., the DBD recognizes a target binding site having a sequence different from the sequence binding site recognized by ^L modified EGR1 or EGR3: (GCG(T/G)GGGCG) (SEQ ID NO: 182).

In other embodiments, an eTF provided herein that activates SCN1A contains a DBD, which is a gRNA/Cas complex. CRISPR (Clustered Regular Intermediate Short Palindromic Repeats)/Cas9 is a genome editing tool that allows site-specific targeting of the genome. The CRISPR/Cas type II system is a prokaryotic adaptive immune response system that uses non-coding RNAs to direct the Cas9 nuclease to induce site-specific DNA cleavage. The CRISPR/Cas9 system has been used to create a simple, RNA-programmable method for genome editing in mammalian cells. A single guide RNA (sgRNA) can be designed to direct Cas9 nuclease to a specific location in the genome, which is then bound by the gRNA/Cas9 complex. gRNA can be designed to bind to a target site of interest using a variety of methods and tools. For example, methods for creating a gRNA to bind to a target DNA sequence of interest are described in Aach, et al. Flexible algorithm for identifying specific Cas9 targets in genomes. BioRxiv, Cold Spring Harbor Labs, doi: http://dx.doi.org/10.1101/005074 (2014); Bae, et al. Cas-OFFinder: a fast and versatile algorithm that searches for potential off-target sites of Cas9 RNA-guided endonucleases. Bioinformatics. 30(10): 1473-1475 (2014); Doench, J. G. et al. Optimized sgRNA design to maximize activity and minimize off-target effects of CRISPRCas9. Nat Biotech 34, 184-191 (2016); Gratz, et al. Highly specific and efficient CRISPR/Cas9-catalyzed homology-directed repair in Drosophila. Genetics. 196(4):961-971 (2014); Heigwer, et al. E-CRISP: fast CRISPR target site identification. Nat Methods. 11(2):122-123 (2014); Ma, et al. A guide RNA sequence design platform for the CRISPR/Cas9 system for model organism genomes. Biomed Res Int. doi:http://doi.org/10.1155/2013/270805 (2013); Montague, et al. CHOPCHOP: a CRISPR/Cas9 and TALEN web tool for genome editing. Nucleic Acids Res. 42(W1):W401-W407 (2014); Liu, et al. CRISPR-ERA: a comprehensive design tool for CRISPR-mediated gene editing, repression and activation. Bioinformatics. 31(22):3676-3678 (2015); Ran, et al. In vivo genome editing using Staphylococcus aureus Cas9. Nature. 520(7546):186-191 (2015); Wu, et al. Target specificity of the CRISPR-Cas9 system. Quant Biol. 2(2):59-70 (2015); Xiao et al. CasOT: a genome-wide Cas9/gRNA off-target searching tool. Bioinformatics. 30(8):11801182 (2014); Zetsche, et al. Cpf1 is a single RNA-guided endonuclease of a Class 2 CRISPR-Cas System. Cell. 163(3):759–771 (2105). In addition, various web-based tools are publicly available to design gRNAs to bind to a target DNA sequence of interest, see for example the CRISPR gRNA design tool available from AUTM online at atum.bio/eCommerce/cas9/input?multipleContacts=false; CRISPRa/i gRNA design tool, access

- 21 046157 ny from the Broad Institute on the Internet at portals.broadinstitute.org/gpp/public/analysis-tools/sgrnadesign-crisprai; the E-CRISP development tool available from the German Cancer Research Center DKFZ, accessible online at e-crisp.org/E-CRISP/; and the Knockout Guide Design tool, available from Synthego online at design.synthego.com/#/. In addition, various commercially available services are available to design a gRNA to bind to a target DNA sequence of interest, see for example the commercially available services offered by IDT (online at idtdna.com/site/order/designtool/index/CRISPR_SEQUENCE). ThermoFisher (online at thermofisher.com/order/custom-oligo/crispr) and GenScript (online at genscript.com/gRNA-design-tool.html).

In exemplary embodiments, a DBD that is a gRNA/Cas complex comprises a nuclease-deactivated Cas or dCas protein, such as, for example, dCas9, such as Staphylococcus aureus nuclease-deactivated (dSaCas9) or Streptococcus pyogenes nuclease-deactivated Cas9 (dSpCas9). The gRNA is presented as a sequence containing a targeting region that targets the gRNA/Cas complex to the desired target site and a scaffold region that facilitates interaction with the Cas protein. Any suitable gRNA framework may be used in connection with the gRNAs provided herein. In an exemplary embodiment, the gRNA is a single gRNA or sgRNA and contains the following framework sequence: 5'-GTTTTAGTACTCTGGAAACAGAATCTACTAAAACAAGGCA AAATGCCGTGTTTATCTCGTCAACTTGTTGGCGAGA-3' (SEQ ID NO: 183). The targeting region of the guide RNA is added to the 5' end of the scaffold sequence to form the complete sgRNA. In certain embodiments, the gRNA and the dCas protein may be expressed from the same expression cassette. In certain embodiments, the U6 promoter is used for gRNA expression. In other embodiments, the gRNA can be expressed in a cell that has been engineered to stably express the dCas-TAD protein, for example, either by stably integrating dCas into the genome or on a plasmid that is stably maintained off chromosomes.

In other embodiments, an eTF provided herein that activates SCN1A may comprise a DBD from TALEN, derived from TALEN, or which is a nuclease-inactivated TALEN. Transcription activator-like effector nucleases (TALENs) are restriction enzymes that contain a DBD and a nuclease domain that can be engineered to cut specific DNA sequences. TALENs are created by conjugating a TAL effector DNA-binding domain to a DNA cleavage domain (e.g., nuclease). Transcription activator-like effectors (TALEs) can be designed to bind to a desired target DNA sequence, thereby targeting the nuclease domain to a specific location.

TAL effectors are bacterial proteins from Xanthomonas bacteria. The DNA binding domain contains a highly conserved repeat sequence of 33-34 amino acids with diverging amino acids 12 and 13. These two positions, called Repeat Variable Diresidue (RVD), are highly variable and show a strong correlation with specific nucleotide recognition. This direct relationship between amino acid sequence and DNA recognition allows the design of DBDs that specifically target the desired sequence by selecting a combination of repeat segments containing the corresponding RVDs.

There are different ways to develop TALE. For example, methods for designing a TALE to bind to a target DNA sequence of interest are described in T. Cermak et al., Nucleic Acids Research. 39 (12):e82 (2011); F. Zhang F et al., Nature Biotechnology. 29 (2):149-53 (2011); R. Morbitzer et al., Nucleic Acids Research. 39 (13):5790-9 (2011); T. Li et al., Nucleic Acids Research. 39 (14):631525 (2011); R. Geissler et al., PLOS One. 6(5): e19509 (2011) and E. Weber et al., PLOS One. 6(5): e19722 (2011). In addition, various web-based tools are publicly available for designing TALEs to bind to a target DNA sequence of interest, see for example E-Talen, available online at e-talen.org/E-TALEN/TAL and the Effector Nucleotide Targeter 2.0 tool. , available online at tall-nt.cac.cornell.edu/node/add/single-tale. In addition, various commercially available services are available for generating TALEs to bind to a target DNA sequence of interest, see for example commercially available services offered by OmicX (online at omictools.com/), Addgene (online at addgene.org /talen/guide/ or ThermoFisher (online: thermofisher.com/us/en/home/life-science/genome-editing/geneart-tals/tal-designtool.html) Alternatively, a publicly available program (DNAWorks) can be used for designing oligonucleotides suitable for TALE assembly, see, for example, D. Hoover D Methods in Molecular Biology.852:215-23 (2012).

Transcription modulation domains.

eTFs provided herein that activate SCN1A may contain any suitable domain that is capable of recruiting one or more protein factors that

- 22 046157 can modulate transcription (eg, RNA polymerase II, CBP/p300, CREB or KRAB) or gene expression level from a gene of interest when the eTF is bound to a target site via a DBD (eg, zinc finger DBD, gRNA/Cas DBD or TALE DBD). In certain embodiments, such a domain recruits protein factors that increase the level of transcription or expression of a gene of interest, and is a transcription activation domain (TAD). In other embodiments, such a domain recruits protein factors that reduce the level of transcription or expression of a gene of interest, and is a transcriptional repressor domain (TRD). In certain embodiments, the transcription modulation domain (TAD or TRD) may be a synthetically engineered domain. In other embodiments, the transcription modulation domain (TAD or TRD) may be derived from a naturally occurring protein, such as a transcription factor, transcription coactivator, transcription corepressor, or silencer protein. In various embodiments, the transcription modulation domain (TAD or TRD) can be derived from any type of protein, such as mouse, rat, monkey, virus, or human.

In one exemplary embodiment, a TAD suitable for use in eTFs provided herein that activate SNC1A is derived from a viral protein. Exemplary TADs derived from viral proteins include, for example, the TAD domain of VP64 (SEQ ID NO: 133), VPR (SEQ ID NO: 132), VP16, VP128, p65, p300, or any functional fragment or variant thereof, or a sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to it .

In another exemplary embodiment, a TAD suitable for use in eTFs provided herein that activate SCN1A is derived from a human protein. Exemplary TADs derived from human proteins include, for example, CBP/p300-interacting transactivator 2 (CITED2) TAD domain (SEQ ID NO: 134), CBP/p300-interacting transactivator 4 (CITED4) TAD (SEQ ID NO: 135), EGR1 (SEQ ID NO: 176), CREB3 (SEQ ID NO: 224) or EGR3 (SEQ ID NO: 175), or any functional fragment or variant thereof, or a sequence having at least 80% identity , 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, in relation to it.

In certain embodiments, the eTF that activates SCN1A contains a zinc finger DBD that is conjugated to a transcription activation domain or TAD. In various embodiments, the zinc finger DBD may be conjugated to a TAD from a viral protein, such as VP64 or VPR, or to a TAD from a human protein, such as CITED2, CITED4, or CREB3. In certain embodiments, a zinc finger DBD derived from a human protein, such as EGR1 or EGR3, is conjugated to a TAD derived from a human protein, such as CITED2, CITED4, or CREB3. In certain embodiments, a zinc finger DBD derived from a human protein, such as EGR1 or EGR3, is conjugated to a VP64 or VPR TAD. In certain embodiments, a synthetic zinc finger DBD or zinc finger DBD having less than 75% sequence identity to a human protein, e.g., EGR1 or EGR3, is conjugated to a TAD derived from a human protein, e.g., CITED2, CITED4 or CREB3. In certain embodiments, a synthetic zinc finger DBD or zinc finger DBD having less than 75% sequence identity to a human protein, such as EGR1 or EGR3, is conjugated to a VP64 or VPR TAD.

In certain embodiments, the dCas protein is conjugated to a TAD. In various embodiments, dCas9 may be conjugated to a TAD from a viral protein, such as VP64 or VPR, or to a TAD from a human protein, such as CITED2, CITED4, or CREB3. In exemplary embodiments, dCas9 is conjugated to a VP64 or VPR TAD.

In certain embodiments, the TALE protein is conjugated to a TAD. In various embodiments, the TALE may be conjugated to a TAD from a viral protein, such as VP64 or VPR, or to a TAD from a human protein, such as CITED2, CITED4, or CREB3. In exemplary embodiments, TALE is conjugated to VP64 or VPR TAD.

eTFs that activate SCN1A and are highly homologous to human proteins.

In certain embodiments, an eTF disclosed herein that activates SCN1A has a high percentage of identity to one or more human proteins (as further described below). In certain embodiments, such eTFs have an overall sequence identity of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94% , 95%, 96%, 97%, 98%, or 99%, relative to one or more human proteins. In certain embodiments, such eTFs exhibit reduced immunogenicity compared to eTFs having a lower overall percentage of sequence identity to one or more human proteins. In various embodiments, the reduction in immunogenicity can be achieved by

- 23 046157 measure using elispot assay, immunoassay or in silico method. In certain embodiments, such eTFs may comprise a DBD derived from human EGR1 or EGR3 and a TAD derived from human EGR1, EGR3, CITED2, CITED4 or CREB3. Such eTFs have little or no immunogenicity when administered to a subject compared to eTFs that have a lower percentage of identity to human protein sequences.

In certain embodiments, an eTF provided herein that activates SNC1A has a sequence identity of at least 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84 %, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, relative to one or more human proteins. When an eTF provided herein that activates SCN1A contains a DBD and a TAD derived from the same protein, the percentage identity with the human protein can be determined by calculating the total number of amino acid residues in the eTF that correspond to the human protein from which it was derived obtained (eg EGR1 or EGR3) divided by the total number of amino acid residues in the eTF. When the eTF conferring active regulation of SCN1A contains a DBD from one human protein and a TAD derived from another human protein, the percent human identity can be determined by separately calculating the percent human identity for each domain and summing the two together, e.g. i) calculating the total number of amino acid residues in the DBD that correspond to the human protein from which it was derived (eg, EGR1 or EGR3) divided by the total number of amino acid residues in the eTF; (ii) calculating the total number of amino acid residues in the TAD that correspond to the human protein from which it was derived (eg, CITED2, CITED4, or CREB3) divided by the total number of amino acid residues in the eTF; and (iii) summing the sum of (i) and (ii). In such an embodiment, the domains are divided as follows: a first domain extends from the N-terminus of the eTF to the beginning of the coding sequence for the second domain, and a second domain extends from the beginning of the coding sequence for the second domain through the C-terminus of the eTF (for example, for an eTF having an NLS configuration -DBD-linker-NLS-TAD, the first domain will be NLS-DBD-linker and the second domain will be NLS-TAD). When the eTF provided herein that activates SNC1A contains a DBD from one human protein and two TADs derived from one or more different human proteins, the percent identity to human can be determined by separately calculating the percent identity to human for each domain and summing all three together, for example, (i) calculating the total number of amino acid residues in the DBD that correspond to the human protein from which it was derived (eg, EGR1 or EGR3) divided by the total number of amino acid residues in the eTF; (ii) calculating the total number of amino acid residues in the first TAD that correspond to the human protein from which it was derived (eg, CITED2, CITED4, or CREB3) divided by the total number of amino acid residues in the eTF; (iii) calculating the total number of amino acid residues in the second TAD that correspond to the human protein from which it was derived (eg, CITED2, CITED4, or CREB3) divided by the total number of amino acid residues in the eTF; and (iv) summing the sum of (i), (ii) and (iii). In such an embodiment, the domains are divided as follows: a first domain extends from the N-terminus of the eTF to the start of the coding sequence for the second domain, a second domain extends from the start of the coding sequence for the second domain through the start of the coding sequence for the third domain, and a third domain extends from the start of the coding sequence sequences for the third domain through the C-terminus of the eTF (for example, for an eTF having the configuration NLSTAD1-linker-NLS-DBD-linker-NLS-TAD2, the first domain would be ^S-TADI^uh^p, the second domain would be NLS-DBD- linker, and the third domain will be NLS-TAD2). Percent identity to one or more human proteins, as described in this section, can be determined using the percent identity output obtained from the standard protein BLAST tool available from NCBI (e.g., the blastp set alignment tool using the blastp algorithm ( protein -> protein) with default parameters) is available online at the NCBI website at blast. ncbi. nlm. nih. gov/.

In certain embodiments, an eTF provided herein that activates SCN1A has the advantage of eliciting little, minimal, or no adverse immune response in humans due to high sequence identity to naturally occurring human proteins. In certain embodiments, an eTF provided herein that activates SCN1A causes decreased immunogenicity, e.g., a decrease in immunogenicity of at least 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4. 4.5, 5, 10, 15, 20, 25, 30, 35, 40, 45 or 50 times or more the immunogenicity observed with an eTF having a lower percentage identity to one or more human proteins, e.g. , an eTF having less than 50%, 55%, 65%, or 70% sequence identity to one or more human proteins. In some cases, reduction in immunogenicity can be measured using an elispot assay, an immunoassay, or an in

- 24 046157 silico. Gene therapy that is low or minimally immunogenic has several advantages, including improved patient tolerability, reduced dosage required to achieve therapeutic effect, prolonged therapeutic effects after a single administration, ability to be administered multiple times or in multiple doses if necessary, sustained therapeutic benefit. efficacy over a longer period of time per administration, increased safety and/or increased efficacy of gene therapy.

In certain embodiments, eTFs provided herein that activate SCN1A and have a high percentage of sequence identity to one or more human proteins include DBDs and TADs derived from one or more naturally occurring human proteins. In certain embodiments, such eTFs may comprise a DBD derived from any naturally occurring human protein containing a DBD. In illustrative embodiments, an eTF provided herein that activates SCN1A and has a high percentage of sequence identity to one or more human proteins includes a DBD derived from a naturally occurring zinc finger protein, such as, for example, any from designs 5-27, 36-41 or 44-53, listed in table. 1. In certain embodiments, an eTF provided herein that activates SCN1A and has a high percentage of sequence identity to one or more human proteins includes a DBD derived from a human EGR protein, such as EGR1, EGR2, EGR3, or EGR4 . In illustrative embodiments, an eTF provided herein that activates SCN1A and has a high percentage of sequence identity to one or more human proteins includes a DBD derived from human EGR1 or EGR3. In various embodiments, an eTF provided herein that activates SCN1A and has a high percentage of sequence identity to one or more human proteins includes a DBD derived from a human zinc finger protein in which there are minimal amino acid changes (e.g., 1 , 2, 3, 4, 5, 6, 7 or 1-2, 1-3, 1-4, 1-5, 1-6, 1-7, 2-3, 2-4, 2-5, 2 -6, 27, 3-4, 3-5, 36, or 3-7 amino acid substitutions) were made in one or more zinc finger domains of the DBD to alter the binding specificity of the DBD to recognize the target binding site of interest. Such sequence modifications are preferably made in the recognition helices of the DBD zinc finger domains, while the rest of the DBD or human zinc finger protein (including the TADs) remains unmodified to maintain as much sequence identity with the native human protein as possible.

In certain embodiments, eTFs provided herein that activate SCN1A and have a high percentage of sequence identity to one or more human proteins contain one or more transcriptional modulation domains (e.g., TADs) derived from a human protein conjugated to a DBD, derived from human protein. In various embodiments, the transcription modulation domain may be derived from any naturally occurring human protein having a domain capable of recruiting one or more protein factors that can modulate transcription (e.g., RNA polymerase II, coactivator protein, or co-repressor protein) or level gene expression from the gene of interest when the eTF binds to the target site through the DBD. In exemplary embodiments, the TAD is derived from a human EGR protein, such as, for example, human EGR1, EGR2, EGR3, or EGR4, or from a human protein, such as, for example, human CITED2 or CITED4 protein. In an exemplary embodiment, an eTF provided herein that activates SCN1A and has a high percentage of sequence identity to one or more human proteins includes a TAD from a human EGR1 or EGR3 protein. In another exemplary embodiment, an eTF provided herein that activates SCN1A and has a high percentage of sequence identity to one or more human proteins includes a TAD from a human CITED2 or CITED4 protein.

In one embodiment, an eTF provided herein that activates SCN1A and has a high percentage of sequence identity to one or more human proteins may comprise a human DBD (hDBD) and a human TAD (hTAD) (e.g., hTAD-hDBD or hDBD- hTAD), and hDBD and hTAD can be derived from the same human protein or from different human proteins. In another embodiment, an eTF provided herein having a high percentage of sequence identity to one or more human proteins may comprise an hDBD and two hTADs, wherein the hDBD and hTAD are derived from the same human protein, the hDBD being derived from the first human protein and both hTADs are derived from a second human protein, hDBD and one hTAD are derived from the first human protein, and the second hTAD is derived from

- 25 046157 a second human protein, or hDBD is derived from a first human protein, one hTAD is derived from a second human protein, and a second hTAD is derived from a third human protein (for example, hTAD1-hDBD-hTAD1, hTAD1-hDBD-hTAD2, hTAD1-hTAD1- hDBD, hTAD1-hTAD2-hDBD, hDBD-hTAD1-hTAD1 or hDBD-hTAD1-hTAD2).

In exemplary embodiments, an eTF provided herein having a high percentage of sequence identity to one or more human proteins comprises any of the following configurations: (i) hDBD and hTAD, both derived from human EGR1; (ii) hDBD and hTAD, both derived from human EGR3; (iii) hDBD derived from human EGR1 and hTAD derived from CITED2 (eg, hEGR1 DBD-hCITED2 TAD or hCITED2 TAD-hEGR1 DBD); (iv) hDBD derived from human EGR1 and hTAD derived from human CITED4 (eg, hEGR1 DBD-hCITED4 TAD or hCITED4 TAD-hEGR1 DBD); (v) hDBD derived from human EGR3 and hTAD derived from CITED2 (eg, hEGR3 DBD-hCITED2 TAD or hCITED2 TAD-hEGR3 DBD); (vi) hDBD derived from human EGR3 and hTAD derived from human CITED4 (eg, hEGR3 DBD-hCITED4 TAD or hCITED4 TAD-hEGR3 DBD); (vii) hDBD derived from human EGR1 and two hTADs derived from CITED2 (e.g., hCITED2 TAD-hEGR1 DBDhCITED2 TAD, hCITED2 TAD-hCITED2 TAD-hEGR1 DBD, or hEGR1 DBD-hCITED2 TAD-hCITED2 TAD); (viii) hDBD derived from human EGR1 and two hTADs derived from human CITED4 (eg, hCITED4 TAD-hEGR1 DBD-hCITED4 TAD, hCITED4 TAD-hCITED4 TAD-hEGR1 DBD, or hEGR1 DBD-hCITED4 TAD-hCITED4 TAD); (ix) hDBD derived from human EGR3 and two hTADs derived from human CITED2 (eg, hCITED2 TAD-hEGR3 DBD-hCITED2 TAD, hCITED2 TADhCITED2 TAD-hEGR3 DBD, or hEGR3 DBD-hCITED2 TAD-hCITED2 TAD); (x) hDBD derived from human EGR3 and two hTADs derived from human CITED4 (eg, hCITED4 TAD-hEGR3 DBDhCITED4 TAD, hCITED4 TAD-hCITED4 TAD-hEGR3 DBD, or hEGR3 DBD-hCITED4 TAD-hCITED4 TAD); (xi) hDBD derived from human EGR1, first hTAD derived from human CITED2, second hTAD derived from human CITED4 (e.g. hCITED2 TAD-hEGR1 DBD-hCITED4 TAD, hCITED4 TAD-hEGR1 DBD-hCITED2 TAD, hCITED2 TAD-hCITED4 TAD-hEGR1 DBD, hCITED4 TADhCITED2 TAD-hEGR1 DBD, hEGR1 DBD-hCITED4 TAD-hCITED2 TAD or hEGR1 DBD-hCITED2 TADhCITED4 TAD); or (xii) an hDBD derived from human EGR3, a first hTAD derived from human CITED2, a second hTAD derived from human CITED4 (e.g., hCITED2 TAD-hEGR3 DBDhCITED4 TAD, hCITED4 TAD-hEGR3 DBD-hCITED2 TAD, hCITED2 TAD-hCITED4 TAD -hEGR3 DBD, hCITED4 TAD-hCITED2 TAD-hEGR3 DBD, hEGR3 DBD-hCITED4 TAD-hCITED2 TAD or hEGR3 DBDhCITED2 TAD-hCITED4 TAD).

In certain embodiments, an eTF provided herein that activates SCN1A and has a high percentage of sequence identity to one or more human proteins comprises any of the following: (i) a sequence comprising any of SEQ ID NOs: 103-124, 128 -131, 205, 207, 209, 213, 217, 219, 221 or 223; (ii) a sequence comprising any of SEQ ID NOs: 92-98; (iii) a sequence having a sequence identity of at least 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96 %, 97%, 98% or 99%, relative to any of the sequences (i) or (ii); or (iv) a functional fragment or variant of any of sequences (i), (ii) or (iii). In exemplary embodiments, such eTFs are capable of increasing SCN1A expression by at least 2-fold, 5-fold, 10-fold, 15-fold, 20-fold, 25-fold, 30-fold, 40-fold, 50-fold, or greater than control, or at least at least 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 90%, 100%, 125%, 150%, 200%, 250%, 300%, 400% or 500% or more compared to control. In exemplary embodiments, such eTFs have an overall sequence identity of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94% , 95%, 96%, 97%, 98% or 99% relative to one or more human proteins. In certain embodiments, such eTFs exhibit reduced immunogenicity compared to eTFs having a lower overall percentage of sequence identity to one or more human proteins. In various embodiments, the reduction in immunogenicity can be measured using an elispot assay, an immunoassay, or an in silico method.

In certain embodiments, an eTF provided herein that activates SCN1A and has a high percentage of sequence identity to one or more human proteins may further comprise one or more amino acid sequences or domains in addition to the DBD and TAD domains, e.g., a nuclear signal localization or linker, etc. In addition, a polynucleotide encoding an eTF provided herein that has a high percentage of sequence identity to one or more human proteins may further comprise one or more nucleic acid sequences in addition to the coding sequence for the eTF, e.g., a promoter, an enhancer, a polyA -tail, etc. In such embodiments, one or more additional amino acid sequences and/or following

- 26 046157 nucleic acid sequences are preferably human sequences derived from human sequences, or have a sequence identity of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% relative to human protein.

Exemplary SCN1A eTFs.

In certain embodiments, an eTF disclosed herein that activates SCN1A comprises a DNA binding domain comprising one or more zinc finger domains containing a recognition helix comprising any of SEQ ID NO: 152167. In certain embodiments, disclosed herein The eTF that activates SCN1A contains a DNA binding domain containing at least one, two, three, four, five, six, seven, eight, nine, ten, eleven, or twelve zinc finger domains, each zinc finger domain independently contains a recognition helix containing any of SEQ ID NOs: 152-167. In certain embodiments, an eTF disclosed herein that activates SCN1A comprises a DNA binding domain comprising six zinc finger domains, with each zinc finger domain independently comprising a recognition helix comprising any of SEQ ID NOs: 152-167. In certain embodiments, an eTF disclosed herein that activates SCN1A comprises a DNA binding domain comprising nine zinc finger domains, with each zinc finger domain independently comprising a recognition helix comprising any of SEQ ID NOs: 152-167. In exemplary embodiments, such eTFs comprise a DNA binding domain comprising SEQ ID NO: 147, wherein each X is independently selected from any of SEQ ID NOs: 152-167, and n is 6 or 9.

In certain embodiments, an eTF disclosed herein that activates SCN1A comprises a DNA binding domain comprising any of the following: (i) a sequence comprising RSDNLVR x REDNLHT x RSDELVR x QSGNLTE x TSGHLVR x QNSTLTE (SEQ ID NO: 148), where x may be a linker of 1-50 amino acids, (ii) a sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity, according to with respect to SEQ ID NO: 148, or (ii) a functional fragment from (i) or (ii). In certain embodiments, such an eTF further comprises one or more TADs selected from VP64, VPR, CITED2, CITED4, or CREB3. In one embodiment, such an eTF comprises a VPR TAD domain conjugated to the C-terminus of a DBD. In certain embodiments, such an eTF comprises a CITED2 TAD conjugated to the N-terminus, the C-terminus, or the N-terminus and the C-terminus of a DBD. In certain embodiments, such an eTF comprises a CITED4 TAD conjugated to the N-terminus, the C-terminus, or the N-terminus and the C-terminus of a DBD. In certain embodiments, such an eTF comprises two CITED4 TADs conjugated to the N-terminus or C-terminus of a DBD. In certain embodiments, such eTF is capable of binding to the target site comprising SEQ ID NO: 18 and increasing SCN1A expression by at least 2-fold, 5-fold, 10-fold, 15-fold, 20-fold, 25-fold, 30-fold, 40-fold , 50 times or more times compared to the control, or at least 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 90%, 100%, 125%, 150 %, 200%, 250%, 300%, 400% or 500% or more compared to control.

In certain embodiments, an eTF disclosed herein that activates SCN1A comprises a DNA binding domain comprising any of the following: (i) a sequence comprising RSDNLVR x HRTTLTN x REDNLHT x TSHSLTE x QSSSLVR x REDNLHT (SEQ ID NO: 149), where x may be a linker of 1-50 amino acids, (ii) a sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity, according to with respect to SEQ ID NO: 149, or (ii) a functional fragment from (i) or (ii). In certain embodiments, such an eTF further comprises one or more TADs selected from VP64, VPR, CITED2, CITED4, or CREB3. In one embodiment, such an eTF comprises a VPR TAD domain conjugated to the C-terminus of a DBD. In certain embodiments, such an eTF comprises a CITED2 TAD conjugated to the N-terminus, the C-terminus, or the N-terminus and the C-terminus of a DBD. In certain embodiments, such an eTF comprises a CITED4 TAD conjugated to the N-terminus, the C-terminus, or the N-terminus and the C-terminus of a DBD. In certain embodiments, such an eTF comprises two CITED4 TADs conjugated to the N-terminus or C-terminus of a DBD. In certain embodiments, such eTF is capable of binding to the target site comprising SEQ ID NO: 30 and increasing SCN1A expression by at least 2-fold, 5-fold, 10-fold, 15-fold, 20-fold, 25-fold, 30-fold, 40-fold , 50 times or more times compared to the control, or at least 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 90%, 100%, 125%, 150 %, 200%, 250%, 300%, 400% or 500% or more compared to control.

In certain embodiments, an eTF disclosed herein that activates SCN1A comprises a DNA binding domain comprising any of the following: (i) a sequence comprising RRDELNV x RSDHLTN x RSDDLVR x RSDNLVR x HRTTLTN x REDNLHT x TSHSLTE x QSSSLVR x REDNLHT (SEQ ID NO: 151), (ii) sequence, characterize

- 27 046157 having at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 151, or (ii) a functional fragment (i) or (ii). In certain embodiments, such an eTF further comprises one or more TADs selected from VP64, VPR, CITED2, CITED4, or CREB3. In one embodiment, such an eTF comprises a VPR TAD domain conjugated to the C terminus of a DBD. In certain embodiments, such an eTF comprises a CITED2 TAD conjugated to the N-terminus, the C-terminus, or the N-terminus and the C-terminus of a DBD. In certain embodiments, such an eTF comprises a CITED4 TAD conjugated to the N-terminus, the C-terminus, or the N-terminus and the C-terminus of a DBD. In certain embodiments, such an eTF comprises two CITED4 TADs conjugated to the N-terminus or C-terminus of a DBD. In certain embodiments, such eTF is capable of binding to the target site comprising SEQ ID NO: 32 and increasing SCN1A expression by at least 2-fold, 5-fold, 10-fold, 15-fold, 20-fold, 25-fold, 30-fold, 40-fold , 50 times or more compared to control, or at least 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 90%, 100%, 125%, 150% , 200%, 250%, 300%, 400% or 500% or more compared to control.

In certain embodiments, an eTF disclosed herein that activates SCN1A comprises a DNA binding domain comprising any of the following: (i) a sequence comprising DPGALVR x RSDNLVR x QSGDLRR x THLDLIR x TSGNLVR x RSDNLVR (SEQ ID NO: 150), (ii) a sequence having at least 89%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 150, or (ii) functional fragment (i) or (ii). In certain embodiments, such an eTF further comprises one or more TADs selected from VP64, VPR, CITED2, CITED4, or CREB3. In one embodiment, such an eTF comprises a VPR TAD domain conjugated to the C-terminus of a DBD. In certain embodiments, such an eTF comprises a CITED2 TAD conjugated to the N-terminus, the C-terminus, or the N-terminus and the C-terminus of a DBD. In certain embodiments, such an eTF comprises a CITED4 TAD conjugated to the N-terminus, the C-terminus, or the N-terminus and the C-terminus of a DBD. In certain embodiments, such an eTF comprises two CITED4 TADs conjugated to the N-terminus or C-terminus of a DBD. In certain embodiments, such eTF is capable of binding to the target site comprising SEQ ID NO: 31 and increasing SCN1A expression by at least 2-fold, 5-fold, 10-fold, 15-fold, 20-fold, 25-fold, 30-fold, 40-fold , 50 times or more compared to the control, or at least 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 90%, 100%, 125%, 150% , 200%, 250%, 300%, 400% or 500% or more compared to control.

In certain embodiments, an eTF disclosed herein that activates SCN1A comprises any of the following: (i) a sequence comprising any of SEQ ID NOs: 99-131, 205, 207, 209, 213, 217, 219, 221, or 223 ; (ii) a sequence comprising any of SEQ ID NOs: 77-98; (iii) a sequence having a sequence identity of at least 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96 %, 97%, 98% or 99%, with respect to any of the sequences (i) or (ii); or (iv) a functional fragment or variant of any of sequences (i), (ii) or (iii). In exemplary embodiments, such eTFs are capable of increasing SCN1A expression by at least 2-fold, 5-fold, 10-fold, 15-fold, 20-fold, 25-fold, 30-fold, 40-fold, 50-fold or more relative to control or at least by 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 90%, 100%, 125%, 150%, 200%, 250%, 300%, 400% or 500 % or more compared to control.

In certain embodiments, an eTF disclosed herein that activates SCN1A comprises any of the following: (i) a sequence comprising any of SEQ ID NOs: 99-102 or 125-127; (ii) a sequence comprising any of SEQ ID NOs: 77-91; (iii) a sequence having a sequence identity of at least 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96 %, 97%, 98% or 99%, with respect to any of the sequences (i) or (ii); or (iv) a functional fragment or variant of any of sequences (i), (ii) or (iii). In exemplary embodiments, such eTFs are capable of increasing SCN1A expression by at least 2-fold, 5-fold, 10-fold, 15-fold, 20-fold, 25-fold, 30-fold, 40-fold, 50-fold or more compared to control, or at least at least 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 90%, 100%, 125%, 150%, 200%, 250%, 300%, 400% or 500% or more compared to control.

In certain embodiments, an eTF disclosed herein that activates SCN1A comprises any of the following: (i) a sequence comprising any of SEQ ID NOs: 103-124, 128-131, 205, 207, 209, 213, 217, 219 , 221 or 223; (ii) a sequence containing any of SEQ ID NO: 92-98; (iii) a sequence having a sequence identity of at least 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96 %, 97%, 98% or 99%, relative to any of the sequences (i) or (ii); or (iv) a functional fragment or variant of any of sequences (i), (ii) or (iii). In exemplary embodiments, such eTFs are capable of increasing SCN1A expression by at least

- 28 046157 at least 2 times, 5 times, 10 times, 15 times, 20 times, 25 times, 30 times, 40 times, 50 times or more compared to the control, or at least 20%, 30%, 40 %, 50%, 60%, 70%, 75%, 80%, 90%, 100%, 125%, 150%, 200%, 250%, 300%, 400% or 500% or more compared to control. In exemplary embodiments, such eTFs have an overall sequence identity of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94% , 95%, 96%, 97%, 98%, or 99%, relative to one or more human proteins. In certain embodiments, such eTFs exhibit reduced immunogenicity compared to eTFs having a lower overall percentage of sequence identity to one or more human proteins. In various embodiments, the reduction in immunogenicity can be measured using an elispot assay, an immunoassay, or an in silico method.

In certain embodiments, an eTF disclosed herein that activates SCN1A comprises any of the following: (i) a sequence comprising SEQ ID NO: 127; (ii) a sequence having a sequence identity of at least 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96 %, 97%, 98% or 99%, relative to SEQ ID NO: 127; or (iii) a functional fragment or variant of any of sequences (i) or (ii). In exemplary embodiments, such eTFs contain SEQ ID NO: 77 and bind to a target site containing SEQ ID NO: 18. In exemplary embodiments, such eTFs are capable of increasing SCNIA expression by at least 2-fold, 5-fold, 10-fold, 15-fold , 20 times, 25 times, 30 times, 40 times, 50 times or more compared to the control, or at least 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80% , 90%, 100%, 125%, 150%, 200%, 250%, 300%, 400% or 500% or more compared to control.

In certain embodiments, an eTF disclosed herein that activates SCN1A comprises any of the following: (i) a sequence comprising SEQ ID NO: 128; (ii) a sequence having a sequence identity of at least 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96 %, 97%, 98% or 99% relative to SEQ ID NO: 128; or (iii) a functional fragment or variant of any of sequences (i) or (ii). In exemplary embodiments, such eTFs contain SEQ ID NO: 92 and bind to a target site containing SEQ ID NO: 18. In exemplary embodiments, such eTFs are capable of increasing SCNIA expression by at least 2-fold, 5-fold, 10-fold, 15-fold , 20 times, 25 times, 30 times, 40 times, 50 times or more compared to the control, or at least 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80% , 90%, 100%, 125%, 150%, 200%, 250%, 300%, 400% or 500% or more compared to control. In exemplary embodiments, such eTFs have an overall sequence identity of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94% , 95%, 96%, 97%, 98%, or 99%, relative to one or more human proteins. In certain embodiments, such eTFs exhibit reduced immunogenicity compared to eTFs having a lower overall percentage of sequence identity to one or more human proteins. In various embodiments, the reduction in immunogenicity can be measured using an elispot assay, an immunoassay, or an in silico method.

In certain embodiments, an eTF disclosed herein that activates SCN1A comprises any of the following: (i) a sequence comprising SEQ ID NO: 129; (ii) a sequence having a sequence identity of at least 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96 %, 97%, 98% or 99%, relative to SEQ ID NO: 129; or (iii) a functional fragment or variant of any of sequences (i) or (ii). In exemplary embodiments, such eTFs contain SEQ ID NO: 92 and bind to a target site containing SEQ ID NO: 18. In exemplary embodiments, such eTFs are capable of increasing SCNIA expression by at least 2-fold, 5-fold, 10-fold, 15-fold , 20 times, 25 times, 30 times, 40 times, 50 times or more compared to the control, or at least 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80% , 90%, 100%, 125%, 150%, 200%, 250%, 300%, 400% or 500% or more compared to control. In exemplary embodiments, such eTFs have an overall sequence identity of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94% , 95%, 96%, 97%, 98%, or 99%, relative to one or more human proteins. In certain embodiments, such eTFs exhibit reduced immunogenicity compared to eTFs having a lower overall percentage of sequence identity to one or more human proteins. In various embodiments, the reduction in immunogenicity can be measured using an elispot assay, an immunoassay, or an in silico method.

In certain embodiments, an eTF disclosed herein that activates SCN1A comprises any of the following: (i) a sequence comprising SEQ ID NO: 130; (ii) a sequence having a sequence identity of at least 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96 %, 97%, 98% or 99%, relative to SEQ ID NO: 130; or (iii) a functional fragment or variant of any of

- 29 046157 sequences (i) or (ii). In exemplary embodiments, such eTFs contain SEQ ID NO: 92 and bind to a target site containing SEQ ID NO: 18. In exemplary embodiments, such eTFs are capable of increasing SCNIA expression by at least 2-fold, 5-fold, 10-fold, 15-fold , 20 times, 25 times, 30 times, 40 times, 50 times or more compared to the control, or at least 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80% , 90%, 100%, 125%, 150%, 200%, 250%, 300%, 400% or 500% or more compared to control. In exemplary embodiments, such eTFs have an overall sequence identity of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94% , 95%, 96%, 97%, 98%, or 99%, relative to one or more human proteins. In certain embodiments, such eTFs exhibit reduced immunogenicity compared to eTFs having a lower overall percentage of sequence identity to one or more human proteins. In various embodiments, the reduction in immunogenicity can be measured using an elispot assay, an immunoassay, or an in silico method.

In certain embodiments, an eTF disclosed herein that activates SCN1A comprises any of the following: (i) a sequence comprising SEQ ID NO: 131; (ii) a sequence having a sequence identity of at least 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96 %, 97%, 98% or 99%, relative to SEQ ID NO: 131; or (iii) a functional fragment or variant of any of sequences (i) or (ii). In exemplary embodiments, such eTFs contain SEQ ID NO: 92 and bind to a target site containing SEQ ID NO: 18. In exemplary embodiments, such eTFs are capable of increasing SCNIA expression by at least 2-fold, 5-fold, 10-fold, 15-fold , 20 times, 25 times, 30 times, 40 times, 50 times or more compared to the control, or at least 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80% , 90%, 100%, 125%, 150%, 200%, 250%, 300%, 400% or 500% or more compared to control. In exemplary embodiments, such eTFs have an overall sequence identity of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94% , 95%, 96%, 97%, 98%, or 99%, relative to one or more human proteins. In certain embodiments, such eTFs exhibit reduced immunogenicity compared to eTFs having a lower overall percentage of sequence identity to one or more human proteins. In various embodiments, the reduction in immunogenicity can be measured using an elispot assay, an immunoassay, or an in silico method.

In certain embodiments, an eTF disclosed herein that activates SCN1A comprises a DBD containing a gRNA/Cas complex, wherein the gRNA comprises a targeting sequence comprising any of SEQ ID NOs: 35-66. The gRNA target sequence is attached to the 5' end of the scaffold sequence containing the sequence: 5'GTTTTAGTACTCTGGAAACAGAATCTACTAAAACAAGGCAAAATGCCGTGTTTATCTCGTCAACT TGTTGGCGAGA-3' (SEQ ID NO: 183). In exemplary embodiments, the Cas protein is a nuclease-deactivated Cas9 protein. In certain embodiments, such eTF further comprises one or more TADs conjugated to a Cas protein, wherein the TAD is selected from VP64, VPR, CITED2, CITED4, or CREB3. In one embodiment, such an eTF comprises a VPR TAD domain conjugated to the C terminus of the Cas protein. In certain embodiments, such an eTF comprises a CITED2 TAD conjugated to the N terminus, C terminus, or N terminus and C terminus of the Cas protein. In certain embodiments, such an eTF comprises a CITED4 TAD conjugated to the N-terminus, C-terminus, or N-terminus and C-terminus of a Cas protein. In exemplary embodiments, such eTFs are capable of increasing SCN1A expression by at least 2-fold, 5-fold, 10-fold, 15-fold, 20-fold, 25-fold, 30-fold, 40-fold, 50-fold, or more compared to a control.

Polynucleotides.

In another aspect, provided herein are polynucleotides encoding any of the eTFs that activate SNC1A disclosed herein. In another aspect, the present application provides polynucleotides comprising a PV-selective miRNA binding site. In certain embodiments, the present application provides polynucleotides comprising a PV-selective regulatory element operably linked to a transgene and a PV-selective microRNA binding site. In certain embodiments, the present application provides polynucleotides comprising a sequence encoding an eTF that activates SCN1A as disclosed herein and a PV-selective miRNA binding site. In certain embodiments, the present application provides a PV-selective regulatory element operably linked to a transgene encoding an eTF that activates SCN1A and a PV-selective regulatory element.

Polynucleotides encoding eTFs that activate SCN1A.

In certain embodiments, the present application provides a polynucleotide

- 30 046157 containing any of the following: (i) a nucleic acid sequence encoding an eTF that activates SCN1A containing any of SEQ ID NO: 77-131, 205, 207, 209, 213, 217, 219, 221 or 223, or their variant or functional fragment; (ii) a nucleic acid encoding a functional eTF fragment that activates SCN1A containing any of SEQ ID NO: 77-131, 205, 207, 209, 213, 217, 219, 221 or 223; or (iii) a nucleic acid encoding an eTF that activates SCN1A having a sequence identity of at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more, relative to an eTF that activates SCN1A containing any of SEQ ID NO: 77-131, 205, 207, 209, 213, 217, 219, 221 or 223, or a variant or functional fragment thereof.

In certain embodiments, the present application provides a polynucleotide comprising any of the following: (i) a nucleic acid sequence encoding a DBD containing any of SEQ ID NOs: 92-98, or a variant or functional fragment thereof; (ii) a nucleic acid encoding a functional DBD fragment containing any of SEQ ID NOs: 92-98; or (iii) a nucleic acid encoding a DBD having a sequence identity of at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92% , 93%, 94%, 95%, 96%, 97%, 98%, 99% or more, with respect to a DBD containing any of SEQ ID NOs: 92-98, or a variant or functional fragment thereof, wherein the DBD is capable of contact the target site associated with any of SEQ ID NO: 92-98.

In certain embodiments, the present application provides a polynucleotide encoding an eTF that activates endogenous SCN1A, wherein the polynucleotide comprises any of the following: (i) a nucleic acid sequence encoding an eTF comprising any of SEQ ID NOs: 103-124, 128-131, 205, 207, 209, 213, 217, 219, 221 or 223; (ii) a nucleic acid encoding a functional eTF fragment containing any of SEQ ID NOs: 103-124, 128-131, 205, 207, 209, 213, 217, 219, 221 or 223; or (iii) a nucleic acid encoding an eTF having a sequence identity of at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92% , 93%, 94%, 95%, 96%, 97%, 98%, 99% or more, with respect to eTF containing any of SEQ ID NO: 103-124, 128-131, 205, 207, 209, 213, 217, 219, 221 or 223, with eTF being able to activate SCN1A.

In certain embodiments, provided herein is a polynucleotide encoding a DBD that binds to a genomic target site capable of upregulating endogenous SCN1A when bound by an eTF disclosed herein, wherein the polynucleotide comprises any of the following: (i) a nucleic acid sequence, encoding a DBD containing any of SEQ ID NO: 77-98; (ii) a nucleic acid encoding a functional DBD fragment containing any of SEQ ID NOs: 77-98; or (iii) a nucleic acid encoding an eTF having a sequence identity of at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92% , 93%, 94%, 95%, 96%, 97%, 98%, 99% or more, with respect to a DBD containing any of SEQ ID NOs: 77-98, wherein the DBD is capable of binding to a target site bound by any from SEQ ID NO: 77-98.

In certain embodiments, provided herein is a polynucleotide encoding a DBD that binds to a genomic target site capable of activating endogenous SCN1A upon binding by an eTF disclosed herein, wherein the polynucleotide comprises any of the following: (i) a nucleic acid sequence encoding the DBD, containing any of SEQ ID NO: 148-151; (ii) a nucleic acid encoding a functional DBD fragment containing any of SEQ ID NOs: 148-151; or (iii) a nucleic acid encoding an eTF having a sequence identity of at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92% , 93%, 94%, 95%, 96%, 97%, 98%, 99% or more, with respect to a DBD containing any of SEQ ID NOs: 148-151, wherein the DBD is capable of binding to a target site bound by any from SEQ ID NO: 92-98.

In certain embodiments, the present application provides a polynucleotide encoding an eTF capable of regulating endogenous SCN1A, the polynucleotide comprising any of the following: (i) a nucleic acid sequence comprising any of SEQ ID NOs: 70-76 or 184; (ii) a nucleic acid containing a functional fragment of any of the sequences (i); or (iii) a nucleic acid having a sequence identity of at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93% , 94%, 95%, 96%, 97%, 98%, 99% or more, with respect to any of sequences (i) or (ii), wherein the polynucleotide encodes an eTF that is capable of activating SCN1A.

In certain embodiments, the present application provides a polynucleotide encoding an eTF capable of regulating endogenous SCN1A, the polynucleotide comprising any of the following: (i) a nucleic acid sequence comprising SEQ ID NO: 70; (ii) a nucleic acid containing a functional fragment of SEQ ID NO: 70; or (iii) a nucleic acid having a sequence identity of at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93% , 94%, 95%, 96%, 97%, 98%, 99% or more, relative to

- 31 046157 connection to any of the sequences (i) or (ii). In exemplary embodiments, such polynucleotides encode an eTF comprising SEQ ID NO: 127, or a functional fragment or variant thereof, that is capable of activating SCN1A.

In certain embodiments, the present application provides a polynucleotide encoding an eTF capable of regulating endogenous SCN1A, the polynucleotide comprising any of the following: (i) a nucleic acid sequence comprising SEQ ID NO: 71; (ii) a nucleic acid containing a functional fragment of SEQ ID NO: 71; or (iii) a nucleic acid having a sequence identity of at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93% , 94%, 95%, 96%, 97%, 98%, 99% or more, with respect to any of the sequences (i) or (ii). In exemplary embodiments, such polynucleotides encode an eTF comprising SEQ ID NO: 127, or a functional fragment or variant thereof, that is capable of activating SCN1A.

In certain embodiments, the present application provides a polynucleotide encoding an eTF capable of regulating endogenous SCN1A, the polynucleotide comprising any of the following: (i) a nucleic acid sequence comprising SEQ ID NO: 72; (ii) a nucleic acid containing a functional fragment of SEQ ID NO: 72; or (iii) a nucleic acid having a sequence identity of at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93% , 94%, 95%, 96%, 97%, 98%, 99% or more, with respect to any of the sequences (i) or (ii). In exemplary embodiments, such polynucleotides encode an eTF comprising SEQ ID NO: 130, or a functional fragment or variant thereof, that is capable of activating SCN1A.

In certain embodiments, the present application provides a polynucleotide encoding an eTF capable of regulating endogenous SCN1A, the polynucleotide comprising any of the following: (i) a nucleic acid sequence comprising SEQ ID NO: 73; (ii) a nucleic acid sequence containing the functional fragment of SEQ ID NO: 73; or (iii) a nucleic acid having a sequence identity of at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93% , 94%, 95%, 96%, 97%, 98%, 99% or more, with respect to any of the sequences (i) or (ii). In exemplary embodiments, such polynucleotides encode an eTF comprising SEQ ID NO: 131, or a functional fragment or variant thereof, that is capable of activating SCN1A.

In certain embodiments, the present application provides a polynucleotide encoding an eTF capable of regulating endogenous SCN1A, the polynucleotide comprising any of the following: (i) a nucleic acid sequence comprising SEQ ID NO: 74; (ii) a nucleic acid sequence containing the functional fragment of SEQ ID NO: 74; or (iii) a nucleic acid having a sequence identity of at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93% , 94%, 95%, 96%, 97%, 98%, 99% or more, with respect to any of the sequences (i) or (ii). In exemplary embodiments, such polynucleotides encode an eTF comprising SEQ ID NO: 127, or a functional fragment or variant thereof, that is capable of activating SCN1A.

In certain embodiments, the present application provides a polynucleotide encoding an eTF capable of regulating endogenous SCN1A, the polynucleotide comprising any of the following: (i) a nucleic acid sequence comprising SEQ ID NO: 75; (ii) a nucleic acid sequence containing the functional fragment of SEQ ID NO: 75; or (iii) a nucleic acid having a sequence identity of at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93% , 94%, 95%, 96%, 97%, 98%, 99% or more, with respect to any of the sequences (i) or (ii). In exemplary embodiments, such polynucleotides encode an eTF comprising SEQ ID NO: 127, or a functional fragment or variant thereof, that is capable of activating SCN1A.

In certain embodiments, the present application provides a polynucleotide encoding an eTF capable of regulating endogenous SCN1A, the polynucleotide comprising any of the following: (i) a nucleic acid sequence comprising SEQ ID NO: 76; (ii) a nucleic acid sequence containing the functional fragment of SEQ ID NO: 76; or (iii) a nucleic acid having a sequence identity of at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93% , 94%, 95%, 96%, 97%, 98%, 99% or more, with respect to any of the sequences (i) or (ii). In exemplary embodiments, such polynucleotides encode an eTF comprising SEQ ID NO: 106, or a functional fragment or variant thereof, that is capable of activating SCN1A.

In certain embodiments, the present application provides a polynucleotide encoding an eTF capable of regulating endogenous SCN1A, the polynucleotide comprising any of the following: (i) a nucleic acid sequence comprising SEQ ID NO: 184; (ii) a nucleic acid sequence containing the functional fragment of SEQ ID NO: 184; or (iii) a nucleic acid having sequence identity equal to or less than

- 32 046157 at least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more, with respect to any of the sequences (i) or (ii). In exemplary embodiments, such polynucleotides encode an eTF comprising SEQ ID NO: 106, or a functional fragment or variant thereof, that is capable of activating SCN1A.

Polynucleotides containing microRNA binding sites for selective expression in PV-containing neurons.

In another aspect, the present application provides polynucleotides containing microRNA binding sites that lead to selective expression of a gene of interest in parvalbumin-containing neurons (PV). MicroRNAs or miRNAs are small non-coding RNAs (~20 nucleotides) that regulate gene expression post-transcriptionally by hybridizing to complementary recognition sites in the mRNA molecule and lead to inhibition of gene expression by promoting the degradation of the mRNA transcript or inhibiting the translation of the protein encoded by the mRNA. MicroRNA binding sites provided herein inhibit expression of a gene of interest in excitatory neurons, thereby promoting selective expression of the gene of interest in PV-containing neurons (eg, PV-selective miRNA binding sites). In certain embodiments, excitatory neurons are neurons that express one or more of STAC, Slc17a7, Car12, Syt17, ITPKA, Col6a1, CamKII, Sv2b, INHBA, and/or DKK3. In an exemplary embodiment, the excitatory neurons are CamKII-expressing neurons.

In certain embodiments, the present application provides polynucleotides comprising one or more miRNA binding sites for one or more miRNAs that promote selective PV expression, for example, promoting the degradation of miRNA binding site-containing mRNA in excitatory neurons. Examples of miRNAs that promote selective PV expression include, for example, miR-128, miR-221 and miR-222. In certain embodiments, the present application provides polynucleotides containing at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more PV-selective miRNA binding sites. In one embodiment, the present application provides polynucleotides containing at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more miR-128 binding sites (SEQ ID NO: 9). In one embodiment, the present application provides polynucleotides containing at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more miR-221 binding sites (SEQ ID NO: 11). In one embodiment, the present application provides polynucleotides containing at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more miR-222 binding sites (SEQ ID NO: 13). In one embodiment, the present application provides polynucleotides comprising at least 1 miR-128 binding site, at least one miR-221 binding site, and at least one miR-222 binding site. In one embodiment, the present application provides polynucleotides comprising at least one miR-128 binding site and at least one miR-222 binding site. In one embodiment, the present application provides polynucleotides comprising at least one miR-221 binding site and at least one miR-222 binding site. In an exemplary embodiment, the present application provides polynucleotides comprising at least one miR-128 binding site (SEQ ID NO: 9) and at least one miR-221 binding site (SEQ ID NO: 11). In one embodiment, the present application provides polynucleotides containing at least 2 miR-128 binding sites (SEQ ID NO: 9) and at least 2 miR-221 binding sites (SEQ ID NO: 11). In one embodiment, the present application provides polynucleotides containing at least 3 miR-128 binding sites (SEQ ID NO: 9) and at least 3 miR-221 binding sites (SEQ ID NO: 11). In one embodiment, the present application provides polynucleotides containing at least 4 miR-128 binding sites (SEQ ID NO: 9) and at least 4 miR-221 binding sites (SEQ ID NO: 11). In one embodiment, the present application provides polynucleotides containing at least 5 miR-128 binding sites (SEQ ID NO: 9) and at least 5 miR-221 binding sites (SEQ ID NO: 11). In such embodiments, the binding sites may be arranged in any order. For example, for a construct containing 2 miR-128 binding sites and 2 miR-221 binding sites, the binding sites can be located in any of the following configurations: miR-128 - miR-128 - miR-221 - miR221, miR-128 - miR -221 - miR-128 - miR-221, miR-128 - miR221 - miR221 - miR-128, miR-221 - miR128 - miR221 - miR128, miR-221 - miR128 - miR128 - miR221 or miR221 - miR221 - miR128 - miR128 . In an exemplary embodiment, the polynucleotides provided herein comprise a sequence comprising 4 miR-128 binding sites (SEQ ID NO: 9) followed by four miR-221 binding sites (SEQ ID NO: 11), for example, miR-128 - miR-128 - miR128 - miR-128 - miR221 - miR221 - miR-221 - miR221. In another exemplary embodiment, the polynucleotides provided herein comprise a sequence comprising 1 miR-221 sequence (SEQ ID NO: 11), 1 miR-222 sequence (SEQ ID NO: 13), and 1 miR-128 binding site (SEQ ID NO : 9), for example miR

- 33 046157

221 - miR222 - miR128. In another exemplary embodiment, the polynucleotides provided herein comprise a sequence comprising 2 miR-221 sequences (SEQ ID NO: 11), 2 miR-222 sequences (SEQ ID NO: 13), and 2 miR128 binding sites (SEQ ID NO: 9 ) are arranged in the following order: miR-221 - miR222 - miR128 - miR-221 - miR222 miR128.

In polynucleotides containing more than one miRNA binding site, the binding sites may be immediately adjacent to each other in the polynucleotide sequence (e.g., without a linker or intervening sequence between the binding sites) or may be separated from each other by at least 1, 2, 3. 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more nucleotides or 1-20, 1-15, 1-10, 1 -9, 1-8, 1-7, 1-6, 1-5, 1-4, 1-3 or 1-2 nucleotides. In exemplary embodiments, miRNA binding sites are separated by approximately 5 nucleotides or 5 nucleotides. In exemplary embodiments, the sequences separating miRNA binding sites (as well as junctions formed between miRNA binding sites, or junctions formed between miRNA binding sites and sequences separating miRNA binding sites) are not complementary to any other miRNA or any other neuronal miRNA.

In certain embodiments, polynucleotides provided herein comprise a microRNA binding site having an identity of at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95% , 96%, 97%, 98%, 99%, or 100%, relative to SEQ ID NO: 7. In an exemplary embodiment, the polynucleotides provided herein comprise a microRNA binding site comprising SEQ ID NO: 7.

In certain embodiments, polynucleotides provided herein comprise a miRNA binding site having an identity of at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95% , 96%, 97%, 98%, 99%, or 100%, relative to SEQ ID NO: 14. In an exemplary embodiment, the polynucleotides provided herein comprise a microRNA binding site comprising SEQ ID NO: 14.

In certain embodiments, polynucleotides provided herein comprise a miRNA binding site having an identity of at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95% , 96%, 97%, 98%, 99%, or 100%, relative to SEQ ID NO: 15. In an exemplary embodiment, the polynucleotides provided herein comprise a microRNA binding site comprising SEQ ID NO: 15.

In certain embodiments, microRNA binding sites as defined herein are located in the 3' untranslated region of the mRNA transcript, for example, after a translation termination codon (i.e., TAA, TGA, or TAG) and before the polyA tail. The miRNA binding site may be located directly adjacent to the translation termination codon or may be separated from the translation termination codon by at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 , 15, 16, 17, 18, 19, 20 or more nucleotides or 1-20, 1-15, 1-10, 1-9, 1-8, 1-7, 1-6, 1-5, 1- 4, 1-3 or 1-2 nucleotides, and/or may be located adjacent to the polyA tail or may be separated from the polyA tail by at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 , 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more nucleotides or 1-20, 1-15, 1-10, 1-9, 1-8, 1-7, 1- 6, 1-5, 1-4, 1-3 or 1-2 nucleotides.

In certain embodiments, a microRNA binding site provided herein results in selective gene expression in a PV-containing cell compared to non-target cell types. In some cases, non-target cell types include, but are not limited to, excitatory neurons, non-PV CNS cell types, and non-neuronal CNS cell types. In certain embodiments, PV-selective miRNA binding sites result in selective gene expression in PV-containing neurons for at least one, two, three, four, five, or more non-PV CNS cell types. In some cases, the non-PV CNS cell is an excitatory neuron, dopaminergic neuron, astrocyte, microglia, motor neuron, vascular cell, or non-GABAergic neuron (e.g., a cell that does not express one or more of GAD2, GAD1, NKX2.1, DLX1, DLX5, SST, and VIP), non-PV neuron (eg, a GABAergic neuron that does not express parvalbumin), or other CNS cells (eg, CNS cell types that never express any of PV, GAD2, GAD1, NKX2.1, DLX1 , DLX5, SST and VIP). In an exemplary embodiment, the PV-selective miRNA binding site provided herein results in increased selectivity of gene expression in PV-containing neurons compared to excitatory neurons (e.g., neurons that express one or more of STAC, Slc17a7, Car12, Syt17, ITPKA, Col6a1, CamKII, Sv2b, INHBA and/or DKK3) by decreasing expression in excitatory neurons. In some cases, cell types are distinguished by different cellular markers, morphology, phenotype, genotype, function, and/or any other means of classifying cell types.

Expression selectivity driven by the PV-selective miRNA binding site can be measured in several ways. In one embodiment, the selectivity of gene expression in a PV cell compared to non-PV cells can be varied

- 34 046157 rena by comparing the number of PV cells that express a detectable level of transcript from a gene that contains a PV-selective miRNA binding site with the total number of cells that express the gene (for example, the ratio of PV-containing to total cells (PV + non-PV cells) expressing the gene). For example, selectivity for PV-containing neurons can be determined using an immunohistochemistry-based colocalization assay using an expression cassette containing a gene encoding a fluorescent protein (e.g., eGFP) and a PV-selective microRNA binding site to measure expression of the gene and antibody that identifies PV-containing cells (eg, an anti-PV antibody that specifically interacts with PV-containing neurons) coupled to a second fluorescent tag (eg, red fluorescent protein). Expression selectivity in PV-containing cells can be calculated by dividing the number of cells that express both PV and eGFP (eg, PV-containing cells) by the total number of cells expressing eGFP (eg, PV-containing cells and non-PV-containing cells). and multiplying by 100 to convert to a percentage. In another example, selectivity for PV-containing neurons can be determined using an immunohistochemistry-based colocalization assay using an expression cassette containing a gene encoding a fluorescent protein (e.g., eGFP) and a PV-selective microRNA binding site to measure gene expression. and a first antibody that identifies PV-containing cells (e.g., an anti-PV antibody that specifically interacts with PV-containing neurons) coupled to a second fluorescent tag (e.g., red fluorescent protein) and a second antibody that identifies excitatory cells (e.g., antibody to CamKII, which specifically interacts with excitatory neurons). Expression selectivity in PV-containing cells can be calculated by dividing the number of cells expressing both PV and eGFP (e.g., PV cells) by the number of cells expressing eGFP + PV and eGFP + CamKII (e.g., PV-containing cells and excitatory cells), and multiplying by 100 to convert to a percentage. The higher the percentage of PV-containing cells expressing the transgene, the more selective the microRNA binding site is for PV-containing cells. In certain embodiments, a PV-selective miRNA binding site provided herein can be highly selective for expression in PV-containing cells. For example, a PV-selective miRNA binding site provided herein may have a selectivity of approximately 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92 %, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or greater than about 99%, for PV-containing neurons (e.g., PV-containing neurons/all cells x 100 or PV-containing neurons/ PV + excitatory neurons x 100).

In some cases, the PV-selective miRNA binding site presented herein is short. In some cases, the size of the PV-selective miRNA binding site is compatible with the cloning ability of a vector, such as a viral vector or rAAV, such that the combined size of the transgene, promoter (and optional enhancer), and miRNA binding site does not exceed the cloning ability of the vector. In some cases, the PV-selective miRNA binding site is up to approximately 500 bp, 400 bp, 300 bp, 250 bp, 225 bp, 215 bp in length. , 210 bp, 200 bp, 150 bp, 140 bp, 135 bp, 130 bp, 125 bp, 120 bp, 115 bp, 110 bp, 100 bp, 90 bp, 80 bp, 75 bp, 70 bp, 65 bp, 60 bp. n. or 50 bp. In some cases, the PV selective miRNA binding site is approximately 50-500 bp, 50-400 bp, 50-300 bp, 50-250 bp. , 50-200 bp, 50-100 bp, 50-75 bp, 50-70 bp, 100-500 bp, 100-400 bp, 100 -300 bp, 100-250 bp, 100-200 bp, 100-150 bp, 100-140 bp, 100-135 bp, 200-500 bp, 200-400 bp, 200-300 bp. or 200-250 bp.

In exemplary embodiments, a polynucleotide provided herein that contains one or more PV-selective miRNA binding sites does not contain SEQ ID NO: 67.

Expression cassettes.

In another aspect, the present application provides expression cassettes comprising a polynucleotide provided herein (e.g., a polynucleotide containing a sequence encoding an eTF that activates SCNIA and/or contains a PV-selective miRNA binding site) and one or more regulatory elements. In certain embodiments, provided herein are expression cassettes comprising a polynucleotide provided herein (e.g., a polynucleotide containing a sequence encoding an eTF that activates SCN1A and/or contains a PV-selective microRNA binding site) and a PV-selective promoter.

In certain embodiments, a polynucleotide provided herein (e.g., a polynucleotide containing a sequence encoding an eTF that activates SCN1A and/or contains a PV-selective miRNA binding site) is part of an expression cassette containing one or more regulatory elements in addition to

- 35 046157 eTF coding sequence. In exemplary embodiments, a polynucleotide provided herein (e.g., a polynucleotide containing a coding sequence for an eTF that activates SCN1A and/or contains a PV-selective miRNA binding site) is part of an expression cassette comprising an upstream promoter from a transgene sequence to be capable of driving the expression of a transgene (eg, an eTF that selectively activates SCN1A) in a cell.

In certain embodiments, an expression cassette disclosed herein comprises a polynucleotide provided herein (e.g., a polynucleotide containing a sequence encoding an eTF that activates SCNIA regulation and/or contains a PV-selective miRNA binding site) and a constitutive promoter located upstream of transcriptional sequence from the transgene coding sequence to be able to direct the expression of the transgene (eg, eTF that selectively activates SCN1A) in the cell. Examples of constitutive promoters include the GAD2 promoter, the human synapsin promoter, the CBA promoter, the CMV promoter, the minCMV promoter, the TATA box, the super basic promoter or the EF1a promoter, or a combination thereof. In certain embodiments, an expression cassette disclosed herein comprises a polynucleotide provided herein (e.g., a polynucleotide containing a sequence encoding an eTF that activates SCN1A and/or contains a PV-selective miRNA binding site) and a short promoter capable of driving expression, transgene (for example, an eTF that selectively activates SCN1A) in a cell. In certain embodiments, a short promoter suitable for use with the nucleic acid molecules described herein comprises less than 500 bp, 450 bp, 400 bp, 350 bp, 300 bp .b., 250 bp., 225 bp., 200 bp., 175 bp., 150 bp., 145 bp., 140 bp., 135 bp. ., 130 bp, 125 bp, 120 bp, 115 bp, 110 bp, 105 bp, 100 bp, 95 bp, 90 bp, 85 bp, 80 bp or 75 bp or approximately 80-300 bp, 80-275 bp, 80-250 bp, 80-200 bp, 80-150 bp, 80-125 bp, 80-120 bp, 80-115 bp, 80-110 bp, 80-105 bp, 80-100 bp, 85-300 bp, 85- 275 bp, 85-250 bp, 85-200 bp, 85-150 bp, 85-125 bp, 85-120 bp, 85-115 p .n., 85-110 bp, 85-105 bp, 85100 bp, 90-300 bp, 90-275 bp, 90-250 bp, 90-200 bp, 90-150 bp, 90-125 bp, 90-120 bp, 90-115 bp, 90-110 bp, 90- 105 bp, 90-100 bp, 95-300 bp, 95-275 bp, 95-250 bp, 95-200 bp, 95-150 p .n., 95-125 bp, 95-120 bp, 95-115 bp, 95-110 bp, 95-105 bp, 95-100 bp ., 100-300 bp, 100-275 bp, 100-250 bp, 100-200 bp, 100-150 bp, 100-125 bp, 100-120 bp, 100-115 bp, 100-110 bp. or 100-105 bp. In exemplary embodiments, a short promoter suitable for use in accordance with the expression cassettes described herein comprises approximately 100-120 bp, approximately 117 bp. or approximately 100 bp.

In certain embodiments, an expression cassette disclosed herein comprises a short promoter comprising or consisting of the following: (i) SEQ ID NO: 1; (ii) a variant or functional fragment thereof; or (iii) a nucleic acid sequence having sequence identity of at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88% , 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, with respect to any of (i) or (ii), functionally related to a polynucleotide encoding any of the eTFs that selectively activate SCN1A disclosed herein and optionally comprising a miRNA binding site disclosed herein. Other examples of short promoter sequences can be found in PCT Publication No. WO 2018/213786.

In certain embodiments, an expression cassette disclosed herein comprises a polynucleotide provided herein (e.g., a polynucleotide containing a sequence encoding an eTF that activates SCN1A and/or contains a PV-selective microRNA binding site) and a cell type-selective promoter located upstream upstream of the sequence encoding the transgene (eg, an eTF that selectively activates SCN1A) to be able to selectively control the expression of the transgene in the cell of interest. In certain embodiments, a cell type selective promoter may be selective (eg, selectively drive expression) for any cell type of interest, such as, for example, a heart cell, a liver cell, a muscle cell, a bone cell, a neuron, or subpopulations thereof. In an exemplary embodiment, an expression cassette disclosed herein comprises a polynucleotide encoding an eTF that selectively activates SCN1A, and a PV-selective regulatory element (e.g., a promoter, enhancer, and/or promoter and enhancer) located upstream of the sequence encoding eTF so as to be capable of selectively driving eTF expression in a PV cell, and optionally a PV-selective miRNA binding site. A PV-selective regulatory element refers to a regulatory element that specifically modulates gene expression in a PV-containing neuron. In certain embodiments, the PV-selective regulatory elements enhance expression in a PV-containing neuron relative to one or more other CNS cell types. In certain embodiments, the PV-selective regulatory element inhibits transcription and/or translation processes in non-target cell types.

In certain embodiments, a PV-selective regulatory element provided herein results in selective gene expression in a PV cell over non-target cell types. In some cases, non-target cell types include, but are not limited to, excitatory neurons, non-PV CNS cell types, and non-neuronal CNS cell types. In certain embodiments, the PV-selective regulatory elements result in selective gene expression in PV-containing neurons of at least one, two, three, four, five, or more non-PV cell types of the CNS. In some cases, the non-PV CNS cell is an excitatory neuron, dopaminergic neuron, astrocyte, microglia, motoneuron, vascular cell, or non-GABAergic neuron (e.g., a cell that does not express one or more of GAD2, GAD1, NKX2.1, DLX1, DLX5, SST, and VIP), a non-PV neuron (eg, a GABAergic neuron that does not express parvalbumin) or other CNS cells (eg, CNS cell types that never express any of PV, GAD2, GAD1, NKX2.1 , DLX1, DLX5, SST and VIP). In some cases, the PV-selective regulatory element presented herein results in increased selectivity of gene expression in PV-containing neurons compared to non-PV GABAergic cells. In some cases, cell types are distinguished by different cellular markers, morphology, phenotype, genotype, function, and/or any other means of classifying cell types.

Expression selectivity controlled by a PV-selective regulatory element can be measured in several ways. In one embodiment, the selectivity of gene expression in a PV-containing cell compared to non-PV cells can be measured by comparing the number of PV-containing cells that express a detectable level of transcript from a gene that is operably linked to the PV-selective regulatory element with the total number of cells that express the gene (eg, the ratio of PV to the total number of cells (PV + non-PV cells) expressing the gene). For example, selectivity for PV-containing neurons can be determined using an immunohistochemistry-based colocalization assay using a gene encoding a fluorescent protein (e.g., eGFP) operably linked to a PV-selective regulatory element to measure gene expression and an antibody that identifies PV-containing cells (eg, an anti-PV antibody that specifically interacts with PV-containing neurons) bound to a second fluorescent tag (eg, red fluorescent protein). Expression selectivity in PV-containing cells can be calculated by dividing the number of cells that express both PV and eGFP (eg, PV-containing cells) by the total number of cells expressing eGFP (eg, PV-containing cells and non-PV-containing cells). and multiplying by 100 to convert to a percentage. The higher the percentage of PV-containing cells expressing the transgene, the more selective the regulatory element is for PV-containing cells. In certain embodiments, a PV-selective regulatory element provided herein may be highly selective for expression in PV-containing cells. For example, a PV selective regulatory element provided herein may have selectivity of approximately 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92% , 93%, 94%, 95%, 96%, 97%, 98%, 99%, or greater than about 99%, with respect to PV-containing neurons (eg, PV-containing neurons/total cells x 100).

In some cases, the PV-selective regulatory element presented herein is short. In some cases, the size of the PV-selective regulatory element is compatible with the cloning ability of the vector, such as a viral vector or rAAV, such that the combined size of the transgene and one or more PV-selective regulatory elements does not exceed the cloning ability of the vector. In some cases, the PV selective regulatory element is up to approximately 2050 bp, 2000 bp, 1900 bp, 1800 bp, 1700 bp, 1600 bp in length. 1500 bp, 1400 bp, 1300 bp, 1200 bp, 1100 bp, 1000 bp, 900 bp, 800 bp, 700 bp .b., 600 bp, 500 bp, 400 bp, 300 bp, 200 bp. or 100 bp In some cases, the PV selective regulatory element is approximately 500-600 bp, 500-700 bp, 500-800 bp, 500-900 bp, 500-1000 bp. , 500-1500 bp, 500-2000 bp or 500-2050 bp.

In certain embodiments, a PV selective regulatory element provided herein comprises or consists of any of the following: (i) SEQ ID NO: 2-4; (ii) a variant, functional fragment, or combination thereof, or (iii) a nucleic acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87% sequence identity, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% in relation to any of (i) or (ii). In some cases, the regulatory element includes any of SEQ ID NO: 2-4. Further examples of PV-selective regulatory elements can be found in PCT Publication No. WO 2018/187363.

According to illustrative embodiments, this application provides cassettes

- 37 046157 expressions containing a nucleic acid sequence encoding an eTF that selectively activates SCN1A under the control of a PV-selective regulatory element. In certain embodiments, provided herein are expression cassettes comprising a nucleic acid sequence encoding an eTF that selectively activates SCN1A containing a DBD containing any of the following sequences: SEQ ID NO: 77-98 under the control of a PV-selective regulatory element comprising any one of SEQ ID NO: 2-4. In certain embodiments, provided herein are expression cassettes containing a nucleic acid sequence encoding an eTF that selectively activates SCN1A, comprising any of the following sequences: SEQ ID NO: 99-131, 205, 207, 209, 213, 217, 219, 221 or 223 under the control of a PV-selective regulatory element comprising any of SEQ ID NOs: 2-4. In certain embodiments, provided herein are expression cassettes comprising a nucleic acid sequence comprising any of the following sequences: SEQ ID NO: 67-73 under the control of a PV selective regulatory element comprising any of SEQ ID NO: 2-4. In certain embodiments, provided herein are expression cassettes comprising a nucleic acid sequence encoding an eTF that selectively activates SCN1A containing a DBD comprising any of the following sequences: SEQ ID NO: 148-151 under the control of a PV-selective regulatory element comprising any of SEQ ID NO: 2. In certain embodiments, provided herein are expression cassettes containing a nucleic acid sequence encoding an eTF that selectively activates SCN1A, comprising any of the following sequences: SEQ ID NO: 99-131, 205, 207, 209, 213, 217, 219, 221, or 223 under the control of a PV-selective regulatory element comprising any of SEQ ID NO: 2. In certain embodiments, provided herein are expression cassettes comprising a nucleic acid sequence comprising any of the following sequences : SEQ ID NO: 67-76 or 316 under the control of a PV-selective regulatory element containing any of SEQ ID NO: 2.

In certain embodiments, the present application provides expression cassettes comprising a PV-selective miRNA binding site and a promoter and/or enhancer sequence. Any of the promoters described herein may be included in an expression cassette. In an exemplary embodiment, an expression cassette provided herein comprises a PV-selective miRNA binding site and a PV-selective regulatory element. In certain embodiments, an expression cassette provided herein comprises a PV-selective miRNA binding site and a PV-selective regulatory element comprising (i) any of SEQ ID NO: 2-4; (ii) a variant, functional fragment, or combination thereof, or (iii) a nucleic acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87% sequence identity, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, in relation to any of (i) or (ii). In certain embodiments, an expression cassette provided herein comprises (1) a PV-selective miRNA binding site comprising (i) any of SEQ ID NO: 7, 14, or 15; (ii) a variant, functional fragment or combination of (i) or (iii) a nucleic acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87 sequence identity %, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, in relation to any of (i) or (ii) , and (2) a PV-selective regulatory element comprising (i) any of SEQ ID NOs: 2-4; (ii) a variant, functional fragment, or combination thereof; (i) or (iii) a nucleic acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86% sequence identity, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, in relation to any of (i) or (ii ). In certain embodiments, an expression cassette provided herein comprises (1) a PV-selective miRNA binding site comprising (i) SEQ ID NO: 7; (ii) a variant, functional fragment or combination of (i) or (iii) a nucleic acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87 sequence identity %, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, in relation to any of (i) or (ii) , and (2) a PV-selective regulatory element comprising (i) SEQ ID NO: 2; (ii) a variant, functional fragment or combination of (i) or (iii) a nucleic acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87 sequence identity %, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, in relation to any of (i) or (ii) . In one embodiment, an expression cassette provided herein comprises a microRNA binding site comprising SEQ ID NO: 7 and a PV selective regulatory element comprising SEQ ID NO: 2. In certain embodiments, a polynucleotide provided herein comprises (1 ) PV-selective miRNA binding site,

- 38 046157 containing (i) SEQ ID NO: 14; (ii) a variant, functional fragment or combination of (i) or (iii) a nucleic acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87 sequence identity %, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, in relation to any of (i) or (ii) , and (2) a PV-selective regulatory element comprising (i) SEQ ID NO: 2; (ii) a variant, functional fragment or combination of (i) or (iii) a nucleic acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87 sequence identity %, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, in relation to any of (i) or (ii) . In one embodiment, an expression cassette provided herein comprises a microRNA binding site comprising SEQ ID NO: 15 and a PV selective regulatory element comprising SEQ ID NO: 2. In certain embodiments, an expression cassette provided herein comprises (1 ) a PV-selective miRNA binding site containing (i) SEQ ID NO: 15; (ii) a variant, functional fragment or combination of (i) or (iii) a nucleic acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87 sequence identity %, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, in relation to any of (i) or (ii) , and (2) a PV-selective regulatory element comprising (i) SEQ ID NO: 2; (ii) a variant, functional fragment or combination of (i) or (iii) a nucleic acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87 sequence identity %, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, in relation to any of (i) or (ii) . In one embodiment, an expression cassette provided herein comprises a miRNA binding site comprising SEQ ID NO: 15 and a PV selective regulatory element comprising SEQ ID NO: 2.

In certain embodiments, an expression cassette provided herein comprises a PV-selective miRNA binding site and a sequence encoding an eTF that promotes expression of SCN1A as provided herein. In an exemplary embodiment, the present application provides an expression cassette comprising a PV-selective regulatory element, an eTF, that activates the expression of SCN1A as provided herein, and a PV-selective microRNA binding site. According to an illustrative embodiment, the present application provides an expression cassette comprising (1) a PV selective regulatory element comprising (i) any of SEQ ID NO: 2-4; (ii) a variant, functional fragment or combination of (i) or (iii) a nucleic acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87 sequence identity %, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, in relation to any of (i) or (ii) (2) a sequence encoding an eTF that activates SCN1A, comprising (i) any of SEQ ID NO: 77-131, 205, 207, 209, 213, 217, 219, 221, or 223; (ii) a variant, functional fragment or combination of (i) or (iii) a nucleic acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87 sequence identity %, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, in relation to any of (i) or (ii) , and (3) a PV-selective miRNA binding site comprising (i) any of SEQ ID NO: 7, 14 or 15; (ii) a variant, functional fragment or combination of (i) or (iii) a nucleic acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87 sequence identity %, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, in relation to any of (i) or (ii) . According to another illustrative embodiment, provided herein is an expression cassette comprising (1) a PV selective regulatory element comprising (i) SEQ ID NO: 2; (ii) a variant, functional fragment or combination of (i) or (iii) a nucleic acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87 sequence identity %, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, in relation to any of (i) or (ii) , (2) a sequence encoding an eTF that activates SCN1A, containing (i) any of SEQ ID NO: 77 or 127; (ii) a variant, functional fragment or combination of (i) or (iii) a nucleic acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87 sequence identity %, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, in relation to any of (i) or (ii) , and (3) a PV-selective miRNA binding site comprising (i) SEQ ID NO: 7; (ii) a variant, functional fragment or combination of (i) or (iii) a nucleic acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87 sequence identity %, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, in relation to any of (i) or (ii) . According to another illustrative embodiment, provided herein is an expression cassette comprising (1) a PV selective regulatory element comprising (i) SEQ ID NO: 2; (ii) variant, functional fragment or combination of (i) or (iii) nucleic acid sequence, ha

- 39 046157 rasterizable sequence identity of at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, relative to any of (i) or (ii), (2) a sequence encoding an eTF that activates SCN1A containing (i) any of SEQ ID NO: 77 or 127; (ii) a variant, functional fragment or combination of (i) or (iii) a nucleic acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87 sequence identity %, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, in relation to any of (i) or (ii) , and (3) a PV-selective miRNA binding site comprising (i) SEQ ID NO: 14; (ii) a variant, functional fragment or combination of (i) or (iii) a nucleic acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87 sequence identity %, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, in relation to any of (i) or (ii) . According to another illustrative embodiment, provided herein is an expression cassette comprising (1) a PV selective regulatory element comprising (i) SEQ ID NO: 2; (ii) a variant, functional fragment or combination of (i) or (iii) a nucleic acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87 sequence identity %, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, in relation to any of (i) or (ii) , (2) a sequence encoding an eTF that activates SCN1A, containing (i) any of SEQ ID NO: 77 or 127; (ii) a variant, functional fragment or combination of (i) or (iii) a nucleic acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87 sequence identity %, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, in relation to any of (i) or (ii) , and (3) a PV-selective miRNA binding site comprising (i) SEQ ID NO: 15; (ii) a variant, functional fragment or combination of (i) or (iii) a nucleic acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87 sequence identity %, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, in relation to any of (i) or (ii) . According to another illustrative embodiment, provided herein is an expression cassette comprising (1) a PV selective regulatory element comprising (i) SEQ ID NO: 2; (ii) a variant, functional fragment or combination of (i) or (iii) a nucleic acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87 sequence identity %, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, in relation to any of (i) or (ii) , (2) a sequence encoding an eTF that activates SCN1A, containing (i) any of SEQ ID NO: 92 or 106; (ii) a variant, functional fragment or combination of (i) or (iii) a nucleic acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87 sequence identity %, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, in relation to any of (i) or (ii) , and (3) a PV-selective miRNA binding site comprising (i) SEQ ID NO: 7; (ii) a variant, functional fragment or combination of (i) or (iii) a nucleic acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87 sequence identity %, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, in relation to any of (i) or (ii) . According to another illustrative embodiment, provided herein is an expression cassette comprising (1) a PV selective regulatory element comprising (i) SEQ ID NO: 2; (ii) a variant, functional fragment or combination of (i) or (iii) a nucleic acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87 sequence identity %, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, in relation to any of (i) or (ii) , (2) a sequence encoding an eTF that activates SCN1A, containing (i) any of SEQ ID NO: 92 or 106; (ii) a variant, functional fragment or combination of (i) or (iii) a nucleic acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87 sequence identity %, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, in relation to any of (i) or (ii) , and (3) a PV-selective miRNA binding site comprising (i) SEQ ID NO: 14; (ii) a variant, functional fragment or combination of (i) or (iii) a nucleic acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87 sequence identity %, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, in relation to any of (i) or (ii) . According to another illustrative embodiment, provided herein is an expression cassette comprising (1) a PV selective regulatory element comprising (i) SEQ ID NO: 2; (ii) a variant, functional fragment or combination of (i) or (iii) a nucleic acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87 sequence identity %, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, in relation to any of (i) or (ii) , (2) a sequence encoding an eTF that activates SCN1A, containing (i) any of SEQ ID NO: 92 or 106; (ii) variant, functional fragment or combination

- 40 046157 tion (i) or (iii) a nucleic acid sequence characterized by a sequence identity of at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, in relation to any of (i) or (ii), and (3) a PV-selective miRNA binding site comprising (i) SEQ ID NO: 15; (ii) a variant, functional fragment or combination of (i) or (iii) a nucleic acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87 sequence identity %, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, in relation to any of (i) or (ii) .

In certain embodiments, an expression cassette provided herein comprising a PV-selective regulatory element and a PV-selective miRNA binding site characterized by a size of less than 5 kb, 4.9 kb, 4 ,8 kb, 4.7 kb, 4.6 kb, 4.5 kb, 4.4 kb, 4 ,3 kb, 4.2 kb, 4.1 kb, 4.0 kb, 3.9 kb, 3 ,8 kb, 3.7 kb, 3.6 kb, 3.5 kb, 3.4 kb, 3 ,3 kb, 3.2 kb, 3.1 kb, 3.0 kb, 2.9 kb, 2 ,8 kb, 2.7 kb, 2.6 kb, 2.5 kb, 2.4 kb, 2 ,3 kb, 2.2 kb, 2.1 kb, 2.0 kb, 1.9 kb, 1 ,8 kb, 1.7 kb, 1.6 kb. or 1.5 kb. or less, or about 1.5-5 kb, 1.5-4.7 kb, 1.5-4.5 kb, 1.5-4 ,0 kb, 1.5-3.5 kb, 1.5-3.0 kb, 1.5-2.5 kb. , 1.5-2.0 kb.

In certain embodiments, an expression cassette provided herein may contain further additional regulatory elements in addition to a promoter, such as, for example, sequences associated with transcription initiation or termination, enhancer sequences, and effective RNA processing signals. Exemplary regulatory elements include, for example, an intron, an enhancer, a UTR, a stability element, a WPRE sequence, a Kozak consensus sequence, a post-translational response element, a microRNA binding site, or a polyadenylation (polyA) sequence, or a combination thereof. Regulatory elements can function to modulate gene expression during the transcriptional phase, posttranscriptional phase, or translational phase of gene expression. At the RNA level, regulation can occur at the level of translation (eg, stability elements that stabilize mRNA for translation), RNA cleavage, RNA splicing, and/or transcription termination. In various embodiments, regulatory elements may recruit transcription factors to the coding region that increase the selectivity of gene expression in the cell type of interest, increase the rate at which RNA transcripts are produced, increase the stability of the RNA produced, and/or increase the rate of production of protein synthesis from RNA transcripts. In an exemplary embodiment, an expression cassette provided herein comprises at least one PV-selective miRNA binding site as provided herein.

In certain embodiments, the expression cassettes described herein further comprise a polyA sequence. Suitable polyA sequences include, for example, an artificial polyA of approximately 75 bp in length. (PA75) (see e.g. WO 2018/126116), bovine growth hormone polyA, SV40 polyA early signal, SV40 late polyA signal, rabbit beta-globin polyA, HSV thymidine kinase polyA, protamine gene polyA, adenovirus 5 EIb polyA, polyA growth hormone or polyA PBGD. In exemplary embodiments, a polyA sequence suitable for use in the expression cassettes provided herein is hGH polyA (SEQ ID NO: 17) or synthetic polyA (SEQ ID NO: 16). Typically, the polyA sequence is located downstream of the polynucleotide encoding the eTF in the expression cassettes described herein.

In certain embodiments, expression cassettes provided herein further comprise one or more nucleic acid sequences encoding one or more nuclear localization signals (NLS). Any NLS peptide that facilitates the import of the protein to which it is attached into the cell nucleus can be used. Examples of NLSs include, for example, SV40 large T antigen NLS, nucleoplasmin NLS, EGL-13 NLS, cMyc NLS, and TUS protein NLS. See, for example, C. Dingwall et al., J. Cell Biol. 107: 841-9 (1988); J.P. Makkerh et al., Curr Biol. 6: 1025-7 (1996) and M. Ray et al., Bioconjug. Chem. 26: 1004-7 (2015). The NLS can be located anywhere in the eTF protein sequence, but in preferred embodiments it is conjugated to the N-terminus of the eTF or an eTF domain. In exemplary embodiments, the nucleic acid cassettes provided herein encode an eTF with an NLS fused to the N-terminus of the eTF. In other embodiments, the nucleic acid cassettes provided herein encode an eTF with a first NLS fused to the N-terminus of the eTF and a second NLS located between the DBD and the TAD domain of the eTF.

Expression vectors.

In certain embodiments, expression cassettes described herein may be included in an expression vector. Expression vectors can be used to deliver an expression cassette to a target cell through transfection or transduction. A vector may be an integrating or non-integrating vector, which refers to the vector's ability

- 41 046157 integrate an expression cassette or transgene into the genome of the host cell. Examples of expression vectors include, but are not limited to, (a) non-viral vectors such as nucleic acid vectors, including linear oligonucleotides and circular plasmids; artificial chromosomes such as human artificial chromosomes (HAC), yeast artificial chromosomes (YAC) and bacterial artificial chromosomes (BAC or PAC)); episomal vectors; transposons (eg PiggyBac); and (b) viral vectors, such as retroviral vectors, lentiviral vectors, adenoviral vectors and adeno-associated viral vectors.

Expression vectors can be linear oligonucleotides or circular plasmids and can be delivered into cells using a variety of transfection methods, including physical and chemical methods. Physical methods generally refer to delivery methods that use physical force to counteract a cell membrane barrier to facilitate intracellular delivery of genetic material. Examples of physical methods include the use of a needle, ballistic DNA, electroporation, sonoporation, photoporation, magnetofection and hydroporation. Chemical methods generally refer to methods in which chemical vehicles deliver a nucleic acid molecule into a cell, and may include inorganic particles, lipid-based vectors, polymer-based vectors and peptide-based vectors.

In certain embodiments, the expression vector is introduced into the target cell using a cationic lipid (eg, a cationic liposome). Various types of lipids have been explored for gene delivery, such as lipid nanoemulsion (eg, which is a dispersion of one immiscible liquid in another, stabilized by an emulsifying agent) or solid lipid nanoparticles.

In certain embodiments, the expression vector is introduced into a target cell using a peptide-based delivery vehicle. Peptide-based carriers may have the advantages of protecting the delivered genetic material, targeting specific cellular receptors, disrupting endosomal membranes, and delivering genetic material to the nucleus. In certain embodiments, the expression vector is introduced into the target cell using a polymer-based delivery vehicle. Polymer-based carriers may contain natural proteins, peptides and/or polysaccharides or synthetic polymers. In one embodiment, the polymer-based delivery vehicle comprises polyethylenimine (PEI). PEI can condense DNA into positively charged particles that bind to anionic cell surface remnants and enter the cell through endocytosis. In other embodiments, the polymer-based delivery vehicle may comprise poly-L-lysine (PLL), poly(DL-lactic acid) (PLA), poly-L-lactide-co-glycoside (PLGA), polyornithine, polyarginine, histones, protamines, dendrimers, chitosans, synthetic amine derivatives of dextran and/or cationic acrylic polymers. In certain embodiments, polymer-based delivery vehicles may include a mixture of polymers, such as, for example, PEG and PLL.

In certain embodiments, the expression vector may be a viral vector suitable for gene therapy. Preferred characteristics of viral gene therapy vectors or gene delivery vectors may include the ability to be reproducibly and stably propagated and purified to high titers; to mediate targeted delivery (eg, to deliver a transgene specifically to a tissue or organ of interest without widespread distribution of the vector elsewhere); and mediate gene delivery and transgene expression without causing harmful side effects.

Several types of viruses, such as a non-pathogenic parvovirus called adeno-associated virus, have been engineered for gene therapy purposes, exploiting the viral infection pathway but avoiding subsequent viral gene expression, which can lead to replication and toxicity. Such viral vectors can be produced by removing all or some of the coding regions from the viral genome but leaving intact those sequences (e.g., terminal repeat sequences) that may be required for functions such as packaging the vector genome into the viral capsid or integrating the vector nucleic acid ( e.g. DNA) into host chromatin.

In various embodiments, suitable viral vectors include retroviruses (eg, A-type, B-type, C-type, and D-type viruses), adenovirus, parvovirus (eg, adeno-associated viruses or AAV), coronavirus, negative-infective viruses polar strand RNA such as orthomyxovirus (eg, influenza virus), rhabdovirus (eg, rabies and vesicular stomatitis virus), paramyxovirus (eg, measles and Sendai), positive strand RNA viruses such as picornavirus and alphavirus, and double-stranded DNA viruses, including adenovirus, herpes virus (eg, herpes simplex virus types 1 and 2, Epstein-Barr virus, cytomegalovirus), and poxvirus (eg, cowpox, fowlpox, and canarypox). Examples of retroviruses include avian leukemia-sarcoma virus, human T-lymphotrophic virus type 1 (HTLV-1), bovine leukemia virus (BLV), lentivirus, and spumavirus. Other viruses include, for example, Norwalk virus, togavirus, flavivirus, reoviruses, papovavirus, hepadnavirus, and

- 42 046157 hepatitis. Viral vectors can be divided into two groups depending on their ability to integrate into the host genome - integrating and non-integrating. Oncoretroviruses and lentiviruses can integrate into the chromatin of the host cell, while adenoviruses, adeno-associated viruses, and herpes viruses are predominantly retained in the cell nucleus as extrachromosomal episomes.

In certain embodiments, a suitable viral vector is a retroviral vector. Retroviruses belong to the viruses of the Retroviridae family. Examples of retroviruses include oncoretroviruses such as murine leukemia virus (MLV) and lentiviruses such as human immunodeficiency virus 1 (HIV-1). Retroviral genomes are single-stranded (ss) RNAs and contain various genes that can be in cis or trans form. For example, a retroviral genome may contain cis-acting sequences, such as two long terminal repeats (LTRs), with elements for gene expression, reverse transcription, and integration into host chromosomes. Other components include the packaging signal (psi or Ψ) for the specific packaging of RNA into newly formed virions and the polypurine tract (PPT), the site of initiation of positive-strand DNA synthesis during reverse transcription. In addition, the retroviral genome may contain gag, pol and env genes. The gag gene encodes structural proteins, the pol gene encodes enzymes that accompany ssRNA and reverse transcription of viral RNA into DNA, and the env gene encodes the viral envelope. Typically, gag, pol, and env are provided in trans for viral replication and packaging.

In certain embodiments, a retroviral vector provided herein may be a lentiviral vector. At least five serogroups or serotypes of lentiviruses are recognized. Viruses of different serotypes may infect certain types of cells and/or hosts differently. Lentiviruses, for example, include primate retroviruses and non-primate retroviruses. Primate retroviruses include HIV and simian immunodeficiency virus (SIV). Non-primate retroviruses include feline immunodeficiency virus (FIV), bovine immunodeficiency virus (BIV), caprine arthritis-encephalitis virus (CAEV), equine infectious anemia virus (EIAV), and visnavirus. Lentiviruses or lentivectors may be capable of transducing resting cells. As with oncoretroviral vectors, the design of lentivectors can be based on the separation of cis- and trans-acting sequences.

In certain embodiments, the present application provides expression vectors that have been designed for delivery using an optimized therapeutic retroviral vector. The retroviral vector may be a lentivirus containing a left-handed (5') LTR; sequences that promote packaging and/or nuclear import of the virus; promoter; optionally, one or more additional regulatory elements (such as, for example, an enhancer or polyA sequence); optionally a lentiviral reverse response element (RRE); a construct containing a PV selective regulatory element operably linked to an eTF coding sequence; optionally an insulator and a right (3') retroviral LTR.

In exemplary embodiments, the viral vector provided herein is an adeno-associated virus (AAV). AAV is a small, non-enveloped, replication-defective animal virus that infects humans and several other primate species. AAV is known to cause human disease and induce a mild immune response. AAV vectors can also infect both dividing and quiescent cells without integrating into the host cell genome.

The AAV genome consists of linear single-stranded DNA ∼4.7 kb in length. The genome consists of two open reading frames (ORFs) flanked by an approximately 145-bp inverted terminal repeat (ITR) sequence. The ITR consists of a nucleotide sequence at the 5' end (5' ITR) and a nucleotide sequence located at the 3' end (3' ITR), which contain palindromic sequences. ITRs function in cis by folding to form T-shaped hairpin structures through complementary base pairing, which act as primers during the initiation of DNA replication for second-strand synthesis. Two open reading frames encode the rep and cap genes, which are involved in virion replication and packaging. In an exemplary embodiment, the AAV vector provided herein does not contain rep or cap genes. Such genes can be presented in trans to produce virions, as described below.

In certain embodiments, the AAV vector may include excess nucleic acid. In certain embodiments, the excess nucleic acid may encode a green fluorescent protein or an antibiotic resistance gene such as kanamycin or ampicillin. In certain embodiments, the excess nucleic acid may be located outside of the ITR sequences (eg, compared to the eTF transgene sequence and regulatory sequences that are located between the 5' and 3' ITR sequences).

There are different serotypes of AAV, including AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, AAV12, and AAV13. These serotypes differ in their tropism

- 43 046157 or by the types of cells they infect. AAVs may contain genomes and capsids from multiple serotypes (eg, pseudotypes). For example, AAV may contain a serotype 2 genome (eg, ITR) packaged within a serotype 5 or serotype 9 capsid. Pseudotypes may increase transduction efficiency as well as alter tropism.

In some cases, the AAV serotype is favored and can cross the blood-brain barrier or infect CNS cells. In some cases, AAV9 or a variant thereof is used to deliver an expression cassette of the present disclosure containing a PV selective regulatory element operably linked to a transgene encoding an eTF that selectively activates SCN1A. In some cases, AAV9 or a variant thereof is used to deliver an expression cassette of the present disclosure containing a PV-selective miRNA binding site. In some cases, AAV9 or a variant thereof is used to deliver an expression cassette of the present disclosure comprising a PV-selective regulatory element operably linked to a transgene encoding an eTF that selectively activates SCN1A and a PV-selective miRNA binding site.

In exemplary embodiments, the present application provides expression vectors that have been designed for delivery by AAV. AAV can be of any serotype, for example, AAV1, AAV2, AAV3, AAV3b, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, AAV12, AAV-DJ, or a chimeric, hybrid, or variant AAV. AAV may also be a self-complementary AAV (scAAV). In certain embodiments, an expression vector designed for delivery by AAV contains a 5' ITR and a 3' ITR. In certain embodiments, an expression vector designed for delivery by AAV comprises a 5' ITR, a promoter, a transgene encoding an eTF, and a 3' ITR. In certain embodiments, an expression vector designed for delivery by AAV comprises a 5' ITR, an enhancer, a promoter, a transgene encoding an eTF, a polyA sequence, and a 3' ITR.

Host cells.

In another aspect, the present invention relates to a host cell containing an expression cassette or expression vector as disclosed herein. The host cell may be a bacterial cell, a yeast cell, an insect cell, or a mammalian cell. In an exemplary embodiment, a host cell is any cell line that is susceptible to infection by the virus of interest and can be cultured in vitro.

In certain embodiments, a host cell provided herein can be used for ex vivo gene therapy purposes. In such embodiments, cells are transfected with a nucleic acid molecule or expression vector containing a PV-selective miRNA binding site and/or an eTF coding sequence that selectively activates SCN1A, as described herein, and then transplanted into a patient or subject. The transplanted cells can be of autologous, allogeneic or heterologous origin. For clinical applications, cell isolation is typically performed under good manufacturing practice (GMP) conditions. Before transplantation, cell quality and absence of microbes or other contaminants are typically checked, and preconditioning such as radiation and/or immunosuppressive treatment may be performed. In addition, host cells can be transplanted along with growth factors to stimulate cell proliferation and/or differentiation.

In certain embodiments, the host cell can be used for ex vivo gene therapy. Preferably, said cells are eukaryotic cells, such as mammalian cells, these include, without limitation, humans other than human primates such as monkeys; chimpanzee; monkeys and orangutans, domestic animals, including dogs and cats, and livestock such as horses, cattle, pigs, sheep and goats, or other species of mammals, including, but not limited to, mice, rats, sea pigs, rabbits, hamsters, etc. One skilled in the art will select more appropriate cells according to the patient or transplant recipient.

In certain embodiments, a host cell provided herein may be a cell with self-renewal and pluripotency properties, such as stem cells or induced pluripotent stem cells. The stem cells are preferably mesenchymal stem cells. Mesenchymal stem cells (MSCs) are capable of differentiating into at least one of osteoblasts, chondrocytes, adipocytes or myocytes and can be isolated from any tissue type. Typically, MSCs are isolated from bone marrow, adipose tissue, umbilical cord or peripheral blood. Methods for their preparation are well known to one skilled in the art. Induced pluripotent stem cells (also known as iPS cells or iPSCs) are a type of pluripotent stem cells that can be derived directly from adult cells. Yamanaka et al. induced iPS cells by transferring the Oct3/4, Sox2, Klf4 and c-Myc genes into mouse and human fibroblasts and forcing

- 44 046157 cells to express genes (WO 2007/069666). Thomson et al. subsequently generated human iPS cells using Nanog and Lin28 instead of Klf4 and c-Myc (WO 2008/118820).

In an exemplary embodiment, the host cell provided herein is a packaging cell. These cells may be adherent or suspension cells. The packaging cell and the helper vector or virus or DNA construct(s) provide together in trans all the missing functions that are required for complete replication and packaging of the viral vector.

Preferably, said packaging cells are eukaryotic cells, such as mammalian cells, including monkey, human, canine and rodent cells. Examples of human cells are PER.C6 cells (WO01/38362), MRC-5 (ATCC CCL-171), WI-38 (ATCC CCL-75), HEK-293 cells (ATCC CRL-1573), HeLa cells (ATCC CCL2 ) and fetal rhesus lung cells (ATCC CL-160). Examples of non-human primate cells are Vero cells (ATCC CCL81), COS-1 cells (ATCC CRL-1650) or COS-7 cells (ATCC CRL-1651). Examples of canine cells are MDCK cells (ATCC CCL-34). Examples of rodent cells are hamster cells such as BHK21-F cells, HKCC cells or CHO cells.

As an alternative to mammalian sources, cell lines for use in the present invention can be obtained from avian sources, such as chicken, duck, goose, quail or pheasant. Examples of avian cell lines include avian embryonic stem cells (WO01/85938 and WO03/076601), immortalized duck retinal cells (WO2005/042728) and cells derived from avian embryonic stem cells, including chicken cells (WO2006/108846) or duck cells such as the EB66 cell line (WO2008/129058 and WO2008/142124).

In another embodiment, said host cells are insect cells, such as SF9 cells (ATCC CRL-1711), Sf21 cells (IPLB-Sf21), MG1 cells (BTI-TNMG1) or High Five™ cells (BTI-TN-5B1 -4).

In certain embodiments, host cells provided herein containing a recombinant AAV vector/genome of the present invention (e.g., containing a PV-selective miRNA binding site and/or a sequence encoding an eTF that selectively activates SCN1A) may further comprise one or more nucleic acid constructs, such as, for example, (i) a nucleic acid construct (eg, an AAV helper plasmid) that encodes the rep and cap genes but does not carry ITR sequences; and/or (ii) a nucleic acid construct (eg, a plasmid) providing the adenoviral functions required for AAV replication. In an exemplary embodiment, a host cell provided herein comprises: i) an expression vector comprising a PV-selective miRNA binding site and/or a sequence encoding an eTF that selectively activates SCN1A as provided herein (i.e., recombinant AAV genome); ii) a nucleic acid construct encoding the AAV rep and cap genes that does not carry an ITR sequence; and iii) a nucleic acid construct containing adenoviral helper genes (as described below).

In certain embodiments, rep, cap, and adenovirus helper genes can be combined in a single plasmid (Blouin V et al. J Gene Med. 2004; 6(suppl): S223-S228; Grimm D. et al. Hum. Gene Ther. 2003;7:839-850). Thus, in another exemplary embodiment, a host cell provided herein comprises: i) an expression vector comprising a PV-selective miRNA binding site and/or an eTF coding sequence that selectively activates SCN1A as disclosed herein (i.e. i.e. recombinant AAV genome); and ii) a plasmid encoding the rep and cap genes of AAV, which does not carry the ITR sequence and additionally contains adenovirus helper genes.

In another embodiment, a host cell provided herein comprises: a) an expression vector containing a PV-selective miRNA binding site and/or a sequence encoding an eTF that selectively activates SCN1A as described herein (i.e., a genome recombinant AAV); b) a plasmid encoding the rep and cap genes of AAV, which does not carry the ITR sequence; and c) a plasmid containing adenoviral RNA helper genes E2a, E4 and VA; wherein the cotransfection is performed in cells, preferably mammalian cells, that constitutively express and are transcomplemented to the adenoviral E1 gene, such as HEK-293 cells (ATCC CRL-1573).

In certain embodiments, a host cell suitable for large-scale production of AAV vectors is insect cells that can be infected with a combination of recombinant baculoviruses (Urabe et al. Hum. Gene Ther. 2002; 13: 1935-1943). For example, SF9 cells can be co-infected with three baculovirus vectors, respectively expressing AAV rep, AAV cap and AAV packaging vector. Recombinant baculovirus vectors will provide viral helper gene functions necessary for viral replication and/or packaging.

Additional guidance on the design and production of virions for gene therapy

- 45 046157 according to the present invention can be found in: Viral Vectors for Gene Therapy, Methods and Protocols. Series: Methods in Molecular Biology, Vol. 737. Merten and Al-Rubeai (Eds.); 2011 Humana Press (Springer); Gene Therapy. M. Giacca. 2010 Springer-Verlag; Heilbronn R. and Weger S. Viral Vectors for Gene Transfer: Current Status of Gene Therapeutics. In: Drug Delivery, Handbook of Experimental Pharmacology 197; M. Schafer-Korting (Ed.). 2010 Springer-Verlag; pp. 143-170; Adeno-Associated Virus: Methods and Protocols. R. O. Snyder and P. Moullier (Eds). 2011 Humana Press (Springer); Bunning H. et al. Recent developments in adeno-associated virus technology. J. Gene Med. 2008; 10:717-733 and Adenovirus: Methods and Protocols. M. Chillon and A. Bosch (Eds.); Third. Edition. 2014 Humana Press (Springer).

Virions and methods for obtaining virions.

In certain embodiments, the present application provides viral particles comprising a viral vector comprising a PV-selective miRNA binding site and/or a sequence encoding an eTF that selectively activates SCN1A, as disclosed herein. The terms viral particle and virion are used interchangeably herein and refer to an infectious and typically replication-defective viral particle containing a viral genome (e.g., a viral expression vector) packaged within a capsid and, as appropriate, e.g. retroviruses - a lipid membrane surrounding the capsid. Capsid refers to the structure in which the viral genome is packaged. The capsid consists of several oligomeric structural subunits consisting of proteins. For example, AAV has an icosahedral capsid formed by the interaction of three capsid proteins: VP1, VP2, and VP3. In one embodiment, the virion provided herein is a recombinant AAV virion or rAAV virion produced by packaging an AAV vector containing a PV-selective regulatory element and a PV-selective miRNA binding site. In another embodiment, the virion provided herein is a recombinant AAV virion or rAAV virion produced by packaging an AAV vector containing a PV-selective regulatory element operably linked to an eTF coding sequence that selectively activates SCN1A, as described herein , in a protein shell. In another embodiment, the virion provided herein is a recombinant AAV virion or rAAV virion produced by packaging an AAV vector containing a PV-selective regulatory element operably linked to an eTF coding sequence that selectively activates SCN1A, as described herein , and a PV-selective miRNA binding site in the protein shell.

In certain embodiments, a recombinant AAV virion provided herein can be produced by encapsidating an AAV genome derived from a specific AAV serotype into a viral particle formed by naturally occurring Cap proteins corresponding to AAVs of the same specific serotype. In other embodiments, an AAV viral particle provided herein comprises a viral vector containing the ITR of a given AAV serotype packaged into proteins from a different serotype. See for example Bunning H et al. J Gene Med 2008; 10: 717-733. For example, a viral vector having an ITR from a given AAV serotype can be packaged into: a) a viral particle consisting of capsid proteins derived from the same or a different AAV serotype (e.g., AAV2 ITR and AAV9 capsid proteins; AAV2 ITR and capsid proteins AAV8, etc.); b) a mosaic viral particle consisting of a mixture of capsid proteins from different AAV serotypes or mutants (eg, ITR AAV2 with capsid proteins AAV1 and AAV9); c) a chimeric viral particle consisting of capsid proteins that have been truncated by domain shuffling between different AAV serotypes or variants (eg, AAV2 ITR with AAV8 capsid proteins with AAV9 domains); or d) a targeted viral particle designed to display selective binding domains that ensure strict interaction with specific target cell receptors (e.g., AAV5 ITR with AAV9 capsid proteins genetically truncated by insertion of a peptide ligand; or AAV9 capsid proteins not genetically modified by binding a peptide ligand on the surface of the capsid).

One of ordinary skill in the art will appreciate that the AAV virion provided herein may contain capsid proteins of any AAV serotype. In one embodiment, the viral particle contains capsid proteins from an AAV serotype selected from the group consisting of AAV1, AAV2, AAV5, AAV8 and AAV9, which are more suitable for delivery to the CNS (M. Hocquemiller et al., Hum Gene Ther 27(7 ): 478-496 (2016)). In a specific embodiment, the viral particle comprises an expression cassette of the present invention, wherein the 5'ITR and 3'ITR sequences of the expression cassette are serotype AAV2 and the capsid proteins are serotype AAV9.

Numerous methods for producing rAAV virions are known in the art, including transfection, production of stable cell lines, and infectious hybrid virus production systems that include adenovirus-AAV hybrids, herpesvirus-AAV hybrids (Conway, J E et al., (1997) ) J. Virology 71(11):8780–8789) and baculovirus-AAV hybrids. Everything is cool

- 46 046157 rAAV production rounds for the production of rAAV viral particles require: 1) suitable host cells, including, for example, cell lines of human origin, such as HeLa, A549 or 293 cells, or cell lines derived from insects, such as SF- 9, in the case of baculovirus production systems; 2) a suitable helper virus function provided by a wild-type or mutant adenovirus (such as a temperature-sensitive adenovirus), a herpes virus, a baculovirus, or a plasmid construct providing helper functions; 3) AAV rep and cap genes and gene products; 4) a transgene (eg, containing one or more of: a PV-selective microRNA binding site, a sequence encoding an eTF that selectively activates SCN1A as described herein, and/or a PV-selective promoter) flanked by AAV ITR sequences ; and 5) suitable media and media components to support rAAV production.

In various embodiments, the host cells described herein contain the following three components: (1) a rep gene and a cap gene, (2) genes providing accessory functions, and (3) a transgene (for example, containing one or more of: a selective regarding a PV miRNA binding site, a sequence encoding an eTF that selectively activates SCN1A as described herein, and/or a PV-selective promoter) flanked by the ITR. The AAV rep gene, the AAV cap gene, and genes providing accessory functions can be introduced into a cell by incorporating the genes into a vector, such as a plasmid, and introducing the vector into a host cell. The rep, cap, and accessory function genes may be included in the same plasmid or in different plasmids. In a preferred embodiment, the AAV rep and cap genes are included on one plasmid, and the genes providing accessory functions are included on another plasmid. Various plasmids to create a host cell for virion production (eg, containing AAV rep and cap genes, accessory functions, or a transgene) can be introduced into the cell using any suitable method well known in the art. Examples of transfection methods include, but are not limited to, coprecipitation with calcium phosphate, DEAE-dextran, polybrene, electroporation, microinjection, liposome-mediated fusion, lipofection, retroviral infection, and biolistic transfection. In certain embodiments, plasmids providing rep and cap genes, accessory functions, and an ITR-flanked transgene can be introduced into a cell simultaneously. In another embodiment, plasmids providing rep and cap genes and accessory functions can be introduced into the cell before or after introduction of the plasmid containing the transgene. In an exemplary embodiment, cells are simultaneously transfected with three plasmids (e.g., in a triple transfection manner): (1) a plasmid containing a transgene (e.g., containing one or more of: a PV-selective microRNA binding site, a sequence encoding an eTF that selectively activates SCN1A , as described herein, and/or a PV-selective promoter) flanked by an ITR, (2) a plasmid containing the AAV rep and cap genes, and (3) a plasmid containing genes providing accessory functions. Exemplary host cells may be 293, A549, or HeLa cells.

In other embodiments, one or more of (1) the AAV rep and cap genes, (2) genes providing accessory functions, and (3) a transgene (e.g., comprising one or more of: a PV-selective miRNA binding site, a sequence, encoding an eTF that selectively activates SCN1A as described herein and/or a PV-selective promoter) flanked by an ITR can be carried by the packaging cell either episomally or integrated into the genome of the packaging cell. In one embodiment, the host cells may be packaging cells in which the AAV rep and cap genes and genes providing accessory functions are stably maintained in the host cell, and the host cell is transiently transfected with a plasmid containing the transgene (e.g., containing one or more of : a PV-selective miRNA binding site, a sequence encoding an eTF that selectively activates SCN1A as described herein, and/or a PV-selective promoter) flanked by an ITR. In another embodiment, the host cells are packaging cells in which the AAV rep and cap genes are stably maintained in the host cell, and the host cell is transiently transfected with a plasmid containing the transgene (e.g., containing one or more of the following: PV-selective a microRNA binding site, a sequence encoding an eTF that selectively activates SCN1A as described herein, and/or a PV-selective promoter) flanked by an ITR and a plasmid containing accessory functions. In another embodiment, the host cells may be packaging cells in which accessory functions are stably maintained in the host cell, and the host cell is transiently transfected with a plasmid containing a transgene (e.g., containing one or more of the following: a PV-selective binding site miRNA, sequence encoding an eTF that selectively activates SCN1A, as described herein, and/or selective

- 47 046157 in relation to the PV promoter), flanked by ITR, and a plasmid containing the rep and cap genes. In another embodiment, the host cells may be producer cell lines that are stably transfected with rep and cap genes, genes providing accessory functions, and a transgene sequence (e.g., containing one or more of the following: a PV-selective miRNA binding site, a sequence encoding an eTF that selectively activates SCN1A, as described herein, and/or a PV-selective promoter) flanked by an ITR. Exemplary packaging and production cells may be derived from 293, A549, or HeLa cells.

In another embodiment, the producer cell line is an insect cell line (typically Sf9 cells) that is infected with baculovirus expression vectors that provide Rep and Cap proteins. This system does not require adenovirus helper genes (Ayuso E, et al., Curr. Gene Ther. 2010, 10:423-436).

As used herein, the term cap protein refers to a polypeptide having at least one functional activity of the native AAV cap protein (eg, VP1, VP2, VP3). Examples of the functional activities of cap proteins include the ability to induce capsid formation, promote the accumulation of single-stranded DNA, facilitate packaging of AAV DNA into capsids (ie, encapsidation), bind to cellular receptors, and facilitate virion entry into host cells. In principle, any Cap protein can be used in the context of the present invention.

Cap proteins have been reported to influence host tropism, cell, tissue, or organ specificity, receptor utilization, infection efficiency, and immunogenicity of AAVs. Accordingly, the AAV cap for use in rAAV may be selected based on, for example, the species of the subject (eg, human or non-human), the immunological condition of the subject, its suitability for long-term or short-term treatment, or the particular therapeutic use (eg, treatment of a particular disease or disorder or delivery to specific cells, tissues or organs). In certain embodiments, the cap protein is from the AAV group consisting of serotypes AAV1, AAV2, AAV5, AAV8, and AAV9. In an exemplary embodiment, the cap protein is derived from AAV9.

In some embodiments, a cap AAV for use in a method of the present invention can be produced by mutagenesis (ie, insertions, deletions, or substitutions) of one of the above cap AAVs or the nucleic acid encoding it. In certain embodiments, the cap AAV is at least 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% or more similar to one or more of the aforementioned cap AAV.

In some embodiments, cap AAV is chimeric and contains domains from two, three, four, or more of the above cap AAVs. In certain embodiments, cap AAV is a mosaic of VP1, VP2, and VP3 monomers derived from two or three different AAVs or recombinant AAV. In certain embodiments, the rAAV composition contains more than one of the above-mentioned caps.

In some embodiments, the AAV cap for use in an rAAV virion is designed to contain a heterologous sequence or other modification. For example, a peptide or protein sequence that provides selective targeting or immune evasion can be converted into a cap protein. Alternatively or in addition, cap can be chemically modified so that the surface of rAAV is polyethylene glycolated (i.e., PEGylated), which may facilitate immune evasion. The cap protein can also be mutagenized (eg, to remove its natural receptor binding or to mask an immunogenic epitope).

As used herein, the term rep protein refers to a polypeptide having at least one of the functional activities of the native AAV rep protein (eg, rep 40, 52, 68, 78). Examples of the functional activity of rep protein include any activity associated with the physiological function of the protein, including facilitating DNA replication by recognizing, binding and terminating the AAV origin of DNA replication, as well as DNA helicase activity. Additional functions include modulation of transcription from AAV (or other heterologous) promoters and site-specific integration of AAV DNA into the host chromosome. In a specific embodiment, the AAV rep genes may be from serotypes AAV1, AAV2, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, or AAVrh10; more preferably from an AAV serotype selected from the group consisting of AAV1, AAV2, AAV5, AAV8 and AAV9.

In certain embodiments, the rep AAV protein for use in the method of the present invention can be obtained by mutagenesis (ie, insertions, deletions, or substitutions) of one of the above rep AAVs or its encoding nucleic acid. In certain embodiments, the rep AAV is at least 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% or more similar to one or more of the above rep AAVs.

The expressions used herein for auxiliary functions or genes

- 48 046157 helpers refer to viral proteins whose replication depends on AAV. Accessory functions include those proteins that are required for AAV replication, including, but not limited to, those proteins that are involved in AAV gene transcription activation, stage-specific AAV mRNA splicing, AAV DNA replication, synthesis of cap expression products, and AAV capsid assembly. Virus-based helper functions can be derived from any of the known helper viruses, such as adenovirus, herpes virus (other than herpes simplex virus type 1), and vaccinia virus. Accessory functions include, but are not limited to, adenovirus E1, E2a, VA and E4 or herpesvirus UL5, ULB, UL52 and UL29 and herpesvirus polymerase. In a preferred embodiment, the proteins on which AAV replication depends are derived from the adenovirus.

In certain embodiments, a viral protein dependent on AAV for replication for use in a method of the present invention may be produced by mutagenesis (ie, insertions, deletions, or substitutions) of one of the aforementioned viral proteins or the nucleic acid encoding the same. In certain embodiments, the viral protein is at least 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% or more similar to one or more of the aforementioned viral proteins.

Methods for analyzing the functions of cap proteins, rep proteins, and viral proteins on which AAV replication depends are well known in the art.

Host cells for expression of the transgene of interest (e.g., containing one or more of the following: a PV-selective microRNA binding site, a sequence encoding an eTF that selectively activates SCN1A as described herein, and/or a PV-selective promoter ) can be grown under conditions suitable for the assembly of AAV virions. In certain embodiments, host cells are grown for a suitable period of time to facilitate the assembly of AAV virions and the release of virions into the medium. Typically, cells can be grown for about 24 hours, about 36 hours, about 48 hours, about 72 hours, about 4 days, about 5 days, about 6 days, about 7 days, about 8 days, about 9 days, or up to about 10 days. After approximately 10 days (or sooner, depending on culture conditions and the specific host cell used), production levels typically decrease significantly. Typically, culture time is measured from the time the virus is produced. For example, in the case of AAV, virus production typically begins after the viral helper function has been established in the appropriate host cell, as described herein. Typically, cells are harvested from about 48 to about 100, preferably from about 48 to about 96, preferably from about 72 to about 96, preferably from about 68 to about 72 hours after infection with the helper virus (or after the start of virus production).

rAAV-producing cultures can be grown under a variety of conditions (a wide range of temperatures, for varying periods of time, etc.) suitable for the particular host cell used. rAAV production cultures include attachment-dependent cultures that can be cultured in suitable attachment-dependent containers, such as, for example, spinner flasks, hollow fiber filters, microcarriers, and packed-bed or fluidized-bed bioreactors. rAAV vector production cultures may also include suspension-adapted host cells, such as HeLa, 293, and SF-9 cells, which can be cultured in a variety of ways, including, for example, spinner flasks, stirred tank bioreactors, and single-use systems such as as a system of disposable reactors from Wave.

Suitable media known in the art can be used to produce rAAV virions. These media include, without limitation, media manufactured by Hyclone Laboratories and JRH, including modified Eagle's medium (MEM), Dulbecco's modified Eagle's medium (DMEM), each of which is incorporated herein by reference in its entirety. In certain embodiments, the rAAV production culture medium may be supplemented with serum or serum-derived recombinant proteins at a level of 0.5-20% (vol/vol or w/v). Alternatively, rAAV vectors can be produced under serum-free conditions, which may also be referred to as animal product-free environments.

After culturing the host cells to allow production of AAV virions, the resulting virions can be collected and purified. In certain embodiments, AAV virions can be obtained from (1) production culture host cells by lysis of the host cells and/or (2) the culture medium of said cells over a period of time after transfection, preferably 72 hours. rAAV virions can be collected from waste medium from a production culture, provided that the cells are cultured under conditions that cause rAAV virions to be released into the medium from intact cells (see, for example, US Pat. No. 6,566,118). Suitable methods for lysing cells are also known in the art and include, for example, multiple freeze/thaw cycles, sonication, microfluidization, and chemical treatment.

- 49 046157 chemical substances such as detergents and/or proteases.

Once harvested, rAAV virions can be purified. As used herein, the term purified includes a preparation of rAAV virions that is devoid of at least some other components that may also be present where rAAV virions occur naturally or from which they are originally derived. Thus, for example, purified rAAV virions can be obtained using an isolation method to enrich them from a starting mixture such as a culture lysate or production culture supernatant. Enrichment can be measured in a variety of ways, for example by the proportion of DNase-resistant particles (DRPs) or genome copies (gc) present in a solution, or by infectivity, or it can be measured relative to a second, potentially interfering substance present in the original mixture. such as pollutants, including industrial culture pollutants or technological pollutants, including a helper virus, environmental components, etc.

In certain embodiments, the rAAV production culture yield can be clarified to remove host cell debris. In some embodiments, the production culture yield can be clarified using a variety of standard technologies, such as centrifugation or filtration through a filter with a pore size of 0.2 μm or larger (eg, a cellulose acetate filter or a series of depth filters).

In certain embodiments, the rAAV production culture yield is further treated with Benzonase™ to digest any high molecular weight DNA present in the production culture. In some embodiments, the benzonase™ digestion is performed under standard conditions, for example, at a final concentration of 1-2.5 units/ml benzonase™ at a temperature between ambient temperature and 37°C for a period of 30 minutes to several hours.

In certain embodiments, rAAV virions can be isolated or purified using one or more of the following purification steps: equilibrium centrifugation; flow anion exchange filtration; tangential flow filtration (TFF) to concentrate rAAV particles; rAAV capture by apatite chromatography; heat inactivation of the helper virus; capture of rAAV by hydrophobic interaction chromatography; buffer exchange using size exclusion chromatography (SEC); nanofiltration and capture of rAAV using anion exchange chromatography, cation exchange chromatography or affinity chromatography. These steps can be used individually, in different combinations, or in different orders. Methods for purifying rAAV particles can be found, for example, in Xiao et al., (1998) Journal of Virology 72:2224-2232; US patents No. 6989264 and 8137948 and WO 2010/148143.

In certain embodiments, purified AAV virions can be dialyzed against PBS, filtered, and stored at -80°C. Viral genome titers can be determined by quantitative PCR using linearized plasmid DNA as a standard curve (see, for example, Lock M, et al., Hum. Gene Ther. 2010; 21:1273-1285).

Pharmaceutical compositions.

In certain embodiments, the present application provides compositions comprising a PV-selective miRNA binding site and/or a sequence encoding an eTF that selectively activates SCN1A, and a pharmaceutically acceptable carrier. In other embodiments, the present application provides virions comprising a PV-selective miRNA binding site and/or a sequence encoding an eTF that selectively activates SCN1A, and a pharmaceutically acceptable carrier. In exemplary embodiments, such compositions are suitable for use in gene therapy. The pharmaceutical compositions are preferably sterile and stable under conditions of manufacture and storage. Sterile solutions can be prepared, for example, by filtration through sterile filter membranes.

Acceptable carriers and excipients in pharmaceutical compositions are preferably non-toxic to recipients at the dosages and concentrations used. Acceptable carriers and excipients may include buffers such as phosphate, citrate, HEPES and TAE, antioxidants such as ascorbic acid and methionine, preservatives such as hexamethonium chloride, octadecyldimethylbenzylammonium chloride, resorcinol and benzalkonium chloride, proteins such as serum albumin human, gelatin, dextran and immunoglobulins, hydrophilic polymers such as polyvinylpyrrolidone, amino acids such as glycine, glutamine, histidine and lysine, and carbohydrates such as glucose, mannose, sucrose and sorbitol. The pharmaceutical compositions of the present disclosure can be administered parenterally in the form of an injectable composition. Pharmaceutical compositions for injection can be formulated using a sterile solution or any pharmaceutically acceptable liquid as a carrier. Pharmaceutically acceptable carriers include, but are not limited to, sterile water and saline.

The pharmaceutical compositions of the present disclosure can be formulated in microcaps

- 50 046157 sulakh, such as hydroxyl methylcellulose or gelatin microcapsules and polymethyl methacrylate microcapsules. The pharmaceutical compositions of the present disclosure can also be formulated in other drug delivery systems such as liposomes, albumin microspheres, microemulsions, nanoparticles and nanocapsules. The pharmaceutical composition for gene therapy may be in a suitable diluent or may comprise a slow release matrix into which a gene delivery vehicle is embedded.

The pharmaceutical compositions provided herein may be formulated for parenteral administration, subcutaneous administration, intravenous administration, intramuscular administration, intraarterial administration, intraparenchymal administration, intrathecal administration, intracerebellopontine administration, intracerebroventricular administration, or intraperitoneal administration. The pharmaceutical composition may also be prepared or administered by nasal, spray, oral, aerosol, rectal or vaginal administration. In one embodiment, a pharmaceutical composition provided herein is administered into the CNS or cerebrospinal fluid (CSF), i.e. by intraparenchymal injection, intrathecal injection, injection into the cerebellopontine cistern, or intracerebroventricular injection. The tissue target may be specific, such as the CNS, or may be a combination of several tissues, such as muscle and CNS tissues. Exemplary tissue or other targets may include liver, skeletal muscle, cardiac muscle, fat, kidney, lung, vascular endothelium, epithelial cells, hematopoietic cells, CNS and/or CSF. In a preferred embodiment, a pharmaceutical composition provided herein comprising a PV-selective miRNA and/or eTF binding site that selectively activates SCN1A is injected into the CNS or cerebrospinal fluid, i.e. by intraparenchymal injection, intrathecal injection, injection into the cerebellopontine cistern, or intracerebroventricular injection. One or more of these methods can be used to administer the pharmaceutical composition of the present disclosure.

In certain embodiments, a pharmaceutical composition provided herein comprises an effective amount or a therapeutically effective amount. As used herein, such amounts refer to an amount effective in dosages and for periods of time necessary to achieve the desired therapeutic effect, such as increasing the level of SCN1A expression and/or reducing the frequency and/or duration of seizures.

The dosage of the pharmaceutical compositions of the present disclosure depends on factors including the route of administration, the disease to be treated, and the physical characteristics (eg, age, weight, general health) of the subject. Dosage may be adjusted to ensure optimal therapeutic response. Typically, the dosage may be an amount that effectively treats the disease without causing significant toxicity. In one embodiment, the AAV vector provided herein can be administered to a patient for the treatment of SCNIA deficiency (including, for example, Dravet syndrome) in an amount or dose ranging from 5 x 10 ¹¹ to 1 x 10 ¹⁴ gc/kg (number of genomic copies per kilogram of patient's body weight (gk/kg)). In a more specific embodiment, the AAV vector is administered in an amount that ranges from about 5 x 10 ¹¹ gc/kg to about 3 x 10 ¹³ gc/kg, or from about 1 x 10 ¹² to about 1 x 10 ¹⁴ gc/kg, or from about 1 x 10 ¹² to approximately 1x10 ¹³ gk/kg, or approximately 5x10 ¹¹ gk/kg, 1x10 ¹² gk/kg, 1.5x10 ¹² gk/kg, 2.0x10 ¹² gk/kg, 2.5x10 ¹² gk/kg, 3x10 ¹² gk/kg, 3.5x10 ¹² gk/kg, 4x10 ¹² gk/kg, 4.5x10 ¹² gk/kg, 5x10 ¹² gk/kg, 5.5x10 ¹² gk/kg, 6x10 ¹² gk/kg, 6.5x10 ¹² gk/kg, 7x10 ¹² gk/kg, 7.5x10 ¹² gk/kg, 8x10 ¹² gk/kg, 8.5x10 ¹² gk/kg, 9x10 ¹² gk/kg or 9.5x10 ¹² gk/kg. gc/kg can be determined, for example, using qPCR or digital droplet PCR (ddPCR) (see, for example, M. Lock et al, Hum Gene Ther Methods. 2014 Apr;25(2): 115-25). In another embodiment, the AAV vector provided herein can be administered to a patient for the treatment of SCNIA deficiency (including, for example, Dravet syndrome) in an amount or dose ranging from 1x 109 to 1x 10 ¹¹ iu/kg (infectious vector units (v) /body weight of the subject or patient (kg)). In certain embodiments, the pharmaceutical composition can be formulated as a unit dose as needed. Such unit dosage units may contain from about 1 x 10 ⁹ gc to about 1 x 10 ¹⁵ gc.

The pharmaceutical compositions of the present disclosure can be administered to a subject in need thereof, for example, one or more times (eg, 1-10 times or more) daily, weekly, monthly, biannually, annually, or as medically indicated. In an exemplary embodiment, a single administration is sufficient. In one embodiment, a pharmaceutical composition comprising an expression cassette encoding a PV-selective miRNA and/or eTF binding site that selectively activates SCN1A is suitable for use in humans and is administered by intraparenchymal injection, intrathecal injection, pontocerebellar cistern injection, or intracerebroventricular injection. . In one embodiment, the pharmaceutical composition is delivered via a peripheral vein by bolus injection. In other embodiments, the pharmaceutical composition is delivered via a peripheral vein by infusion over about 10 minutes (±5 minutes), over about 20 minutes (±5 minutes), over about 30 minutes (±5 minutes), over about 60 minutes (±5 minutes) or for more than 90 minutes (±10 minutes).

In another aspect, the present application further provides a kit comprising a nucleic acid molecule, vector, host cell, virion, or pharmaceutical composition as described herein in one or more containers. The kit may contain instructions or packaging materials that describe how to administer to a patient the nucleic acid molecule, vector, host cell, or virion contained in the kit. The kit containers may be of any suitable material, such as glass, plastic, metal, etc., and be of any suitable size, shape or configuration. In certain embodiments, kits may contain one or more vials or syringes that contain a nucleic acid molecule, vector, host cell, virion, or pharmaceutical composition in a suitable liquid or solution form.

Methods of treatment.

In one aspect, the present application provides methods of using eTFs that selectively activate SCN1A, as disclosed herein. In certain embodiments, provided herein are methods of introducing an expression cassette, expression vector, or viral particle containing a polynucleotide encoding an eTF that selectively activates SCN1A, as disclosed herein, to enhance expression of SCNIA in a cell. In various embodiments, an eTF that selectively activates SCN1A, as disclosed herein, can be used to modulate SCNIA expression in a cell in vitro, in vivo, or ex vivo.

In certain embodiments, provided herein are methods of treating a disease or disorder associated with SCN1A by administering to a subject an expression cassette, expression vector, or viral particle containing a polynucleotide encoding an eTF that selectively activates SCN1A, as disclosed herein, as appropriate. In certain embodiments, the disorder is a disorder of the central nervous system. In exemplary embodiments, the disease or disorder is associated with SCN1A haploinsufficiency. In certain embodiments, the disorder is epilepsy associated with SCN1A haploinsufficiency. In certain embodiments, the haploinsufficiency results from the subject being heterozygous for a loss of function mutation in the SCN1A gene. In certain embodiments, the disorder is epilepsy associated with an insertion, deletion, or substitution in the SCN1A gene. In certain embodiments, the disorder is epilepsy associated with a point mutation in the SCN1A gene. In certain embodiments, a method of treating a disease or disorder comprises administering an expression cassette, expression vector, or viral particle comprising a polynucleotide encoding an eTF that selectively activates SCN1A, as disclosed herein, such that underexpression of SCN1A is corrected to normal levels. or brought within the normal range as defined by the standard of care. In certain embodiments, the methods disclosed herein are used to treat a disease or disorder associated with endogenous SCN1A containing one or more mutations that result in abnormal expression of SCN1A.

In certain embodiments, this application provides methods for alleviating a symptom associated with a disease or disorder by administering an expression cassette, expression vector, or viral particle containing a polynucleotide encoding an eTF that selectively activates SCN1A, as disclosed herein, to a subject in need thereof. .

In an exemplary embodiment, the present application provides methods for treating a disease, disorder, or symptom associated with a mutation in SCN1A (e.g., point mutation, substitution, deletion, inversion, etc.), deficiency in Nav1.1, and/or reduced Nav1 activity .1 by administering to a subject in need thereof an expression cassette, expression vector or viral particle containing a polynucleotide encoding an eTF that selectively enhances the expression of the SCN1A gene or its protein product Nav1.1. Voltage-gated sodium ion channels are important for the generation and propagation of action potentials in striated muscle and neural tissues. Voltage-gated sodium ion channels are heteromeric complexes consisting of a large central pore-forming glycosylated alpha subunit and two smaller accessory beta subunits. The large alpha subunit Nav1.1 subunit, encoded by the SCN1A gene, is relevant for various diseases or disorders such as Dravet syndrome. Nav1.1 is expressed in neurons and can be assembled with various beta 52 046157 subunits, including the Νανβΐ expressed by the SCN1B gene.

In certain embodiments, provided herein are methods of treating diseases associated with a mutation in SCN1A (eg, deletion, insertion, inversion, point mutation (eg, nonsense mutation, missense mutation), etc.) or reduced Nav1 activity. 1 using an eTF that selectively increases the expression of the endogenous SCN1A gene. Diseases and disorders associated with SCN1A mutations include, but are not limited to: Dravet syndrome, Ohtahara syndrome, epilepsy, early childhood epileptic encephalopathy 6 (EIEE6), familial febrile seizures 3A (FEB3A), intractable childhood epilepsy with generalized tonic-clonic seizures (ICEGTC ), migraine, familial hemiplegia 3 (FHM3), Panagiotopoulos syndrome, familial atrial fibrillation 13 (ATFB13), generalized epilepsy with febrile seizures plus type 1 (gefs + type 1), Brugada syndrome, nonspecific cardiac conduction defect, generalized epilepsy plus, benign early infantile epileptic syndrome, early childhood epileptic encephalopathy (EIEE11), benign familial childhood epilepsy, neurodegeneration, tauopathies and Alzheimer's disease. In some cases, the neurological disorder is Dravet syndrome. Mutations or abnormalities in SCN1A have also been associated with seizure disorders, epilepsy, autism, familial hemiplegic migraine type 3 (FHM3), genetic epilepsy with febrile seizures plus (GEFS+), and the effectiveness of certain anticonvulsant drugs. For example, the ICS5N+5G>A mutation in SCN1A is associated with the maximum safe amount (dose) of the anticonvulsant drugs phenytoin and carbamazepine.

In certain embodiments, the present application provides a method of treating a subject with or at risk of developing Dravet syndrome by administering an expression cassette, expression vector, or viral particle containing a polynucleotide encoding an eTF that selectively activates SCN1A. Dravet syndrome is characterized by prolonged febrile and nonfebrile seizures during the first year of a child's life. This disease progresses to other types of seizures such as myoclonic and partial seizures, psychomotor retardation, and ataxia. It is characterized by cognitive impairment, behavioral disturbances and movement disorders. Behavioral disorders often include hyperactivity and impulsivity, and in rarer cases, autistic behavior. Dravet syndrome is also associated with sleep disturbances, including drowsiness and insomnia. In many patients, Dravet syndrome is caused by genetic mutations that lead to the production of nonfunctional proteins. There are many challenges in treating diseases associated with genetic causes. Thus, most existing treatments have been aimed at preventing seizures and other symptoms.

In 70-90% of patients, Dravet syndrome is caused by nonsense mutations in the SCN1A gene, which results in a premature stop codon and therefore a nonfunctional protein. Typically, a missense mutation in the S5 or S6 segment of the sodium channel pore results in loss of channel function and the development of Dravet syndrome. Heterozygous inheritance of SCN1A mutation, such as nonsense mutation, missense mutation, deletion, insertion, inversion, etc. - this is all that is necessary for the development of a defective sodium channel; Patients with Dravet syndrome will have one normal copy of the gene. Thus, the disease is characterized as haploinsufficiency, and thus increasing the expression of a functional copy of SCN1A may restore normal levels of Nav1.1 production.

Symptoms associated with Dravet syndrome include seizures, memory defects, developmental delays, low muscle tone and/or cognitive problems. Treatment with an expression cassette, expression vector, or virial particle described herein may result in improvement in one or more symptoms, such as a reduction in the number, duration, and/or intensity of seizures. Administration of gene therapy, as described herein, to a subject at risk of developing Dravet syndrome may prevent the development or slow the progression of one or more Dravet symptoms.

In certain embodiments, treatment with an expression cassette, expression vector, or virial particle containing a polynucleotide encoding an eTF that selectively activates SCN1A, as described herein, reduces the duration and/or frequency of seizures, such as seizures associated with Dravet syndrome, by at least at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33% , 34%, 35%, 36%, 37%, 38%, 39%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90 % 95% or more compared to untreated controls or compared to pre-treatment levels.

In some patients with Alzheimer's disease, amyloid β (Ae) production involving many peptides and proteases can influence neuronal excitability, causing seizures and downregulating the Nav1.1 sodium channel in PV neurons. In another embodiment, the present application provides methods for treating a subject suffering from Alzheimer's disease by administering the described

- 53 046157 herein an expression cassette, expression vector or viral particle that contains a polynucleotide encoding an eTF that selectively activates SCN1A. Symptoms associated with Alzheimer's disease include short-term memory loss, cognitive difficulties, seizures, and difficulties with speech, executive function, perception (agnosia), and motor performance (apraxia). Treatment with an expression cassette, expression vector, or viral particle containing a polynucleotide encoding an eTF that selectively activates SCN1A may result in improvement of one or more symptoms of Alzheimer's disease, such as reducing the progression of memory loss or preventing one or more symptoms. In some cases, treatment can lead to correction of brain activity with high gamma radiation output. Treatment may result in a reduction in seizure frequency and/or severity of seizures or a reduction in high-power gamma radiation activity by at least 10%, 20%, 30%, 40%, 50%, 60%, 70% or more, as compared with no treatment. In some cases, treatment may lead to improvements in cognitive function. Learning and/or memory may be improved by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, or greater than 100% compared to without treatment or before treatment with a polynucleotide encoding an eTF that selectively activates SCN1A, as disclosed herein.

In some cases, treatment with an expression cassette, expression vector, or viral particle containing a polynucleotide encoding an eTF that selectively activates SCN1A reduces high-gamma activity (eg, high-gamma activity associated with Alzheimer's disease) by at least 1% , 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18 %, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or 95% compared to untreated controls or compared to pre-treatment levels.

Parkinsonism refers to the collection of signs and symptoms found in Parkinson's disease (PD), including slowness (bradykinesia), stiffness (rigidity), tremors, and imbalance (postural instability). In some cases, administration of an expression cassette, expression vector, or viral particle containing a polynucleotide encoding an eTF that selectively activates SCN1A, as described herein, to a subject at risk for or suffering from Parkinson's disease may prevent the development of one or more symptoms or slow the progression of Parkinson's disease. by at least 10%, by at least 20%, by at least 30%, by at least 40%, by at least 50%, by at least 60%, by at least 70%, according to at least 80% or at least 90% compared to no treatment.

In certain embodiments, this application provides methods that can be used to treat a subject who is at risk of developing a disease. The subject may be known to be predisposed to a disease, such as a neurological disease or a disease associated with epilepsy, seizures and/or encephalopathy. A subject may be predisposed to a disease due to a genetic event or known risk factors. For example, a subject may carry a mutation in SCN1A that is associated with epilepsy (such as Dravet syndrome). Any mutation in the SCN1A gene that reduces its activity (by reducing expression levels, disrupting protein function, or a combination of both) may predispose a subject to the disease, including any one or more of insertions, deletions, inversions, translocations, or substitutions (eg , point mutations, including nonsense mutations and/or missense mutations) in the SCN1A gene. In some cases, a subject may be predisposed to a disease such as Alzheimer's disease due to the age of the subject. In some cases, a subject may have insufficient amounts of SCN1A protein, and treatment of the SCN1A-associated disease involves administration of an expression cassette, expression vector, or viral particle containing a polynucleotide encoding an eTF that selectively activates endogenous SCN1A, as described herein.

In certain embodiments, treatment using an expression cassette, expression vector, or viral particle containing a polynucleotide encoding an eTF that selectively activates endogenous SCN1A as provided herein may result in a reduction or resolution of symptoms associated with Dravet syndrome or other disease associated with with SCN1A, or a disorder such as epilepsy associated with SCN1A haploinsufficiency. For example, treatment may improve learning, memory, cognitive and/or motor function; reduce the frequency and/or duration of seizures; and/or reduce temperature sensitivity (or increase the temperature threshold for triggering a seizure).

In another aspect, the present application provides methods for selectively expressing a transgene in PV-containing neurons by introducing an expression cassette, expression vector, or viral particle containing at least one PV-selective miRNA binding site. In certain embodiments, the present application provides methods for selectively expressing a transgene in primate PV-containing neurons by introducing an ex cassette.

- 54 046157 press, expression vector or viral particle containing a transgene and at least one PV-selective microRNA binding site. In certain embodiments, the present application provides methods for selectively expressing a transgene in PV-containing neurons by introducing an expression cassette, expression vector, or viral particle containing a PV-selective regulatory element operably linked to the transgene and at least one PV-selective regulatory element. microRNA binding site. In exemplary embodiments, the transgene comprises a sequence encoding any of the eTFs that selectively activate SCN1A as described herein.

In certain embodiments, the present application provides a method of gene therapy comprising administering to a subject an expression cassette, expression vector, or viral particle containing a transgene and at least one PV-selective miRNA binding site. In certain embodiments, the application provides gene therapy methods comprising administering to a subject an expression cassette, expression vector, or viral particle containing a PV-selective regulatory element operably linked to a transgene and at least one PV-selective miRNA binding site. In exemplary embodiments, the transgene comprises a sequence encoding any of the eTFs that selectively activate SCN1A as described herein.

In certain embodiments, the present application provides a method of treating a disease or disorder comprising administering an expression cassette, expression vector, or viral particle containing a transgene and at least one PV-selective miRNA binding site. In certain embodiments, the application provides methods of treating a disease or disorder comprising administering to a subject an expression cassette, expression vector, or viral particle comprising a PV-selective regulatory element operably linked to a transgene and at least one PV-selective microRNA binding site . In exemplary embodiments, the transgene comprises a sequence encoding any of the eTFs that selectively activate SCN1A as described herein. In certain embodiments, an expression cassette, expression vector, or viral particle containing a transgene and a PV-selective miRNA binding site, and optionally a PV-selective regulatory element, can be used to treat a disease or disorder that involves PV-containing neurons. In certain embodiments, an expression cassette, expression vector, or viral particle containing a transgene and a PV-selective miRNA binding site and optionally a PV-selective regulatory element is used to treat a neuronal condition. Neuronal diseases or disorders suitable for treatment include, but are not limited to, Dravet syndrome, Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis (ALS), spinal muscular atrophy (SMA), epilepsy, neurodegenerative disorders, movement disorders, mood disorders, motor neuron diseases, progressive muscular atrophy (PMA), progressive bulbar palsy, pseudobulbar palsy, primary lateral sclerosis, neurological effects of AIDS, developmental disorders, multiple sclerosis, neurogenetic disorders, stroke, spinal cord injury, traumatic brain injury, tauopathy , neuronal hypoexcitability and/or seizures. In some embodiments, a viral vector, viral particle, or pharmaceutical composition comprising a transgene and a PV-selective microRNA binding site and optionally a PV-selective regulatory element is used to treat a mental disorder (e.g., schizophrenia, obsessive-compulsive disorder, drug addiction, depression , anxiety, psychosis); autism spectrum disorders (eg, fragile X syndrome, Rett syndrome); epilepsy (eg, Dravet syndrome, chronic traumatic encephalopathy, generalized epilepsy with febrile seizures plus (GEFS+), epileptic encephalopathy, temporal lobe epilepsy, focal epilepsy, tuberous sclerosis, epilepsy associated with SCN1A haploinsufficiency); and/or neurodegeneration (eg, Alzheimer's disease, Parkinson's disease). Diseases associated with dysfunctional PV neurons, such as those caused by loss-of-function mutations in SCN1A or Nav1.1, include: Dravet syndrome, Ohtahara syndrome, epilepsy, early childhood epileptic encephalopathy 6 (EIEE6), familial febrile seizures 3A (FEB3A), intractable childhood epilepsy with generalized tonic-clonic seizures (ICEGTC), migraine, familial hemiplegia 3 (FHM3), Panagiotopoulos syndrome, familial atrial fibrillation 13 (ATFB13), generalized epilepsy with febrile seizures plus type 1 (gefs + type 1), nonspecific cardiac syndrome, nonspecific Brugada syndrome conduction defect, generalized epilepsy with febrile seizures plus, benign early infantile epileptic syndrome, early childhood epileptic encephalopathy 11 (EIEE11), benign familial childhood epilepsy, neurodegeneration, tauopathies and Alzheimer's disease.

In certain embodiments, treatment using an expression cassette, expression vector, or viral particle containing a transgene and a PV selective site

- 55 046157 microRNA binding, and possibly the PV-selective regulatory element described herein, leads to improvement of symptoms associated with a neuronal disease or disorder. For example, a patient with Parkinson's disease may be monitored symptomatically for improvement in motor function, indicating a positive response to treatment. Administration of therapy using the method described herein to a subject at risk of developing a neuronal disorder may prevent the development or slow the progression of one or more symptoms.

In certain embodiments, an expression cassette, expression vector, or viral particle comprising a transgene and a PV-selective miRNA binding site, and optionally a PV-selective regulatory element, provided herein can be used to treat a subject who has A neuronal disease has been diagnosed, such as epilepsy associated with SCN1A haploinsufficiency, such as Dravet syndrome. In various embodiments, any of the neuronal diseases or disorders described herein are caused by a known genetic event (eg, any of the SCN1A mutations known in the art) or have an unknown cause.

In certain embodiments, an expression cassette, expression vector, or viral particle containing a transgene and a PV-selective miRNA binding site, and optionally a PV-selective regulatory element provided herein, can be used to treat a subject who is in group at risk of developing a disease or disorder. In certain embodiments, it may be known that the subject is predisposed to a disease, such as a neuronal disease (eg, epilepsy associated with SCN1A haploinsufficiency, such as, for example, Dravet syndrome). In certain embodiments, the subject may be predisposed to a disease due to a genetic event or due to known risk factors. For example, a subject may carry a mutation in SCN1A that is associated with epilepsy or Dravet syndrome, such as an insertion, deletion, inversion, translocation or substitution (eg, a point mutation, including a nonsense mutation and/or a missense mutation).

In certain embodiments, an expression cassette, expression vector, or viral particle containing a transgene and a PV-selective miRNA binding site, and optionally a PV selective regulatory element, provided herein can be used to reduce one or more symptoms associated with disease or disorder. For example, symptoms associated with Dravet syndrome include seizures, memory defects, developmental delays, poor muscle tone, and/or cognitive problems. Treatment with a viral vector, viral particle, or pharmaceutical composition comprising a transgene and a PV-selective miRNA binding site and, optionally, a PV-selective regulatory element provided herein may result in improvement of one or more symptoms, such as a decrease in the number, duration and/or intensity of seizures.

In certain embodiments, the methods described herein are used to increase transgene expression in a PV neuron, gene therapy, or treat a disease or disorder in primates. In certain embodiments, the primate is a human. In certain embodiments, the primate is a non-human primate. In certain embodiments, the non-human primate is an old world monkey, an orangutan, a gorilla, a chimpanzee, a cynomolgus monkey, a rhesus monkey, or a pig-tailed monkey.

The terms subject and individual are used interchangeably herein to refer to a vertebrate animal, preferably a mammal, more preferably a human. The methods described herein may be useful for human therapeutic purposes, veterinary medicine, and/or preclinical studies in animal models of a disease or condition. In various embodiments, the subject that can be treated in accordance with the methods described herein is a mammal, such as, for example, a mouse, rat, hamster, guinea pig, gerbil, cow, sheep, pig, goat, donkey, horse, dog, cat, llama, monkey (such as an old world monkey, a marmoset, or a macaque such as a rhesus macaque, pig-tailed macaque, or cynomolgus macaque (such as a cynomolgus macaque)), an ape (such as an orangutan, a gorilla, or a chimpanzee), or a human . In an exemplary embodiment, the subject is a human.

The following tables present the sequences disclosed herein.

Table 1. Exemplary engineered transcription factors disclosed herein. The sequences of regulatory elements (RE) are disclosed below in table. 2 and 8. For RE, when m1 is specified, this means that the miRNA binding site m1 (SEQ ID NO: 7, Table 8) is included between the coding region and the polyA tail. The DNA binding domain (DBD) sequences are disclosed in Table 1 below. 3. For DBD, the engineered zinc finger (eZF) indicates that the design has the formula set forth in SEQ ID NO: 147 (Table 10); EGR1 indicates that DBD is occurring

- 56046157 dit from wild type human EGR1 (SEQ ID NO: 176; Table 12); and EGR3 indicates that the DBD is derived from wild-type human EGR3 (SEQ ID NO: 175, Table 12). Sequences for target sites (eg, sequences associated with the DBD) are presented in Table. 4 below. The sequences for transcription activation domains (TADs) are disclosed below in table. 5. For TAD (c) indicates that the TAD is located at the c-terminus of the DBD, (n) indicates that the TAD is located at the n-terminus of the DBD, (n/c) indicates that there is a TAD located both at the n-terminus and the c-terminus of the DBD, and 2x CITED4 (n) indicates that there are 2 copies of the CITED4 TAD located at the n-terminus of the DBD. The sequences of the designed full-length transcription factors (DBD + TAD) are presented in Table. 6 below.

Table 1

Design RE DBD Target site TAD (position) SEQ ID NO (DBD) SEQ ID NO (DBD+TAD) 1 RE 1 eZF Z13 VPR (c) 89 99 2 RE 1 eZF Z1 VPR (c) 77 100 3 RE 1 eZF Z13 VP64(c) 89 101 4 RE 1 eZF Z1 VP64(c) 77 102 5 RE 1 EGR1 Z13 CITED4 (n/c) 93 103 6 RE 1 EGR1 Z13 CITED4(n) 93 104 7 RE 1 EGR1 Z1 CITED4 (n/c) 92 105 8 RE 1 EGR1 Z1 CITED4(n) 92 106 9 RE 1 EGR1 Z1 CITED4 (c) 92 107 10 RE 1 EGR1 Z1 CITED2 (c) 92 108 AND RE 1 EGR1 Z1 CITED2(n) 92 109 12 RE 1 EGR3 Z1 CITED4 (n/c) 96 110 13 RE 1 EGR3 Z1 CITED4 (c) 96 111 14 RE 1 EGR3 Z1 CITED2 (c) 96 112 15 RE 1 EGR3 Z1 CITED2(n) 96 113 16 NVA EGR3 Z15 N/A 98 114 17 RE 1 EGR1 Z13 N/A 93 115 18 RE 1 EGR1 Z15 N/A 94 116 19 RE 1 EGR1 Z13 N/A 93 117 20 RE 1 EGR1 Z13 N/A 93 115 21 RE 1 EGR1 Z1 N/A 92 118 22 NVA EGR3 Z13 N/A 97 119 23 RE 1 EGR1 Z17 N/A 95 120 24 RE 1 EGR1 Z13 N/A 93 121 25 NVA EGR3 Z1 N/A 96 122 26 RE 1 EGR1 Z1 N/A 92 123 27 RE 1 EGR1 Z1 N/A 92 124 28 RE 1 eZF Z8 VP64(c) 84 125 29 RE 1 eZF Z14 VP64(c) 90 126 thirty NVA eZF Z13 VPR (c) 89 99 31 RE2(ml) eZF Z1 VP64(c) 77 102 32 RE2 eZF Z1 VP64(c) 77 102 33 RE2 eZF Z1 VPR (c) 77 100 34 RE2 eZF Z1 VP64(c) 77 127 35 RE 2 (ml) eZF Z1 VP64(c) 77 127 36 RE2 EGR1 Z1 CITED4(n) 92 128 37 RE 2 (ml) EGR1 Z1 CITED4(n) 92 106 38 RE 2 (ml) EGR1 Z1 CITED4(n) 92 129 39 RE 2 (ml) EGR1 Z1 CITED4 (n/c) 92 105 40 RE 2 (ml) EGR1 Z1 2x CITED4 (n) 92 130

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41 RE2(ml) EGR1 Z1 2x CITED4 (n) 92 131 42 RE 2 (m2) eZF Z1 VP64(c) 77 102 43 RE 2 (m3) eZF Z1 VP64(c) 77 102 44 RE 2 (m2) EGR1 Z1 CITED4(n) 92 106 45 RE 2 (m3) EGR1 Z1 CITED4(n) 92 106 46 RE 1 EGR1 Z1 CITED4(n) 92 205 47 RE 1 EGR1 Z1 2x CITED4 (n) 92 207 48 RE 1 EGR1 Z1 2x CITED4 (n) 92 209 49 C.B.A. eZF Z1 CREB3(n) 77 213 50 C.B.A. EGR1 Z1 CREB3(n) 92 (without Conc. Lys) 217 51 C.B.A. EGR1 Z13 CREB3(n) 93 (without Conc. Lys) 219 52 C.B.A. eZF Z1 CREB3(n); without TM domain on (c) 77 221 53 C.B.A. EGR1 Z1 CREB3(n); without TM domain on (c) 92 (without Conc. Lys) 223

Table 2 Nucleic acid sequences for various regulatory elements (RE) ____________________disclosed herein____________________

RE Subsequence SEQ ID NO RE 1 GTGATGCGGTTTTGGCAGTACATCAATGGGCGTGGATAG CGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATT GACGTCAATGGGAGTTTGTTTTGGCACCAAAATCAACGG GACTTTCCAAAATGTCGTAACAACTCCGCCCCATTGACG CAAATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATA AGCAGAGCTGGTACCGTGTGTATGCTCAGGGGCTGGGAA A GGAGGGGAGGGAGCTCCGGCTCAG 1 RE2 ggaggaagccatcaactaaactacaatgactgtaagatacaaaattgggaatggtaacatatttt gaagttctgttgacataaagaatcatgattaatgcccatggaaatgaaagggcgatcaacac tatggtttgaaaagggggaaattgtagagcacagatgtgttcgtgtggcagtgtgctgtctctag caatactcagaga agagagagaacaatgaaattctgattggccccagtgtgagcccagatga ggttcagctgccaactttctctttcacatcttatgaaagtcatttaagcacaactaactttttttttttttt tttttttttgagacagagtcttgctctgttgcccaggacagagtgcagtagtgactcaatctcggct cactgcagcct ccacctcctaggctcaaacggtcctcctgcatcagcctcccaagtagctgga attacaggagtggcccaccatgcccagctaatttttgtatttttaatagatacgggggtttcaccat atcacccaggctggtctcgaactcctggcctcaagtgatccacctgcctcggcctcccaaagt gctgggattataggcgtcagccactatgcccaacc cgaccaaccttttttaaaataaatatttaaa aaattggtatttcacatatatactagtatttacatttatccacacaaaacggacgggcctccgctg aaccagtgaggccccagacgtgcgcataaataacccctgcgtgctgcaccacctggggaga gggggaggaccacggtaaatggagcgagcgcatagcaaaagggacgcggggtcctt ttctc tgccggtggcactgggtagctgtggccaggtgtggtactttgatggggcccagggctggagc 2

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tcaaggaagcgtcgcagggtcacagatctgggggaaccccggggaaaagcactgaggcaa aaccgccgctcgtctcctacaatatatgggagggggaggttgagtacgttctggattactcata agacctttttttttccttccgggcgcaaaaccgtgagctggatttataatcgccctataaagctcc agaggcggtca ggcacctgcagaggagccccgccgctccgccgactagctgcccccgcga gcaacggcctcgtgatttccccgccgatccggtccccgcctccccactctgcccccgcctacc ccggagccgtgcagccgcctctccgaatctctcttctcctggcgctcgcgtgcgagaggga actagcgagaacgaggaagcagctggaggt gacgccgggcagattacgcctgtcagggcc gagccgagcggatcgctgggcgctgtgcagaggaaaggcgggagtgcccggctcgctgtc gcagagccgaggtgggtaagctagcgaccacctggacttcccagcgcccaaccgtggctttt cagccaggtcctctcctcccgcggcttctcaaccaaccccatcccag cgccggccacccaac ctcccgaaatgagtgcttcctgccccagcagccgaaggcgctactaggaacggtaacctgtta cttttccaggggccgtagtcgacccgctgcccgagttgctgtgcgactgcgcgcgcggggct agagtgcaaggtgactgtggttcttctctggccaagtccgagggagaacgtaaaga tatgggc ctttttccccctctcaccttgtctcaccaaagtccctagtccccggagcagttagcctctttctttcc agggaattagccagacacaacaacgggaaccagacaccgaaccagacatgcccgccccgt gcgccctccccgctcgctgcctttcctccctcttgtctctccagagccggatcttcaaggggag cctccgtgc ccccggctgctcagtccctccggtgtgcaggaccccggaagtcctccccgcac agctctcgcttctctttgcagcctgtttctgcgccggaccagtcgaggactctggacagtagag gccccgggacgaccgagctg RE 3 TCAACAGGGGGACACTTGGGAAAGAAGGATGGGGACAG AGCCGAGAGGACTGTTACACATTAGAGAAACATCAGTG ACTGTGCCAGCTTTGGGGTAGACTGCACAAAAGCCCTGA GGCAGCACAGGCAGGATCCAGTCTGCTGGTCCCAGGAA GCTAACCGTCTCAGACAGAGCACAAAGCACCGAGACAT GTGCCACAAGGCTTGTGTAGAGAGGTCAGAGGACAGCG TACAGGTCC CAGAGATCAAACTCAACCTCACCAGGCTTG GCAGCAAGCCTTTACCAACCCACCCCCACCCCACCCACC CTGCACGCGCCCCTCTCCCCTCCCCATGGTCTCCCATGGC TATCTCACTTGGCCCTAAAATGTTTAAGGATGACACTGG CTGCTGAGTGGAAATGAGACAGCAGAAGTCAACAGTAG ATTTTAGGAAAGCCAGAGAAAAAGGCTTGTGCTGTTTTTT AGAAAGCCAAGGGACAA GCTAAGATAGGGCCCAAGTAA TGCTAGTATTTACATTTATCCACACAAAACGGACGGGCC TCCGCTGAACCAGTGAGGCCCCAGACGTGCGCATAAATA ACCCCTGCGTGCTGCACCACCTGGGGAGAGGGGGAGGA CCACGGTAAATGGAGCGAGCGCATAGCAAAAGGGACGC GGGGTCCTTTTCTCTGCCGGTGGCACTGGGTAGCTGTGG CCAGGTGTGGTACTTTGATGGGGCCCAGGGCT GGAGCTC AAGGAAGCGTCGCAGGGTCACAGATCTGGGGGAACCCC GGGGAAAAGCACTGAGGCAAAACCGCCGCTCGTCTCCT ACAATATATGGGAGGGGGAGGTTGAGTACGTTCTGGATT ACTCATAAGACCTTTTTTTTTTCCTTCCGGGCGCAAAACC GTGAGCTGGATTTATAATCGCCCTATAAAGCTCCAGAGG CGGTCAGGCACCTGCAGAGGAGCCCCGCCGCTCCGCCGA CTAGCT GCCCCCGCGAGCAACGGCCTCGTGATTTCCCCG CCGATCCGGTCCCCGCCTCCCCACTCTGCCCCCGCCTACC CCGGAGCCGTGCAGCCGCCTCTCCGAATCTCTCTCTTCTC CTGGCGCTCGCGTGCGAGAGGGAACTAGCGAGAACGAG GAAGCAGCTGGAGGTGACGCCGGGCAGATTACGCCTGT 3

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CAGGGCCGAGCCGAGCGGATCGCTGGGCGCTGTGCAGA GGAAAGGCGGGAGTGCCCGGCTCGCTGTCGCAGAGCCG AGGTGGGTAAGCTAGGCGACCACCTGGACTTCCCAGCGCC CAACCGTGGCTTTTCAGCCAGGTCCTCTCCTCCCGCGGCT TCTCAACCAACCCCATCCCAGCGCCGGCCACCCAACCTC CCGAAATGAGTGCTTCCTGCCCCAGCAGCCGAAGGCGCT ACTAGGAAC GGTAACCTGTTACTTTTCCAGGGGCCGTAG TCGACCCGCTGCCCGAGTTGCTGTGCGACTGCGCGCGCG GGGCTAGAGTGCAAGGTGACTGTGGTTCTTCTCTGGCCA AGTCCGAGGGAGAACGTAAAGATATGGGCCTTTTTCCCC CTCTCACCTTGTCTCACCAAAGTCCCTAGTCCCCGGAGC AGTTAGCCTCTTTCTTTCCAGGGAATTAGCCAGACACAA CAACGGGAAC CAGACACCGAACCAGACATGCCCGCCCC GTGCGCCTCCCCGCTCGCTGCCTTTCCTCCCTCTTGTCT CTCCAGAGCCGGATCTTCAAGGGGAGCCTCCGTGCCCCC GGCTGCTCAGTCCCTCCGGTGTGCAGGACCCCGGAAGTC CTCCCCGCACAGCTCTCGCTTCTCTTTGCAGCCTGTTTCT GGCGCCGGACCAGTCGAGGACTCTGGACAGTAGAGGCCC CGGGACGACCGA GCTG RE 4 GCCCTCTAGGCCACCTGACCAGGTCCCCTCAGTCCCCCCC CTTCCCACACTCCCACACTCAGCCCCCCTCCCCCCCCCCC GACCCCTGCAGGATTATCCTGTCTGTGTTCCTGACTCAGC CTGGGAGCCACCTGGGCAGCAGGGGCCAAGGGTGTCCT AGAAGGGACCTGGAGTCCACGCTGGGCCAAGCCTGCCCT TTCTCCCTCTGTCTTCCGTCCCTGCTTGCGGTTCTGCTGA ATGTGGTTATTTCTCTGGCTCCTTTTACAGAGAATGCTGC TGCTAATTTTATGTGGAGCTCTGAGGCAGTGTAATTGGA AGCCAGACACCCTGTCAGCAGTGGGCTCCCGTCCTGAGC TGCCATGCTTCCTGCTCTCCTCCCGTCCCGGCTCCTCATT TCATGCAGCCACCTGTCCCAGGGAGAGAGGAGTCACCCA GGCCCCTCAGTCCGCCCCTTAAATAAGAAAGCCTCCGTT GCTCGGCACACATACCAAGCAGCCGCTGGTGCAATCT 4 CBA (CMV enhancer + chicken betaactin promoter) CGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACC GCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTA TGTTCCCATAGTAACGCCAATAGGGACTTTCCATTGACG TCAATGGTGGAGTATTTACGGTAAACTGCCCACTTGGC AGTACATCAAGTGTATCATATGCCAAGTACGCCCCCTAT TGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGC CCAGTA CATGACCTTATGGGACTTTCCTACTTGGCAGTA CATCTACGTATTAGTCATCGCTATTACCATGggtcgaggtgagc cccacgttctgcttcactctccccatctcccccccctccccacccccaattttgtatttatttatttttt aattattttgtgcagcgatggggggcggggggggggggcgcgcgccaggcggggcggg gcggggc gaggggcggggcggggcgaggcggagaggtgcggcggcagccaatcagag cggcgcgctccgaaagtttccttttatggcgaggcggcggcggcggcggccctataaaaagc gaagcgcgcggcgggcgggagtcgctgcgttgccttcgccccgtgccccgctccgcgccg cctcgcgccg cccgccccggctctgactgaccgcgttactcccacaggtgagcgggcggga cggcccttctcctccgggctgtaattagcgcttggtttaatgacggctcgtttcttttctgtggctg cgtgaaagccttaaagggctccgggagggccctttgtgcgggggggagcggctcgggggg tgcgtgcgtgtg tgtgtgcgtggggagcgccgcgtgcggcccgcgctgcccggcggctgtg agcgctgcgggcgcggcgcggggctttgtgcgctccgcgtgtgcgcgaggggagcgcgg 5

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ccggggggcggtgccccgcggtgcggggggggctgcgaggggaacaaaggctgcgtgcgg ggtgtgtgcgtgggggggtgagcagggggtgtgggcgcggcggtcgggctgtaacccccc cctgcacccccctccccgagttgctgagcacggcccggcttcgggtgcggggctccgtgc g gggcgtggcgcggggctcgccgtgccggggcggggggtggcggcaggtgggggtgccgg gcggggcggggccgcctcgggccggggagggctcgggggaggggcgcggcggccccg gagcgccggcggctgtcgaggcgcggcgagccgcagccattgccttttatggtaatcgtgcg agagggcgca gggacttcctttgtcccaaatctggcggagccgaaatctgggaggcgccgc cgcaccccctctagcgggcgcgggcgaagcggtgcggcgccggcaggaaggaaatggg cggggagggccttcgtgcgtcgccgcgccgccgtccccttctccatctccagcctcggggct gccgcaggggacggctgcct tcgggggggacggggcagggcggggttcggcttctggc gtgtgaccggcggctctagagcctctgctaaccatgttcatgccttcttctttttcctacagctcct gggcaacgtgctggttgttgtgctgtctcatcattttggcaaagaatt EF1alpha GAGTAATTCATACAAAAGGACTCGCCCCTGCCTTGGGGA ATCCCAGGGACCGTCGTTAAACTCCCACTAACGTAGAAC CCAGAGATCGCTGCGTTCCCGCCCCCTCACCCGCCCGCT CTCGTCATCACTGAGGTGGAGAAGAGCATGCGTGAGGCT CCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAG TCCCCGAGAAGTTGGGGGGAGGGGTCGGCAATTGAACC GGTGCC TAGAGAAGGTGGCGCGGGGTAAACTGGGAAAG TGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTGGG GGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTT CTTTTTCGCAACGGGTTTGCCGCCAGAACACAGGTAAGT GCCGTGTGTGGTTCCCGCGGGCCTGGCCTCTTTACGGGTT ATGGCCCTTGCGTGCCTTGAATTACTTCCACGCCCCTGGC TGCAG TACGTGATTCTTGATCCCGAGCTTCGGGTTGGAA GTGGGTGGGAGAGTTCGAGGCCTTGCGCTTAAGGAGCCC CTTCGCCTCGTGCTTGAGTTGAGGCCTGGCTTGGGCGCT GGGGCCGCCGCGTGCGAATCTGGTGGCACCTTCGCGCCT GTCTCGCTGCTTTCGATAAGTCTCTAGCCATTTAAAATTT TTGATGACCTGCTGCGACGCTTTTTTTCTGGCAAGATAGT CTTGTAAAT GCGGGCCAAGATCTGCACACTGGTATTTCG GTTTTTGGGGCCGCGGGCGGCGACGGGGCCCGTGCGTCC CAGCGCACATGTTCGGCGAGGCGGGGCCTGCGAGCGCG GCCACCGAGAATCGGACGGGGGTAGTCTCAAGCTGGCC GGCCTGCTCTGGTGCCTGGCCTCGCCGCCGTGTATCG CCCCGCCCTGGGCGGCAAGGCTGGCCCGGTCGGCACCAG TTGCGTGAGCGGAAA GATGGCCGCTTCCCGGCCCTGCTG CAGGGAGCTCAAAATGGAGGACGCGGCTCGGGAGAG CGGGCGGGTGAGTCACCCACACAAAGGAAAAGGGCCTT TCCGTCCTCAGCCGTCGCTTCATGTGACTCCACGGAGTA CCGGGCGCCGTCCAGGCACCTCGATTAGTTCTCGAGCTT TTGGAGTACGTCGTCTTTAGGTTGGGGGGAGGGGTTTTA TGCGATGGAGTTTCCCCACACTGA GTGGGTGGAGACTGA AGTTAGGCCAGCTTGGCACTTGATGTAATTCTCCTTGGA ATTTGCCCTTTTTGAGTTTGGATCTTGGTTCATTCTCAAG CCTCAGACAGTGGTTCAAAGTTTTTTTCTTCCATTTCAGG TGTCGTGA 6

Table 3. Amino acid sequences for exemplary DNA binding domains (DBDs) presented herein. For DBD, the engineered zinc finger (eZF) indicates that the design has the formula set forth in SEQ ID NO: 147 (Table 10); EGR1 indicates that the DBD is derived from wild-type human EGR1 (SEQ ID NO: 176; Table 12); and EGR3 indicates that the DBD is derived from wild-type human EGR3 (SEQ ID NO: 175, Table 12). The target sites are sequences associated with the DBD and are presented in Table. 4 below.

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Table 3

DBD/target site Subsequence SEQ ID NO eZF/zl LEPGEKPYKCPECGKSFSRSDNLVRHQRTHTGEKPYKCPE CGKSFSREDNLHTHQRTHTGEKPYKCPECGKSFSRSDELV RHQRTHTGEKPYKCPECGKSFSQSGNLTEHQRTHTGEKPY KCPECGKSFSTSGHLVRHQRTHTGEKPYKCPECGKSFSQN STLTEHQRTHTTGKKTS 77 eZF/z2 LEPGEKPYKCPECGKSFSTKNSLTEHQRTHTGEKP YKCPEC GKSF SRADNLTEHQRTHTGEKP YKCPEC GKSF SQL AHLR A HQRTHTGEKPYKCPECGKSFSTKNSLTEHQRTHTGEKPYK CPECGKSFSQAGHLASHQRTHTGEKPYKCPECGKSFSTHL DLIRHQRTHTGKKTS 78 eZF/z3 LEPGEKPYKCPECGKSFSQAGHLASHQRTHTGEKPYKCPE CGKSFSREDNLHTHQRTHTGEKPYKCPECGKSFSTSGNLTE HQRTHTGEKPYKCPECGKSFSTHLDLIRHQRTHTGEKPYK CPECGKSFSQKSSLIAHQRTHTGEKPYKCPECGKSFSQAGH LASHQRTHTTGKKTS 79 eZF/z4 LEPGEKPYKCPECGKSFSTTGNLTVHQRTHTGEKPYKCPE CGKSFSTSGELVRHQRTHTGEKPYKCPECGKSFSREDNLH THQRTHTGEKPYKCPECGKSFSTSGNLTEHQRTHTGEKPY KCPECGKSFSQS S SLVRHQRTHTGEKPYKCPECGKSF SQRA NLRAHQRTHTGKKT S 80 eZF/z5 LEPGEKPYKCPECGKSF S SRRTCRAHQRTHTGEKP YKCPEC GKSFSTTGALTEHQRTHTGEKPYKCPECGKSFSRSDELVRH QRTHTGEKP YKCPEC GKSFSRNDALTEHQRTHTGEKPYKC PECGKSFSQSGDLRRHQRTHTGEKPYKCPECGKSFSTSHSL TEHQRTHTGKKTS 81 eZF/z6 LEPGEKPYKCPECGKSFSRKDNLKNHQRTHTGEKPYKCPE CGKSFSDPGALVRHQRTHTGEKPYKCPECGKSFSREDNLH THQRTHTGEKPYKCPECGKSFSDPGALVRHQRTHTGEKPY KCPECGKSFSTSGELVRHQRTHTGEKPYKCPECGKSFSRK DNLKNHQRTHTGKKTS 82 eZF/z7 LEPGEKPYKCPECGKSF S SKKALTEHQRTHTGEKP YKCPEC GKSF S SP ADLTRHQRTHTGEKP YKCPEC GKSF SRSDNLVR HQRTHTGEKPYKCPECGKSFSREDNLHTHQRTHTGEKPYK CPECGKSFSRSDELVRHQRTHTGEKPYKCPECGKSFSQSGN LTEHQRTHTGEKPYKCPECGKSF STSGHLVRHQRTHTGEK PYKCPECGKSFSQNSTLTEHQRTHTTGKKTS 83

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eZF/z8 LEPGEKPYKCPECGKSF S SPADLTRHQRTHTGEKPYKCPEC GKSF SRSDNLVRHQRTHTGEKPYKCPECGKSF SREDNLHT HQRTHTGEKPYKCPECGKSFSRSDELVRHQRTHTGEKPYK CPECGKSFSQSGNLTEHQRTHTGEKPYKCPECGKSFSTSGH LVRHQRTHTTGKKTS 84 eZF/z9 LEPGEKPYKCPECGKSF S SKKALTEHQRTHTGEKPYKCPEC GKSF S SPADLTRHQRTHTGEKPYKCPEC GKSF SRSDNLVR HQRTHTGEKPYKCPECGKSFSREDNLHTHQRTHTGEKPYK CPECGKSFSRSDELVRHQRTHTGEKPYKCPECGKSFSQSGN LTEHQRTHTGKKTS 85 eZF/zlO LEPGEKPYKCPECGKSFSDCRDLARHQRTHTGEKPYKCPE CGKSFSRNDALTEHQRTHTGEKPYKCPECGKSFSRNDALT EHQRTHTGEKPYKCPECGKSFSSPADLTRHQRTHTGEKPY KCPECGKSFSDPGNLVRHQRTHTGEKPYKCPECGKSFSQR AHLERHQRTHTGEKPYKCPECGKSFSQSS SLVRHQRTHTG EKPYKCPECGKSFSHRTTLTNHQRTHTGKKTS 86 eZF/zll LEPGEKPYKCPECGKSFSRNDALTEHQRTHTGEKPYKCPE CGKSFSSPADLTRHQRTHTGEKPYKCPECGKSFSDPGNLV RHQRTHTGEKPYKCPECGKSFSQRAHLERHQRTHTGEKPY KCPECGKSFSQS S SLVRHQRTHTGEKPYKCPECGKSF SHRT TLTNHQRTHTGKKTS 87 eZF/zl2 LEPGEKPYKCPECGKSFSRNDALTEHQRTHTGEKPYKCPE CGKSFSDPGHLVRHQRTHTGEKPYKCPECGKSFSTSGELV RHQRTHTGEKPYKCPECGKSFSTHLDLIRHQRTHTGEKPY KCPECGKSFS SKKALTEHQRTHTGEKPYKCPEC GKSF SQL AHLRAHQRTHTGEKPYKCPECGKSFSR SDHLTNHQRTHTG KKTS 88 eZF/zl3 LEPGEKPYKCPECGKSFSRSDNLVRHQRTHTGEKPYKCPE CGKSFSHRTTLTNHQRTHTGEKPYKCPECGKSFSREDNLH THQRTHTGEKPYKCPECGKSFSTSHSLTEHQRTHTGEKPY KCPECGKSFSQS S SLVRHQRTHTGEKPYKCPECGKSF SRED NLHTHQRTHTGKKTS 89 eZF/zl4 LEPGEKPYKCPECGKSFSDPGALVRHQRTHTGEKPYKCPE CGKSFSRSDNLVRHQRTHTGEKPYKCPECGKSFSQSGDLR RHQRTHTGEKPYKCPECGKSFSTHLDLIRHQRTHTGEKPY KCPECGKSFSTSGNLVRHQRTHTGEKPYKCPECGKSFSRSD NLVRHQRTHTGKKTS 90 eZF/zl5 LEPGEKPYKCPECGKSFSRRDELNVHQRTHTGEKPYKCPE CGKSFSRSDHLTNHQRTHTGEKPYKCPECGKSFSRSDDLV RHQRTHTGEKPYKCPECGKSFSRSDNLVRHQRTHTGEKPY KCPECGKSFSHRTTLTNHQRTHTGEKPYKCPECGKSFSRED NLHTHQRTHTGEKPYKCPECGKSFSTSHSL TEHQRTHTGE KPYKCPECGKSFSQSSSLVRHQRTHTGEKPYKCPECGKSFS REDNLHTHQRTHTGKKTS 91 EGRl/zl RPYACPVESCDRRFSRSDNLVRHIRIHTGQKPFQCRICMRN FSREDNLHTHIRTHTGEKPFACDICGRKFARSDELVRHTKI HLRQKDRPYACPVESCDRRFSQSGNLTEHIRIHTGQKPFQC RICMRNFSTSGHLVRHIRTHTGEKPFACDICGRKFAQNSTL TEHTKIHLRQKDK 92

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EGR1/Z13 RPYACPVESCDRRFSRSDNLVRHIRIHTGQKPFQCRICMRN FSHRTTLTNHIRTHTGEKPFACDICGRKFAREDNLHTHTKI HLRQKDRPYACPVESCDRRFSTSHSLTEHIRIHTGQKPFQC RICMRNFSQSSSLVRHIRTHTGEKPFACDICGRKFAREDNL HTHTKIHLRQKDK 93 EGR1/Z15 RPYACPVESCDRRFSRRDELNVHIRIHTGQKPFQCRICMRN FSRSDHLTNHIRTHTGEKPFACDICGRKFARSDDLVRHTKI HLRQKDRP YACPVESCDRRF SRSDNLVRHIRIHTGQKPFQC RICMRNFSHRTTLTNHIRTHTGEKPFACDICGRKFAREDNL HTHTKIHLRQKDRPYACPVESCDRRFSTSHSLTEHIRIHTG Q KPFQCRICMRNFSQSSSLVRHIRTHTGEKPFACDICGRKFA REDNLHTHTKIHLRQKD 94 EGrl/zl7 RPYACPVESCDRRFSDPGALVRHIRIHTGQKPFQCRICMRN FSRSDNLVRHIRTHTGEKPFACDICGRKFAQSGDLRRHTKI HLRQKDRPYACPVESCDRRFSTHLDLIRHIRIHTGQKPFQC RICMRNFSTSGNLVRHIRTHTGEKPFACDICGRKFARSDNL VRHTKIHLRQKDRPYACPVESCDRRFSQSGHLTEHIRIHTG QKPFQCRICMRNFSERSHLREHIRTHTGEKPFACDICGRKF AQAGHLASHTKIHLRQKD 95 EGR3/zl RPHACPAEGCDRRFSRSDNLVRHLRIHTGHKPFQCRICMRS FSREDNLHTHIRTHTGEKPFACEFCGRKFARSDELVRHAKI HLKQKEHACPAEGCDRRFSQSGNLTEHLRIHTGHKPFQCRI CMRSFSTSGHLVRHIRTHTGEKPFACEFCGRKFAQNSTLTE HAKIHLKQKEK 96 EGR3/zl3 RPHACPAEGCDRRFSRSDNLVRHLRIHTGHKPFQCRICMRS FSHRTTLTNHIRTHTGEKPFACEFCGRKFAREDNLHTHAKI HLKQKEHACPAEGCDRRFSTSHSLTEHLRIHTGHKPFQCRI CMRSFSQSSSLVRHIRTHTGEKPFACEFCGRKFAREDNLHT HAKIHLKQKEK 97 EGR3/zl5 RPHACPAEGCDRRFSRRDELNVHLRIHTGHKPFQCRICMRS FSRSDHLTNHIRTHTGEKPFACEFCGRKFARSDDLVRHAKI HLKQKEHACPAEGCDRRF SRSDNLVRHLRIHTGHKPFQCR ICMRSFSHRTTLTNHIRTHTGEKPFACEFCGRKFAREDNLH THAKIHLKQKEHACPAEGCDRRFSTSHSLTEHLRIHTGHKP FQCRIC MRSF SQS S SLVRHIRTHTGEKPF ACEFCGRKFARE DNLHTHAKIHLKQKEK 98

Table 4 Target site sequences and chromosomal locations of exemplary target sites associated with DNA binding domains disclosed herein

Initial position e chrome. 2 Target Site Sequence Target site SEQ ID NO for sequential target site 166149168 CTAGGTCAAGTGTAGGAG zl 18 166149158 ACTTGACCTAGACAGCCT z2 19 166073978 TGAATAACTCATTAGTGA z3 20

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166073933 AAAGTACATTAGGCTAAT z4 21 166149199 CCAGCACTGGTGCTTCGT z5 22 166149176 AAGGCTGTCTAGGTCAAG z6 23 166149168 CTAGGTCAAGTGTAGGAGACACAC z7 24 166149165 GGTCAAGTGTAGGAGACA z8 25 166149162 CAAGTGTAGGAGACACAC z9 26 166149160 AGTGTAGGAGACACACTGCTGGCC zlO 27 166149160 AGTGTAGGAGACACACTG zll 28 166149155 AGGAGACACACTGCTGGCCTG zl2 29 166128025 TAGGTACCATAGAGTGAG zl3 thirty 166127991 GAGGATACTGCAGAGGTC zl4 31 166127999 TAGGTACCATAGAGTGAGGCGAGGA TG zl5 32 166127991 ATAGAGTGAGGCGAGGATGAAGCCG AG zl6 33 166127974 TGAAGCCGAGAGGATACTGCAGAGG TS zl7 34

Table 5 Amino acid sequences for exemplary transcription activation domains (TADs) disclosed herein______________________________

TAD Subsequence SEQ ID NO VPR DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDA LDDFDLDMLINSRSSGSPKKKRKVGSQYLPDTDDRHRIEEKRK RTYETFKSIMKKSPFSGPTDPRPPPRRIAVPSRSSASVPKPAPQPY PFTSSLSTINYDEFPTMVFPSGQISQASALAPAPPQVLPQAPAPA PAPAMVSALAQAPAPVPVLAPGPPQAVAPPAPK PTQAGEGTLS EALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNSEFQQLLNQ GIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLP NGLLSGDEDFSSIADMDFSALLGSGSGSRDSREGMFLPKPEAGS AISDVFEGREVCQPKRIRPFHPPGSPWANRPLPASLAPTPTGPVH EPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPD EETSQAVKALR EMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDL NLDSPLTPELNEILDTFLNDECLLHAMHISTGLSIFDTSLF 132 VP64 DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDA LDDFDLDML 133 CITED 2 MSGLEMADHMMAMNHGRFPDGTNGLHHHPAHRMGMGQFP S PHHHQQQQPQHAFNALMGEHIHYGAGNMNATSGIRHAMGPG TVNGGHPPSALAPAARFNNSQFMGPPVASQGGSLPASMQLQKL NNQYFNHHPYPHNHYMPDLHPAAGHQMNGTNQHFRDCNPKH SGGSSTPGGSGGSSTPGGSGSSSGGGAGSSNSG GGSGSGNMPAS VAHVPAAMLPPNVIDTDFIDEEVLMSLVIEMGLDRIKELPELWL GQNEFDFMTDFVCKQQPSRVSC 134 CITED 4 ADHLMLAEGYRLVQRPPSAAAAHGPHALRTLPPYAGPGLDSG LRPRGAPLGPPPPRQPGALAYGAFGPPSSFQPFPAVPPPAAGIAH LQPVATPYPGRAAAAPPNAPGGPPGPQPAPSAAAPPPPAHALGG MDAELIDEEALTSLELELGLHRVRELPELFLGQSEFDDCFSDLGS APPAGSVSC 135 CREB3 MELELDAGDQDLLAFLLEESGDLGTAPDEAVRAPLDWALPLSE VPSDWEVDDLLCSLLSPPASLNILSSSNPCLVHHDHTYSLPRET VSMDLESESCRKEGTQMTPQHMEELAEQEIARLVLTDEEKSLL EKEGLILPETLPLTKTEEQILKRVR 224

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Table 6

Amino acid sequences for exemplary designed transcription factors (DBD + TAD) disclosed herein __________

Design Subsequence SEQ ID NO 1 MAPKKKRKVGIHGVPAALEPGEKPYKCPECGKSFS RSDNLVRHQRTHTGEKPYKCPECGKSF SHRTTLTNH QRTHTGEKPYKCPECGKSFSREDNLHTHQRTHTGE KPYKCPECGKSFSTSHSLTEHQRTHTGEKPYKCPEC GKSFSQSSSLVRHQRTHTGEKPYKCPECGKSFSRED NLHTHQRTHTTGK KTSKRPAATKKAGQAKKKKGSY PYDVPDYALEEASGSGRADALDDFDLDMLGSDALD DFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLIN SRS SGSPKKKRKVGSQ YLPDTDDRHRIEEKRKRTYE TFKSIMKKSPFSGPTDPRPPPRRIAVPSRSSASVPKPA PQPYPFTSSLSTINYDEFPTMVFPSGQISQASALAP AP PQVLPQAPAPAPAPAMVSALAQAPAPVPVLAPGPPQ AVAPPAPKPTQAGEGTLSEALLQLQFDDEDLGALLG NSTDPAVFTDLASVDNSEFQQLLNQGIPVAPHTTEP MLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGL LSGDEDFSSIADMDFSALLGSGSGSRDSREGMFLPK PEAGSAISDVFEGREVCQPKRIRPFHPPGSPWA NRPL PASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPE ASHLLEDPDEETSQAVKALREMADTVIPQKEEAAIC GQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTP ELNEILDTFLNDECLLHAMHISTGLSIFDTSLF 99 2 MAPKKKRKVGIHGVPAALEPGEKPYKCPECGKSFS RSDNLVRHQRTHTGEKPYKCPECGKSF SREDNLHT HQRTHTGEKPYKCPECGKSFSRSDELVRHQRTHTGE KPYKCPECGKSFSQSGNLTEHQRTHTGEKPYKCPEC GKSFSTSGHLVRHQRTHTGEKPYKCPECGKSFSQNS TLTEHQRTH TGKKTSKRPAATKKAGQAKKKKGSYP YDVPDYALEEASGSGRADALDDFDLDMLGSDALD DFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLIN SRS SGSPKKKRKVGSQ YLPDTDDRHRIEEKRKRTYE TFKSIMKKSPFSGPTDPRPPPRRIAVPSRSSASVPKPA PQPYPFTSSLSTINYDEFPTMVFPSGQISQAS ALAPAP PQVLPQAPAPAPAPAMVSALAQAPAPPVVLAPGPPQ AVAPPAPKPTQAGEGTLSEALLQLQFDDEDLGALLG 100

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NSTDPAVFTDLASVDNSEFQQLLNQGIPVAPHTTEP MLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGL LSGDEDFSSIADMDFSALLGSGSGSRDSREGMFLPK PEAGSAISDVFEGREVCQPKRIRPFHPPGSPWANRPL PASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPE ASHLLEDPDEETSQAVKALREMADTVIPQKEEAAIC GQ MDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTP ELNEILDTFLNDECLLHAMHISTGLSIFDTSLF 3 MAPKKKRKVGIHGVPAALEPGEKPYKCPECGKSFS RSDNLVRHQRTHTGEKPYKCPECGKSF SHRTTLTNH QRTHTGEKPYKCPECGKSFSREDNLHTHQRTHTGE KPYKCPECGKSFSTSHSLTEHQRTHTGEKPYKCPEC GKSFSQSSSLVRHQRTHTGEKPYKCPECGKSFSRED NLHTHQRTHTTGK KTSKRPAATKKAGQAKKKKGSY PYDVPDYALEDALDDFDLDMLGSDALDDFDLDML GSDALDDFDLDMLGSDALDDFDLDML 101 4 MAPKKKRKVGIHGVPAALEPGEKPYKCPECGKSFS RSDNLVRHQRTHTGEKPYKCPECGKSF SREDNLHT HQRTHTGEKPYKCPECGKSFSRSDELVRHQRTHTGE KPYKCPECGKSFSQSGNLTEHQRTHTGEKPYKCPEC GKSFSTSGHLVRHQRTHTGEKPYKCPECGKSFSQNS TLTEHQRTH TGKKTSKRPAATKKAGQAKKKKGSYP YDVPDYALEDALDDFDLDMLGSDALDDFDLDMLG SDALDDFDLDMLGSDALDDFDLDML 102 5 MAADHLMLAEGYRLVQRPPSAAAAHGPHALRTLPP YAGPGLDSGLRPRGAPLGPPPPRQPGALAYGAFGPP SSFQPFPAVPPPAAGIAHLQPVATPYPGRAAAAPPNAP GGPPGPQPAPSAAAPPPPAHALGGMDAELIDEEALT SLELELGLHRVRELPELFLGQSEFDDCFSDLGSAPPAG SVSCGGSGGGSGQSQLIKPSRMRKYPNRPSKTPP HE RPYACPVESCDRRFSRSDNLVRHIRIHTGQKPFQCRI CMRNFSHRTTLTNHIRTHTGEKPFACDICGRKFARE DNLHTHTKIHLRQKDRPYACPVESCDRRFSTSHSLT EHIRIHTGQKPFQCRICMRNFSQSSSLVRHIRTHTGE KPFACDICGRKFAREDNLHTHTKIHLRQKDKLEMA DHLMLAEGYRLVQRPPSAAAAHGPHA C 103 6 MAADHLMLAEGYRLVQRPPSAAAAHGPHALRTLPP YAGPGLDSGLRPRGAPLGPPPPRQPGALAYGAFGPP SSFQPFPAVPPPAAGIAHLQPVATPYPGRAAAAPPNAP GGPPGPQPAPSAAAPPPPAHALGGMDAELIDEEALT SLELELGLHRVRELPELFLGQSEFDDCFSDLGSAPPAG SVSCGGSGGGSGQSQLIKPSRMRKYPNRPSKTPP HE RPYACPVESCDRRFSRSDNLVRHIRIHTGQKPFQCRI CMRNFSHRTTLTNHIRTHTGEKPFACDICGRKFARE DNLHTHTKIHLRQKDRPYACPVESCDRRFSTSHSLT 104

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EHIRIHTGQKPFQCRICMRNFSQSSSLVRHIRTHTGE KPF ACDICGRKF AREDNLHTHTKIHLRQKDK 7 MAADHLMLAEGYRLVQRPPSAAAAHGPHALRTLPP YAGPGLDSGLRPRGAPLGPPPPRQPGALAYGAFGPP SSFQPFPAVPPPAAGIAHLQPVATPYPGRAAAAPPNAP GGPPGPQPAPSAAAPPPPAHALGGMDAELIDEEALT SLELELGLHRVRELPELFLGQSEFDDCFSDLGSAPPAG SVSCGGSGGGSGQSQLIKPSRMRKYPNRPSKTPP HE RPYACPVESCDRRFSQSGNLT EHIRIHTGQKPFQCRICMRNFSTSGHLVRHIRTHTGE KPF ACDICGRKF AQNSTLTEHTKIHLRQKDKLEMAD HLMLAEGYRLVQRPPSAAA AHGPHALRTLPPYAGP GLDSGLRPRGAPLGPPPPRQPGALAYGAFGPPSSFQP FPAVPPPAAGIAHLQPVATPYPGRAAAAPPNAPGGPP GPQPAPSAAAPPPPAHALGGMDAELIDEEALTSLEL ELGLHRVRELPELFLGQSEFDCFSDDLGSAPPAGSVSC 105 8 MAADHLMLAEGYRLVQRPPSAAAAHGPHALRTLPP YAGPGLDSGLRPRGAPLGPPPPRQPGALAYGAFGPP SSFQPFPAVPPPAAGIAHLQPVATPYPGRAAAAPPNAP GGPPGPQPAPSAAAPPPPAHALGGMDAELIDEEALT SLELELGLHRVRELPELFLGQSEFDDCFSDLGSAPPAG SVSCGGSGGGSGQSQLIKPSRMRKYPNRPSKTPP HE RPYACPVESCDRRFSQSGNLT EHIRIHTGQKPFQCRICMRNFSTSGHLVRHIRTHTGE KPFACDICGRKFAQNSTLTEHTKIHLRQKDK 106 9 MQSQLIKPSRMRKYPNRPSKTPPHERPYACPVESCD RRFSRSDNLVRHIRIHTGQKPFQCRICMRNFSREDNL HTHIRTHTGEKPFACDICGRKFARSDELVRHTKIHLR QKDRPYACPVESCDRRFSQSGNLTEHIRIHTGQKPF QCRICMRNF STSGHLVRHIRTHTGEKPF ACDICGRKF AQNSTLTEHTKIHLRQKDK LEMADHLMLAEGYRLV QRPPSAAAAHGPHALRTLPPYAGPGLDSGLRPRGAP LGPPPPRQPGALAYGAFGPPSSFQPPFAVPPPAAGIA HLQPVATPYPGRAAAAPPNAPGGPPGPQPAPSAAAPP PPAHALGGMDAELIDEEALTSLELELGLHRVRELPE LFLGQSEFDDCFSDLGSAPPAGSVSC 107 10 MQSQLIKPSRMRKYPNRPSKTPPHERPYACPVESCD RRFSRSDNLVRHIRIHTGQKPFQCRICMRNFSREDNL HTHIRTHTGEKPF ACDICGRKF ARSDELVRHTKIHLR QKDRPYACPVESCDRRFSQSGNLTEHIRIHTGQKPF QCRICMRNF STSGHLVRHIRTHTGEKPF ACDICGRKF AQNSTLTEHTKIHLRQKDK LEMSGLEMADHMMAM NHGRFPDGTNGLHHHPAHRMGMGQFPSPHHHQQQ QPQHAFNALMGEHIHYGAGNMNATSGIRHAMGPG TVNGGHPPSALAPAARFNNSQFMGPPVASQGGSLP ASMQLQKLNNQYFNHHPYPHNHYMPDLHPAAGHQ MNGTNQHFRDCNPKHSGGS STPGGSGGS STPGGSG 108

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S S SGGGAGS SNSGGGSGSGNMP AS VAHVP AAMLPP NVIDTDFIDEEVLMSLVIEMGLDRIKELPELWLGQN EFDFMTDFVCKQQPSRVSC AND MSGLEMADHMMAMNHGRFPDGTNGLHHHPAHRM GMGQFPSPHHHQQQPQHAFNALMGEHIHYGAGN MNATSGVRHAMGPGTVNGGHPPSALAPAARFNNS QFMGPPVASQGGSLPASMQLQKLNNQYFNHHPYPH NHYMPDLHP AAGHQMNGTNQHFRDCNPKHSGGS S TPGGSGGSSTPGGSSSGGGAGSSN SGGGSGSGNM PASVAHVPAAMLPPNVIDTDFIDEEVLMSLVIEMGL DRIKELPELWLGQNEFDFMTDFVCKQQPSRVSCQSQ LII<PSRMRI<YPNRPSI<TPPHERPYACPVESCDRRFSR SDNLVRHIRIHTGQKPFQCRICMRNFSREDNLHTHIR THTGEKPFACDICGRKFARSDELVRHTKIHLRQKDR PYACPVESCDRRFSQ SGNLTEHIRIHTGQKPFQCRIC MRNFSTSGHLVRHIRTHTGEKPFACDICGRKFAQNS TLTEHTKIHLRQKDK 109 12 MAADHLMLAEGYRLVQRPPSAAAAHGPHALRTLPP YAGPGLDSGLRPRGAPLGPPPPRQPGALAYGAFGPP SSFQPPFAVPPPAAGIAHLQPVATPYPGRAAAAPPNAP GGPPGPQPAPSAAAPPPPAHALGGMDAELIDEEALT SLELELGLHRVRELPELFLGQSEFDDCFSDLGSAPPAG SVSCGGSGGGSGRPHACPAEGCDRRFSRSDNLVRH LRI HTGHKPFQCRICMRFSREDNLHTHIRTHTGEKP FACEFCGRKFARSDELVRHAKIHLKQKEHACPAEG CDRRFSQSGNLTEHLRIHTGHKPFQCRICMRSFSTSG HLVRHIRTHTGEKPFACEFCGRKFAQNSTLTEHAKI HLKQKEKLEMADHLMLAEGYRLVQRPPSAAAAHG PHALRTLPPYAGPGLDSGLRPRGAPLGPPPPRQPGA L AYGAFGPPSSFQPPFAVPPPAAGIAHLQPVATPYPG RAAAPPNAPGGPPGPQPAPSAAAPPPPAHALGGMD AELIDEEALTSLELELGLHRVRELPELFLGQSEFDCF SDLGSAPPAGSVSC 110 13 MRPHACP AEGCDRRF SRSDNLVRHLRIHTGHKPFQC RICMRSFSREDNLHTHIRTHTGEKPFACEFCGRKFAR SDELVRHAKIHLKQKEHACPAEGCDRRFSQSGNLTE HLRIHTGHKPFQCRICMRSFSTSGHLVRHIRTHTGEK PFACEFCGRKFAQNSTLTEHAKIHLKQKEKLEMAD HLMLAEGYRLVQRPPSAAAAHGPHALRTL PPYAGP GLDSGLRPRGAPLGPPPPRQPGALAYGAFGPPSSFQP FPAVPPPAAGIAHLQPVATPYPGRAAAAPPNAPGGPP GPQPAPSAAAPPPPAHALGGMDAELIDEEALTSLEL ELGLHRVRELPELFLGQSEFDCFDSDLGSAPPAGSVSC 111 14 MRPHACP AEGCDRRF SRSDNLVRHLRIHTGHKPFQC RICMRSFSREDNLHTHIRTHTGEKPFACEFCGRKFAR SDELVRHAKIHLKQKEHACPAEGCDRRFSQSGNLTE HLRIHTGHKPFQCRICMRSFSTSGHLVRHIRTHTGEK PFACEFCGRKFAQNSTLTEHAKIHLKQKEKKAEKG GAPSASSAPPVSLAPVVTTCALEMSGL EMADHMMA MNHGRFPDGTNGLHHHPAHRMGMGQFPSPHHHQQ QQPQHAFNALMGEHIHYGAGNMNATSGIRHAMGP 112

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GTVNGGHPPSALAPAARFNNSQFMGPPVASQGGSL PASMQLQKLNNQYFNHHPYPHNHYMPDLHPAAGH QMNGTNQHFRDCNPKHSGGS STPGGSGGS STPGGS GS S SGGGAGS SNSGGGSGSGNMP AS VAHVP AAMLP PNVIDTDFIDEEVLMSLVIEMGLDRIKELPELWLGQN EFDFMTDFVCKQQPSRVSC 15 MSGLEMADHMMAMNHGRFPDGTNGLHHHPAHRM GMGQFPSPHHHQQQPQHAFNALMGEHIHYGAGN MNATSGVRHAMGPGTVNGGHPPSALAPAARFNNS QFMGPPVASQGGSLPASMQLQKLNNQYFNHHPYPH NHYMPDLHP AAGHQMNGTNQHFRDCNPKHSGGS S TPGGSGGSSTPGGSSSGGGAGSSN SGGGSGSGNM PASVAHVPAAMLPPNVIDTDFIDEEVLMSLVIEMGL DRIKELPELWLGQNEFDFMTDFVCKQQPSRVSCRPH ACP AEGCDRRF SRSDNLVRHLRIHTGHKPFQCRICM RSFSREDNLHTHIRTHTGEKPFACEFCGRKFARSDEL VRHAKIHLKQKEHACPAEGCDRRFSQSGNLTEHLRI HTGHKPFQCRICMRSFSTSGHL VRHIRTHTGEKPFA CEFCGRKFAQNSTLTEHAKIHLKQKEKKAEKGGAP SAS S APP VSL AP VVTTC A 113 16 MTGKLAEKLPVTMSSLLNQLPDNLYPEEIPSALNLF SGSSDSVVHYNQMATENVMDIGLTNEKPNPELSYS GSFQPAPGNKTVTYLGKFAFDSPSNWCQDNIISLMS AGILGVPPASGALSTQTSTASMVQPPQGDVEAMYP ALPPYSNCGDLYSEPVSFHDPQGNPGLAYSPQDYQS AKPALDSNLFPMIPDYNLYH HPNDMGSIPEHKPFQG MDPIRVNPPPPITPLETIKAFKDKQIHPGFGSLPQPPLT LI<PIRPRI<YPNRPSI<TPLHERPHACPAEGCDRRFSRR DELNVHLRIHTGHKPFQCRICMRSFSRSDHLTNHIRT HTGEKPFACEFCGRKFARSDDLVRHAKIHLKQKEH ACP AEGCDRRF SRSDNLVRHLRIHTGHKPFQCRICM RSF SH RTTLTNHIRTHTGEKPF ACEFCGRKF AREDNL HTHAKIHLKQKEHACP AEGCDRRF STSHSLTEHLRI HTGHKPFQCRICMRFSQSSSLVRHIRTHTGEKPFAC EFCGRKFAREDNLHTHAKIHLKQKEKKAEKGGAPS AS SAPP VSLAP VVTTC A 114 17 MAAAKAEMQLMSPLQISDPFGSFPHSPTMDNYPKL EEMMLLSNGAPQFLGAAGAPEGSGSNS S S S S SGGGG GGGGGSNS S S S S STFNPQ ADTGEQPYEHLTAESFPDI SLNNEKVLVETSYPSQTTRLPPITYTGRFSLEPAPNS GNTLWPEPLF SL VSGL VSMTNPP AS S S SAP SPAAS SA SASQSPPLSCAVPSNDSSPIY SAAPTFPTPNTDIFPEPQ SQAFPGSAGTALQYPPPAYPAAKGGFQVPMIPDYLF PQQQGDLGLGTPDQKPFQGLESRTQQPSLTPLSTIKA FATQSGSQDLKALNTSYQSQLIKPSRMRKYPNRPSK TPPHERP YACPVESCDRRF SRSDNLVRHIRIHTGQKP FQCRICMRNF SHRTTLTNHIRTHTGEKPF ACDIC GRK FAREDNLHTHTKIHLRQKDRP YACPVESCDRRF STS HSLTEHIRIHTGQKPFQCRICMRNF SQS S SLVRHIRTH TGEKPFACDICGRKFAREDNLHTHTKIHLRQKDKKA DKSVVASSATSSLSSYPSPVATSYPSPVTTSYPSPATT 115

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SYPSPVPTSFSSPGSSTYPSPVHSGFPSPSVATTYSSV PP AFP AQ VS SFP S S AVTNSF S ASTGLSDMT ATF SPRTI EIC 18 MAAAKAEMQLMSPLQISDPFGSFPHSPTMDNYPKL EEMMLLSNGAPQFLGAAGAPEGSGSNS S S S SSGGGG GGGGGSNS S S S S STFNPQ ADTGEQPYEHLTAESFPDI SLNNEKVLVETSYPSQTTRLPPITYTGRFSLEPAPNS GNTLWPEPLF SL VSGL VSMTNPP AS S S SAP SPAAS SA SASQSPPLSCAVPSNDSSPIY SAAPTFPTPNTDIFPEPQ SQAFPGSAGTALQYPPPAYPAAKGGFQVPMIPDYLF PQQQGDLGLGTPDQKPFQGLESRTQQPSLTPLSTIKA FATQSGSQDLKALNTSYQSQLIKPSRMRKYPNRPSK TPPHERP YACPVESCDRRF SRRDELN VH IRIHTGQKP FQCRICMRNF SRSDHLTNHIRTHTGEKPF ACDICGRK FARSDDLVRHTKIHLRQKDRPYACPVESCDRRFSRS DNLVRHIRIHTGQKPFQCRICMRNFSHRTTLTNHIRT HTGEKPF ACDICGRKF AREDNLHTHTKIHLRQKDRP YACPVESCDRRFSTSHSLTEHIRIHTGQKPFQCRICM RNF SQ S S SL VRHIRTHTGEKPF ACDICGRKF AREDNL HTKIHL RQKDKK ADKS WAS SATS SLS S YP SP VA TSYPSPVTTSYPSPATTSYPSPVPTSFSSPGSSTYPSPV HSGFP SP S VATT YS S VPP AFP AQ VS SFP S S AVTNSF SA STGLSDMTATFSPRTIEIC 116 19 MAAAKAEMQLMSPLQISDPFGSFPHSPTMDNYPKL EEMMLLSNGAPQFLGAAGAPEGSGSNS S S S SSGGGG GGGGGSNS S S S S STFNPQ ADTGEQPYEHLTAESFPDI SLNNEKVLVETSYPSQTTRLPPITYTGRFSLEPAPNS GNTLWPEPLF SL VSGL VSMTNPP AS S S SAP SPAAS SA SASQSPPLSCAVPSNDSSPIY SAAPTFPTPNTDIFPEPQ SQAFPGSAGTALQYPPPAYPAAKGGFQVPMIPDYLF PQQQGDLGLGTPDQKPFQGLESRTQQPSLTPLSTIKA FATQSGSQDLKALNTSYQSQLIKPSRMRKYPNRPSK TPPHERP YACPVESCDRRF SRSDNLVRHIRIHTGQKP FQCRICMRNF SHRTTLTNHIRTHTGEKPF ACDIC GRK FAREDNLHTHIRTHTGEKPFACDICGRKFSTSHSLTE HIRIHTGQKPFQCRICMRNFSQSSSLVRHIRTHTGEK PF ACDICGRKF AREDNLHTHTKIHLRQKDKKADKS VVASSATSSLSSYPSPVATSYPSPVTTSYPSPATTSYP SPVPTSF S SPGS STYPSPVHSGFPSPS VATTYS S VPP A FP AQ VS SFP S S AVTN SF S ASTGLSDMT ATF SPRTIEIC 117 20 MAAAKAEMQLMSPLQISDPFGSFPHSPTMDNYPKL EEMMLLSNGAPQFLGAAGAPEGSGSNS S S S SSGGGG GGGGGSNS S S S S STFNPQ ADTGEQPYEHLTAESFPDI SLNNEKVLVETSYPSQTTRLPPITYTGRFSLEPAPNS GNTLWPEPLF SL VSGL VSMTNPP AS S S SAP SPAAS SA SASQSPPLSCAVPSNDSSPIY SAAPTFPTPNTDIFPEPQ SQAFPGSAGTALQYPPPAYPAAKGGFQVPMIPDYLF PQQQGDLGLGTPDQKPFQGLESRTQQPSLTPLSTIKA FATQSGSQDLKALNTSYQSQLIKPSRMRKYPNRPSK TPPHERP YACPVESCDRRF SRSDNLVRHIRIHTGQKP FQCRICMRNF SHRTTLTNHIRTHTGEKPF ACDIC GRK 115

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FAREDNLHTHTKIHLRQKDRP YACPVESCDRRF STS HSLTEHIRIHTGQKPFQCRICMRNF SQS S SLVRHIRTH TGEKPFACDICGRKFAREDNLHTHTKIHLRQKDKKA DKSVVASSATSSLSSYPSPVATSYPSPVTTSYPSPATT SYPSPVPTSFSSPGSSTYPSPVHSGFPSPSVATTYSSV PP AFP AQ VS SFP S S AV TNSF S ASTGLSDMT ATF SPRTI EIC 21 MAAAKAEMQLMSPLQISDPFGSFPHSPTMDNYPKL EEMMLLSNGAPQFLGAAGAPEGSGSNS S S S SSGGGG GGGGGSNS S S S S STFNPQ ADTGEQPYEHLTAESFPDI SLNNEKVLVETSYPSQTTRLPPITYTGRFSLEPAPNS GNTLWPEPLF SL VSGL VSMTNPP AS S S SAP SPAAS SA SASQSPPLSCAVPSNDSSPIY SAAPTFPTPNTDIFPEPQ SQAFPGSAGTALQYPPPAYPAAKGGFQVPMIPDYLF PQQQGDLGLGTPDQKPFQGLESRTQQPSLTPLSTIKA FATQSGSQDLKALNTSYQSQLIKPSRMRKYPNRPSK TPPHERP YACPVESCDRRF SRSDNLVRHIRIHTGQKP FQCRICMRNF SREDNLHTHIRTHTGEKPF ACDIC GR KFARSDELVRHTKIHLRQKDRPYACPVESCDRRFSQ SGNLTEHIRIHTGQKPFQCRICMRNF STSGHLVRHIR THTGEKPFACDICGRKFAQNSTLTEHTKIHLRQKDK KADKSVVASSATSSLSSYPSPVATSYPSPVTTSYPSP ATTSYPSPVPTSFSSPGSSTYPSPVHSGFPSPSVATTY SSVPPAFPAQVSSFPSSAVTNSFSASTGL SDMTATFS PRTIIEIC 118 22 MTGKLAEKLPVTMSSLLNQLPDNLYPEEIPSALNLF SGSSDSVVHYNQMATENVMDIGLTNEKPNPELSYS GSFQPAPGNKTVTYLGKFAFDSPSNWCQDNIISLMS AGILGVPPASGALSTQTSTASMVQPPQGDVEAMYP ALPPYSNCGDLYSEPVSFHDPQGNPGLAYSPQDYQS AKPALDSNLFPMIPDYNLYH HPNDMGSIPEHKPFQG MDPIRVNPPPPITPLETIKAFKDKQIHPGFGSLPQPPLT LI<PIRPRI<YPNRPSI<TPLHERPHACPAEGCDRRFSRS DNLVRHLRIHTGHKPFQCRICMRSF SHRTTLTNHIRT HTGEKPFACEFCGRKFAREDNLHTHAKIHLKQKEH ACPAEGCDRRFSTSHSLTEHLRIHTGHKPFQCRICMR SFSQSSSLVR HIRTHTGEKPFACEFCGRKFAREDNLH THAKIHLKQKEKKAEKGGAPSASSAPPVSLAPVVTT CA 119 23 MAAAKAEMQLMSPLQISDPFGSFPHSPTMDNYPKL EEMMLLSNGAPQFLGAAGAPEGSGSNS S S S SSGGGG GGGGGSNS S S S S STFNPQADTGEQPYEHLTAESFPDI SLNNEKVLVETSYPSQTTRLPPITYTGRFSLEPAPNS GNTLWPEPLF SL VSGL VSMTNPP AS S S SAP SPAAS SA SASQSPPLSCAVPSNDSSPIY SAAPTFPTPNTDIFPEPQ SQAFPGSAGTALQYPPPAYPAAKGGFQVPMIPDYLF PQQQGDLGLGTPDQKPFQGLESRTQQPSLTPLSTIKA FATQSGSQDLKALNTSYQSQLIKPSRMRKYPNRPSK TPPHERP YACPVESCDRRF SDPG ALVRHIRIHTGQKP FQCRICMRNF SRSDNLVRHIRTHTGEKPF ACDIC GR 120

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KF AQSGDLRRHTKIHLRQKDRP YACPVESCDRRF ST HLDLIRHIRIHTGQKPFQC RIC MRNF STSGNLVRHIR THTGEKPFACDICGRKFARSDNLVRHTKIHLRQKDR PYACPVESCDRRFSQSGHLTEHIRIHTGQKPFQCRIC MRNFSERSHLREHIRTHTGEKPFACDICGRKFAQAG HL ASHTKH4LRQKD KK ADKS WAS S AT S SLS S YP SP VATSYPSPVTTSYPSPATTSYPSPVPTSFSSPGSSTYP SPVHSGFPSPSVATTYSSVPPAFPAQVSSFPSSAVTNS FSASTGLSDMTATFSPRTIEIC 24 MAAAKAEMQLMSPLQISDPFGSFPHSPTMDNYPKL EEMMLLSNGAPQFLGAAGAPEGSGSNS S S S SSGGGG GGGGGSNS S S S S STFNPQADTGEQPYEHLTAESFPDI SLNNEKVLVETSYPSQTTRLPPITYTGRFSLEPAPNS GNTLWPEPLF SL VSGL VSMTNPP AS S S SAP SPAAS SA SASQSPPLSCAVPSNDSSPIY SAAPTFPTPNTDIFPEPQ SQAFPGSAGTALQYPPPAYPAAKGGFQVPMIPDYLF PQQQGDLGLGTPDQKPFQGLESRTQQPSLTPLSTIKA FATQSGSQDLKALNTSYQSQLIKPSRMRKYPNRPSK TPPHERP YACPVESCDRRF SRSDNLTRHIRIHTGQKP FQCRICMRNFSHSTTLTNHIRTHTGEKPFACDIC GRK FARSDNRKTHIRTHTGEKPFACDICGRKFSTSHSLTE HIRIHTGQKPFQCRICMRNFSQSSSLTRHIRTHTGEKP FACDICGRKFARSDNRKTHTKIHLRQKDKKADKSV VASSATSSLSSYPSPVATSYPSPVTTSYPSPATTSYPS PVPTSFSSPGSSTYPSPVHSGFPSPSVATTYSSVPPAF PAQVSSFPSSAVTNSFSASTGLSDM TATFSPRTIEIC 121 25 MTGKLAEKLPVTMSSLLNQLPDNLYPEEIPSALNLF SGSSDSVVHYNQMATENVMDIGLTNEKPNPELSYS GSFQPAPGNKTVTYLGKFAFDSPSNWCQDNIISLMS AGILGVPPASGALSTQTSTASMVQPPQGDVEAMYP ALPPYSNCGDLYSEPVSFHDPQGNPGLAYSPQDYQS AKPALDSNLFPMIPDYNLYH HPNDMGSIPEHKPFQG MDPIRVNPPPPITPLETIKAFKDKQIHPGFGSLPQPPLT LI<PIRPRI<YPNRPSI<TPLHERPHACPAEGCDRRFSRS DNLVRHLRIHTGHKPFQCRICMRSF SREDNLHTHIRT HTGEKPFACEFCGRKFARSDELVRHAKIHLKQKEH ACPAEGCDRRFSQSGNLTEHLRIHTGHKPFQCRICM RSFSTSGHL VRHIRTHTGEKPFACEFCGRKFAQNSTL TEHAKIHLKQKEKKAEKGGAPSASSAPPVSLAPWT TCA 122 26 MAAAKAEMQLMSPLQISDPFGSFPHSPTMDNYPKL EEMMLLSNGAPQFLGAAGAPEGSGSNS S S S SSGGGG GGGGGSNS S S S S STFNPQADTGEQPYEHLTAESFPDI SLNNEKVLVETSYPSQTTRLPPITYTGRFSLEPAPNS GNTLWPEPLF SL VSGL VSMTNPP AS S S SAP SPAAS SA SASQSPPLSCAVPSNDSSPIY SAAPTFPTPNTDIFPEPQ SQAFPGSAGTALQYPPPAYPAAKGGFQVPMIPDYLF PQQQGDLGLGTPDQKPFQGLESRTQQPSLTPLSTIKA FATQSGSQDLKALNTSYQSQLIKPSRMRKYPNRPSK TPPHERP YACPVESCDRRF SRSDNLTRHIRIHTGQKP FQCRICMRNFSRSDNLTTHIRTHTGEKPFACDIC GRK 123

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FARSDERKRHIRTHTGEKPFACDICGRKFSQSGNLTE HIRIHTGQKPFQCRICMRNFSTSGHLTRHIRTHTGEK PFACDICGRKFAQSSTRKEHTKIHLRQKDKKADKSV VASSATSSLSSYPSPVATSYPSPVTTSYPSPATTSYPS PVPTSFSSPGSSTYPSPVHSGFPSPSVATTYSSVPPAF PAQVSSFPSSAVTNSFSASTGLSDMTATFSP RTIEIC 27 MAAAKAEMQLMSPLQISDPFGSFPHSPTMDNYPKL EEMMLLSNGAPQFLGAAGAPEGSGSNS S S S SSGGGG GGGGGSNS S S S S STFNPQ ADTGEQPYEHLTAESFPDI SLNNEKVLVETSYPSQTTRLPPITYTGRFSLEPAPNS GNTLWPEPLF SL VSGL VSMTNPP AS S S SAP SPAAS SA SASQSPPLSCAVPSNDSSPIY SAAPTFPTPNTDIFPEPQ SQAFPGSAGTALQYPPPAYPAAKGGFQVPMIPDYLF PQQQGDLGLGTPDQKPFQGLESRTQQPSLTPLSTIKA FATQSGSQDLKALNTSYQSQLIKPSRMRKYPNRPSK TPPHERP YACPVESCDRRF SRSDNLVRHIRIHTGQKP FQCRICMRNF SREDNLHTHIRTHTGEKPF ACDIC GR KFARSDELVRHIRTHTGEKPFACDICGRKFSQSGNLT EHIRIHTGQKPFQCRICMRNFSTSGHLVRHIRTHTGE KPFACDICGRKFAQNSTLTEHTKIHLRQKDKKADKS VVASSATSSLSSYPSPVATSYPSPVTTSYPSPATTSYP SPVPTSF S SPGS STYPSPVHSGFPSPS VATTYS S VPP A FP AQ VS SFP S S AVTNSF S ASTGLSDMT ATF SPRTIEIC 124 28 MAPKKKRKVGIHGVPAALEPGEKPYKCPECGKSFSS P ADLTRHQRTHTGEKPYKCPECGKSF SRSDNLVRH QRTHTGEKPYKCPECGKSFSREDNLHTHQRTHTGE KPYKCPECGKSFSRSDELVRHQRTHTGEKPYKCPEC GKSFSQSGNLTEHQRTHTGEKPYKCPECGKSFSTSG HLVRHQR THTGKKTSKRPAATKKAGQAKKKKGSY PYDVPDYALEDALDDFDLDMLGSDALDDFDLDML GSDALDDFDLDMLGSDALDDFDLDML 125 29 MAPKKKRKVGIHGVPAALEPGEKPYKCPECGKSFS DPGALVRHQRTHTGEKPYKCPECGKSFSRSDNLVR HQRTHTGEKPYKCPECGKSFSQSGDLRRHQRTHTG EKPYKCPECGKSFSTHLDLIRHQRTHTGEKPYKCPE CGKSF STSGNLVRHQRTHTGEKPYKCPECGKSF SRS DNLVRHQ RTHTGKKTSKRPAATKKAGQAKKKKGS YPYDVPDYALEDALDDFDLDMLGSDALDDFDLDM LGSDALDDFDLDMLGSDALDDFDLDML 126 thirty MAPKKKRKVGIHGVPAALEPGEKPYKCPECGKSFS RSDNLVRHQRTHTGEKPYKCPECGKSF SHRTTLTNH QRTHTGEKPYKCPECGKSFSREDNLHTHQRTHTGE KPYKCPECGKSFSTSHSLTEHQRTHTGEKPYKCPEC GKSFSQSSSLVRHQRTHTGEKPYKCPECGKSFSRED NLHTHQRTHTTGK KTSKRPAATKKAGQAKKKKGSY PYDVPDYALEEASGSGRADALDDFDLDMLGSDALD DFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLIN SRS SGSPKKKRKVGSQ YLPDTDDRHRIEEKRKRTYE TFKSIMKKSPFSGPTDPRPPPRRIAVPSRSSASVPKPA PQPYPFTSSLSTINYDEFPTMVFPSGQISQASALAP AP PQVLPQAPAPAPAPAMVSALAQAPAPPVVLAPGPPQ 99

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AVAPPAPKPTQAGEGTLSEALLQLQFDDEDLGALLG NSTDPAVFTDLASVDNSEFQQLLNQGIPVAPHTTEP MLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGL LSGDEDFFSSIADMDFSALLGSGSGSRDSREGMFLPK PEAGSAISDVFEGREVCQPKRIRPFHPPGSPWANRPL PASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPE ASHLLEDPDEETSQAVKALREMADTVIPQKEEAAIC GQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTP ELNEILDTFLNDECLLHAMHISTGLSIFDTSLF 31 MAPKKKRKVGIHGVPAALEPGEKPYKCPECGKSFS RSDNLVRHQRTHTGEKPYKCPECGKSF SREDNLHT HQRTHTGEKPYKCPECGKSFSRSDELVRHQRTHTGE KPYKCPECGKSFSQSGNLTEHQRTHTGEKPYKCPEC GKSFSTSGHLVRHQRTHTGEKPYKCPECGKSFSQNS TLTEHQRTH TGKKTSKRPAATKKAGQAKKKKGSYP YDVPDYALEDALDDFDLDMLGSDALDDFDLDMLG SDALDDFDLDMLGSDALDDFDLDML 102 32 MAPKKKRKVGIHGVPAALEPGEKPYKCPECGKSFS RSDNLVRHQRTHTGEKPYKCPECGKSF SREDNLHT HQRTHTGEKPYKCPECGKSFSRSDELVRHQRTHTGE KPYKCPECGKSFSQSGNLTEHQRTHTGEKPYKCPEC GKSFSTSGHLVRHQRTHTGEKPYKCPECGKSFSQNS TLTEHQRTH TGKKTSKRPAATKKAGQAKKKKGSYP YDVPDYALEDALDDFDLDMLGSDALDDFDLDMLG SDALDDFDLDMLGSDALDDFDLDML 102 33 MAPKKKRKVGIHGVPAALEPGEKPYKCPECGKSFS RSDNLVRHQRTHTGEKPYKCPECGKSF SREDNLHT HQRTHTGEKPYKCPECGKSFSRSDELVRHQRTHTGE KPYKCPECGKSFSQSGNLTEHQRTHTGEKPYKCPEC GKSFSTSGHLVRHQRTHTGEKPYKCPECGKSFSQNS TLTEHQRTH TGKKTSKRPAATKKAGQAKKKKGSYP YDVPDYALEEASGSGRADALDDFDLDMLGSDALD DFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLIN SRS SGSPKKKRKVGSQ YLPDTDDRHRIEEKRKRTYE TFKSIMKKSPFSGPTDPRPPPRRIAVPSRSSASVPKPA PQPYPFTSSLSTINYDEFPTMVFPSGQISQAS ALAPAP PQVLPQAPAPAPAPAMVSALAQAPAPVPVLAPGPPQ AVAPPAPKPTQAGEGTLSEALLQLQFDDEDLGALLG NSTDPAVFTDLASVDNSEFQQLLNQGIPVAPHTTEP MLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGL LSGDEDFSIADMDFSALLGSGSGSRDSREGMFLPK PEAGSAISDVFEGREVCQPKRIRPFHPPG SPWANRPL PASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPE ASHLLEDPDEETSQAVKALREMADTVIPQKEEAAIC GQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTP ELNEILDTFLNDECLLHAMHISTGLSIFDTSLF 100 34 MAPKKKRKVGIHGVPAALEPGEKPYKCPECGKSFS RSDNLVRHQRTHTGEKPYKCPECGKSF SREDNLHT HQRTHTGEKPYKCPECGKSFSRSDELVRHQRTHTGE KPYKCPECGKSFSQSGNLTEHQRTHTGEKPYKCPEC GKSFSTSGHLVRHQRTHTGEKPYKCPECGKSFSQNS 127

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TLTEHQRTHTGKKTSKRPAATKKAGQAKKKKGSD ALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLD MLGSDALDDFDLDML 35 MAPKKKRKVGIHGVPAALEPGEKPYKCPECGKSFS RSDNLVRHQRTHTGEKPYKCPECGKSF SREDNLHT HQRTHTGEKPYKCPECGKSFSRSDELVRHQRTHTGE KPYKCPECGKSFSQSGNLTEHQRTHTGEKPYKCPEC GKSFSTSGHLVRHQRTHTGEKPYKCPECGKSFSQNS TLTEHQRTH TGKKTSKRPAATKKAGQAKKKKGSD ALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLD MLGSDALDDFDLDML 127 36 MAADHLMLAEGYRLVQRPPSAAAAHGPHALRTLPP YAGPGLDSGLRPRGAPLGPPPPRQPGALAYGAFGPP SSFQPPFAVPPPAAGIAHLQPVATPYPGRAAAAPPNAP GGPPGPQPAPSAAAPPPPAHALGGMDAELIDEEALT SLELELGLHRVRELPELFLGQSEFDDCFSDLGSAPPAG SVSCQSQLIKPSRMRKYPNRPSKTPPHERPYAC PVES CDRRF SRSDNL VRHIRIHTGQK P FQC R IC MRNF SRED NLHTHIRTHTGEKPFACDICGRKFARSDELVRHTKIH LRQKDRPYACPVESCDRRFSQSGNLTEHIRIHTGQK PFQCRICMRNFSTSGHLVRHIRTHTGEKPFACDICGR KFAQNSTLTEHTKIHLRQKDK 128 37 MAADHLMLAEGYRLVQRPPSAAAAHGPHALRTLPP YAGPGLDSGLRPRGAPLGPPPPRQPGALAYGAFGPP SSFQPFPAVPPPAAGIAHLQPVATPYPGRAAAAPPNAP GGPPGPQPAPSAAAPPPPAHALGGMDAELIDEEALT SLELELGLHRVRELPELFLGQSEFDDCFSDLGSAPPAG SVSCGGSGGGSGQSQLIKPSRMRKYPNRPSKTPP HE RPYACPVESCDRRF SRSDNL VRHIRIHTGQKPFQCRI CMRNFSREDNLHTHIRTHTGEKPFACDICGRKFARS DELVRHTKIHLRQKDRPYACPVESCDRRFSQSGNLT EHIRIHTGQKPFQCRICMRNFSTSGHLVRHIRTHTGE KPFACDICGRKFAQNSTLTEHTKIHLRQKDK 106 38 MAADHLMLAEGYRLVQRPPSAAAAHGPHALRTLPP YAGPGLDSGLRPRGAPLGPPPPRQPGALAYGAFGPP SSFQPPFAVPPPAAGIAHLQPVATPYPGRAAAAPPNAP GGPPGPQPAPSAAAPPPPAHALGGMDAELIDEEALT SLELELGLHRVRELPELFLGQSEFDDCFSDLGSAPPAG SVSCGGSGGGSGGGSGGGSGQSQLIKPSRMRKYPN R PSKTPPHERPYACPVESCDRRFSRSDNLVRHIRIHT GQKPFQCRICMRNFSREDNLHTHIRTHTGEKPFACDI CGRKFARSDELVRHTKIHLRQKDRPYACPVESCDRR FSQSGNLTEHIRIHTGQKPFQCRICMRNFSTSGHLVR HIRTHTGEKPFACDICGRKFAQNSTLTEHTKIHLRQK DK 129 39 MAADHLMLAEGYRLVQRPPSAAAAHGPHALRTLPP YAGPGLDSGLRPRGAPLGPPPPRQPGALAYGAFGPP SSFQPFPAVPPPAAGIAHLQPVATPYPGRAAAAPPNAP GGPPGPQPAPSAAAPPPPAHALGGMDAELIDEEALT SLELELGLHRVRELPELFLGQSEFDDCFSDLGSAPPAG SVSCGGSGGGSGQSQLIKPSRMRKYPNRPSKTPP HE 105

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RPYACPVESCDRRFSQSGNLT EHIRIHTGQKPFQCRICMRNFSTSGHLVRHIRTHTGE KPFACDICGRKFAQNSTLTEHTKIHLRQKDKLEMAD HLMLAEGYRLVQRPPSAAAAHGP HALRTLPYAGP GLDSGLRPRGAPLGPPPPRQPGALAYGAFGPPSSFQP FPAVPPPAAGIAHLQPVATPYPGRAAAAPPNAPGGPP GPQPAPSAAAPPPPAHALGGMDAELIDEEALTSLEL ELGLHRVRELPELFLGQSEFDDCFSDLGSAPPAGSVSC 40 MAADHLMLAEGYRLVQRPPSAAAAHGPHALRTLPP YAGPGLDSGLRPRGAPLGPPPPRQPGALAYGAFGPP SSFQPFPAVPPPAAGIAHLQPVATPYPGRAAAAPPNAP GGPPGPQPAPSAAAPPPPAHALGGMDAELIDEEALT SLELELGLHRVRELPELFLGQSEFDDCFSDLGSAPPAG SVSCADHLMLAEGYRLVQRPPSAAAAHGPHALRTL PP YAGPGLDSGLRPRGAPLGPPPPRQPGALAYGAFG PPSSFQPPFAVPPPAAGIAHLQPVATPYPGRAAAAPPN APGGPPGPQPAPSAAAPPPPAHALGGMDAELIDEEA LTSLELELGLHRVRELPELFLGQSEFDDCFSDLGSAPP AGSVSCQSQLIKPSRMRKYPNRPSKTPPHERPYACP VESCDRRFSRSDNLVRHIRIHTGQKPFQCRICMR NFS REDNLHTHIRTHTGEKPFACDICGRKFARSDELVRH TKIHLRQKDRPYACPVESCDRRFSQSGNLTEHIRIHT GQKPFQCRICMRNFSTSGHLVRHIRTHTGEKPFACDI CGRKFAQNSTLTEHTKIHLRQKDK 130 41 MAADHLMLAEGYRLVQRPPSAAAAHGPHALRTLPP YAGPGLDSGLRPRGAPLGPPPPRQPGALAYGAFGPP SSFQPFPAVPPPAAGIAHLQPVATPYPGRAAAAPPNAP GGPPGPQPAPSAAAPPPPAHALGGMDAELIDEEALT SLELELGLHRVRELPELFLGQSEFDDCFSDLGSAPPAG SVSCGGSGGGSGADHLMLAEGYRLVQRPPSAAAAH GP HALRTLPPYAGPGLDSGLRPRGAPLGPPPPRQPG ALAYGAFGPPSSFQPPFAVPPPAAGIAHLQPVATPYP GRAAAPPNAPGGPPGPQPAPSAAAPPPPAHALGGM DAELIDEEALTSLELELGLHRVRELPELFLGQSEFDC FSDLGSAPPAGSVSCGGSGGGSGQSQLIKPSRMRKY PNRPSKTPPHERPYACPVESCDRRFSRSDNLVRHIRI HTG QKPFQCRICMRNF SREDNLHTHIRTHTGEKPF A CDICGRKFARSDELVRHTKIHLRQKDRPYACPVESC DRRFSQSGNLTEHIRIHTGQKPFQCRICMRNFSTSGH LVRHIRTHTGEKPFACDICGRKFAQNSTLTEHTKIHL RQKDK 131 42 MAPI<I<I<RI<VGIHGVPAALEPGEI<P YKCPECGKSFS RSDNLVRHQRTHTGEKPYKCPECGKSF SREDNLHT HQRTHTGEKPYKCPECGKSFSRSDELVRHQRTHTGE KPYKCPECGKSFSQSGNLTEHQRTHTGEKPYKCPEC GKSFSTSGHLVRHQRTHTGEKPYKCPECGKSFSQNS TLTEH QRTHTTGKKTSKRPAATKKAGQAKKKKGSYP YDVPDYALEDALDDFDLDMLGSDALDDFDLDMLG SDALDDFDLDMLGSDALDDFDLDML 102

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43 MAPKKKRKVGIHGVPAALEPGEKP YKCPECGKSFS RSDNLVRHQRTHTGEKPYKCPECGKSF SREDNLHT HQRTHTGEKPYKCPECGKSFSRSDELVRHQRTHTGE KPYKCPECGKSFSQSGNLTEHQRTHTGEKPYKCPEC GKSFSTSGHLVRHQRTHTGEKPYKCPECGKSFSQNS TLTEHQRTH TGKKTSKRPAATKKAGQAKKKKGSYP YDVPDYALEDALDDFDLDMLGSDALDDFDLDMLG SDALDDFDLDMLGSDALDDFDLDML 102 44 MAADHLMLAEGYRLVQRPPSAAAAHGPHALRTLPP YAGPGLDSGLRPRGAPLGPPPPRQPGALAYGAFGPP SSFQPPFAVPPPAAGIAHLQPVATPYPGRAAAAPPNAP GGPPGPQP AP S AAAPPPP AHALGGMD A ELID EE A LT SLELELGLHRVRELPELFLGQSEFDCFSDDLGSAPPAG SVSCGGSGGGSGQSQLIKPSRMRK YPNRPSKTPPHE RPYACPVESCDRRFSRSDNLVRHIRIHTGQKPFQCRI CMRNFSREDNLHTHIRTHTGEKPFACDICGRKFARS DELVRHTKIHLRQKDRPYACPVESCDRRFSQSGNLT EHIRIHTGQKPFQCRICMRNFSTSGHLVRHIRTHTGE KPFACDICGRKFAQNSTLTEHTKIHLRQKDK 106 45 MAADHLMLAEGYRLVQRPPSAAAAHGPHALRTLPP YAGPGLDSGLRPRGAPLGPPPPRQPGALAYGAFGPP SSFQPPFAVPPPAAGIAHLQPVATPYPGRAAAAPPNAP GGPPGPQP AP S AAAPPPP AHALGGMD A ELID EE A LT SLELELGLHRVRELPELFLGQSEFDCFSDDLGSAPPAG SVSCGGSGGGSGQSQLIKPSRMRK YPNRPSKTPPHE RPYACPVESCDRRFSRSDNLVRHIRIHTGQKPFQCRI CMRNFSREDNLHTHIRTHTGEKPFACDICGRKFARS DELVRHTKIHLRQKDRPYACPVESCDRRFSQSGNLT EHIRIHTGQKPFQCRICMRNFSTSGHLVRHIRTHTGE KPFACDICGRKFAQNSTLTEHTKIHLRQKDK 106 46 MAADHLMLAEGYRLVQRPPSAAAAHGPHALRTLPP YAGPGLDSGLRPRGAPLGPPPPRQPGALAYGAFGPP SSFQPPFAVPPPAAGIAHLQPVATPYPGRAAAAPPNAP GGPPGPQP AP S AAAPPPP AHALGGMD A ELID EE A LT SLELELGLHRVRELPELFLGQSEFDDCFSDLGSAPPAG SVSCGGSGGGSGGGSGGGSGQSQLIK PSRMRKYPN RPSKTPPHERPYACPVESCDRRFSRSDNLVRHIRIHT GQKPFQCRICMRNFSREDNLHTHIRTHTGEKPFACDI CGRKFARSDELVRHTKIHLRQKDRPYACPVESCDRR FSQSGNLTEHIRIHTGQKPFQCRICMRNFSTSGHLVR HIRTHTGEKPFACDICGRKFAQNSTLTEHTKIHLRQK DK 205 47 MAADHLMLAEGYRLVQRPPSAAAAHGPHALRTLPP YAGPGLDSGLRPRGAPLGPPPPRQPGALAYGAFGPP SSFQPFPAVPPPAAGIAHLQPVATPYPGRAAAAPPNAP GGPPGPQP AP S AAAPPPP AHALGGMD A ELID EE A LT SLELELGLHRVRELPELFLGQSEFDCFSDDLGSAPPAG SVSCADHLMLAEGYRLVQRPPSAAA AHGPHALRTL PPYAGPGLDSGLRPRGAPLGPPPPRQPGALAYGAFG PPSSFQPPFAVPPPAAGIAHLQPVATPYPGRAAAAPPN APGGPPGPQPAPSAAAPPPP AHALGGMD AELIDEEA 207

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LTSLELELGLHRVRELPELFLGQSEFDDCFSDLGSAPP AGSVSCQSQLIKPSRMRKYPNRPSKTPPHERPYACP VESCDRRFSRSDNLVRHIRIHTGQKPFQCRICMRNFS REDNLHTHIRTHTGEKPFACDICGRKFARSDELVRH TKIHLRQKDRPYACPVESCDRRFSQSGNLTEHIRIHT GQKPFQCRICMRNFSTSGHL VRHIRTHTGEKPFACDI CGRKFAQNSTLTEHTKIHLRQKDK 48 MAADHLMLAEGYRLVQRPPSAAAAHGPHALRTLPP YAGPGLDSGLRPRGAPLGPPPPRQPGALAYGAFGPP SSFQPFPAVPPPAAGIAHLQPVATPYPGRAAAAPPNAP GGPPGPQPAPSAAAPPPPAHALGGMDAELIDEEALT SLELELGLHRVRELPELFLGQSEFDDCFSDLGSAPPAG SVSCGGSGGGSGADHLMLAEGYRLVQRPPSAAAAH GP HALRTLPPYAGPGLDSGLRPRGAPLGPPPPRQPG ALAYGAFGPPSSFQPPFAVPPPAAGIAHLQPVATPYP GRAAAPPNAPGGPPGPQPAPSAAAPPPPAHALGGM DAELIDEEALTSLELELGLHRVRELPELFLGQSEFDC FSDLGSAPPAGSVSCGGSGGGSGQSQLIKPSRMRKY PNRPSKTPPHERPYACPVESCDRRFSRSDNLVRHIRI HTG QKPFQCRICMRNF SREDNLHTHIRTHTGEKPF A CDICGRKFARSDELVRHTKIHLRQKDRPYACPVESC DRRFSQSGNLTEHIRIHTGQKPFQCRICMRNFSTSGH LVRHIRTHTGEKPFACDICGRKFAQNSTLTEHTKIHL RQKDK 209 49 MELELDAGDQDLLAFLLEESGDLGTAPDEAVRAPL DWALPLSEVPSDWEVDDLLCSLLSPPASLNILSSSNP CLVHHDHTYSLPRETVSMDLESESCRKEGTQMTPQ HMEELAEQEIARLVLTDEEKSLLEKEGLILPETLPLT KTEEQILKRVRLEPGEKPYKCPECGKSFSRSDNLVR HQRTHTGEKPYKCPECGKSFSRED S S STCILV LL VSFCLLLVPAMYSSDTRGSLPAEHGVLSRQLRALPS EDPYQLELPALQSEVPKDSTHQWLDGSDCVLQAPG NT SCLLHYMPQ AP S AEPPLEWPFPDLF SEPLCRGPIL PLQANLTRKGGWLPTGSPSVILQDRYSG 213 50 MELELDAGDQDLLAFLLEESGDLGTAPDEAVRAPL DWALPLSEVPSDWEVDDLLCSLLSPPASLNILSSSNP CLVHHDHTYSLPRETVSMDLESESCRKEGTQMTPQ HMEELAEQEIARLVLTDEEKSLLEKEGLILPETLPLT KTEEQILKRVRRPYACPVESCDRRFSRSDNLVRHIRI HTGQKPFQHTCRICMRNF SREDNLHIRTHT GEKPF A CDICGRKFARSDELVRHTKIHLRQKDRPYACPVESC DRRFSQSGNLTEHIRIHTGQKPFQCRICMRNFSTSGH LVRHIRTHTGEKPFACDICGRKFAQNSTLTEHTKIHL RQKDVYVGGLESRVLKYTAQNMELQNKVQLLEEQ NLSLLDQLRKLQ AMVIEISNKT S S S STCILVLL VSFCL LLVP AM Y S SDTRGSLP AEHGVLSRQLRALP SEDP YQ 217

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LELPALQSEVPKDSTHQWLDGSDCVLQAPGNTSCL LHYMPQAPSAEPPLEWPFPDLFSEPLCRGPILPLQAN LTRKGGWLPTGSPSVILQDRYSG 51 MELELDAGDQDLLAFLLEESGDLGTAPDEAVRAPL DWALPLSEVPSDWEVDDLLCSLLSPPASLNILSSSNP CLVHHDHTYSLPRETVSMDLESESCRKEGTQMTPQ HMEELAEQEIARLVLTDEEKSLLEKEGLILPETLPLT KTEEQILKRVRRPYACPVESCDRRFSRSDNLVRHIRI HTGQKPFQCRICMRNF SHRTTLTNHIRTHT GEKPF A CDICGRKFAREDNLHTHTKIHLRQKDRPYACPVESC DRRFSTSHSLTEHIRIHTGQKPFQCRICMRNFSQSSSL VRHIRTHTGEKPFACDICGRKFAREDNLHTHTKIHL RQKDVYVGGLESRVLKYTAQNMELQNKVQLLEEQ NLSLLDQLRKLQ AMVIEISNKT S S S STCIL VLL VSFCL LLVP AM Y S SDTRGSLP AEHGVLSRQLRALP SEDP YQ LELPALQSEVPKDSTHQWLDGSDCVLQAPGNTSCL LHYMPQAPSAEPPLEWPFPDLFSEPLCRGPILPLQAN LTRKGGWLPTGSPSVILQDRYSG 219 52 MELELDAGDQDLLAFLLEESGDLGTAPDEAVRAPL DWALPLSEVPSDWEVDDLLCSLLSPPASLNILSSSNP CLVHHDHTYSLPRETVSMDLESESCRKEGTQMTPQ HMEELAEQEIARLVLTDEEKSLLEKEGLILPETLPLT KTEEQILKRVRLEPGEKPYKCPECGKSFSRSDNLVR HQRTHTGEKPYKCPECGKSFSRED NLHTHQRTHTG EKPYKCPECGKSFSRSDELVRHQRTHTGEKPYKCPE CGKSFSQSGNLTEHQRTHTGEKPYKCPECGKSFSTS GHLVRHQRTHTGEKPYKCPECGKSFSQNSTLTEHQ RTHTGKKTSVYVGGLESRVLKYTAQNMELQNKVQ LLEEQNLSLLDQLRKLQAMVIEIS 221 53 MELELDAGDQDLLAFLLEESGDLGTAPDEAVRAPL DWALPLSEVPSDWEVDDLLCSLLSPPASLNILSSSNP CLVHHDHTYSLPRETVSMDLESESCRKEGTQMTPQ HMEELAEQEIARLVLTDEEKSLLEKEGLILPETLPLT KTEEQILKRVRRPYACPVESCDRRFSRSDNLVRHIRI HTGQKPFQHTCRICMRNF SREDNLHIRTHT GEKPF A CDICGRKFARSDELVRHTKIHLRQKDRPYACPVESC DRRFSQSGNLTEHIRIHTGQKPFQCRICMRNFSTSGH LVRHIRTHTGEKPFACDICGRKFAQNSTLTEHTKIHL RQKDVYVGGLESRVLKYTAQNMELQNKVQLLEEQ NLSLLDQLRKLQ AMVIEIS 223

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Table 7

Nucleic acid sequences encoding exemplary engineered transcription factors disclosed herein

Design Subsequence SEQ ID NO 31 (RE + encoding) ggaggaagccatcaactaaactacaatgactgtaagatacaaaattgggaatggt aacatattttgaagttctgttgacataaagaatcatgattaatgcccatggaaatg aaagggcgatcaacactatggtttgaaaagggggaaattgtagagcacagatgt 67 gttcgtgtggcagtgtgctgtctctagcaatactcagagaagagagagaacaatg aaattctgattggccccagtgtgagcccagatgaggttcagctgccaactttctctt tcacatcttatgaaagtcatttaagcacaactaactttttttttttttttttttttttttgagaca gagtcttgctctgtt gcccaggacagagtgcagtagtgactcaatctcggctcact gcagcctccacctcctaggctcaaacggtcctcctgcatcagcctcccaagtagc tggaattacaggagtggcccaccatgcccagctaatttttgtatttttaatagatacg ggggtttcaccatatcacccaggctggtctcgaactcctggcctcaagtgatcca cctg cctcggcctcccaaagtgctgggattataggcgtcagccactatgcccaac ccgaccaaccttttttaaaataaatatttaaaaaattggtatttcacatatatactagta tttacatttatccacacaaaacggacgggcctccgctgaaccagtgaggcccca gacgtgcgcataaataacccctgcgtgctgcaccacctggggagaggggga g gaccacggtaaatggagcgagcgcatagcaaaagggacgcggggtccttttct ctgccggtggcactgggtagctgtggccaggtgtggtactttgatggggcccag ggctggagctcaaggaagcgtcgcaggtcacagatctgggggaaccccgg ggaaaagcactgaggcaaaaccgccgctcgtctcctacaatatatgg gagggg gaggttgagtacgttctggattactcataagacctttttttttccttccgggcgcaaa accgtgagctggatttataatcgccctataaagctccagaggcggtcaggcacct gcagaggagccccgccgctccgccgactagctgcccccgcgagcaacggcct cgtgatttccccgccgatccggtccccgcc tccccactctgcccccgcctacccc ggagccgtgcagccgcctctccgaatctctctcttctcctggcgctcgcgtgcga gagggaactagcgagaacgaggaagcagctggaggtgacgccgggcagatt acgcctgtcagggccgagccgagcggatcgctgggcgctgtgcagaggaaa ggcgggagtgc ccggctcgctgtcgcagagccgaggtgggtaagctagcgac cacctggacttcccagcgcccaaccgtggcttttcagccaggtcctctcctcccg cggcttctcaaccaaccccatcccagcgccggccacccaacctcccgaaatga gtgcttcctgccccagcagccgaaggcgctactaggaacggtaacctgttacttt tcca ggggccgtagtcgacccgctgcccgagttgctgtgcgactgcgcgcgcg gggctagagtgcaaggtgactgtggttcttctctggccaagtccgagggagaac gtaaagatatgggcctttttccccctctcaccttgtctcaccaaagtccctagtccc cggagcagttagcctctttcttccagggaattag gacacaacaacgggaac cagacaccgaaccagacatgcccgccccgtgcgccctccccgctcgctgccttt cctccctcttgtctctccagagccggatcttcaaggggagcctccgtgcccccgg ctgctcagtccctccggtgtgcaggaccccggaagtcctccccgcacagctctc gcttctctttgcagcctgttt ctgcgccggaccagtcgaggactctggacagtaga ggccccgggacgaccgagctgGAATTCGCCACCATGGCCCC AAAGAAGAAGCGGAAGGTCGGTATCCACGGAGT CCCAGCAGCCCTCGAACCAGGTGAAAAACCTTA CAAATGTCCTGAATGTGGGAAATCATTCAGTCGC AGCGACAACCTGGTGAGACATCAACGCACCCAT ACAGGAGAAAACCTTATAAATGTCCAGA ATGT GGAAAGTCCTTCACGAGAGGATAACTTGCAC ACTCATCAACGAACACATACTGGTGAAAAACCA TACAAGTGTCCCGAATGTGGTAAAAGTTTTAGCC GGAGCGATGAACTTGTCCGACACCAACGAACCC ATACAGGCGAGAAGCCTTACAAATGTCCCGAGT GTGGCAAGAGCTTCTCAATCAGGGAATCTGA CTGAGCATCAACGAACTCATACCGGGGAAAAAC CTTACA AGTGTCCAGAGTGTGGGAAGAGCTTTTC CACAAGTGGACATCTGGTACGCCACCAGAGGAC ACATACAGGGGAGAAGCCCTACAAATGCCCCGA

- 81 046157

ATGCGGTAAAAGTTTCTCTCAGAATAGTACCCTG ACCGAACACCAGCGAACACACACTGGGAAAAAA ACGAGTAAAAGGCCGGCGGCCACGAAAAAGGCC GGCCAGGCAAAAAAGAAAAAGGGATCCTACCCA TACGACGTACCAGATTACGCTCTCGAGGACGCGC TGGACGATTTCGATCTCGACATGCTGGGTTCTGA TGCCCTCGATGACTTTGACCTGGATATGTTGGGA AGCGACGCATTGGA TGACTTTGATCTGGACATGC TCGGCTCCGATGCTCTGGACGATTTCGATCTCGA T AT GTT AT A A AC T AGT aaagagaccggttcactgtgacagtaaaa gagaccggttcactgtgagaatgaaagagaccggttcactgtgatcggaaaaga gaccggttcactgtgagcggccttgaaacccagcagacaatgtagctcagtaga aacc cagcagacaatgtagctgaatggaaacccagcagacaatgtagcttcgg agaaacccagcagacaatgtagctAAGCTTGGGTGGCATCCC TGTGACCCCTCCCCAGTGCCTCTCCTGGCCCTGG AAGTTGCCACTCCAGTGCCCACCAGCCTTGTCCT AATAAAATTAAGTTGCATCATTTTGTCTGACTAG GTGTCCTTCTATAATATTATGGGGTGGAGGGGGG TGGTATGGAGC AAGGGGCAAGTTGGGAAGACAA CCTGTAGGGCCTGCGGGGTCTATTGGGAACCAA GCTGGAGTGCAGTGGCACAATCTTGGCTCACTGC AATCTCCGCCTCCTGGGTTCAAGCGATTCTCCTG CCTCAGCCTCCCGAGTTGTTGGGATTCCAGGCAT GCATGACCAGGCTCAGCTAATTTTTGTTTTTTTGG TAGAGACGGGGTTTCACCATATTGGCCAGGCTGG TCTCCAACTCCTAAT CTCAGGTGATCTACCCACC TTGGCCTCCCAAATTGCTGGGATTACAGGCGTGA ACCACTGCTCCCTTCCCTGTCCTT 32 (RE + encoding) ggaggaagccatcaactaaactacaatgactgtaagatacaaaattgggaatggt aacatattttgaagttctgttgacataaagaatcatgattaatgcccatggaaatg aaagggcgatcaacactatggtttgaaaagggggaaattgtagagcacagatgt gttcgtgtggcagtgtgctgtctctagcaatactcagaga agagagagaacaatg aaattctgattggccccagtgtgagcccagatgaggttcagctgccaactttctctt tcacatcttatgaaagtcatttaagcacaactaactttttttttttttttttttttttgagaca gagtcttgctctgttgcccaggacagagtgcagtagtgactcaatctcggctcact gcagcct ccacctcctaggctcaaacggtcctcctgcatcagcctcccaagtagc tggaattacaggagtggcccaccatgcccagctaatttttgtatttttaatagatacg ggggtttcaccatatcacccaggctggtctcgaactcctggcctcaagtgatcca cctgcctcggcctcccaaagtgctgggattataggcgtcagccactatgc ccaac ccgaccaaccttttttaaaataaatatttaaaaaattggtatttcacatatatactagta tttacatttatccacacaaaacggacgggcctccgctgaaccagtgaggcccca gacgtgcgcataaataacccctgcgtgctgcaccacctggggagagggggag gaccacggtaaatggagcgagcgcatagcaaaagggacgcggggt ccttttct ctgccggtggcactgggtagctgtggccaggtgtggtactttgatggggcccag ggctggagctcaaggaagcgtcgcagggtcacagatctgggggaaccccgg ggaaaagcactgaggcaaaaccgccgctcgtctcctacaatatatgggagggg gaggttgagtacgttctggattactcataagacctttttt tttttccttccgggcgcaaa accgtgagctggatttataatcgccctataaagctccagaggcggtcaggcacct gcagaggagccccgccgctccgccgactagctgcccccgcgagcaacggcct cgtgatttccccgccgatccggtccccgcctccccactctgcccccgcctacccc 68

- 82 046157

ggagccgtgcagccgcctctctccgaatctctctcttctcctggcgctcgcgtgcga gagggaactagcgagaacgaggaagcagctggaggtgacgccgggcagatt acgcctgtcagggccgagccgagcggatcgctgggcgctgtgcagaggaaa ggcgggagtgcccggctcgctgtcgcagag ccgaggtgggtaagctagcgac cacctggacttcccagcgcccaaccgtggcttttcagccaggtcctctcctcccg cggcttctcaaccaaccccatcccagcgccggccacccaacctcccgaaatga gtgcttcctgccccagcagccgaaggcgctactaggaacggtaacctgttacttt tccaggggccgtagtcgacccgctg cccgagttgctgtgcgactgcgcgcgcg gggctagagtgcaaggtgactgtggttcttctctggccaagtccgagggaac gtaaagatatgggcctttttccccctctcaccttgtctcaccaaagtccctagtccc cggagcagttagcctctttctttccagggaattagccagacacaacaacgggaac cagacacc gaaccagacatgcccgccccgtgcgccctccccgctcgctgccttt cctccctcttgtctctccagagccggatcttcaaggggagcctccgtgcccccgg ctgctcagtccctccggtgcaggaccccggaagtcctccccgcacagctctc gcttctctttgcagcctgtttctgcgccggaccagtcgaggact ctggacagtaga ggccccgggacgaccgagctgGAATTCGCCACCATGGCCCC AAAGAAGAAGCGGAAGGTCGGTATCCACGGAGT CCCAGCAGCCCTCGAACCAGGTGAAAAACCTTA CAAATGTCCTGAATGTGGGAAATCATTCAGTCGC AGCGACAACCTGGTGAGACATCAACGCACCCAT ACAGGAGAAAAACCTTATAAATGTCCAGAATGT GGAAAGTCCTTCTCACGA GAGGATAACTTGCAC ACTCATCAACGAACACATACTGGTGAAAAACCA TACAAGTGTCCCGAATGTGGTAAAAGTTTTAGCC GGAGCGATGAACTTGTCCGACACCAACGAACCC ATACAGGCGAGAAGCCTTACAAATGTCCCGAGT GTGGCAAGAGCTTCTCACAATCAGGGAATCTGA CTGAGCATCAACGAACTCATACCGGGGAAAAAC CTTACAAGTGTCCAGAGTGTGGGAAGAGCT TTTC CACAAGTGGACATCTGGTACGCCACCAGAGGAC ACATACAGGGGAGAAGCCCTACAAATGCCCCGA ATGCGGTAAAAGTTTCTCTCAGAATAGTACCCTG ACCGAACACCAGCGAACACACACTGGGAAAAAA ACGAGTAAAAGGCCGGCGGCCACGAAAAAGGCC GGCCAGGCAAAAAAGAAAAAGGGATCCTACCCA TACGACGTACCAGATTACGCTCTCGAGGACGCGC TGGACGATTTCGATC TCGACATGCTGGGTTCTGA TGCCCTCGATGACTTTGACCTGGATATGTTGGGA AGCGACGCATTGGATGACTTTGATCTGGACATGC TCGGCTCCGATGCTCTGGACGATTTCGATCTCGA TATGTTATAAACTAGTAAGCTTGGGTGGCATCCC TGTGACCCCTCCCAGTGCCTCTCCTGGCCCTGG AAGTTGCCACTCCAGTGCCCACCAGCCTTGTCCT AATAAAATTAAGTTGCATCATTT TGTCTGACTAG GTGTCCTTCTATAATATTATGGGGTGGAGGGGGG TGGTATGGAGCAAGGGGCAAGTTGGGAAGACAA CCTGTAGGGCCTGCGGGGTCTATTGGGAACCAA GCTGGAGTGCAGTGGCACAATCTTGGCTCACTGC AATCTCCGCCTCCTGGGTTCAAGCGATTCTCCTG CCTCAGCCTCCCGAGTTGTTGGGATTCCAGGCAT

- 83 046157

GCATGACCAGGCTCAGCTAATTTTTGTTTTTTTGG TAGAGACGGGGTTTCACCATATTGGCCAGGCTGG TCTCCAACTCCTAATCTCAGGTGATCTACCCACC TTGGCCTCCCAAATTGCTGGGATTACAGGCGTGA ACCACTGCTCCCTTCCCTGTCCTT 33 (RE + encoding) ggaggaagccatcaactaaactacaatgactgtaagatacaaaattgggaatggt aacatattttgaagttctgttgacataaagaatcatgattaatgcccatggaaatg aaagggcgatcaacactatggtttgaaaagggggaaattgtagagcacagatgt gttcgtgtggcagtgtgctgtctctagcaatactcagaga agagagagaacaatg aaattctgattggccccagtgtgagcccagatgaggttcagctgccaactttctctt tcacatcttatgaaagtcatttaagcacaactaactttttttttttttttttttttttgagaca gagtcttgctctgttgcccaggacagagtgcagtagtgactcaatctcggctcact gcagcct ccacctcctaggctcaaacggtcctcctgcatcagcctcccaagtagc tggaattacaggagtggcccaccatgcccagctaatttttgtatttttaatagatacg ggggtttcaccatatcacccaggctggtctcgaactcctggcctcaagtgatcca cctgcctcggcctcccaaagtgctgggattataggcgtcagccactatgc ccaac ccgaccaaccttttttaaaataaatatttaaaaaattggtatttcacatatatactagta tttacatttatccacacaaaacggacgggcctccgctgaaccagtgaggcccca gacgtgcgcataaataacccctgcgtgctgcaccacctggggagagggggag gaccacggtaaatggagcgagcgcatagcaaaagggacgcggggt ccttttct ctgccggtggcactgggtagctgtggccaggtgtggtactttgatggggcccag ggctggagctcaaggaagcgtcgcagggtcacagatctgggggaaccccgg ggaaaagcactgaggcaaaaccgccgctcgtctcctacaatatatgggagggg gaggttgagtacgttctggattactcataagaccttttt tttttccttccgggcgcaaa accgtgagctggatttataatcgccctataaagctccagaggcggtcaggcacct gcagaggagccccgccgctccgccgactagctgcccccgcgagcaacggcct cgtgatttccccgccgatccggtccccgcctccccactctgcccccgcctacccc ggagccgtgcagccgcctct ccgaatctctctcttctcctggcgctcgcgtgcga gagggaactagcgagaacgaggaagcagctggaggtgacgccgggcagatt acgcctgtcagggccgagccgagcggatcgctgggcgctgtgcagaggaaa ggcgggagtgcccggctcgctgtcgcagagccgaggtgggtaagctagcgac cacc tggacttcccagcgcccaaccgtggcttttcagccaggtcctctcctcccg cggcttctcaaccaaccccatcccagcgccggccacccaacctcccgaaatga gtgcttcctgccccagcagccgaaggcgctactaggaacggtaacctgttacttt tccaggggccgtagtcgacccgctgcccgagttgctgtgcgactg cgcgcgcg gggctagagtgcaaggtgactgtggttcttctctggccaagtccgagggagaac gtaaagatatgggcctttttccccctctcaccttgtctcaccaaagtccctagtccc cggagcagttagcctctttctttccagggaattagccagacacaacaacgggaac cagacaccgaaccagacatgcccgccccgtgcg ccctccccgctcgctgccttt cctccctcttgtctctccagagccggatcttcaaggggagcctccgtgcccccgg ctgctcagtccctccggtgtgcaggaccccggaagtcctccccgcacagctctc gcttctctttgcagcctgtttctgcgccggaccagtcgaggactctggacagtaga ggccccgggacgacc gagctgGAATTCGCCACCATGGCCCC AAAGAAGAAGCGGAAGGTCGGTATCCACGGAGT CCCAGCAGCCCTCGAACCAGGTGAAAAACCTTA CAAATGTCCTGAATGTGGGAAATCATTCAGTCGC AGCGACAACCTGGTGAGACATCAACGCACCCAT ACAGGAGAAAAAAACCTTATAAATGTCCAGAATGT GGAAAGTCCTTCTCACGAGAGGATAACTTGCAC ACTCATCAACGAACA CATACTGGTGAAAAACCA 69

- 84 046157

TACAAGTGTCCCGAATGTGGTAAAAGTTTTAGCC GGAGCGATGAACTTGTCCGACACCAACGAACCC ATACAGGCGAGAAGCCTTACAAATGTCCCGAGT GTGGCAAGAGCTTCTCAATCAGGGAATCTGA CTGAGCATCAACGAACTCATACCGGGGAAAAAC CTTACAAGTGTCCAGAGTGTGGGAAGAGCTTTTC CACAAGTGGACATCTGGTACGCCACCAGAGGAC ACATACAG GGGAGAAGCCCTACAAATGCCCCGA ATGCGGTAAAAGTTTCTCTCAGAATAGTACCCTG ACCGAACACCAGCGAACACACACTGGGAAAAAA ACGAGTAAAAGGCCGGCGGCCACGAAAAAGGCC GGCCAGGCAAAAAAGAAAAAGGGATCCTACCCA TACGACGTACCAGATTACGCTCTCGAGGAGGCC AGCGGTTCCGGACGGGCTGACGCATTGGACGAT TTTGATCTGGATATGCTGGGAAGTGAC GCCCTCG ATGATTTTGACCTTGACATGCTTGGTTCGGATGC CCTTGATGACTTTGACCTCGACATGCTCGGCAGT GACGCCCTTGATGATTTCGACCTGGACATGCTGA TTAACTCTAGAAGTTCCGGATCTCCGAAAAAGAA ACGCAAAGTTGGTAGCCAGTACCTGCCCGACAC CGACGACCGGCACCGGATCGAGGAAAAGCGGAA GCGGACCTACGAGACATTCAAGAGCATCATGAA GAAGTCCCCCTTCAGCGGCCCCACCGACCCTAGA CCTCCACCTAGAAGAATCGCCGTGCCCAGCAGAT CCAGCGCCAGCGTGCCAAAACCTGCCCCCCAGC CTTACCCCTTCACCAGCAGCCTGAGCACCATCAA CTACGACGAGTTCCCTACCATGGTGTTCCCCAGC GGCCAGATCTCTCAGGCCTCTGCTCTGGCTCCAG CCCCTCCTCAGGTGCTGCCTCAGGCTCCTGCTCC TGCACCA GCTCCAGCCATGGTGTCTGCACTGGCT CAGGCACCAGCACCCGTGCCTGTGCTGGCTCCTG GACCTCCACAGGCTGTGGCTCCACCAGCCCCTAA ACCTACACAGGCCGGCGAGGGCACACTGTCTGA AGCTCTGCTGCAGCTGCAGTTCGACGACGAGGAT CTGGGAGCCCTGCTGGGAAACAGCACCGATCCT GCCGTGTTCACCGACCTGGCCAGCTGGACAAC AGCGAGTTCCAG CAGCTGCTGAACCAGGGCATC CCTGTGGCCCCTCACACCACCGAGCCCATGCTGA TGGAATACCCCGAGGCCATCACCCGGCTCGTGAC AGGCGCTCAGAGGCCTCCTGATCCAGCTCCTGCC CCTCTGGGAGCACCAGGCCTGCCTAATGGACTGC TGTCTGGCGACGAGGACTTCAGCTCTATCGCCGA TATGGATTTCTCAGCCTTGCTGGGCTCTGGCAGC GGCAGCCGGGATTCC AGGGAAGGGATGTTTTTG CCGAAGGCCTGAGGCCGGCTCCGCTATTAGTGACG TGTTTGAGGGCCGCGAGGTGTGCCAGCCAAAAC GAATCCGGCCATTTCATCCTCCAGGAAGTCCATG GGCCAACCGCCCACTCCCCGCCAGCCTCGCACCA ACACCAACGGTCCAGTACATGAGCCAGTCGGG TCACTGACCCCGGCACCAGTCCCTCAGCCACTGG

- 85 046157

ATCCAGCGCCCGCAGTGACTCCCGAGGCCAGTC ACCTGTTGGAGGATCCCGATGAAGAGACGAGCC AGGCTGTCAAAGCCCTTCGGGAGATGGCCGATA CTGTGATTCCCCAGAAGGAAGAGGCTGCAATCT GTGGCCAAATGGACCTTTCCCATCCGCCCCCAAG GGGCCATCTGGATGAGCTGACAACCACACTTGA GTCCATGACCGAGGATCTGAACCTGGACTCACCC CTGACCCCGG AATTGAACGAGATTCTGGATACCT TCCTGAACGACGAGTGCCTCTTGCATGCCATGCA TATCAGCACAGGACTGTCCATCTTCGACACATCT CTGTTTTAAACTAGTaataaaagatctttattttcattagatctgtgtgt tggttttttgtgtg 34 (RE + encoding) ggaggaagccatcaactaaactacaatgactgtaagatacaaaattgggaatggt aacatattttgaagttctgttgacataaagaatcatgattaatgcccatggaaatg aaagggcgatcaacactatggtttgaaaagggggaaattgtagagcacagatgt gttcgtgtggcagtgtgctgtctctagcaatactcagaga agagagagaacaatg aaattctgattggccccagtgtgagcccagatgaggttcagctgccaactttctctt tcacatcttatgaaagtcatttaagcacaactaactttttttttttttttttttttttgagaca gagtcttgctctgttgcccaggacagagtgcagtagtgactcaatctcggctcact gcagcct ccacctcctaggctcaaacggtcctcctgcatcagcctcccaagtagc tggaattacaggagtggcccaccatgcccagctaatttttgtatttttaatagatacg ggggtttcaccatatcacccaggctggtctcgaactcctggcctcaagtgatcca cctgcctcggcctcccaaagtgctgggattataggcgtcagccactatgc ccaac ccgaccaaccttttttaaaataaatatttaaaaaattggtatttcacatatatactagta tttacatttatccacacaaaacggacgggcctccgctgaaccagtgaggcccca gacgtgcgcataaataacccctgcgtgctgcaccacctggggagagggggag gaccacggtaaatggagcgagcgcatagcaaaagggacgcggggt ccttttct ctgccggtggcactgggtagctgtggccaggtgtggtactttgatggggcccag ggctggagctcaaggaagcgtcgcagggtcacagatctgggggaaccccgg ggaaaagcactgaggcaaaaccgccgctcgtctcctacaatatatgggagggg gaggttgagtacgttctggattactcataagacctttttt tttttccttccgggcgcaaa accgtgagctggatttataatcgccctataaagctccagaggcggtcaggcacct gcagaggagccccgccgctccgccgactagctgcccccgcgagcaacggcct cgtgatttccccgccgatccggtccccgcctccccactctgcccccgcctacccc ggagccgtgcagccgcctct ccgaatctctctcttctcctggcgctcgcgtgcga gagggaactagcgagaacgaggaagcagctggaggtgacgccgggcagatt acgcctgtcagggccgagccgagcggatcgctgggcgctgtgcagaggaaa ggcgggagtgcccggctcgctgtcgcagagccgaggtgggtaagctagcgac cacc tggacttcccagcgcccaaccgtggcttttcagccaggtcctctcctcccg cggcttctcaaccaaccccatcccagcgccggccacccaacctcccgaaatga gtgcttcctgccccagcagccgaaggcgctactaggaacggtaacctgttacttt tccaggggccgtagtcgacccgctgcccgagttgctgtgcgactg cgcgcgcg gggctagagtgcaaggtgactgtggttcttctctggccaagtccgagggagaac gtaaagatatgggcctttttccccctctcaccttgtctcaccaaagtccctagtccc cggagcagttagcctctttctttccagggaattagccagacacaacaacgggaac cagacaccgaaccagacatgcccgccccgtgcg ccctccccgctcgctgccttt cctccctcttgtctctccagagccggatcttcaaggggagcctccgtgcccccgg ctgctcagtccctccggtgtgcaggaccccggaagtcctccccgcacagctctc gcttctctttgcagcctgtttctgcgccggaccagtcgaggactctggacagtaga ggccccgggacgacc gagctgGAATTCGCCACCATGGCCCC 70

- 86 046157

AAAGAAGAAGCGGAAGGTCGGTATCCACGGAGT CCCAGCAGCCCTCGAACCAGGTGAAAAACCTTA CAAATGTCCTGAATGTGGGAAATCATTCAGTCGC AGCGACAACCTGGTGAGACATCAACGCACCCAT ACAGGAAAAAAACCTTATAAATGTCCAGAATGT GGAAAGTCCTTCTCACGAGAGGATAACTTGCAC ACTCATCAACGAACACATACTGGTGAAAAACCA TACAAGTG TCCCGAATGTGGTAAAAGTTTTAGCC GGAGCGATGAACTTGTCCGACACCAACGAACCC ATACAGGCGAGAAGCCTTACAAATGTCCCGAGT GTGGCAAGAGCTTCTCACAATCAGGGAATCTGA CTGAGCATCAACGAACTCATACCGGGGAAAAAC CTTACAAGTGTCCAGAGTGTGGGAAGAGCTTTTC CACAAGTGGACATCTGGTACGCCACCAGAGGAC ACATACAGGGGAGAAGCCCT ACAAATGCCCCGA ATGCGGTAAAAGTTTCTCTCAGAATAGTACCCTG ACCGAACACCAGCGAACACACACTGGGAAAAAA ACGAGTAAAAGGCCGGCGGCCACGAAAAAGGCC GGCCAGGCAAAAAAGAAAAAGGGATCCGACGC GCTGGACGATTTCGATCTCGACATGCTGGGTTCT GATGCCCTCGATGACTTTGACCTGGATATGTTGG GAAGCGACGCATTGGATGACTTTGATCTGGACAT GCTCGG CTCCGATGCTCTGGACGATTTCGATCTC GATATGTTATAAAAGCTTGGGTGGCATCCCTGTG ACCCCTCCCCAGTGCCTCTCCTGGCCCTGGAAGT TGCCACTCCAGTGCCCACCAGCCTTGTCCTAATA AAATTAAGTTGCATCATTTTGTCTGACTAGGTGT CCTTCTATAATATTATGGGGTGGAGGGGGGTGGT ATGGAGCAAGGGGCAAGTTGGGAAGACAACCTG TAGGGCCTGC GGGGTCTATTGGGAACCAAGCTG GAGTGCAGTGGCACAATCTTGGCTCACTGCAATC TCCGCCTCCTGGGTTCAAGCGATTCTCCTGCCTC AGCCTCCCGAGTTGTTGGGATTCCAGGCATGCAT GACCAGGCTCAGCTAATTTTTGTTTTTTTGGTAG AGACGGGGTTTCACCATATTGGCCAGGCTGGTCT CCAACTCCTAATCTCAGGTGATCTACCCACCTTG GCCTCCCAAA TTGCTGGGATTACAGGCGTGAACC ACTGCTCCCTTCCCTGTCCTT 26 (RE + encoding) ggaggaagccatcaactaaactacaatgactgtaagatacaaaattgggaatggt aacatattttgaagttctgttgacataaagaatcatgattaatgcccatggaaatg aaagggcgatcaacactatggtttgaaaagggggaaattgtagagcacagatgt gttcgtgtggcagtgtgctgtctctagcaatactcagaga agagagagaacaatg aaattctgattggccccagtgtgagcccagatgaggttcagctgccaactttctctt tcacatcttatgaaagtcatttaagcacaactaactttttttttttttttttttttttgagaca gagtcttgctctgttgcccaggacagagtgcagtagtgactcaatctcggctcact gcagcct ccacctcctaggctcaaacggtcctcctgcatcagcctcccaagtagc tggaattacaggagtggcccaccatgcccagctaatttttgtatttttaatagatacg ggggtttcaccatatcacccaggctggtctcgaactcctggcctcaagtgatcca cctgcctcggcctcccaaagtgctgggattataggcgtcagccactatgc ccaac ccgaccaaccttttttaaaataaatatttaaaaaattggtatttcacatatatactagta 71

- 87 046157

tttacattttccacacaaaacggacgggcctccgctgaaccagtgaggcccca gacgtgcgcataaataacccctgcgtgctgcaccacctggggagagggggag gaccacggtaaatggagcgagcgcatagcaaaagggacgcggggtccttttct ctgccggtggcactgggtagctgtggccaggtgtggtactttgatgg ggcccag ggctggagctcaaggaagcgtcgcagggtcacagatctgggggaaccccgg ggaaaagcactgaggcaaaaccgccgctcgtctcctacaatatatgggagggg gaggttgagtacgttctggattactcataagacctttttttttccttccgggcgcaaa accgtgagctggatttataatcgccctataaag ctccagaggcggtcaggcacct gcagaggagccccgccgctccgccgactagctgcccccgcgagcaacggcct cgtgatttccccgccgatccggtccccgcctccccactctgcccccgcctacccc ggagccgtgcagccgcctctccgaatctctctcttctcctggcgctcgcgtgcga gagggaactagcgagaacgagg aagcagctggaggtgacgccggggcagatt acgcctgtcagggccgagccgagcggatcgctgggcgctgtgcagaggaaa ggcgggagtgcccggctcgctgtcgcagagccgaggtgggtaagctagcgac cacctggacttcccagcgcccaaccgtggcttttcagccaggtcctctcctcccg cggct tctcaaccaaccccatcccagcgccggccacccaacctcccgaaatga gtgcttcctgccccagcagccgaaggcgctactaggaacggtaacctgttacttt tccaggggccgtagtcgacccgctgcccgagttgctgtgcgactgcgcgcgcg gggctagagtgcaaggtgactgtggttcttctctggccaagtcc gagggagaac gtaaagatatgggcctttttccccctctcaccttgtctcaccaaagtccctagtccc cggagcagttagcctctttctttccagggaattagccagacacaacaacgggaac cagacaccgaaccagacatgcccgccccgtgcgccctccccgctcgctgccttt cctccctcttgtctctccagagccggatcttcaagg ggagcctccgtgcccccgg ctgctcagtccctccggtgtgcaggaccccggaagtcctccccgcacagctctc gcttctctttgcagcctgtttctgcgccggaccagtcgaggactctggacagtaga ggccccgggacgaccgagctgGAATTCGCCACCATGGCCCC AAAGAAGAAGCGGAAGGTCGGTATCCACGGAGT CCCAG CAGCCCTCGAACCAGGTGAAAAACCTTA CAAATGTCCTGAATGTGGGAAATCATTCAGTCGC AGCGACAACCTGGTGAGACATCAACGCACCCAT ACAGGAGAAAAAACCTTATAAATGTCCAGAATGT GGAAAGTCCTTCTCACGAGAGGATAACTTGCAC ACTCATCAACGAACACATACTGGTGAAAAACCA TACAAGTGTCCCGAATGTGGTAAAAGTTTTAGCC GGAGCGATGAACTTG TCCGACACCAACGAACCC ATACAGGCGAGAAGCCTTACAAATGTCCCGAGT GTGGCAAGAGCTTCTCAATCAGGGAATCTGA CTGAGCATCAACGAACTCATACCGGGGAAAAAC CTTACAAGTGTCCAGAGTGTGGGAAGAGCTTTTC CACAAGTGGACATCTGGTACGCCACCAGAGGAC ACATACAGGGGAGAAGCCCTACAAATGCCCCGA ATGCGGTAAAAGTTTCTCTCAGAATAGTACC CTG ACCGAACACCAGCGAACACACACTGGGAAAAAA ACGAGTAAAAGGCCGGCGGCCACGAAAAAGGCC GGCCAGGCAAAAAAGAAAAAGGGATCCGACGC GCTGGACGATTTCGATCTCGACATGCTGGGTTCT GATGCCCTCGATGACTTTGACCTGGATATGTTGG GAAGCGACGCATTGGATGACTTTGATCTGGACAT GCTCGGCTCCGATGCTCTGGACGATTTCGATCTC GATATGTTATAAaa agagaccggttcactgtgacagtaaaagac

- 88 046157

cggttcactgtgagaatgaaagagaccggttcactgtgatcggaaaagagaccg gttcactgtgagcggccttgaaacccagcagacaatgtagctcagtagaaaccc agcagacaatgtagctgaatggaaacccagcagacaatgtagcttcggagaaa cccagcagacaatgtagctAAGCTTGGTGGCATCCCTGTG ACCCCTCCCCA GTGCCTCTCCTGGCCCTGGAAGT TGCCACTCCAGTGCCCACCAGCCTTGTCCTAATA AAATTAAGTTGCATCATTTTGTCTGACTAGGTGT CCTTCTATAATATTATGGGGTGGAGGGGGGTGGT ATGGAGCAAGGGGCAAGTTGGGAAGACAACCTG TAGGGCCTGCGGGGTCTATTGGGAACCAAGCTG GAGTGCAGTGGCACAATCTTGGCTCACTGCAATC TCCGCCTCCTG GGTTCAAGCGATTCTCCTGCCTC AGCCTCCCGAGTTGTTGGGATTCCAGGCATGCAT GACCAGGCTCAGCTAATTTTTGTTTTTTTGGTAG AGACGGGGTTTCACCATATTGGCCAGGCTGGTCT CCAACTCCTAATCTCAGGTGATCTACCCACCTTG GCCTCCCAAATTGCTGGGATTACAGGCGTGAACC ACTGCTCCCTTCCCTGTCCTT 40 (encoding) ATGGCCGCAGATCACCTGATGCTGGCTGAAGGCT ACAGACTGGTGCAGCGGCCTCCATCTGCCGCTGC CGCCCACGGCCCCCACGCCCTGAGAACACTGCCC CCCTACGCCGGCCCTGGTCTTGATAGCGGACTCA GACCTAGAGGCGCCCCTCTGGGCCCTCCACCTCC AAGACAGCCTGGAGCCCTGGCCTACGGCGCCTTC GGCCCTCCTTCTAGCTTCCAGCCCTTCCCCGCCGT GC CTCCTCCAGCcGCTGGCATCGCCCACCTGCAG CCTGTGGCCACCCCTTACCCCGGAAGAGCCGCCG CCCCTCCAAACGCCCCTGGCGGACCTCCTGGCCC CCAGCCTGCTCCAAGCGCCGCTGCCCCTCCACCT CCTGCTCATGCCCTGGGCGGCATGGACGCCGAGC TGATCGACGAGGAAGCCCTGACCAGCCTGGAAC TGGAACTGGGCCTGCACAGAGTGCGGGAACTGC CTGAGCTGTT CCTGGGACAGAGCGAGTTCGACTG CTTCAGCGACCTGGGCAGCGCCCCTCCTGCCGGC TCTGTGTCCTGCgccgaccacctgatgctcgccgagggctaccgcct ggtgcagaggccgccgtccgccgccgccgcccatggccctcatgcgctccgg actctgccgccgtacgcgggcccgggcctggacagtgggctgaggccgcgg gggg ctccgctggggccgccgccgccccgccaacccggggccctggcgtac ggggccttcgggccgccgtcctccttccagccctttccggccgtgcctccgccg gccgcgggcatcgcgcacctgcagcctgtggcgacgccgtaccccggccgc gcCgccgcgccccccaacgctccgggaggccccggggcccg cagccggc cccaagcgccgcagccccgccgccgcccgcgcacgccctgggcggcatgga cgccgaactcatcgacgaggaggcgctgacgtcgctggagctggagctgggg ctgcaccgcgtgcgcgagctgcccgagctgttcctgggccagagcgagttcga ctgcttctcggacttggggtccgcg ccgcccgccggctccgtgagctgccagtc ccagctcatcaaacccagccgcatgcgcaagtaccccaaccggcccagcaag acgcccccccacgaacgcccttacgcttgcccagtggagtcctgtgatcgccgc ttctccCGCAGCGACAACCTGGTGAGAcacatccgcatccac acaggccagaagcccttccagtgccgcatctgcatg AGAaacttcagcCG AGAGGAT AACTT GC AC ACTcacatccgcacccacacaggcg 72

- 89 046157

aaaagcccttcgcctgcgacatctgtggaagaaagtttgccCGGAGCGA TGAACTTGTCCGAcataccaagatccacttgcggcagaaggaccgc ccttacgcttgcccagtggagtcctgtgatcgccgcttctccCAATCAGG GAATCTGACTGAGcacatccgcatccacacaggccagaagcccttc cagtgccgcatctgcatgA GAaacttcagcACAAGTGGACATCT GGTACGCcacatccgcacccacacaggcgaaaagcccttcgcctgcgac atctgtggaagaaagtttgccC AGA AT AGT AC CC T GAC CGA A cataccaagatccacttgcggcagaaggacaag 41 (encoding) ATGGCCGCAGATCACCTGATGCTGGCTGAAGGCT ACAGACTGGTGCAGCGGCCTCCATCTGCCGCTGC CGCCCACGGCCCCCACGCCCTGAGAACACTGCCC CCCTACGCCGGCCCTGGTCTTGATAGCGGACTCA GACCTAGAGGCGCCCCTCTGGGCCCTCCACCTCC AAGACAGCCTGGAGCCCTGGCCTACGGCGCCTTC GGCCCTCCTTCTAGCTTCCAGCCCTTCCCCGCCGT GCC TCCTCCAGCTGCTGGCATCGCCCACCTGCAG CCTGTGGCCACCCCTTACCCCGGAAGAGCCGCCG CCCCTCCAAACGCCCCTGGCGGACCTCCTGGCCC CCAGCCTGCTCCAAGCGCCGCTGCCCCTCCACCT CCTGCTCATGCCCTGGGCGGCATGGACGCCGAGC TGATCGACGAGGAAGCCCTGACCAGCCTGGAAC TGGAACTGGGCCTGCACAGAGTGCGGGAACTGC CTGAGCTGTTCC TGGGACAGAGCGAGTTCGACTG CTTCAGCGACCTGGGCAGCGCCCCTCCTGCCGGC TCTGTGTCCTGCGGCGGCAGCGGCGGCGGAAGC GGCgccgaccacctgatgctcgccgagggctaccgcctggtgcagaggcc gccgtccgccgccgccgcccatggccctcatgcgctccggactctgccgccgt acgcgggcccgggcctggacagtg ggctgaggccgcggggggctccgctgg ggccgccgccgccccgccaacccggggccctggcgtacggggccttcgggc cgccgtcctccttccagccctttccggccgtgcctccgccggccgcgggcatcg cgcacctgcagcctgtggcgacgcctaccccggccgcgcggccgcgcccc ccaacgctccgg gaggccccccgggcccgcagccggccccaagcgccgca gccccgccgccgcccgcgcacgccctgggcggcatggacgccgaactcatc gacgaggaggcgctgacgtcgctggagctggagctggggctgcaccgcgtgc gcgagctgcccgagctgttcctgggccagagcgagttcgactgcttctcggactt c gcatccacacaggccagaagcccttccagtgccgcatctgcatgAGAaact tcagcCGAGAGGATAACTTGCACACTcacatccgcacccac acaggcgaaaagcccttcgcctgcgacatctgtggaagaaagtttgccCGG AGCGATGAACTTGTCCGAcataccaagatccacttgcggcaga aggaccgcccttacgcttgcccagtggagtcc tgtgatcgccgcttctccCAA TCAGGGAATCTGACTGAGcacatccgcatccacacaggccag aagcccttccagtgccgcatctgcatgAGAaacttcagcACAAGTGG ACATCTGGTACGCcacatccgcacccacacaggcgaaaagcccttc gcctgcgacatctgtggaagaaagtttgccC AGA AT AGT ACC C TG ACCGAAc ataccaagatccacttgcggcagaaggacaag 73

- 90 046157 (RE + encoding) ggaggaagccatcaactaaactacaatgactgtaagatacaaaattgggaatggt aacatattttgaagttctgttgacataaagaatcatgattaatgcccatggaaatg aaagggcgatcaacactatggtttgaaaaagggggaaattgtagagcacagatgt gttcgtgtggca gtgtgctgtctctagcaatactcagagaagagagagaacaatg aaattctgattggccccagtgtgagcccagatgaggttcagctgccaactttctctt tcacatcttatgaaagtcatttaagcacaactaacttttttttttttttttttttttttgagaca gagtcttgctctgttgcccaggacagagtgcag tagtgactcaatctcggctcact gcagcctccacctcctaggctcaaacggtcctcctgcatcagcctcccaagtagc tggaattacaggagtggcccaccatgcccagctaatttttgtatttttaatagatacg ggggtttcaccatatcacccaggctggtctcgaactcctggcctcaagtgatcca cctgcctcggcctcccaaagt gctgggattataggcgtcagccactatgcccaac ccgaccaaccttttttaaaataaatatttaaaaaattggtatttcacatatatactagta tttacatttatccacacaaaacggacgggcctccgctgaaccagtgaggcccca gacgtgcgcataaataacccctgcgtgctgcaccacctggggagagggggag gaccacggtaaatggagc gagcgcatagcaaaagggacgcggggtccttttct ctgccggtggcactgggtagctgtggccaggtgtggtactttgatggggcccag ggctggagctcaaggaagcgtcgcagggtcacagatctgggggaaccccgg ggaaaagcactgaggcaaaaccgccgctcgtctcctacaatatatggagagggg gaggttgagt acgttctggattactcataagacctttttttttccttccgggcgcaaa accgtgagctggatttataatcgccctataaagctccagaggcggtcaggcacct gcagaggagccccgccgctccgccgactagctgcccccgcgagcaacggcct cgtgatttccccgccgatccggtccccgcctccccactctgcccccgcc tacccc ggagccgtgcagccgcctctccgaatctctctcttctcctggcgctcgcgtgcga gagggaactagcgagaacgaggaagcagctggaggtgacgccgggcagatt acgcctgtcagggccgagccgagcggatcgctgggcgctgtgcagaggaaa ggcgggagtgcccggctcgctgtcgc agagccgaggtgggtaagctagcgac cacctggacttcccagcgcccaaccgtggcttttcagccaggtcctctcctcccg cggcttctcaaccaaccccatcccagcgccggccacccaacctcccgaaatga gtgcttcctgccccagcagccgaaggcgctactaggaacggtaacctgttacttt tccaggggccgtagtcgacccg ctgcccgagttgctgtgcgactgcgcgcgcg gggctagagtgcaaggtgactgtggttcttctctggccaagtccgagggaac gtaaagatatgggcctttttccccctctcaccttgtctcaccaaagtccctagtccc cggagcagttagcctcttttctttccagggaattagccagacacaacaacgggaac caga caccgaaccagacatgcccgccccgtgcgccctccccgctcgctgccttt cctccctcttgtctctccagagccggatcttcaaggggagcctccgtgcccccgg ctgctcagtccctccggtgtgcaggaccccggaagtcctccccgcacagctctc gcttctctttgcagcctgtttctgcgccggaccagtcga ggactctggacagtaga ggccccgggacgaccgagctgGAATTCGCCACCATGGCCCC AAAGAAGAAGCGGAAGGTCGGTATCCACGGAGT CCCAGCAGCCCTCGAACCAGGTGAAAAACCTTA CAAATGTCCTGAATGTGGGAAATCATTCAGTCGC AGCGACAACCTGGTGAGACATCAACGCACCCAT ACAGGAGAAAACCTTATAAATGTCCAGAATGT GGAAAGTCCTTCTCACGAGAGGATAACTTGCAC ACTCATCAACGAACACATACTGGTGAAAAACCA TACAAGTGTCCCGAATGTGGTAAAAGTTTTAGCC GGAGCGATGAACTTGTCCGACACCAACGAACCC ATACAGGCGAGAAGCCTTACAAATGTCCCGAGT GTGGCAAGAGCTTCTCACAATCAGGGAATCTGA CTGAGCAT CAACGAACTCATACCGGGGAAAAAC

- 91 046157

CTTACAAGTGTCCAGAGTGTGGGAAGAGCTTTTC CACAAGTGGACATCTGGTACGCCACCAGAGGAC ACATACAGGGGAGAAGCCCTACAAATGCCCCGA ATGCGGTAAAAGTTTCTCTCAGAATAGTACCCTG ACCGAACACCAGCGAACACACACTGGGAAAAAA ACGAGTAAAAGGCCGGCGGCCACGAAAAAGGCC GGCCAGGCAAAAAAGAAAAGGATCCTACCCA TACGACGTACCAGAT TACGCTCTCGAGGACGCGC TGGACGATTTCGATCTCGACATGCTGGGTTCTGA TGCCCTCGATGACTTTGACCTGGATATGTTGGGA AGCGACGCATTGGATGACTTTGATCTGGACATGC TCGGCTCCGATGCTCTGGACGATTTCGATCTCGA TATGTTATAAACTAGTGAAACCCAGCAGACAAT GTAGCTAGACCCAGTAGCCAGATGTAGCTAAAG AGACCGGTTCACTGTGAAAGCTTG GGTGGCATCC CTGTGACCCCTCCCCAGTGCCTCTCCTGGCCCTG GAAGTTGCCACTCCAGTGCCCACCAGCCTTGTCC TAATAAAATTAAGTTGCATCATTTTGTCTGACTA GGTGTCCTTCTATAATATTATGGGGTGGAGGGGG GTGGTATGGAGCAAGGGGCAAGTTGGGAAGACA ACCTGTAGGGCCTGCGGGGTCTATTGGGAACCA AGCTGGAGTGCAGTGGCACAATCTTG GCTCACTG CAATCTCCGCCTCCTGGGTTCAAGCGATTCTCCT GCCTCAGCCTCCCGAGTTGTTGGGATTCCAGGCA TGCATGACCAGGCTCAGCTAATTTTTGTTTTTTTG GTAGAGACGGGGTTTCACCATATTGGCCAGGCTG GTCTCCAACTCCTAATCTCAGGTGATCTACCCAC CTTGGCCTCCCAAATTGCTGGGATTACAGGCGTG AACCACTGCTCCCTTCCCTGTCCTT 43 (RE + encoding) ggaggaagccatcaactaaactacaatgactgtaagatacaaaattgggaatggt aacatattttgaagttctgttgacataaagaatcatgattaatgcccatggaaatg aaagggcgatcaacactatggtttgaaaagggggaaattgtagagcacagatgt gttcgtgtggcagtgtgctgtctctagcaatactcagaga agagagagaacaatg aaattctgattggccccagtgtgagcccagatgaggttcagctgccaactttctctt tcacatcttatgaaagtcatttaagcacaactaactttttttttttttttttttttttgagaca gagtcttgctctgttgcccaggacagagtgcagtagtgactcaatctcggctcact gcagcct ccacctcctaggctcaaacggtcctcctgcatcagcctcccaagtagc tggaattacaggagtggcccaccatgcccagctaatttttgtatttttaatagatacg ggggtttcaccatatcacccaggctggtctcgaactcctggcctcaagtgatcca cctgcctcggcctcccaaagtgctgggattataggcgtcagccactatgc ccaac ccgaccaaccttttttaaaataaatatttaaaaaattggtatttcacatatatactagta tttacatttatccacacaaaacggacgggcctccgctgaaccagtgaggcccca gacgtgcgcataaataacccctgcgtgctgcaccacctggggagagggggag gaccacggtaaatggagcgagcgcatagcaaaagggacgcggggt ccttttct ctgccggtggcactgggtagctgtggccaggtgtggtactttgatggggcccag ggctggagctcaaggaagcgtcgcagggtcacagatctgggggaaccccgg ggaaaagcactgaggcaaaaccgccgctcgtctcctacaatatatgggagggg gaggttgagtacgttctggattactcataagacctttttt tttttccttccgggcgcaaa accgtgagctggatttataatcgccctataaagctccagaggcggtcaggcacct gcagaggagccccgccgctccgccgactagctgcccccgcgagcaacggcct 75

- 92 046157

cgtgatttccccgccgatccggtccccgcctccccactctgcccccgcctacccc ggagccgtgcagccgcctctccgaatctctctcttctcctggcgctcgcgtgcga gagggaactagcgagaacgaggaagcagctggaggtgacgccgggcagatt acgcctgtcagggccgagccgagcggatcgctggggc gctgtgcagaggaaa ggcgggagtgcccggctcgctgtcgcagagccgaggtgggtaagctagcgac cacctggacttcccagcgcccaaccgtggcttttcagccaggtcctctcctcccg cggcttctcaaccaaccccatcccagcgccggcacccaacctcccgaaatga gtgcttcctgccccagcagccgaa c ggagcagttagcctctttctttccagggaattagccagacacaacaacgggaac cagacaccgaaccagacatgcccgccccgtgcgccctccccgctcgctgccttt cctccctcttgtctctccagagccggatcttcaaggggagcctccgtgcccccgg ctgctcagtccctccggtgtgcaggaccccggaagtcctccccgcaca gctctc gcttctctttgcagcctgtttctgcgccggaccagtcgaggactctggacagtaga ggccccgggacgaccgagctgGAATTCGCCACCATGGCCCC AAAGAAGAAGCGGAAGGTCGGTATCCACGGAGT CCCAGCAGCCCTCGAACCAGGTGAAAAACCTTA CAAATGTCCTGAATGTGGGAAATCATTCAGTCGC AGCGACAACCTGGTGAGACAT CAACGCACCCAT ACAGGAAAAAACCTTATAAATGTCCAGAATGT GGAAAGTCCTTCTCACGAGAGGATAACTTGCAC ACTCATCAACGAACACATACTGGTGAAAAACCA TACAAGTGTCCCGAATGTGGTAAAAGTTTTAGCC GGAGCGATGAACTTGTCCGACACCAACGAACCC ATACAGGCGAGAAGCCTTACAAATGTCCCGAGT GTGGCAAGAGCTTCTCAATCAGGGAATCT GA CTGAGCATCAACGAACTCATACCGGGGAAAAAC CTTACAAGTGTCCAGAGTGTGGGAAGAGCTTTTC CACAAGTGGACATCTGGTACGCCACCAGAGGAC ACATACAGGGGAGAAGCCCTACAAATGCCCCGA ATGCGGTAAAAGTTTCTCTCAGAATAGTACCCTG ACCGAACACCAGCGAACACACACTGGGAAAAAA ACGAGTAAAAGGCCGGCGGCCACGAAAAAGGCC GGCCAGGCAAAAAAGA AGGGATCCTACCCA TACGACGTACCAGATTACGCTCTCGAGGACGCGC TGGACGATTTCGATCTCGACATGCTGGGTTCTGA TGCCCTCGATGACTTTGACCTGGATATGTTGGGA AGCGACGCATTGGATGACTTTGATCTGGACATGC TCGGCTCCGATGCTCTGGACGATTTCGATCTCGA TATGTTATAAACTAGTGAAACCCAGCAGACAAT GTAGCTAGACCCAGTAGCCAGATGTAGCT AAAG AGACCGGTTCACTGTGAGAAACCCAGCAGACAA TGTAGCTAGACCCAGTAGCCAGATGTAGCTAAA GAGACCGGTTCACTGTGAAAGCTTGGGTGGCATC CCTGTGACCCCTCCCCAGTGCCTCTCCTGGCCCT GGAAGTTGCCACTCCAGTGCCCACCAGCCTTGTC CTAATAAAATTAAGTTGCATCATTTTGTCTGACT AGGTGTCCTTCTATAATATTATGGGGTGGAGG

- 93 046157

GGTGGTATGGAGCAAGGGGCAAGTTGGGAAGAC AACCTGTAGGGCCTGCGGGGTCTATTGGGAACC AAGCTGGAGTGCAGTGGCACAATCTTGGCTCACT GCAATCTCCGCCTCCTGGGTTCAAGCGATTCTCC TGCCTCAGCCTCCCGAGTTGTTGGGATTCCAGGC ATGCATGACCAGGCTCAGCTAATTTTTGTTTTTTTT GGTAGAGACGGGTTTCACCATATTGGCCA GGCT GGTCTCCAACTCCTAATCTCAGGTGATCTACCCA CCTTGGCCTCCCAAATTGCTGGGATTACAGGCGT GAACCACTGCTCCCTTCCCTGTCCTT 44 (RE + encoding) ggaggaagccatcaactaaactacaatgactgtaagatacaaaattgggaatggt aacatattttgaagttctgttgacataaagaatcatgattaatgcccatggaaatg aaagggcgatcaacactatggtttgaaaagggggaaattgtagagcacagatgt gttcgtgtggcagtgtgctgtctctagcaatactcagaga agagagagaacaatg aaattctgattggccccagtgtgagcccagatgaggttcagctgccaactttctctt tcacatcttatgaaagtcatttaagcacaactaactttttttttttttttttttttttgagaca gagtcttgctctgttgcccaggacagagtgcagtagtgactcaatctcggctcact gcagcct ccacctcctaggctcaaacggtcctcctgcatcagcctcccaagtagc tggaattacaggagtggcccaccatgcccagctaatttttgtatttttaatagatacg ggggtttcaccatatcacccaggctggtctcgaactcctggcctcaagtgatcca cctgcctcggcctcccaaagtgctgggattataggcgtcagccactatgc ccaac ccgaccaaccttttttaaaataaatatttaaaaaattggtatttcacatatatactagta tttacatttatccacacaaaacggacgggcctccgctgaaccagtgaggcccca gacgtgcgcataaataacccctgcgtgctgcaccacctggggagagggggag gaccacggtaaatggagcgagcgcatagcaaaagggacgcggggt ccttttct ctgccggtggcactgggtagctgtggccaggtgtggtactttgatggggcccag ggctggagctcaaggaagcgtcgcagggtcacagatctgggggaaccccgg ggaaaagcactgaggcaaaaccgccgctcgtctcctacaatatatgggagggg gaggttgagtacgttctggattactcataagacctttttt tttttccttccgggcgcaaa accgtgagctggatttataatcgccctataaagctccagaggcggtcaggcacct gcagaggagccccgccgctccgccgactagctgcccccgcgagcaacggcct cgtgatttccccgccgatccggtccccgcctccccactctgcccccgcctacccc ggagccgtgcagccgcctct ccgaatctctctcttctcctggcgctcgcgtgcga gagggaactagcgagaacgaggaagcagctggaggtgacgccgggcagatt acgcctgtcagggccgagccgagcggatcgctgggcgctgtgcagaggaaa ggcgggagtgcccggctcgctgtcgcagagccgaggtgggtaagctagcgac cacc tggacttcccagcgcccaaccgtggcttttcagccaggtcctctcctcccg cggcttctcaaccaaccccatcccagcgccggccacccaacctcccgaaatga gtgcttcctgccccagcagccgaaggcgctactaggaacggtaacctgttacttt tccaggggccgtagtcgacccgctgcccgagttgctgtgcgactg cgcgcgcg gggctagagtgcaaggtgactgtggttcttctctggccaagtccgagggagaac gtaaagatatgggcctttttccccctctcaccttgtctcaccaaagtccctagtccc cggagcagttagcctctttctttccagggaattagccagacacaacaacgggaac cagacaccgaaccagacatgcccgccccgtgcg ccctccccgctcgctgccttt cctccctcttgtctctccagagccggatcttcaaggggagcctccgtgcccccgg ctgctcagtccctccggtgtgcaggaccccggaagtcctccccgcacagctctc gcttctctttgcagcctgtttctgcgccggaccagtcgaggactctggacagtaga ggccccgggacgacc gagctgGAATTCGCCACCATGGCCgc cgaccacctgatgctcgccgagggctaccgcctggtgcagaggccgccgtcc gccgccgccgcccatggccctcatgcgctccggactctgccgccgtacgcggg 76

- 94 046157

cccggggcctggacagtgggctgaggccgcggggggctccgctggggccgcc gccgccccgccaacccggggccctggcgtacggggccttcgggccgccgtc ctccttccagccctttccggccgtgcctccgccggccgcgggcatcgcgcacct gcagcctgtggcgacgccgtaccccggccgcgcggccg cgccccccaacgc tccgggaggccccccgggcccgcagccggccccaagcgccgcagccccgc cgccgcccgcgcacgccctgggcggcatggacgccgaactcatcgacgagg aggcgctgacgtcgctggagctggagctggggctgcaccgcgtgcgcgagct gcccgagctgttcctgggccagagcgagtt cgactgcttctcggacttggggtc cgcgccgcccgccggctccgtgagctgcggtggttctggtggtggttctggtca gtcccagctcatcaaacccagccgcatgcgcaagtaccccaaccggcccagca agacgcccccccacgaacgcccttacgcttgcccagtggagtcctgtgatcgcc gcttctccCGCAGC GACAACCTGGTGAGAcacatccgcatcc acacaggccagaagcccttccagtgccgcatctgcatgAGAaacttcagcC GAGAGGATAACTTGCACACTcacatccgcacccacacaggc gaaaagcccttcgcctgcgacatctgtggaagaaagtttgccCGGAGCG ATGAACTTGTCCGAcataccaagatccacttgcggcagaaggacc gcccttac gcttgcccagtggagtcctgtgatcgccgcttctccCAATCAG GGAATCTGACTGAGcacatccgcatccacacaggccagaagccct tccagtgccgcatctgcatgAGAaacttcagcACAAGTGGACATC TGGTACGCcacatccgcacccacacaggcgaaaagcccttcgcctgcg acatctgtggaagaaagtttgccCAGA ATAGTACCCTGACCG AAcataccaagatccacttgcggcagaaggacaagtaaCTCGAGGA AACCCAGCAGACAATGTAGCTAGACCCAGTAGC CAGATGTAGCTAAAGAGACCGGTTCACTGTGAA AGCTTGGGTGGCATCCCTGTGACCCCTCCCCAGT GCCTCTCCTGGCCCTGGAAGTTGCCACTCCAGTG CCCACCAGCCTTGTCCTAATAAAATTAAGTTGCA TCATTT TGTCTGACTAGGTGTCCTTCTATAATATT ATGGGGTGGAGGGGGGTGGTATGGAGCAAGGGG CAAGTTGGGAAGACAACCTGTAGGGCCTGCGGG GTCTATTGGGAACCAAGCTGGAGTGCAGTGGCA CAATCTTGGCTCACTGCAATCTCCGCCTCCTGGG TTCAAGCGATTCTCCTGCCTCAGCCTCCCGAGTT GTTGGGATTCCAGGCATGCATGACCAGGCTCAGC TAATTTTTGTTTT TTTGGTAGAGACGGGGTTTCAC CATATTGGCCAGGCTGGTCTCCAACTCCTAATCT CAGGTGATCTACCCACCTTGGCCTCCCAAATTGC TGGGATTACAGGCGTGAACCACTGCTCCCTTCCC TGTCCTT 45 (RE + encoding) ggaggaagccatcaactaaactacaatgactgtaagatacaaaattgggaatggt aacatattttgaagttctgttgacataaagaatcatgattaatgcccatggaaatg aaagggcgatcaacactatggtttgaaaagggggaaattgtagagcacagatgt gttcgtgtggcagtgtgctgtctctagcaatactcagaga agagagagaacaatg aaattctgattggccccagtgtgagcccagatgaggttcagctgccaactttctctt tcacatcttatgaaagtcatttaagcacaactaacttttttttttttttttttttttttgagaca gagtcttgctctgttgcccaggacagagtgcagtagtgactcaatctcggctcact gcagcct ccacctcctaggctcaaacggtcctcctgcatcagcctcccaagtagc tggaattacaggagtggcccaccatgcccagctaatttttgtatttttaatagatacg ggggtttcaccatatcacccaggctggtctcgaactcctggcctcaagtgatcca cctgcctcggcctcccaaagtgctgggattataggcgtcagccactatgc ccaac 184

- 95 046157

ccgaccaacctttttaaaataaatatttaaaaaattggtatttcacatatatactagta tttacatttatccacacaaaacggacgggcctccgctgaaccagtgaggcccca gacgtgcgcataaataacccctgcgtgctgcaccacctggggagagggggag gaccacggtaaatggagcgagcgcatagcaaaagggacgcggggtcc ttttct ctgccggtggcactgggtagctgtggccaggtgtggtactttgatggggcccag ggctggagctcaaggaagcgtcgcagggtcacagatctgggggaaccccgg ggaaaagcactgaggcaaaaccgccgctcgtctcctacaatatatgggagggg gaggttgagtacgttctggattactcataagaccttttttt ttttccttccgggcgcaaa accgtgagctggatttataatcgccctataaagctccagaggcggtcaggcacct gcagaggagccccgccgctccgccgactagctgcccccgcgagcaacggcct cgtgatttccccgccgatccggtccccgcctccccactctgcccccgcctacccc ggagccgtgcagccgcctctcc gaatctctctcttctcctggcgctcgcgtgcga gagggaactagcgagaacgaggaagcagctggaggtgacgccgggcagatt acgcctgtcagggccgagccgagcggatcgctgggcgcttgcagaggaaa ggcgggagtgcccggctcgctgtcgcagagccgaggtgggtaagctagcgac cacct ggacttcccagcgcccaaccgtggcttttcagccaggtcctctcctcccg cggcttctcaaccaaccccatcccagcgccggccacccaacctcccgaaatga gtgcttcctgccccagcagccgaaggcgctactaggaacggtaacctgttacttt tccaggggccgtagtcgacccgctgcccgagttgctgtgcgactgc gcgcgcg gggctagagtgcaaggtgactgtggttcttctctggccaagtccgagggaac gtaaagatatgggcctttttccccctctcaccttgtctcaccaaagtccctagtccc cggagcagttagcctctttctttccagggaattagccagacacaacaacgggaac cagacaccgaaccagacatgcccgccccgtgcgcc ctccccgctcgctgccttt cctccctcttgtctctccagagccggatcttcaaggggagcctccgtgcccccgg ctgctcagtccctccggtgtgcaggaccccggaagtcctccccgcacagctctc gcttctctttgcagcctgtttctgcgccggaccagtcgaggactctggacagtaga ggccgggacgaccga gctgGAATTCGCCACCATGGCCgc cgaccacctgatgctcgccgagggctaccgcctggtgcagaggccgccgtcc gccgccgccgcccatggccctcatgcgctccggactctgccgccgtacgcggg cccgggcctggacagtgggctgaggccgcggggggctccgctggggccgcc gccgccccgccaacccggggccctggcgt acggggccttcgggccgccgtc ctccttccagccctttccggccgtgcctccgccggccgcgggcatcgcgcacct gcagcctgtggcgacgccgtaccccggccgcgcggccgcgccccccaacgc tccgggaggccccccgggcccgcagccggccccaagcgccgcagccccgc cgccgcccgcgca cgccctgggcggcatggacgccgaactcatcgacgagg aggcgctgacgtcgctggagctggagctggggctgcaccgcgtgcgcgagct gcccgagctgttcctgggccagagcgagttcgactgcttctcggacttggggtc cgcgccgcccgccggctccgtgagctgcggtggttctggtggtggtt ctggtca gtcccagctcatcaaacccagccgcatgcgcaagtaccccaaccggcccagca agacgcccccccacgaacgcccttacgcttgcccagtggagtcctgtgatcgcc gcttctccCGCAGCGACAACCTGGTGAGAcacatccgcatcc acacaggccagaagcccttccagtgccgcatctgcatgAGAaacttcagcC GAGAGG ATAACTTGCACACTcacatccgcacccacacaggc gaaaagcccttcgcctgcgacatctgtggaagaaagtttgccCGGAGCG ATGAACTTGTCCGAcataccaagatccacttgcggcagaaggacc gcccttacgcttgcccagtggagtcctgtgatcgccgcttctccCAATCAG GGAATCTGACTGAGcacatccgcatccacacaggccaga agccct tccagtgccgcatctgcatgAGAaacttcagcACAAGTGGACATC TGGTACGCcacatccgcacccacacaggcgaaaagcccttcgcctgcg acatctgtggaagaaagtttgccCAGAATAGTACCCTGACCG

- 96 046157

AAcataccaagatccacttgcggcagaaggacaagtaaCTCGAGGA AACCCAGCAGACAATGTAGCTAGACCCAGTAGC CAGATGTAGCTAAAGAGACCGGTTCACTGTGAG AAACCCAGCAGACAATGTAGCTAGACCCAAGTAG CCAGATGTAGCTAAAGAGACCGGTTCACTGTGA AAGCTTGGGTGGCATCCCTGTGACCCCTCCCCAG TGCCTCTCCTGGCCCTGGAAG TTGCCACTCCAGT GCCCACCAGCCTTGTCCTAATAAAATTAAGTTGC ATCATTTTGTCTGACTAGGTGTCCTTCTATAATAT TATGGGGTGGAGGGGGGTGGTATGGAGCAAGGG GCAAGTTGGGAAGACAACCTGTAGGGCCTGCGG GGTCTATTGGGAACCAAGCTGGAGTGCAGTGGC ACAATCTTGGCTCACTGCAATCTCCGCCTCCTGG GTTCAAGCGATTCTCCTGCCTCAGCC TCCCGAGT TGTTGGATTCCAGGCATGCATGACCAGGCTCAG CTAATTTTTGTTTTTTTGGTAGAGACGGGGTTTCA CCATATTGGCCAGGCTGGTCTCCAACTCCTAATC TCAGGTGATCTACCCACCTTGGCCTCCCAAATTG CTGGGATTACAGGCGTGAACCACTGCTCCCTTCC CTGTCCTT 8 (encoding) ATGGCCgccgaccacctgatgctcgccgagggctaccgcctggtgcaga ggccgccgtccgccgccgccgcccatggccctcatgcgctccggactctctgccg ccgtacgcgggcccgggcctggacagtgggctgaggccgcggggggctccg ctggggccgccgccgccccgccaacccggggccctggcgtacggggcctt c gggccgccgtcctccttccagccctttccggccgtgcctccgccggccgcggg catcgcgcacctgcagcctgtggcgacgccgtaccccggccgcgcggccgcg ccccccaacgctccgggaggccccccgggcccgcagccggccccaagcgcc gcagccccgccgccgcccgcgcacgccctgggcgg catggacgccgaactc atcgacgaggaggcgctgacgtcgctggagctggagctggggctgcaccgcg tgcgcgagctgcccgagctgttcctgggccagagcgagttcgactgcttctcgg acttggggtccgcgccgcccgccggctccgtgagctgcggtggttctggtggtg gttctggtcagtc ccagctcatcaaacccagccgcatgcgcaagtaccccaacc ggcccagcaagacgcccccccacgaacgcccttacgcttgcccagtggagtcc tgtgatcgccgcttctccCGCAGCGACAACCTGGTGAGAcac atccgcatccacacaggccaagcccttccagtgccgcatctgcatgAGAa acttcagcCGAGAGGATAACTTGCACACTcacatcc gcacc cacacaggcgaaaagcccttcgcctgcgacatctgtggaagaaagtttgccCG GAGCGATGAACTTGTCCGAcataccaagatccacttgcggca gaaggaccgcccttacgcttgcccagtggagtcctgtgatcgccgcttctccCA AT C AGGGA ATCTGACTGAGcacatccgcatccacacaggcca gaagcccttccagtg ccgcatctgcatgAGAaacttcagcACAAGTG GACATCTGGTACGCcacatccgcacccacacaggcgaaaagccct tcgcctgcgacatctgtggaagaaagtttgccCAGAATAGTACCCT GACCGAAcataccaagatccacttgcggcagaaggacaag 203 46 (encoding) ATGGCCgccgaccacctgatgctcgccgagggctaccgcctggtgcaga ggccgccgtccgccgccgccgcccatggccctcatgcgctccggactctctgccg ccgtacgcgggcccgggcctggacagtgggctgaggccgcggggggctccg ctggggccgccgccgccccgccaacccggggccctggcgtacggggcctt c gggccgccgtcctccttccagccctttccggccgtgcctccgccggccgcggg catcgcgcacctgcagcctgtggcgacgccgtaccccggccgcgcggccgcg 204

- 97 046157

ccccccaacgctccgggaggccccccgggcccgcagccggccccaagcgcc gcagccccgccgccgcccgcgcacgccctgggcggcatggacgccgaactc atcgacgaggaggcgctgacgtcgctggagctggagctggggctgcaccgcg tgcgcgagctgcccgagctgttcctgggccagagcgagttc gactgcttctcgg acttggggtccgcgccgcccgccggctccgtgagctgcggtggttctggtggtg gttctggtGGTGGCAGCGGGGGAGGTTCTGGTcagtccca gctcatcaaacccagccgcatgcgcaagtaccccaaccggcccagcaagacg cccccccacgaacgcccttacgcttgcccagtggagtcctgtgat cgccgcttct ccCGCAGCGACAACCTGGTGAGAcacatccgcatccacaca ggccagaagcccttccagtgccgcatctgcatgAGAaacttcagcCGAG AGGAT AACTTGC AC ACT cacatccgcacccacacaggcgaaaa gcccttcgcctgcgacatctgtggaagaaagtttgccCGGAGCGATGA ACTTGTCCGAcataccaagatccact tgcggcagaaggaccgccctta cgcttgcccagtggagtcctgtgatcgccgcttctccCAATCAGGGAA TCTGACTGAGcacatccgcatccacacaggccagaagcccttccagtg ccgcatctgcatg AGAaacttcagc AC A AGT GGAC AT C T GGT ACGCcacatccgcacccacacaggcgaaaagcccttcgcctg cgacatctg tggaagaaagtttgccC AGAAT AGT ACCCTGACCGAAcatac caagatccacttgcggcagaaggacaag 47 (encoding) ATGGCCGCAGATCACCTGATGCTGGCTGAAGGCT ACAGACTGGTGCAGCGGCCTCCATCTGCCGCTGC CGCCCACGGCCCCCACGCCCTGAGAACACTGCCC CCCTACGCCGGCCCTGGTCTTGATAGCGGACTCA GACCTAGAGGCGCCCCTCTGGGCCCTCCACCTCC AAGACAGCCTGGAGCCCTGGCCTACGGCGCCTTC GGCCCTCCTTCTAGCTTCCAGCCCTTCCCCGCCGT GC CTCCTCCAGCcGCTGGCATCGCCCACCTGCAG CCTGTGGCCACCCCTTACCCCGGAAGAGCCGCCG CCCCTCCAAACGCCCCTGGCGGACCTCCTGGCCC CCAGCCTGCTCCAAGCGCCGCTGCCCCTCCACCT CCTGCTCATGCCCTGGGCGGCATGGACGCCGAGC TGATCGACGAGGAAGCCCTGACCAGCCTGGAAC TGGAACTGGGCCTGCACAGAGTGCGGGAACTGC CTGAGCTGTT CCTGGGACAGAGCGAGTTCGACTG CTTCAGCGACCTGGGCAGCGCCCCTCCTGCCGGC TCTGTGTCCTGCgccgaccacctgatgctcgccgagggctaccgcct ggtgcagaggccgccgtccgccgccgccgcccatggccctcatgcgctccgg actctgccgccgtacgcgggcccgggcctggacagtgggctgaggccgcgg gggg ctccgctggggccgccgccgccccgccaacccggggccctggcgtac ggggccttcgggccgccgtcctccttccagccctttccggccgtgcctccgccg gccgcgggcatcgcgcacctgcagcctgtggcgacgccgtaccccggccgc gcCgccgcgccccccaacgctccgggaggccccggggcccg cagccggc cccaagcgccgcagccccgccgccgcccgcgcacgccctgggcggcatgga cgccgaactcatcgacgaggaggcgctgacgtcgctggagctggagctgggg ctgcaccgcgtgcgcgagctgcccgagctgttcctgggccagagcgagttcga ctgcttctcggacttggggtccgcg ccgcccgccggctccgtgagctgccagtc ccagctcatcaaacccagccgcatgcgcaagtaccccaaccggcccagcaag acgcccccccacgaacgcccttacgcttgcccagtggagtcctgtgatcgccgc ttctccCGCAGCGACAACCTGGTGAGAcacatccgcatccac acaggccagaagcccttccagtgccgcatctgcatg AGAaacttcagcCG 206

- 98 046157

AGAGGAT AACTT GC AC ACTcacatccgcacccacacaggcg aaaagcccttcgcctgcgacatctgtggaagaaagtttgccCGGAGCGA TGAACTTGTCCGAcataccaagatccacttgcggcagaaggaccgc ccttacgcttgcccagtggagtcctgtgatcgccgcttctccCAATCAGG GAATCTGACTGAGcacatccg catccacacaggccagaagcccttc cagtgccgcatctgcatgAGAaacttcagcACAAGTGGACATCT GGTACGCcacatccgcacccacacaggcgaaaagcccttcgcctgcgac atctgtggaagaaagtttgccC AGA AT AGT AC CC T GAC CGA A cataccaagatccacttgcggcagaaggacaag 48 (encoding) ATGGCCGCAGATCACCTGATGCTGGCTGAAGGCT ACAGACTGGTGCAGCGGCCTCCATCTGCCGCTGC CGCCCACGGCCCCCACGCCCTGAGAACACTGCCC CCCTACGCCGGCCCTGGTCTTGATAGCGGACTCA GACCTAGAGGCGCCCCTCTGGGCCCTCCACCTCC AAGACAGCCTGGAGCCCTGGCCTACGGCGCCTTC GGCCCTCCTTCTAGCTTCCAGCCCTTCCCCGCCGT GCC TCCTCCAGCTGCTGGCATCGCCCACCTGCAG CCTGTGGCCACCCCTTACCCCGGAAGAGCCGCCG CCCCTCCAAACGCCCCTGGCGGACCTCCTGGCCC CCAGCCTGCTCCAAGCGCCGCTGCCCCTCCACCT CCTGCTCATGCCCTGGGCGGCATGGACGCCGAGC TGATCGACGAGGAAGCCCTGACCAGCCTGGAAC TGGAACTGGGCCTGCACAGAGTGCGGGAACTGC CTGAGCTGTTCC TGGGACAGAGCGAGTTCGACTG CTTCAGCGACCTGGGCAGCGCCCCTCCTGCCGGC TCTGTGTCCTGCGGCGGCAGCGGCGGCGGAAGC GGCgccgaccacctgatgctcgccgagggctaccgcctggtgcagaggcc gccgtccgccgccgccgcccatggccctcatgcgctccggactctgccgccgt acgcgggcccgggcctggacagtg ggctgaggccgcggggggctccgctgg ggccgccgccgccccgccaacccggggccctggcgtacggggccttcgggc cgccgtcctccttccagccctttccggccgtgcctccgccggccgcgggcatcg cgcacctgcagcctgtggcgacgcctaccccggccgcgcggccgcgcccc ccaacgctccgg gaggccccccgggcccgcagccggccccaagcgccgca gccccgccgccgcccgcgcacgccctgggcggcatggacgccgaactcatc gacgaggaggcgctgacgtcgctggagctggagctggggctgcaccgcgtgc gcgagctgcccgagctgttcctgggccagagcgagttcgactgcttctcggactt c gcatccacacaggccagaagcccttccagtgccgcatctgcatgAGAaact tcagcCGAGAGGATAACTTGCACACTcacatccgcacccac acaggcgaaaagcccttcgcctgcgacatctgtggaagaaagtttgccCGG AGCGATGAACTTGTCCGAcataccaagatccacttgcggcaga aggaccgcccttacgcttgcccagtggagtcc tgtgatcgccgcttctccCAA TCAGGGAATCTGACTGAGcacatccgcatccacacaggccag aagcccttccagtgccgcatctgcatgAGAaacttcagcACAAGTGG ACATCTGGTACGCcacatccgcacccacacaggcgaaaagcccttc gcctgcgacatctgtggaagaaagtttgccC AGA AT AGT ACC C TG ACCGAAc ataccaagatccacttgcggcagaaggacaag 208

- 99 046157

49 (encoding) atggagctggaattggatgctggtgaccaagacctgctggccttcctgctagag gaaagtggagatttggggacggcacccgatgaggccgtgagggccccactgg actgggcgctgccgctttctgaggtGccgagcgactgggaagtagatgatttgc tgtgctccctgctgagtcccccagcgtcgttgaacattctcagctcc tccaacccc tgccttgtccaccatgaccacacctactccctcccacgggaaactgtctctatgga tctagagagtgagagctgtagaaaagaggggacccagatgactccacagcata tggagggagctggcagagcaggagattgctaggctagtactgacagatgaggag aagagtctattggagaaggaggggcttattctgcctgagacacttcctctcactaa gacagaggaacaaattctgaaacgtgtgcggCTCGAACCAGGTGA AAAACCTTACAAATGTCCTGAATGTGGGAAATC ATTCAGTCGCAGCGACAACCTGGTGAGACATCA ACGCACCCATACAGGAGAAAAACCTTATAAATG TCCAGAATGTGGAAAGTCCTTCTCACGAGAGGAT AACTTGCACACTCATCAACGAACACATACTGGTG AAAACCATACAAGTGTCCCGAATGTGG TAAAA GTTTTAGCCGGAGCGATGAACTTGTCCGACACCA ACGAACCCATACAGGCGAGAAGCCTTACAAATG TCCCGAGTGTGGCAAGAGCTTCTCACAATCAGGG AATCTGACTGAGCATCAACGAACTCATACCGGG GAAAAACCTTACAAGTGTCCAGAGTGTGGGAAG AGCTTTTCCACAAGTGGACATCTGGTACGCCACC AGAGGACACATACAGGGGAGAAGCCCTACAAAT GCCCC GAATGCGGTAAAAGTTTCTCTCAGAATAG TACCCTGACCGAACACCAGCGAACACACACTGG GAAAAAAACGAGTgtgtaCgttgggggtttagagagcCgggtctt gaaatacacagcccagaatatggagcttcagaacaaagtacagcttctggagga acagaatttgtcccttctagatcaactgaggaaactccaggccatggtgattgaga tCt caaacaaaaccagcagcagcagcacctgcatcttggtcctGctagtctcctt ctgcctcctccttgtacctgctatgtactcctctgacacaagggggagcctgccag ctgagcatggagtgttgtcccgccagcttcgtgccctccccagtgaggaccctta ccagctggagctgcctgccctgcagtcagaagtg ccgaaagacagcacacacc agtggttggacggctcagactgtgtactccaggcccctggcaacacttcctgcct gctgcattacatgcctcaggctcccagtgcagagcctcccctggagtggccCtt ccctgacctcttctcagagcctctctgccgaggtcccatcctccccctgcaggca aatctcacaaggaagggaggatggcttcct actggtagcccctctgtcattttgca ggacagatactcaggc 212 50 (encoding) atggagctggaattggatgctggtgaccaagacctgctggccttcctgctagag gaaagtggagatttggggacggcacccgatgaggccgtgagggccccactgg actgggcgctgccgctttctgaggtGccgagcgactgggaagtagatgatttgc tgtgctccctgctgagtcccccagcgtcgttgaacattctcagctcc tccaacccc tgccttgtccaccatgaccacacctactccctcccacgggaaactgtctctatgga tctagagagtgagagctgtagaaaagaggggacccagatgactccacagcata tggagggagctggcagagcaggagattgctaggctagtactgacagatgaggag aagagtctattggagaaggaggggcttattctgcctgagacacttcctctcactaa gacagaggaacaaattctgaaacgtgtgcggcgcccttacgcttgcccagtgga gtcctgtgatcgccgcttctccCGCAGCGACAACCTGGTGAG AcacatccgcatccacacaggccagaagcccttccagtgccgcatctgcatgA GAaacttcagcCGAGAGGATAACTTGCACACTcacatccg cacccacacaggcgaaaagccct tcgcctgcgacatctgtggaagaaagtttgc cCGGAGCGATGAACTTGTCCGAcataccaagatccacttgcg 216

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gcagaaggaccgcccttacgcttgcccagtggagtcctgtgatcgccgcttctcc S A AT C AGGGA AT C T GACTGAGcacatccgcatccacacagg ccagaagcccttccagtgccgcatctgcatgAGAaacttcagcACAAGT GGACATCTGGTACGCcacatccgcacccacacaggcgaaaagcc cttcgcctgc gacatctgtggaagaaagtttgccCAGAATAGTACCC TGACCGAAcataccaagatccacttgcggcagaaggacgtgtaCgttg ggggtttagagagcCgggtcttgaaatacacagcccagaatatggagcttcag aacaaagtacagcttctggaggaacagaatttgtcccttctagatcaactgagga aactccaggccatggtgattg agatCtcaaacaaaaccagcagcagcagcacc tgcatcttggtcctGctagtctccttctgcctcctccttgtacctgctatgtactcctc tgacacaagggggagcctgccagctgagcatggagtgttgtcccgccagcttc gtgccctccccagtgaggacccttaccagctggagctgcctgccctgcagtcag a agtgccgaaagacagcacacaccagtggttggacggctcagactgtgtactcc aggcccctggcaacacttcctgcctgctgcattacatgcctcaggctcccagtgc agagcctcccctggagtggccCttccctgacctcttctcagagcctctctgccga ggtcccatcctccccctgcaggcaaatctcacaaggaagggaggatgg cttcct actggtagcccctctgtcattttgcaggacagatactcaggc 51 (encoding) atggagctggaattggatgctggtgaccaagacctgctggccttcctgctagag gaaagtggagatttggggacggcacccgatgaggccgtgagggccccactgg actgggcgctgccgctttctgaggtGccgagcgactgggaagtagatgatttgc tgtgctccctgctgagtcccccagcgtcgttgaacattctcagctcc tccaacccc tgccttgtccaccatgaccacacctactccctcccacgggaaactgtctctatgga tctagagagtgagagctgtagaaaagaggggacccagatgactccacagcata tggagggagctggcagagcaggagattgctaggctagtactgacagatgaggag aagagtctattggagaaggaggggcttattctgcctgagacacttcctctcactaa gacagaggaacaaattctgaaacgtgtgcggcgcccttacgcttgcccagtgga gtcctgtgatcgccgcttctccCGCTCAGACAACCTCGTTCGA cacatccgcatccacacaggccagaagcccttccagtgccgcatctgcatgAG AaacttcagcCACCGGACTACACTCACGAACcacatccgca cccacacaggcgaaaagccctt cgcctgcgacatctgtggaagaaagtttgcc AGAGAAGAC AATCTCC AT ACT cataccaagatccacttgcg gcagaaggaccgcccttacgcttgcccagtggagtcctgtgatcgccgcttctcc AC C AGC C ATTS TC T C AC T GA Acacatccgcatccacacagg ccagaagcccttccagtgccgcatctgcatgA GAaacttcagcCAGTCT AGC T C ACTGGT GAGGcacatccgcacccacacaggcgaaaagc ccttcgcctgcgacatctgtggaagaaagtttgccAGGGAGGATAAC CTGCATACGcataccaagatccacttgcggcagaaggacgtgtaCgtt gggggtttagagagcCgggtcttgaaatacacagcccagaatatggag cttca gaacaaagtacagcttctggaggaacagaatttgtcccttctagatcaactgagg aaactccaggccatggtgattgagatCtcaaacaaaaccagcagcagcagcac ctgcatcttggtcctGctagtctccttctgcctcctccttgtacctgctatgtactcct ctgacacaagggggagcctgccagctgagcatggag tgttgtcccgccagctt cgtgccctccccagtgaggacccttaccagctggagctgcctgccctgcagtca gaagtgccgaaagacagcacacaccagtggttggacggctcagactgtgtact ccaggcccctggcaacacttcctgcctgctgcattacatgcctcaggctcccagt gcagagcctcccctggagtggccCt tccctgacctcttctcagagcctctctgcc gaggtcccatcctccccctgcaggcaaatctcacaaggaagggaggatggcttc ctactggtagcccctctgtcattttgcaggacagatactcaggc 218 52 (encoding) atggagctggaattggatgctggtgaccaagacctgctggccttcctgctagag gaaagtggagattggggacggcacccgatgaggccgtgagggccccactgg 220

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actgggcgctgccgctttctgaggtGccgagcgactgggaagtagatgatttgc tgtgctccctgctgagtcccccagcgtcgttgaacattctcagctcctccaacccc tgccttgtccaccatgaccacacctactccctcccacgggaaactgtctctatgga tctagagagtgagagctgtagaaaagaggggacccagatgact ccacagcata tggaggagctggcagagcaggagattgctaggctagtactgacagatgaggag aagagtctattggagaaggaggggcttattctgcctgagacacttcctctcactaa gacagaggaacaaattctgaaacgtgtgcggCTCGAACCAGGTGA AAAACCTTACAAATGTCCTGAATGTGGGAAATC ATTCAGTCGCAGCGACAACCTGGTGAGACAT CA ACGCACCCATACAGGAGAAAAACCTTATAAATG TCCAGAATGTGGAAAGTCCTTCACGAGAGGAT AACTTGCACACTCATCAACGAACACATACTGGTG AAAAACCATACAAGTGTCCCGAATGTGGTAAAA GTTTTAGCCGGAGCGATGAACTTGTCCGACACCA ACGAACCCATACAGGCGAGAAGCCTTACAAATG TCCCGAGTGTGGCAAGAGCTTCTCACAATCAGGG AATCTGACTGAG CATCAACGAACTCATACCGGG GAAAAACCTTACAAGTGTCCAGAGTGTGGGAAG AGCTTTTCCACAAGTGGACATCTGGTACGCCACC AGAGGACACATACAGGGGAGAAGCCCTACAAAT GCCCCGAATGCGGTAAAAGTTTCTCTCAGAATAG TACCCTGACCGAACACCAGCGAACACACACTGG GAAAAAAACGAGTgtgtaCgttgggggtttagagagcCgggt ctt gaaatacacagcccagaatatggagcttcagaacaaagtacagcttctggagga acagaatttgtcccttctagatcaactgaggaaactccaggccatggtgattgaga tatca 53 (encoding) atggagctggaattggatgctggtgaccaagacctgctggccttcctgctagag gaaagtggagatttggggacggcacccgatgaggccgtgagggccccactgg actgggcgctgccgctttctgaggtGccgagcgactgggaagtagatgatttgc tgtgctccctgctgagtcccccagcgtcgttgaacattctcagctcc tccaacccc tgccttgtccaccatgaccacacctactccctcccacgggaaactgtctctatgga tctagagagtgagagctgtagaaaagaggggacccagatgactccacagcata tggagggagctggcagagcaggagattgctaggctagtactgacagatgaggag aagagtctattggagaaggaggggcttattctgcctgagacacttcctctcactaa gacagaggaacaaattctgaaacgtgtgcggcgcccttacgcttgcccagtgga gtcctgtgatcgccgcttctccCGCAGCGACAACCTGGTGAG AcacatccgcatccacacaggccagaagcccttccagtgccgcatctgcatgA GAaacttcagcCGAGAGGATAACTTGCACACTcacatccg cacccacacaggcgaaaagccct tcgcctgcgacatctgtggaagaaagtttgc cCGGAGCGATGAACTTGTCCGAcataccaagatccacttgcg gcagaaggaccgcccttacgcttgcccagtggagtcctgtgatcgccgcttctcc C A AT C AGGGA AT C T GACTGAGcacatccgcatccacacagg ccagaagcccttccagtgccgcatctgcatgAGAaact tcagcACAAGT GGACATCTGGTACGCcacatccgcacccacacaggcgaaaagcc cttcgcctgcgacatctgtggaagaaagtttgccCAGAATAGTACCC TGACCGAAcataccaagatccacttgcggcagaaggacgtgtaCgttg ggggtttagagagcCgggtcttgaaatacacagcccagaatatggagcttcag a acaaagtacagcttctggaggaacagaatttgtcccttctagatcaactgagga aactccaggccatggtgattgagatCtca 222

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Table 8

Nucleic acid sequences for exemplary miRNA and miRNA binding sites

Description Subsequence SEQ ID NO Ml binding site aaagagaccggttcactgtgacagtaaaagagaccggttcactgtgagaatgaaagag accggttcactgtgatcggaaaagagaccggttcactgtgagcggccttgaaacccagc agacaatgtagctcagtagaaacccagcagacaatgtagctgaatggaaacccagcag acaatgtagcttcggagaaacccagcagacaat gtagct 7 miR128 sequence UCACAGUGAACCGGUCUCUUU 8 miR128 binding site AAAGAGACCGGTTCACTGTGA 9 miR221 sequence AGCUACAUUGUCUGCUGGGUUUC 10 miR221 binding site GAAACCCAGCAGACAATGTAGCT eleven miR222 sequence AGCUACAUCUGGCUACUGGGUCU 12 miR222 binding site AGACCCAGTAGCCAGATGTAGCT 13 M2 binding site GAAACCCAGCAGACAATGTAGCTAGACCCAGTAGCC AGATGTAGCTAAAGAGACCGGTTCACTGTGA 14 M3 binding site GAAACCCAGCAGACAATGTAGCTAGACCCAGTAGCC AGATGTAGCTAAAGAGACCGGTTCACTGTGAGAAAC CCAGCAGACAATGTAGCTAGACCCAGTAGCCAGATG TAGCTAAAGAGACCGGTTCACTGTGA 15

Table 9

Various types of zinc finger structures and illustrative zinc finger proteins for eTF production

Type name ZF SEQ ID NO ZF structure (each x can independently be any residue) Exemplary proteins that can serve as a protein platform for an eTF or DNA binding domain of an eTF disclosed herein Zinc fingers C2H2type (ZNF) 136 S-x-S-x-N-x-N KLF4, KLF5, EGR3, ZFP637, SLUG, ZNF750, ZNF281, ΖΠ.Η.89, GLIS1, GLIS3 Proteins type Ring fingers (RNF) 137 S-x-S-x-S-x-N-xxx-S-xS-x-S-x-S MDM2, BRCA1, ZNF 179 Proteins type "PHD fingers" (PHF) 138 S-x-S-x-S-x-S-xxx-N-xS-x-S-x-S KDM2A, PHF1, ING1

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Containing the LIM domain 139 S-x-S-x-N-x-S-x-S-x-S-x- C-x-(C,H,D) ZNF185, LIMK1, PXN Nuclear hormone receptors (NR) 140 S-x-S-x-S-x-S-xxx-S-x-Sx-S-x-S VDR, ESRI, NR4A1 "Zinc fingers" CCSN type (ZC3H) 141 S-x-S-x-S-x-N RC3H1, HELZ, MBNL1, ZFP36, ZFP36L1 Zinc fingers FYVE type (ZFYVE) 140 S-x-S-x-S-x-S-xxx-S-x-Sx-S-x-S EEA1, HGS, PIKFYVE "Zinc fingers" CCHC type (ZCCHC) 142 S-x-S-x-N-x-S CNH.H.,SF1,LIN28A Zinc fingers DHHC type (ZDHHC) 143 S-x-S-x-N-x-S-xxx-S-xS-x-N-x-S ZDHHC2, ZDHHC8, ZDHHC9 “Zinc fingers” MYNDTHna(ZMYND) 144 S-x-S-x-S-x-S-xxx-S-x-Sx-N-x-S PDCD2, RUNX1T1, SMYD2, SMYD1 “Zinc fingers” RANBP2THna(ZRANB) 145 S-x-S-x-S-x-S YAF2, SHARPIN, EWSR1 "Zinc fingers" ZZ type (ZZZ) 145 S-x-S-x-S-x-S HERC2, NBR1, CREBH.H. “Zinc fingers” C2HC type (ZC2HC) 142 S-x-S-x-N-x-S IKBKG, L3MBTL1, ZNF746 Containing GATA zinc finger domain (GATAD) 145 S-x-S-x-S-x-S GATA4, GATA6, MT Al ZF class homeoboxes and pseudogenes 136 S-x-S-x-N-x-N ADNP, ZEB1, ZHX1 Containing domain TNAR (TNAR) 141 S-x-S-x-S-x-N THAP1, THAP4, THAP11

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"Zinc fingers" CХХСtype (СХХС) 140 S-x-S-x-S-x-S-xxx-S-x-Sx-S-x-S СХХС1, СХХС5, MBD1, DNMT1 Zinc fingers SWIM type (ZSWIM) 141 S-x-S-x-S-x-N MARZK1, ZSWIM5, ZSWIM6 "Zinc fingers" AN1 type (ZFAND) 146 S-x-S-x-S-x-S-xxx-S-xN-x-N-x-S ZFAND3, ZFAND6, IGHMH.H.2 “Zinc fingers” ZСххType (Z3CXXC) 142 S-x-S-x-N-x-S ZAR1, RTP1, RTP4 Zinc fingers CW type (ZCW) 145 S-x-S-x-S-x-S MORC1, ZCWPW1, KDM1B Zinc fingers GRF type (ZGRF) 145 S-x-S-x-S-x-S TTF2, NEIL3, TORZ A "Zinc fingers" MIZ type (ZMIZ) 142 S-x-S-x-N-x-S PIAS1, PIAS3, PIAS4 Zinc pins BED type (ZBED) 136 S-x-S-x-N-x-N ZBED1, ZBED4, ZBED6 Zinc fingers HIT type (ZNHIT) 144 S-x-S-x-S-x-S-xxx-S-x-Sx-N-x-S ZNHIT3, DDX59, INO80B Zinc fingers MYM type (ZMYM) 145 S-x-S-x-S-x-S ZMYM2, ZMYM3, ZMYM4 "Zinc fingers" matrin type (ZMAT) 136 S-x-S-x-N-x-N ZNF638, ZMAT1, ZMAT3, ZMAT5 “Zinc fingers” C2H2C type 136 S-x-S-x-N-x-N MYT1,MYT1L, ST18 Zinc fingers DBF type (ZDBF) 136 S-x-S-x-N-x-N DBF4, DBF4B, ZDBF2 PARP-type zinc fingers 142 S-x-S-x-N-x-S LIG3, PARP1

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Table 10 Amino acid sequences of exemplary zinc finger DNA binding domains

DBD/target site Subsequence SEQID NO eZF LEPGEKP - [YKCPECGKSFS X HQRTH TGEKP]n YKCPECGKSFS X HQRTH - TGKKTS where η is an integer from 1 to 15 and each X is a recognition sequence capable of binding to 3 bp. target sequence 147 Target site Z1 RSDNLVR x REDNLHT x RSDELVR x QSGNLTE x TSGHLVR x QNSTLTE, where each x is a linker containing 1-50 amino acid residues 148 Target site Z13 RSDNLVR x HRTTLTN x REDNLHT x TSHSLTE x QSSSLVR x REDNLHT, where each x is a linker containing 1-50 amino acid residues 149 Target site Z14 DPGALVR x RSDNLVR x QSGDLRR x THLDLIR x TSGNLVR x RSDNLVR, where each x is a linker containing 1-50 amino acid residues 150 Target site Z15 RRDELNV x RSDHLTN x RSDDLVR x RSDNLVR x HRTTLTN x REDNLHT x TSHSLTE x QSSSLVR x REDNLHT, where each x is a linker containing 1-50 amino acid residues 151

Table 11

Amino acid sequences for exemplary zinc finger recognition sequences disclosed herein

Subsequence SEQ ID NO RSDNLVR 152 REDNLHT 153 RSDELVR 154 QSGNLTE 155 TSGHLVR 156 QNSTLTE 157 DPGALVR 158 HRTTLTN 159 QSGDLRR 160 TSHSLTE 161 THLDLIR 162 QSSSLVR 163 TSGNLVR 164 RRDELNV 165 RSDDLVR 166 RSDHLTN 167

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Table 12

Other nucleotide and amino acid sequences disclosed herein

Description Subsequence SEQ ID NO EGR1 NLS domain LH<PSRMRI<YPNRPSI< 168 SV40 NLS PKKKRKV 169 Nucleoplasma hNLS KRPAATKKAGQAKKKK 170 Tag ON YPYDVPDYA 171 spA (synthetic polyA) AATAAAAGATCTTTATTTTCATTAGATCTGTGTGTTGG TTTTTTGTGTGCGGACCGCACGTG 16 hGH (polyA human growth hormone) GGGTGGCATCCCTGTGACCCCTCCCCAGTGCCTCTCCT GGCCCTGGAAGTTGCCACTCCAGTGCCCACCAGCCTT GTCCTAATAAAATTAAGTTGCATCATTTTGTCTGACTA GGTGTCCTTCTATAATATTATGGGGTGGAGGGGGGTG GTATGGAGCAAGGGGCAAGTTGGGAAGACAACCTGT AGGGCCTGCGGGGTCTATTGGGAACCAAGCTGGAGTG CAGTGGCACAATCTTGGC TCACTGCAATCTCCGCCTCC TGGGTTCAAGCGATTCTCCTGCCTCAGCCTCCCGAGTT GTTGGGATTCCAGGCATGCATGACCAGGCTCAGCTAA TTTTTGTTTTTTTGGTAGAGACGGGGTTTCACCATATT GGCCAGGCTGGTCTCCAACTCCTAATCTCAGGTGATCT ACCCACCTTGGCCTCCCAAATTGCTGGGATTACAGGC GTGAACCACTGCTCCCTTCCCTGTCCTT 17 SCN1A protein MEQTVLVPPGPDSFNFFTRESLAAIERRIAEEKAKNPKPD I<I<DDDENGPKPNSDLEAGI<NLPFIYGDIPPEMVSEPLED LDPYYINKKTFIVLNKGKAIFRFSATSALYILTPFNPLRKI AIKILVHSLFSMLIMCTILTNCVFMTMSNPPDWTKNVEY TFTGIYTFESLIKIIARGFCLEDFTFLRDPWNWLDFTVITF AYVT EFVDLGNVSALRTFRVLRALKTISVIPGLKTIVGAL IQSVKKLSDVMILTVFCLSVFALIGLQLFMGNLRNKCIQ WPPTNASLEEHSIEKNITVNYNGTLINETVFEFDWKSYIQ DSRYHYFLEGFLDALLCGNSSDAGQCPEGYMCVKAGRN PNYGYTSFDTFSWAFLSLFRLMTQDFWENLYQLTLRAA GKTYMIFFVLVIFLGSF YLINLILAVVAMAYEEQNQATLE EAEQKEAEFQQMIEQLKKQQEAAQQAATATASEHSREP S AAGRLSDS S SEASKLS SKS AKERRNRRKKRKQKEQSGG EEKDEDEFQKSESEDSIRRKGFRF SIEGNRLTYEKRYS SP HQSLLSIRGSLFSPRRNSRTSLFSFRGRAKDVGSENDFAD DEHSTFEDNESRR DSLFVPRRHGERRNSNLSQTSRSSRM LAVFPANGKMHSTVDCNGVVSLVGGPSVPTSPVGQLLP EVIIDKPATDDNGTTTETEMRKRRSSSFHVSMDFLEDPSQ RQRAMSIASILTNTVEELEESRQKCPPCWYKFSNIFLIWD CSPYWLKVKHVVNLVVMDPFVDLAITICIVLNTLFMAM EHYPMTDHFNNVLTVGN LVFTGIFTAEMFLKIIAMDPYY YFQEGWNIFDGFIVTLSLVELGLANVEGLSVLRSFRLLRV FKLAKSWPTLNMLIKIIGNSVGALGNLTLVLAIIVFIFAVV GMQLFGKSYKDCVCKIASDCQLPRWHMNDFFHSFLIVF RVLCGEWIETMWDCMEVAGQAMCLTVFMMVMVIGNL VVLNLFLALLLSSFSADNLAATDDDNEMNNLQIAV DRM HKGVAYVKRKIYEFIQQSFIRKQKILDEIKPLDDLNNKKD SCMSNHTAEIGKDLDYLKDVNGTTSGIGTGSSVEKYIIDE SDYMSFINNPSLTVTVPIAVGESDFENLNTEDFSSESDLEE 172

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SKEKLNES S S S SEGSTVDIGAPVEEQP VVEPEETLEPEACF TEGCVQRFKCCQINVEEGRGKQWWNLRRTCFRIVEHNW FETFIVFMILLSSGALAFEDIYIDQRI<TH<TMLEYADI<VFT YIFILEMLLKWVAYGYQTYFTNAWCWLDFLIVDVSLVS LTANALGYSELGAIKSLRTLRALRPLRALSRFEGMRVV V NALLGAIPSIMNVLLVCLIFWLIFSIMGVNLFAGKFYHCI NTTTGDRFDIEDVNNHTDCLI<LIERNETARWI<NVI<VNF DNVGFGYLSLLQVATFKGWMDIMYAAVDSRNVELQPK YEESLYMYLYFVIFIIFGSFFTLNLFIGVIIDNFNQQKKKFG GQDIFMTEEQKKYYNAMKKLGSKKPQKPIPR PGNKFQG MVFDFVTRQVFDISIMILICLNMVTMMVETDDQSEYVTT ILSRINLVFIVLFTGECVLKLISLRHYYFTIGWNIFDFVVVI LSIVGMFLAELIEKYFVSPTLFRVIRLARIGRILRLIKGAK GIRTLLFALMMSLPALFNIGLLLFLVMFIYAIFGMSNFAY VKREVGIDDMFNFETFGNSMICLFQITTSAGWDG LLAPIL NSKPPDCDPNKVNPGSSVKGDCGNPSVGIFFFVSYIIISFL VVVNMYIAVILENF S VATEES AEPLSEDDFEMF YEVWEK FDPDATQFMEFEKLSQFAAALEPPLNLPQPNKLQLIAMD LPMVSGDRIHCLDILFAFTKRVLGESGEMDALRIQMEER FMASNPSKVSYQPITTTLKRKQEEVSAVIIQRA YRRHLLK RTVKQASFTYNKNKIKGGANLLIKEDMIIDRINENSITEK TTDLTMSTAACPPSYDRVTKPIVEKHEQEGKDEKAKGK dCAS protein KRNYILGLAIGITSVGYGIIDYETRDVIDAGVRLFKEANV ENNEGRRSKRGARRLKRRRRHRIQRVKKLLFDYNLTTD HSELSGINPYEARVKGLSQKLSEEEF S A ALLHLAKRRG V HNVNEVEEDTGNELSTKEQISRNSKALEEKYVAELQLER LKKDGEVRGSINRFKTSDYVKEAKQLLKVQKAY HQLDQ SFIDTYIDLLETRRTYYEGPGEGSPFGWKDIKEWYEMLM GHCTYFPEELRSVKYAYNADLYNALNDLNNLVITRDEN EKLEYYEKFQIIENVFKQKKKPTLKQIAKEILVNEEDIKG YRVTSTGKPEFTNLKVYHDIKDITARKEIIENAELLDQIA KILTIYQSSEDIQEELTNLNSELTQEE IEQISNLKGYTGTH NLSLKAINLILDELWHTNDNQIAIFNRLKLVPKKVDLSQQ KEIPTTLVDDFILSPVVKRSFIQSIKVINAIIKKYGLPNDIIIE LAREKNSKDAQKMINEMQKRNRQTNERIEEIIRTTGKEN AKYLIEKIKLHDMQEGKCLYSLEAIPLEDLLNNPFNYEV DHIIPRS VSFDNSFNNK VLVK QEE ASKKGNRTPFQ YLS S S DSKISYETFKKHILNLAKGKGRISKTKKEYLLEERDINRF SVQKDFINRNLVDTRYATRGLMNLLRSYFRVNNLDVKV KSINGGFTSFLRRKWKFKKERNKGYKHHAEDALIIANAD FIFKEWKKLDKAKKVMENQMFEEKQAESMPEIETEQEY KEIFITPHQIKHIK DFKDYKYSHRVDKKPNRELINDTLYS TRKDDKGNTLIVNNLNGLYDKDNDKLKKLINKSPEKLL MYHHDPQTYQKLKLIMEQYGDEKNPLYKYYEETGNYL TKYSKKDNGPVIKKIKYYGNKLNAHLDITDDYPNSRNK VVKLSLKPYRFDVYLDNGVYKFVTVKNLDVIKKEN YYE VNSKCYEEAKKLKKISNQAEFIASFYNNDLIKINGELYRV IGVNNDLLNRIEVNMIDITYREYLENMNDKRPPRIIKTIAS KTQSIKKYSTDILGNLYEVKSKKHPQIIKKG 173

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Design dCAS9-VP64 MAPKKKRKVGIHGVPAAKRNYILGLAIGITSVGYGIIDYE TRDVIDAGVRLFKEANVENNEGRRSKRGARRLKRRRRH RIQRVKKLLFDYNLLTDHSELSGINPYEARVKGLSQKLSE EEFSAALLHLAKRRGVHNVNEVEEDTGNELSTKEQISRN SKALEEKYVAELQLERLKKDGEVRGSINRFKTSDYV KEA KQLLKVQKAYHQLDQSFIDTYIDLLETRRTYYEGPGEGS PFGWKDIKEWYEMLMGHCTYFPEELRSVKYAYNADLY NALNDLNNLVITRDENEKLEYYEKFQIIENVFKQKKKPT LKQIAKEILVNEEDIKGYRVTSTGKPEFTNLKVYHDIKDI TARKEIIENAELLDQIAKILTIYQSSED IQEELTNLNSELTQ EEIEQISNLKGYTGTHNLSLKAINLILDELWHTNDNQIAIF NRLKLVPKKVDLSQQKEIPTTLVDDFILSPVVKRSFIQSIK VINAIIKKYGLPNDIIIELAREKNSKDAQKMINEMQKRNR QTNERIEEIIRTTGKENAKYLIEKIKLHDMQEGKCLYSLE AIPLEDLLNNPFNYEVDHIIPR SVSFDNSFNNKVLVKQEE ASKKGNRTPFQ YLS S SD SKIS YETFKKHILNL AKGKGRIS I<TI<I<EYLLEERDINRF S VQKDFINRNLVDTRYATRGLMN LLRSYFRVNNLDVKVKSINGGFTSFLRRKWKFKKERNK GYKHHAEDALIIANADFIFKEWKKLDKAKK VMENQMFE EKQAESMPEIETE QEYKEIFITPHQIKHIKDFKDYKYSHR VDKKPNRELINDTLYSTRKDDKGNTLIVNNLNGLYDKD NDKLKKLINKSPEKLLMYHHDPQTYQKLKLIMEQYGDE KNPLYKYYEETGNYLTKYSKKDNGPVIKKIKYYGNKLN AHLDITTDDYPNSRNKVVKLSLKPYRFDVYLDNGV YKFV TVKNLDVIKKENYYEVNSKCYEEAKKLKKISNQAEFIAS FYNNDLIKINGELYRVIGVNNDLLNRIEVNMIDITYREYL ENMNDKRPPRIIKTIASKTQSIKKYSTDILGNLYEVKSKK HPQIIKKGKRPAATKKAGQAKKKKGSYPYDVPDYALED ALDDFDLDMLGSDALDDFDLDML GSDALDDFDLDMLG SDALDDFDLDML 174 WT protein EGR3 (human) MTGKLAEKLPVTMSSLLNQLPDNLYPEEIPSALNLFSGSS DSVVHYNQMATENVMDIGLTNEKPNPELSYSGSFQPAP GNKTVTYLGKFAFDSPSNWCQDNIISLMSAGILGVPPAS GALSTQTSTASMVQPPQGDVEAMYPALPPYSNCGDLYS EPVSFHDPQGNPGLAYSPQDYQSAKPALDSNLFPMIPDY NLYH HPNDMGSIPEHKPFQGMDPIRVNPPPITPLETIKAF KDKQIHPGFGSLPQPPLTLKPIRPRKYPNRPSKTPLHERPH ACPAEGCDRRFSRSDELTRHLRIHTGHKPFQCRICMRSFS RSDHLTTHIRTHTGEKPFACEFCGRKFARSDERKRHAKI HLKQKEKKAEKGGAPSASSAPVSLAPVVTTCA 175 WT protein EGR1 (human) maaakaemqlmsplqisdpfgsfphsptmdnypkleemmllsngapqflgaagap egsnssssssgggggggggsnsssssstfnpqadtgeqpyehltaesfpdislnnek vlvetsypsqttrlppitytgrfslepapnsgntlwpeplfslvsglvsmtnppassssap spaassasqspplscavps ndsspiysaaptfptpntdifpepqsqafpgsagtalqy pppaypaakggfqvpmipdylfpqqqgdlglgtpdqkpfqglesrtqqpsltplstik afatqsgsqdlkalntsyqsqlikpsrmrkypnrpsktppherpyacpvescdrrfsrs deltrhirihtgqkpfqcricmrnfsrsdh ltthirthtgekpfacdicgrkfarsderkrht kihlrqkdkkadksvvas sats si s syp spvatsyp spvttsyp sp attsyp spvptsfs spgsstypspvhsgfpspsvattyssvppafpaqvssfpssavtnsfsastglsdmtatf sprtieic 176

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Linker GGSGGGSG 177 Linker GGSGGGSGGGSGGGGSG 178 Linker [GGGS]n 179 Linker [GGGGS]n 180 Linker [GGSG]n 181 Recognition site for WT EGR1 or EGR3 GCG(T/G)GGGCG 182 sgRNA scaffold GTTTTAGTACTCTGGAAACAGAATCTACTAAAACAAG GCAAAATGCCGTGTTTATCTCGTCAACTTGTTGGCGAG A 183 Coding sequence of human CREB3 atggagctggaattggatgctggtgaccaagacctgctggccttcctgctagaggaaagt ggagatttggggacggcacccgatgaggccgtgagggccccactggactgggcgctg ccgctttctgaggtGccgagcgactgggaagtagatgatttgctgtgctccctgctgagtc ccccagcgtcgttgaacattctcagct cctccaacccctgccttgtccaccatgaccacac ctactccctcccacgggaaactgtctctatggatctagagagtgagagctgtagaaaaga ggggacccagatgactccacagcatatggaggagctggcagagcaggagattgctagg ctagtactgacagatgaggagaagagtctattggagaaggaggggcttattctgcctgag acacttcctctcactaaga cagaggaacaaattctgaaacgtgtgcggaggaagattcga aataaaagatctgctcaagagagccgcaggaaaaagaaggtgtaCgttgggggtttaga gagcCgggtcttgaaatacacagcccagaatatggagcttcagaacaaagtacagcttct ggaggaacagaatttgtcccttctagatcaactgaggaaactccaggcca tggtgattgag atCtcaaacaaaaccagcagcagcagcacctgcatcttggtcctGctagtctccttctgc ctcctccttgtacctgctatgtactcctctgacacaagggggagcctgccagctgagcatg gagtgttgtcccgccagcttcgtgccctccccagtgaggacccttaccagctggagctgc ctgcc ctgcagtcagaagtgccgaaagacagcacacaccagtggttggacggctcaga ctgtgtactccaggcccctggcaacacttcctgcctgctgcattacatgcctcaggctccca gtgcagagcctcccctggagtggccCttccctgacctcttctcagagcctctctgccgag gtcccatcctccccctgcaggcaaatctcacaagga agggaggatggcttcctactggta gcccctctgtcattttgcaggacagatactcaggc 210 Sequence AA CREB3 person MELELDAGDQDLLAFLLEESGDLGTAPDEAVRAPLDWA LPLSEVP SDWE VDDLLC SLLSPP ASLNILS S SNPCLVHHD HTYSLPRETVSMDLESESCRKEGTQMTPQHMEELAEQEI ARLVLTDEEKSLLEKEGLILPETLPLTKTEEQILKRVRRKI RNKRSAQESRRKKKVYVGGLESRVLKYTAQNMELQNK VQLLEEQNLSLLDQLRKLQAMVIEISNKTS S S STCILVLL VSFCLLLVPAMYSSDTRGSLPAEHGVLSRQLRALPSEDP YQLELPALQSEVPKDSTHQWLDGSDCVLQAPGNTSCLL HYMPQAPSAEPPLEWPFPDLFSEPLCRGPILPLQANLTRK GGWLPTGSPSVILQDRYSG 211 Human C-terminal domain of CREB3 with transmembrane region VYVGGLESRVLKYTAQNMELQNKVQLLEEQNLSLLDQL RKLQAMVIEISNKTS S S STCILVLLVSFCLLLVP AMYS SD TRGSLPAEHGVLSRQLRALPSEDPYQLELPALQSEVPKD STHQWLDGSDCVLQAPGNTSCLLHYMPQAPSAEPPLEW PFPDLFSEPLCRGPILPLQANLTRKGGWLPTGSPSVILQ D RYSG 225 Human C-terminal VYVGGLESRVLKYTAQNMELQNKVQLLEEQNLSLLDQL RKLQAMVIEIS 226

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CREB3 domain without transmembrane region Coding sequence CREB3TRE atggagctggaattggatgctggtgaccaagacctgctggccttcctgctagaggaaagt ggagatttggggacggcacccgatgaggccgtgagggccccactggactgggcgctg ccgctttctgaggtGccgagcgactgggaagtagatgatttgctgtgctccctgctgagtc ccccagcgtcgttgaacattctcagct cctccaacccctgccttgtccaccatgaccacac ctactccctcccacgggaaactgtctctatggatctagagagtgagagctgtagaaaaga ggggacccagatgactccacagcatatggaggagctggcagagcaggagattgctagg ctagtactgacagatgaggagaagagtctattggagaaggaggggcttattctgcctgag acacttcctctcactaaga cagaggaacaaattctgaaacgtgtgcggCTTGAGCC CGGAGAGAAGCCGTACAAGTGCCCTGAGTGCGGCAA GTCTTTTAGCAGAAGAGACGAACTTAATGTCCACCAG CGAACGCATACTGGTGAAAAGCCCTATAAATGTCCTG AATGTGGGAAATCATTCTCCAGCCGCAGAACCTGTAG GGCTCACCAGCGAACACACCGGCGAAAACCATA CAAATGTCCAGAATGC GGGAAATCCTTTTCTCAGTCAT CCAACTTGGTGAGACATCAACGCACGCACACTGGAGA AAAGCCTTACAAATGCCCGGAATGTGGAAAGTCTTTT TCCCAATTGGCCCATTTGCGAGCCCATCAGAGGACTC ACACGGGCGAGAAACCTTACAAATGCCCGGAATGCGG GAAATCTTTTTCAACGAGTGGCAACCTCGTAAGACAC CAAAGAACGCATACAGGCGAAAAGCCATATAAGTGTC CTG AGTGTGGTAAATCATTCTCACACAGGACCACCCT GACAAATCACCAGCGCACGCACACCGGCAAGAAGAC AAGCgtgtaCgttgggggtttagagagcCgggtcttgaaatacacagcccagaatat ggagcttcagaacaaagtacagcttctggaggaacagaatttgtcccttctagatcaactg aggaaactccaggccatggtgattgagatC tcaaacaaaaccagcagcagcagcacct gcatcttggtcctGctagtctccttctgcctcctccttgtacctgctatgtactcctctgacac aagggggagcctgccagctgagcatggagtgttgtcccgccagcttcgtgccctcccca gtgaggacccttaccagctggagctgcctgccctgcagtcagaagtgcc gaaagacagc acacaccagtggttggacggctcagactgtgtactccaggcccctggcaacacttcctgc ctgctgcattacatgcctcaggctcccagtgcagagcctcccctggagtggccCttccct gacctcttctcagagcctctctgccgaggtcccatcctccccctgcaggcaaatctcacaa ggaagggaggatggcttcctact ggtagcccctctgtcattttgcaggacagatactcagg c 214 CREB3-TRE MELELDAGDQDLLAFLLEESGDLGTAPDEAVRAPLDWA LPLSEVP SDWEVDDLLC SLLSPP ASLNILS S SNPCLVHHD HTYSLPRETVSMDLESESCRKEGTQMTPQHMEELAEQEI ARLVLTDEEKSLLEKEGLILPETLPLTKTEEQILKRVRLEP GEKPYKCPECGKSFSRRDELNVHQRTHTGEKPYKCPEC GKS FS SRRTCRAHQRTHTGEKPYKCPECGKSF SQS SNLV RHQRTHTGEKPYKCPECGKSFSQLAHLRAHQRTHTGEK PYKCPECGKSFSTSGNLVRHQRTHTGEKPYKCPECGKSF SHRTTLTNHQRTHTTGKKTSVYVGGLESRVLKYTAQNME LQNK VQLLEEQNLSLLDQLRKLQ AM VIEISNKT S S S STCI LVLLVSFCLLLVPAMYSSDTRGSLPAEHGVLSRQLRALP SEDPYQLELPALQSEVPKDSTHQWLDGSDCVLQAPGNTS 215 CLLHYMPQAPSAEPPLEWPFPDLFSEPLCRGPILPLQANL TRKGGWLPTGSPSVILQDRYSG

Examples

The following examples are included to further describe certain aspects of the present disclosure and should not be used to limit the scope of the present invention.

Example 1.

Identification of target regions capable of activating SCN1A using SCN1A-specific transcriptional activators.

To identify regions of the genome capable of upregulating the expression of endogenous SCN1A, various engineered transcription factors (either zinc finger nucleases or gRNA/daCas9 constructs) have been developed that target different regions of the genome, as listed in Table 1. 4 and 13 above. For gRNA/daCas9 constructs, this gRNA had the same sequence as the target region because the gRNA was designed to target the complementary genomic strand. The dCas9 protein was the construct dCAS9-VP64 (SEQ ID NO: 174). HEK293 cell culture

- 111 046157 were prepared using standard methods and transfected (FugeneHD, Promega) with 3 μg of plasmid carrying the engineered transcription factor or control construct per well of a 6-well plate. Cells were transfected with plasmids expressing the constructs indicated in the table. 13 below. 48 hours after transfection, cells were harvested, RNA was isolated (Qiagen RNeasy Mini kit) and treated with DNase. RNA (3 μg) was reverse transcribed using OligoDT primers (Superscript IV, Invitrogen). cDNA samples were analyzed by qPCR using Phusion Polymerase (New England Biolabs) and SYBR Green I: (30 seconds at 98°C, 40 times [10 seconds at 98°C, 15 seconds at 66°C, 15 seconds at 72°C]). Primers against SCN1A (5'-TGTCTCGGCATTGAGAACATTC-3' (SEQ ID NO: 185); 5'ATTGGTGGGAGGCCATTGTAT-3' (SEQ ID NO: 186)) were used to quantify endogenous SCN1A transcript levels, and relative SCN1A expression levels were determined by the delta method -delta Ct with GAPDH as the reference gene (5'-ACCACAGTCCATGCCATCAC'-3' (SEQ ID NO: 187); 5'-TCCACCACCCTGTTGCTGTA-3' (SEQ ID NO: 188)). Data are presented as fold changes relative to the control condition.

The results are shown in Fig. 1 and in table. 13 below as the fold change in SCN1A transcription relative to control conditions (eg, EGFP-KASH reporter construct). In table Figure 13 shows values for constructs that resulted in at least a 1.5-fold increase in transcription compared to control conditions.

Table 13. Effect of different genomic target sites and corresponding eTFs on transcription. CON indicates the zinc finger design that was used in the experiment (see Table 1). For gRNA constructs, the target site and gRNA sequence are the same because the gRNA was designed to target the complementary strand of DNA.

Table13

CO N Initial position on chromosome 2 Target sequence Target site from Fig. 1 SEQ ID NO for the target site Average meaning 4 166149168 ctaggtcaagtgtaggag zl 18 5.12090848 0.0096628 28 166149165 GGTCAAGTGTAGGAGACA z8 25 1.52068773 0.62403349 3 166128025 taggtaccatagtgag zl3 thirty 25.4730028 0.14942042 29 166127991 gaggatactgcagaggtc zl4 31 8.6766618 0.16432794 166149176 aaggctgtctaggtcaagtgt g9 35 1.36378425 0.18753821 166149118 tgttcctccagattaacactt gio 36 1.63040825 0.46710683 166128002 gatgaagccgagaggatactg gH 37 18.7579211 0.13148732 166128037 gctgatttgtattaggtacca gl2 38 22.4892633 0.09291316 166177299 AGAAAGCTGATACAGATACA A gl5 39 1.7542842 0.34519408 166178880 ggtacgggcaaagatttcttg gl7 40 1.36801947 0.48762102 166177299 AGAAAGCTGATACAGATACA A gl9 41 1.45636874 0.44464045 166177369 ACACAATGAGCCACCTACAAG g20 42 1.31187425 0.42605224 166177362 GTGGCTCATTGTGTGTGTGCC g22 43 1.25217773 0.26572657 166155264 CATATCCCTGCAGGTTCAGAA g24 44 1.75688991 0.28984533 166155099 agagagagagagagagagaga g25 45 2.05701745 0.42409102 166155393 TTCTCAGTTTTGAAAATTAAAA g26 46 1.64498972 0.21705582 166155255 TGGATTCTCTTCTGAACCTGC g27 47 2.27026665 0.43195546 166148361 TGCTGAGGCAGGACACAGTGT g29 48 2.4290169 0.30364553 166148843 ATCATCTGTAACCATCAAGGA g30 49 2.58328006 0.0748197 166148565 TCCTGCCTACTTAGTTTCAAG g31 50 1.97097781 0.25980859 166148953 ATTACAGTTCTGTCAGCATGC g32 51 1.34500323 0.32186367 166149373 TGGTCTCATTCTTTTTGTGGG g33 52 1.71471378 0.32104302 166142239 CGATATTTTCATGGATTCCCTT g34 53 1.7735976 0.21954265

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166142391 CTGACACTTACTTTGTCTAAA g35 54 1.95513108 0.02069095 166142219 AAAACTGGAACCGCATTCCCA g36 55 2.08698135 0.0454403 166142396 ACAAAGTAAGTGTCAGTGTGG g37 56 1.30739959 0.72725347 166142344 ATAATAGTTGTGTCTTTATAA g38 57 1.55783618 0.29846459 166141162 TGTACAAGCAGGGCTGCAAA G g39 58 1.4663605 0.02946062 166140590 GTTAACAAATACACTAAACAC g40 59 1.37399196 0.33638238 166140928 ttcaacaagctcccaagaagt g42 60 1.46929899 0.24465271 166141090 ATGTTCAAGGTGCAGAAGGA A g43 61 2.04547409 0.09880194 165990246 TGTTTTGCTCAAACGTGCACCA g44 62 2.13402102 0.25583999 165989684 AAATAAGACATGAAAACAAG A g45 63 1.27016182 0.32368695 165990193 AAATATGTACCACAAGAAATG g46 64 2.29522738 0.41829497 165990076 TATCTGGTTTCTCTCACTGCT g47 65 1.44542116 0.0947106 165989571 ATTGCAAAGCATAATTTGGAT g48 66 1.42246971 0.18117243

Example 2.

Activation of endogenous SCN1A in HEK293 cells using SCN1A-specific transcription factors.

HEK293 cells were cultured using standard methods and seeded in 6-well plates. Cells in each well were transfected (FugeneHD, Promega) with 3 μg of plasmid carrying either a single engineered transcription factor construct, human CREB3 WT (SEQ ID NO: 211), or a control EGFP construct. The engineered transcription factor constructs studied included: constructs 1-27 and 46-53 (Table 1) and a plasmid expressing CREB3-TRE (SEQ ID NO: 215; CREB3 with the bZIP DNA binding domain replaced by a synthetic ZF targeting domain on the TET promoter) (each was studied separately). 48 hours after transfection, cells were harvested and RNA was isolated (Qiagen RNeasy Mini kit) and treated with DNase. RNA (3 μg) was reverse transcribed using OligoDT primers (Superscript IV, Invitrogen). cDNA samples were analyzed by qPCR using Phusion Polymerase (New England Biolabs) and SYBR Green I: (30 seconds at 98°C, 40 times [10 seconds at 98°C, 15 seconds at 66°C, 15 seconds at 72°C]). Primers against SCN1A (5'-TGTCTCGGCATTGAGAACATTC-3' (SEQ ID NO: 185); 5'-ATTGGTGGGAGGCCATTGTAT-3' (SEQ ID NO: 186)) were used to quantify endogenous SCN1A transcript levels, and relative SCN1A expression levels were determined by the method delta-delta Ct with GAPDH as the reference gene (5'-ACCACAGTCCATGCCATCAC'3' (SEQ ID NO: 187); 5'-TCCACCACCCTGTTGCTGTA3' (SEQ ID NO: 188)). Data are presented as fold changes relative to the control condition (see Fig. 2A, Fig. 2B and Fig. 2C). The control construct consisted of EGFP expressed under the control of RE 1 (SEQ ID NO: 1). Delivery of engineered transcription factors induced varying degrees of activation of the endogenous SCNIA transcript relative to the EGFP state.

Example 3.

Activation of endogenous SCN1A in GABA neurons using SCN1A-specific transcription factors.

iCell GABA neurons (Cellular Dynamics) were plated in a 6-well plate (~1E6 cells/well) and maintained according to the protocol recommended by the manufacturer. 72 hours after seeding, recombinant AAV (serotype AAV-DJ) expressing EGFP or activator construct 30 in FIG. 3A, or structure 25, or structure 16 in FIG. 3B under the control of a universal promoter (CBA promoter) was added to the culture medium at approximately 2E11 genome copies per well. One week (Fig. 3A) or two weeks (Fig. 3B) postinfection, RNA was isolated from cultured cells (Qiagen RNeasy Mini kit) and treated with DCCase. The recovered RNA was reverse transcribed using OligoDT primers (Superscript IV, Invitrogen). cDNA samples were analyzed by qPCR using Phusion Polymerase (New England Biolabs) and SYBR Green I: (30 seconds at 98°C, 40 times [10 seconds at 98°C, 15 seconds at 66°C, 15 seconds at 72°C]). Primers against SCN1A (5'-TGTCTCGGCATTGAGAACATTC-3' (SEQ ID NO: 185); 5'-ATTGGTGGGAGGCCATTGTAT-3' (SEQ ID NO: 186)) were used to quantify endogenous SCN1A transcript levels, and relative SCN1A expression levels were determined by the method delta-delta Ct with GAPDH as the reference gene (5'-ACCACAGTCCATGCCATCAC'3' (SEQ ID NO: 187); 5'TCCACCACCCTGTTGCTGTA-3' (SEQ ID NO: 188)). Data are presented as fold changes

- 113 046157 relative to the control condition (see Fig. 3A and 3B). AAV-driven expression of engineered transcription factors caused significant activation of the endogenous SCNIA transcript in cultured iPS-derived GABA neurons.

Example 4.

Specific activation of endogenous SCN1A in GABA neurons using a SCN1A-specific transcription factor.

iCell GABA neurons (Cellular Dynamics) were plated in a 6-well plate (~1E6 cells/well) and maintained according to the protocol recommended by the manufacturer. At 72 hours post-plating, recombinant AAV (serotype AAV-DJ) expressing EGFP or an activator (construct 30) that contains a zinc finger DBD fused to a VPR TAD driven by approximately 2E11 CBA promoter was added to the culture medium. copies of the genome per well.

One week after infection, RNA was isolated from cultured cells (Qiagen RNeasy Mini kit) and treated with DNase. RNAseq libraries were prepared from recovered RNA using the TruSeq Stranded mRNA Library Kit (Illumina) and sequenced on Illumina NextSeq (2x75 cycle paired-end sequencing). Sequencing reads were mapped to the human genome (RNASTAR) and differential expression analysis was performed using DESeq2. Data are presented as fold change relative to control (AAVDJ-CBA-EGFP) samples (see Fig. 4). The results are shown in table. 14, and in FIG. Figure 4 shows the relative expression of endogenous SCN1A and transcripts of the 40 nearest neighboring genes, presented as fold changes relative to the control condition. Design 30, as described in table. 1 was able to specifically increase the expression of the SCN1A gene or Nav1.1 protein compared to other genes tested. This indicates that the target site recognized by transcription activator construct 30 is specific to the SCN1A gene, resulting in increased expression of the SCNIA gene in GABA neurons.

Table 14

Effects on transcription of endogenous SCN1A and 40 immediate neighboring genes in GABA neurons treated with a SCN1A-specific transcription factor (construct 30)

Gene name Start chrome. 2 The end is chrome. 2 The thread is lame. Fold change vs control PLA2R1 160788518 160919121 - 0.16367458 ITGB6 160956176 161128399 - 0.20679884 RBMS1 161128661 161350305 - 1.63514667 TANK 161993418 162092732 + 0.90946407 PSMD14 162164548 162268228 + 0.92699237 TBR1 162272604 162282381 + 0.53199642 SLC4A10 162280842 162841792 + 1.89407328 DPP4 162848750 162931052 - 2.82345284 FAP 163027193 163101661 - 2.26977379 IFIHI 163123588 163175213 - 1.46146481 G.C.A. 163175349 163228105 + 2.58702426 FIGN 164449905 164592522 - 0.46785861 GRB14 165349321 165478358 - 0.5631965 COBLL1 165510133 165700189 - 0.43199257 SLC38A11 165752695 165812035 - 4.06730119 SCN3A 165944031 166060577 - 1.0807866 SCN2A 166095911 166248818 + 1.24475196 CSRNP3 166326156 166545917 + 0.82971233 GALNT3 166604100 166651192 - 0.33804418

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TTS21V 166713984 166810353 - 1.58661143 SCN1A 166845669 166984523 - 62.9552975 SCN9A 167051694 167232503 - 1.71659087 SCN7A 167260082 167350757 - 0.29331967 B3GALT1 168675181 168730551 + 0.64436013 STK39 168810529 169104651 - 1.19821739 CERS6 169312371 169631644 + 0.86828378 NOSTRIN 169643048 169722024 + 1.82142718 SPC25 169690641 169769881 - 0.86880697 ABCB11 169779447 169887832 - 3.1441368 DHRS9 169921298 169952677 + 1.10381777 BBS5 170335687 170382432 + 0.65476347 KLHL41 170366211 170382772 + 0.87373377 FASTKD1 170386258 170430385 - 1.02786927 P.P.I.G. 170440849 170497916 + 1.09866236 CCDC173 170501934 170550943 - 0.67290779 PHOSPHO2 170550974 170558218 + 0.91339152 KLHL23 170550997 170633499 + 0.73926347 SSB 170648442 170668574 + 1.00631994 METTL5 170666590 170681441 - 1.21271497 UBR3 170683967 170940641 + 1.21350908 MYO3B 171034654 171511681 + 0.52839217

Example 5.

Expression of SCN1A from an expression cassette in vivo.

To test the expression of SCN1A transcriptional activators in vivo, recombinant AAV9 vectors were generated by VectorBiolabs (Malvern, PA). Male C57Bl/6 mice (N=5 per group, 7-8 weeks old) received a bilateral infusion of 1.5 μl of purified AAV vector into the dorsal hippocampus (AP -2.0 mm, lateral ±1.5, DV -1.4 mm from the dura mater) and ventral hippocampus (AP -3.1 mm, lateral ± 2.8, DV -3.8 mm from the dura mater), a total of 4 injection sites. AAV was delivered at a rate of 0.3 μl/min with a rest period of 4 min after each injection. Four weeks after exposure, mice were sacrificed and hippocampal tissue was excised. For each group, tissue from the left and right hippocampus was collected together for homogenization in most animals (N=4), with the exception of one animal in which only the left hippocampus was collected and homogenized. RNA was isolated from the homogenate (Qiagen RNeasy Mini kit) and treated with DNase. RNA (3 μg) was reverse transcribed using OligoDT primers (Superscript IV, Invitrogen). cDNA samples were analyzed by qPCR for mouse SCN1A expression using Phusion Polymerase (New England Biolabs) and SYBR Green I: 30 seconds at 98°C, 40 times [10 seconds at 98°C, 15 seconds at 64°C C, 15 seconds at 72°C]. Primers against mouse SCN1A (5'-CAAAAAAGCCACAAAAGCCT-3' (SEQ ID NO: 189); 5'TTAGCTCCGCAAGAAACATC-3' (SEQ ID NO: 190)) were used to quantify endogenous SCN1A transcript levels, and relative SCN1A expression levels in vivo determined by the delta-delta Ct method with GAPDH as the reference gene (5'-ACCACAGTCCATGCCATCAC'3' (SEQ ID NO: 187); 5'-TCCACCACCCTGTTGCTGTA-3' (SEQ ID NO: 188)).

In fig. 5A and FIG. 5B shows the average results for five animals each treated with the AAV9 construct. The eGFP control construct contained the eGFP reporter transgene. Construct 4 (see Table 1) contained a transcription activator that recognized the target sequence containing SEQ ID NO: 18, as described in Table. 1 above. In fig. 5A shows the relative expression of SCN1A in vivo. In fig. Figure 5B shows the change in SCN1A expression in vivo as a percentage of average eGFP expression. These results indicated that transcriptional activator of SCN1A expression cassette A resulted in an approximately 20-30% increase in SCNIA expression in vivo.

Such expression cassettes may be adapted for use in humans to treat Dravet syndrome, epilepsy, seizures, Alzheimer's disease, Parkinson's disease and/or any other diseases or conditions associated with deficiency and/or impairment of SCN1A activity.

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Example 6.

Analysis of hyperthermic seizures (HTS) in mice with Dravet syndrome.

A. Heterozygous Scn1a knockout mouse model.

Treatment of Dravet syndrome and/or its symptoms using expression cassettes was investigated in the Scn1a ^tm1Kea mouse line. This mouse line is an established mouse model for Dravet syndrome. Scn1a ^tm1Kea mouse lines do not require CRE recombinase. The Scn1a ^tm1Kea mouse (available from the Jackson Laboratory; described in Hawkins et al., Scientific Reports, vol. 7: 15327 (2017)) contains a deletion of the first coding exon of SCN1A. Mice homozygous for the SCN1A knockout allele are characterized by tremors, ataxia, seizures, and die by 16 days after birth. Heterozygous mice on a C57BL/6 background develop spontaneous seizures and most die within a few weeks. This mouse line can be used to study the safety and effectiveness of treatment for epilepsy and Dravet syndrome. See Miller et al., Genes Brain Behav. 2014 Feb;13(2):163-72 for more information.

To examine the efficacy of transcriptional activators in the Scn1a ^tm1Kea mouse line, litters of pups derived from Scn1a +/− males crossed with C57Bl/6J females were injected with the AAV vector via bilateral ICV at P1. Mice were administered doses of constructs 31–34 (Table 1). Mice were left undisturbed with their mother until weaning at P18 and then left undisturbed again until P26–P28, when the hyperthermic seizure (HTS) assay began. Individual litters of mice dosed at P1 were screened out at P18 and monitored for mortality daily. Hyperthermic seizure induction was performed in heterozygous (HET) mice and WT Scn1a mice on a mixed 129Stac X C57BL/6 background at P26-P28. Before the study, mice were inserted with a lubricated rectal temperature probe (Ret-4) and connected to a temperature control module (TCAT 2DF, Physitemp), which was connected in series with a heat lamp (HL-1). The mice were then placed in a large glass beaker and allowed to equilibrate with the environment for a short time. Thereafter, body temperature increased by ~0.5°C every 2 minutes until the onset of the first tonic-clonic seizure accompanied by loss of standing position or until the temperature reached 43°C. If a mouse experienced a seizure with loss of body position, the experiment was terminated and the mouse's core body temperature was recorded. If no postural seizures were detected throughout the experiment, that mouse was considered seizure-free and the analysis was completed. Tissue samples were obtained from mice at P1, and genotyping of mice was performed during the experiment using real-time PCR. Once the analysis was completed, genotyping was not blinded, and the status of the mice as HET or WT was correlated with the data obtained. Data were plotted on a Kaplan-Meier survival curve, and significance was determined using the Mantel-Cox test. The results are shown in table. 15 and 16 and in fig. 6A-E.

Table 15

Summary of conditions used in Example 6

Design Dosage (gene copies/mouse) Design 31 (Fig. 6A and 6E)) 6.0E+10 Design 32 (Fig. 6B, 6D, 6E) 3.1E+11 Design 33 (Fig. 6C) 5.8E+10 Design 34 (Fig. 6D) 4.9E+13

Table 16

Summary of test results for hyperthermic seizures

eTF design (FIG.) Number of control animals (PBS treatment) Number of animals treated % without seizures at temp. 42.6°C Rvalue EGFP reporter 16 N/A 44% Design 31 (Fig. 6A and 6E)) 16 18 95 P<0.0001 Design 32 (Fig. 6B, 6D, 6E) 16 21 76 P<0.05 Design 33 (Fig. 6C) 16 14 93 P<0.001 Design 34 (Fig. 6D) 16 12 83 P<0.05

Additional experiments were performed in Scn1a ^tm1Kea mice as described above to test the effects of constructs 42 and 43 on seizures in the HTS assay. In these experiments, construct 42 was administered at a dose of 9x1010 genome copies/mouse via bilateral ICV in P1, and construct 43 was administered at a dose of 6x1010 genome copies/mouse via bilateral ICV in P1 or P5. The results are shown in Fig. 6F (structure 42) and FIG. 6G (design 43). Both constructs showed a significant reduction in seizures compared to EGFP controls (P<0.0001 for both construct 42 and construct 43).

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B. ScnlaRX heterozygous mutant mouse model.

Treatment of Dravet syndrome and/or its symptoms using the expression cassette of the present disclosure was studied in the Scn1a ^RX mouse line. This mouse strain is an established mouse model for Dravet syndrome. Scn1aRX mouse lines do not require CRE recombinase. The Scn1aRX mouse (available from the Jackson Laboratory; described in Ogiwara et al., J. Neuroscience, vol. 27: 59035914 (2007)) contains a loss-of-function nonsense mutation in exon 21 of the SCN1A gene (CgG to TgA; R1407X). Heterozygous mice on a C57BL/6 background develop spontaneous seizures, and a large percentage of mice die within a few weeks.

To test the effectiveness of transcription activators in the Scn1aRX mouse line, the AAV vector (design 31) was introduced into litters of pups obtained through IVF crossing of Scn1a ^RX/+ male sperm with C57Bl/6J eggs with embryos implanted into CD-1 females at a dose of 5.1x10 ¹⁰ copies of the mouse genome or PBS control via bilateral ICV in P1. Mice were left unexposed with their mother until weaning at P18 and then left unexposed again until P26–P28, when the HTS assay began. Individual litters of mice dosed at P1 were examined at P18 and monitored daily for mortality. Hyperthermic seizure induction was performed in heterozygous (HET) P26-P28 mice and WT Scn1a mice on a C57BL/6 background. Before the study, mice were inserted with a lubricated rectal temperature probe (Ret-4) and connected to a temperature control module (TCAT 2DF, Physitemp), which was connected in series with a heat lamp (HL-1). The mice were then placed in a large glass beaker and allowed to equilibrate with the environment for a short time. Thereafter, body temperature increased by ~0.5°C every 2 minutes until the onset of the first tonic-clonic seizure, accompanied by loss of body position, or until the temperature reached 43°C. If a mouse experienced a seizure with loss of body position, the experiment was terminated and the mouse's core body temperature was recorded. If no postural seizures were detected throughout the experiment, that mouse was considered seizure-free and the analysis was completed. Tissue samples were obtained from mice at P1, and genotyping of mice was performed during the experiment using real-time PCR. Once the analysis was completed, genotyping was not blinded, and the status of the mice as HET or WT was correlated with the data obtained. None of the WT Scna1 mice studied had a seizure. Data for HET mice treated with construct 31 (n=13) and PBS control (n=14) were plotted on a Kaplan-Meier survival curve and significance was determined using the Cox-Mantel test. As shown in FIG. 6H, HET mice treated with construct 31 show a significant reduction in hyperthermic seizure induction compared to HET mice treated with control PBS (P<0.01).

Example 7.

Survival analysis in a mouse model of Dravet syndrome.

A. Heterozygous Scn1a knockout mouse model.

To test the efficacy of transcriptional activators in the Scn1a ^tmlKea mouse line, litters of pups derived from Scn1a +/− males crossed with C57Bl/6J females were injected with the AAV vector via bilateral ICV at P1. Mice were left unexposed with their mother until weaning. Health monitoring of Scn1a +/− mice was performed daily after weaning at P18. Mice that were found dead in their home cage for any reason were dated. Data were plotted on a Kaplan-Meier survival curve and significance was determined using the Cox-Mantel test.

The results are shown in table. 17 and FIG. 7A-D.

Table 17

Summary of conditions and results of survival analysis

SEQ ID Dosage (gc/mouse) Number of control animals (PBS treatment) Number of animals treated % of survivors in P100 (*in P83) Rvalue PBS N/A 53 N/A 49% Design 33 (Figs. 7C and 7D) 5.8E+10 53 29 76% P<0.05 Design 31 (Figs. 7B and 7D) 6.0E+10 53 34 97% P<0.0001

_~.RX

B. Heterozygous Scn1a mutant mouse model.

To test the effectiveness of transcriptional activators in the Scn1a ^RX mouse line, litters of pups generated from males and IVF crosses of Scn1aRX/+ sperm with C57Bl/6J eggs were injected into embryos implanted with CD-1 females with the AAV vector (construct 31) at 5. 1x10 ¹⁰ copies of the genome ^cMouse or PBS control via bilateral ICV in P1. Mice were left unexposed with their mother until weaning. Health Monitoring of Scn1a Mice

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RX/+ was performed daily after weaning at P18. Mice that were found dead in their home cage for any reason were dated. Data for construct 31 treated (n=27) and control treated (n=18) were plotted on a Kaplan-Meier survival curve and significance was determined using the Cox-Mantel test.

As shown in FIG. 7E, Scn1aRX ^/+ mice receiving construct 31 had increased survival compared to Scn1aRX ^/+ mice receiving control PBS (P<0.0001).

Example 8.

Transcription levels of SCN1A in non-human primates following treatment with AAV encoding the specific transcription factor SCN1A.

The study used male cynomolgus monkeys (macaca fascicularis) aged 2 to 3 years. Animals were prescreened for the presence of cross-reactive antibodies to AAV9 using a cellular neutralizing antibody assay before inclusion in the study. AAV9 expressing a transcription factor specific for SCN1A (construct 33) or control was diluted in PBS and injected intraparenchyally at a concentration of 1.2E12 gc/animal. For injections, three different stereotaxic coordinates were determined in each hemisphere, six injection sites per animal. A volume of 10 μl was injected into each point. Injections into the right hemisphere were symmetrical to the left. Two untreated animals were used as controls.

To evaluate Scn1A mRNA expression, reverse transcription followed by qPCR was performed. At necropsy 28 days after dosing, tissue sections from various brain regions (frontal cortex, parietal cortex, temporal cortex, occipital cortex, hippocampus, medulla oblongata, cerebellum; 200 mg each) from control and treated animals were collected in RNAlater. and then frozen. Briefly, 30 mg of tissue was dissected, RNA extracted (using Qiagen Rneasy Lipid Mini Tissue Kit, part number 1023539), converted to cDNA by reverse transcription (using Applied Biosystems High Capacity cDNA Reverse Transcription Kit, part number 4368814 ) and qPCR was performed using a primer/probe set for Scn1A and the housekeeping gene GAPDH (Applied Biosystems, catalog number Rh02621745-gI FAM).

Primer/probe sets for SCNIA are given below.

Table 18

Primer sequences used in example 8

Gene SEQ ID NO Sequence (5’-3’) Note ScnlA 191 CCATGGAACTGGCTCGATTTCAC F primer 192 ATTGGTGGGAGGCCACTGTAT R primer 193 AGGCCTGAAAACCATTGTGGGAGCCCT Probe (FAM)

Scn1A gene expression in each test sample was determined by relative quantitation (RQ) using the comparative Ct (ΔΟ) method. This method measures the difference in Ct(ΔΟ0) between a target gene and a housekeeping gene, and then compares the ΔΟ values of treatment samples with control samples.

ACt = average Ct of target gene - average Ct of housekeeping gene.

ΔΔ(ΐ = Δ(ΐ treatment sample - Δ(ΐ control sample.

Relative expression (treatment sample) = 2 - ΔΔ(ΐ

Data are presented as normalized target mRNA expression in different brain tissues (see Fig. 8). As shown in FIG. 8, brain regions proximal to intraparenchymal injection sites showed the highest levels of SCN1A transcript expression.

Example 9.

Selective transgene expression in PV-containing neurons in non-human primates following treatment with AAV having a PV-selective promoter and microRNA binding site.

The study used six marmosets (Callithrix jacchus), which were pretested for cross-reactive antibodies to AAV9 before inclusion in the study. Two monkeys were administered AAV9 containing the EGFP transgene under the control of the EF1alpha promoter, two monkeys were administered AAV9 containing the EGFP transgene under the control of RE 2 (SEQ ID NO: 2), and two monkeys were administered AAV9 containing the EGFP transgene under the control of RE 2 (SEQ ID NO: 2), and also containing a microRNA binding site (SEQ ID NO: 7), located between the EGFP coding region and the polyTail. AAV9 vectors were diluted in PBS and animals received three intracerebral injections (2 μl each) into the hippocampus/entorhinal cortex of each hemisphere, for a total of 6 injection sites per animal. Two animals received an AAV9 vector containing EF1alpha-EGFP, each receiving a total dose of 5.8E+11 genome copies/animal, two animals received an AAV9 vector containing RE 2-EGFP, each receiving a total dose of 3.0E+11 genome copies/animal , and two animals received the AAV9 vector containing RE 2 + m1-EGFP, each receiving a total dose of 2.3E+11 genome copies/animal.

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Immunohistochemistry was used to evaluate the selective expression of parvalbumin (PV). At autopsy, 28 days after dosing, tissue sections were collected from various areas of the brain. Floating sections of marmoset brains (35 μm) were fixed in 4% paraformaldehyde, blocked with buffer (PBS, 3% BSA, 3% donkey serum, 0.3% Triton-X 100, 0.2% Tween-20), and then stained anti-GFP (Abcam ab290) followed by a secondary antibody conjugated to Alexa488 (Thermo A21206). This was followed by an anti-PV antibody (Swant) and a secondary antibody conjugated to Alexa-647 (Thermo A31571) and 4′,6-diamidino-2-phenylindole (DAPI). Sections were mounted and imaged using a PerkinElmer Vectra3.

The results are shown in Fig. 9A-F and 10A-L.

Example 10.

Biodistribution of eTF ^SCN1A

The aim of this study was to compare the biodistribution of eTF ^SCN1A in the central nervous system (CNS) of juvenile cynomolgus monkeys when administered at a dose of 4.8E+13 via unilateral intracerebroventricular (ICV) injection. Each animal was injected with AAV9 containing an expression cassette encoding eTF ^SCN1A under the control of a GABA selective regulatory element (RE ^GABA -eTF ^SCN1A ). AAV9 particles were formulated in PBS + 0.001% pluronic acid and administered at a dose of 4.8E+13 or 8E+13 vg/animal. Each animal was injected with 2 ml of prepared viral particles. The study design is presented in Table. 19.

Cynomolgus macaques at twenty-four months of age were grouped as indicated in Table. 19. Before the start of the study, animal blood samples were tested for levels of neutralizing antibody titre to AAV9 using the NAb titer assay described above. Animals with low or negative antibody results were selected for the study. Samples were administered via ICV injection using standard surgical procedures. Thawed dispense material was briefly stored on wet ice and warmed to room temperature immediately before dispensing. Animals were anesthetized, prepared for surgery, and placed in an MPT-compatible stereotactic frame (Kopf). Baseline MRI was performed to determine target coordinates. An incision was made and one hole was drilled into the skull above the target area. A 3 mL BD syringe attached to a 36-inch microchannel extension set was prepared with the sample and placed into the infusion pump. The extension line was primed. The dura mater was opened and the dosing needle was advanced to a depth of 13.0 to 18.1 mm from the pia mater. Contrast injection and fluoroscopy were used to confirm insertion of the spinal needle into the right lateral ventricle. Contrast was added to the Quinke BD 3.0 inch 22 g spinal surgery needle to determine its location before attaching the primed extension line and syringe. Pump settings were 0.1 ml/min for 19-20 minutes. Buffer was manually injected after dosing to clear the extension line. The needle was left in place for 1-2 minutes after completion of the infusion, and then the needle was removed. Vehicle and test substance were administered once on day 1, and subjects were maintained during a 27- or 29-day recovery period.

Table 19

Biodistribution Study Design

Group Floor ID Dose (VG/animal) Group 1 (control buffer) M 21001 AND 11501 Group 2 (RE ^GABA -eTF ^SCN1A ) m 2001 4.8E+13 and 2501 4.8E+13 m 3001 4.8E+13 m 3002 8E+13

Following dosing, animals were monitored regularly throughout the study and blood samples were collected periodically. Administration of eTF ^SCN1A was not associated with unexpected mortality, clinical findings, or macroscopic observations. Animals receiving AAV9-RE ^gaba eTFS ^CN1A survived until scheduled necropsy for 28 ± 2 days. No clinical or behavioral signs, fever, or weight loss were observed during daily or weekly medical examinations. A transient increase in hepatic transaminases (ALT and AST) was observed in animals treated with AAV9-RE ^GABA -eTF ^SCN1A , but completely resolved by the end of the study without immunomodulation, and no concomitant increase in serum bilirubin or alkaline phosphatase was noted. None of the other clinical chemistry results measured were outstanding. No microscopic observations were reported in histopathological studies of the liver. CSF leukocytes were elevated at final collection compared to pre-exposure values, but comparable between control and AAV9-RE ^GABA -eTF ^SCN1A -treated animals. No AAV9-RE ^GABA -eTF ^SCN1A- associated pleocytosis was observed. Gross examination and detailed microhistopathological examination of non-neuronal tissues in all animals were unremarkable. Tissues included major peripheral organs (e.g.

- 119 046157 heart, lungs, spleen, liver and gonads). Macroexamination and detailed microhistopathology of neuronal tissues did not show any significant findings. Tissues included the brain, spinal cord, and associated intervertebral nodes (from the cervical, thoracic, and lumbar regions).

The studies were performed by three independent pathologists, including one at a specialized neuropathology center.

ICV administration of AAV9 did not prevent the post-dose immune response in serum, as neutralizing antibodies to the AAV9 capsid were observed four weeks post-dose. However, CSF levels of neutralizing antibodies to AAV9 remained unchanged and comparable to predose levels (Table 20).

Table 20

Serum NAb titer to AAV9

Subject number Serum NAb titer to AAV9 CSF NAb titer to AAV9 Before screening When injected 4 weeks after injections. When injected 4 weeks after injections. 21001 1:5 <1:5 <1:5 <1:5 <1:5 11501 <1:5 <1:5 <1:5 <1:5 <1:5 2001 <1:5 <1:5 1:405 <1:5 1:5 2501 <1:5 <1:5 1:135 <1:5 1:5 3001 <1:5 <1:5 1:1215 <1:5 <1:5 3002 <1:5 <1:5 1:135 <1:5 <1:5

Specimens were collected 27 to 29 days after dosing from major organs (cardiac ventricles, liver lobes, cardiac lobes, lungs, kidneys, spleen, pancreas, and cervical lymph nodes) during routine necroscopy. Targeted biopsies were collected using eight-mm biopsy forceps and processed as described below.

Example 11.

Biodistribution of eTF ^SCN1A in the brain.

ddPCR was used to measure the biodistribution of eTF ^SCN1A in the brain. Samples from different regions of cynomolgus brain tissue (FC: frontal cortex; PC: parietal cortex; TC: temporal cortex; Hip: hippocampus; Med: medulla; OC: occipital cortex) were measured for vector copy number to assess the biodistribution of eTF ^SCN1A under control of the selective regulatory element GABA (RE ^GABA -eTF ^SCN1A ) upon administration of AAV9 via unilateral ICV. Tissue DNA was isolated using DNeasy Blood & Tissues kits (Qiagen). The amount of DNA was determined and normalized using a UV spectrophotometer. 20 nanograms of tissue DNA were added to a 20 microliter reaction mixture along with ddPCR Super Mix for Probes (no dUTP) (Bio-Rad) and TaqMan primers and probes directed against eTF sequence regions ^{of SCN1A} . Droplets were generated and templates were amplified using an automated droplet generator and thermal cycler (BioRad). After the PCR step, the plate was loaded and read using a QX2000 Droplet Reader to determine the vector copy number in tissues. The simian albumin gene (MfAlb) served as an internal control to normalize genomic DNA content and was amplified in the same reaction. Primers and probes for eTF ^SCN1A and MfAlb are presented in Table. 21.

Table 21

Primers and probes for eTF ^SCN1A and MfAlb

Name of primers/probe gTpSCNIA MfAlbumin Forward primer _e ypSCNlA Reverse primer _eTF SCNlA eTF probe ^SCN1A Forward primer MfAlb Reverse primer MfAlb MfAlb probe Sequence (5’-3’) GAATGTGGGAAATCATTCAGTCGC (SEQ ID NO: 194) GC AAGTTATCCTCTCGTGAGAAGG (SEQ ID NO: 195) GCGACAACCTGGTGAGACATCAACGCACC (SEQ ID NO: 196) GCTGTTATCTCTTGTGGGCTGT (SEQ ID NO: 197) AAACTCATGGGAGCTGCCGGTT (SEQ ID NO: 198) CCACACAAATCTCTCCTGGCATTG (SEQ ID NO: 199)

The ^SCN1A eTF was widely distributed throughout the brain at a dose of 4.8E+13 viral genomes per animal with an average of 1.3-3.5 viral genomes/diploid genome (Fig. 11). In addition, when comparing brain-wide gene transfer RE ^GABA -eTF ^SCN1A , following dosing of 120 046157 4.8E+13 viral genomes per animal, with brain-wide gene transfer eGFP induced by ICV at various doses, the increase viral genome/diploid genome was observed with increasing dose. This indicates that gene transfer in the brain occurs in a dose-dependent manner when administered to AAV9 via the ICV.

Example 12.

Transcription of the ^SCN1A eTF in the brain.

Transcription of the ^SCN1A eTF under the control of a selective GABA regulatory element, RE ^GABA (RE ^GABA -eTF ^SCN1A ), was assessed by measuring eTF ^SCN1A mRNA using ddPCR-based gene expression analysis. Tissue RNA was isolated using the RNeasy Plus Mini (Qiagen) or RNeasy Lipid Tissue Mini (Qiagen) brain tissue kits. RNA quantity was determined and normalized using a UV spectrophotometer, and RNA quality (RIN) was checked using a Bioanalyzer RNA Chip. One microgram of tissue RNA was used for DNase treatment and cDNA synthesis using the SuperScript VILO cDNA Synthesis Kit with ezDNase™ Enzyme Kits (Thermo Fisher). 50 micrograms of RNA was converted to cDNA. The cDNA was added to a 20 microliter reaction mixture along with ddPCR Super Mix for Probes (no dUTP) (Bio-Rad) and TaqMan primers and probes directed against the ^SCN1A eTF sequence regions (Table 22). Droplets were generated and templates were amplified using an automated droplet generator and thermal cycler (Bio-Rad). After PCR amplification, the plate was loaded and read using a QX2000 Droplet Reader to obtain tissue gene expression levels. The monkey ARFGAP2 (MfARFGAP2) gene (Thermo Fisher Scientific) served as an endogenous control to normalize gene expression levels and was amplified in the same reaction. The average transcript abundance for ARFGAP2 was 1.85E+6 μg RNA (Fig. 12, upper limit). The detection limit is indicated by the lower limit.

eTF ^SCN1A mRNA was observed throughout the brain in all animals, indicating that the GABA-selective promoter, RE ^GABA , was transcriptionally active in brain tissue for all macaques that received AAV9-RE ^GABA -eTF ^SCN1A (Figure 12) . FC: frontal cortex; PC: parietal cortex; TC: temporal cortex; Hip: hippocampus; Med: medulla oblongata; OC: occipital cortex.

Table 22 and ^SCN1A eTF sequence region probes

Name of primers/probe Description Sequence (5'-3') _e ypSCNlA Forward primer _eTF SCNlA GAATGTGGGAAATCATTCAGTCGC (SEQ Ш NO: 200) Reverse primer _eTF SCNlA GCAAGTTATCCTCTCGTGAGAAGG (SEQ Ш NO: 201) Probe _e TF ^SCN1A GCGACAACCTGGTGAGACATCAACGCACC (SEQIDNO: 202) MSARFGAP2 Direct, reverse primers, probe Thermo Fisher (catalog number: 4448491)

Example 13.

Biodistribution and transcription of the ^SCN1A eTF in peripheral tissues.

Vector copy number was further measured in various organs to assess RE ^GABA -eTF ^SCN1A transduction in tissues throughout the body upon administration of AAV9 via unilateral ICV. Transcription levels of the ^SCN1A eTF were also measured by ddPCR to assess the transcriptional activity of the ^SCN1A eTF under the control of the GABA-selective regulatory element RE ^GABA in tissues throughout the body when introduced into AAV9 via unilateral ICV. Both methods were performed as generally described above. RE ^GABA -eTF ^SCN1A transduction and ^SCN1A eTF transcription in the spinal cord (SC) and intervertebral ganglion (DRG) were comparable to the levels observed in the brain. With the exception of the liver, RE ^GABA -eTF ^SCN1A transduction was lower in peripheral tissues outside the brain (Fig. 13). RE ^GABA -eTF ^SCN1A transduction was higher in liver than in brain. Transcription of the eTF ^SCN1A was not detected in peripheral tissues, including the heart, lungs, and gonads. However, eTF ^SCN1A transcript levels in the liver were comparable to eTF ^SCN1A levels measured in the brain. In addition, eTFSCN1A transcription in the liver is extremely low when normalized to the number of vector copies present (approximately 1000-fold lower compared to eTF ^SCN1A transcription in the brain). Overall, this demonstrated that transcription of the eTF ^SCN1A under the control of the GABA-selective regulatory element RE ^GABA is restricted to the CNS.

- 121 046157

Sequence Listing <110> INCOUDID THERAPEUTICS, INC.

<120> COMPOSITIONS AND METHODS FOR SELECTIVE REGULATION OF GENE EXPRESSION <130> 46482-724.601 <140>

<141>

<150> 63/008,569 <151> 2020-04-10 <150> 62/857,727 <151> 2019-06-05 <150> 62/854,238 <151> 2019-05-29 <160> 226 <170> PatentIn version 3.5 <210> 1 <211> 259 <212> DNA <213> Artificial sequence <220>

<223> Description of the artificial sequence: Synthetic polynucleotide <400> 1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt60 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac120 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg180 tgggaggtct atataagcag agctggtacc gtgtgtatgc tcaggggctg ggaaggagg240 ggagggagct ccggctcag259 <210> 2 <211> 2051 <212> DNA <213> Artificial sequence <220>

<223> Description of the artificial sequence: Synthetic polynucleotide <400> 2 ggaggaagcc atcaactaaa ctacaatgac tgtaagatac aaaattggga atggtaacat60 attttgaagt tctgttgaca taaagaatca tgatattaat gcccatggaa atgaaagggc120 gatcaacact atggtttgaa aagggggaaa ttgtag agca cagatgtgtt cgtgtggcag180 tgtgctgtct ctagcaatac tcagagaaga gagagaacaa tgaaattctg attggcccca240

- 122 046157

gtgtgagccc agatgaggtt cagctgccaa ctttctcttt cacatcttat gaaagtcatt 300 taagcacaac taactttttt tttttttttt tttttttgag acagagtctt gctctgttgc 360 ccaggacaga gtgcagtagt gactcaatct cggctcactg cagcctccac ctcctaggct 420 caaacggtcc tcctgcatca gcctcccaag tagctggaat tacaggagtg gccccacatg 480 cccagctaat ttttgtattt ttaatagata cggggggtttc accatatcac ccaggctggt 540 ctcgaactcc tggcctcaag tgatccacct gcctcggcct cccaaagtgc tggggattata 600 ggcgtcagcc actatgccca acccgaccaa ccttttttaa aataaatatt taaaaaattg 660 gtatttcaca tatatactag tatttacatt tatccacaca aaacggacgg gcctccgctg 720 aaccagtgag gccccagacg tgcgcataaa taacccctgc gtgctgcacc acctggggag 780 agggggga ccacggtaaa tggagcgagc gcatagcaaa agggacgcgg ggtccttttc 840 tctgccggtg gcactgggta gctgtggcca ggtgtggtac tttgatgggg cccagggctg 900 gagctcaagg aagcgtcgca gggtcacaga tctgggggaa ccccggggaa aagcactgag 960 gcaaaaccgc cgctcgtctc ctacaatata tgggggggg aggttgagta cgttctggat 1020 tactcataag accttttttt tttccttccg ggcgcaaaac cgtgagctgg atttataatc 1080 gccctataaa gctccagagg cggtcaggca cctgcagagg agccccgccg ctccgccgac 1140 tagctgcccc cgcgagcaac ggcctcgtga tttccccgcc gatccggtcc ccgcctcccc 1200 actctgcccc cgcctacccc ggagccgtgc agccgcctct ccgaatctct ctcttctcct 1260 ggcgctcgcg tgcgagagg aactagcgag aacgaggaag cagctggagg tgacgccggg 1320 cagattacgc ctgtcagggc cgagccgagc ggatcgctgg gcgctgtgca gaggaaaggc 1380 ggggagtgccc ggctcgctgt cgcagagccg aggtgggtaa gctagcgacc acctggactt 1440 cccagcgccc aaccgtggct tttcagccag gtcctctcct cccgcggctt ctcaaccaac 1500 cccaccag cgccggccac ccaacctccc gaaatgagtg cttcctgccc cagcagccga 1560 aggcgctact aggaacggta acctgttact tttccagggg ccgtagtcga cccgctgccc 1620 gagttgctgt gcgactgcgc gcgcggggct agagtgcaag gtgactgtgg ttcttctctg 1680 gccaagtccg aggggagaacg taaagatatg ggcctttttc cccctctcac cttgtctcac 1740 caaagtccct agtccccgga gcagttagcc tctttctttc cagggaatta gccagacaca 1800 acaacgggaa ccagacaccg aaccagacat gcccgccccg tgcgccctcc ccgctcgctg 1860 cctttcctcc ctcttgtctc tccagagccg gatcttcaag gggagcctcc gtgcccccgg 1920 ctgctcagtc cctccggtgt gcaggacccc ggaagtcctc cccgcacagc tctcgcttct 1980 ctttgcagcc tgtttctgcg ccggaccagt cgaggactct ggacagtaga ggccccggga 2040

cgaccgagct g 2051

- 123 046157 <210> 3 <211> 1878 <212> DNA <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic polynucleotide

<400> 3 tcaacagggg gacacttgggg aaagaaggat ggggacagag ccgagaggac tgttacacat 60 tagagaaaca tcagtgactg tgccagcttt ggggtagact gcacaaaagc cctgaggcag 120 cacaggcagg atccagtctg ctggtcccag gaagctaacc gtctcagaca gagcacaaag 180 caccgagaca tgtgccacaa ggcttgtgta gagaggtcag aggacagcgt acaggtccca 240 gagatcaaac tcaacctcac caggcttggc agcaagcctt taccaaccca cccccacccc 300 acccaccctg cacgcgcccc tctcccctcc ccatggtctc ccatggctat ctcacttggc 360 cctaaaatgt ttaaggatga cactggctgc tgagtggaaa tgagacagca gaagtcaaca 420 gtagattta ggaaagccag agaaaaaggc ttgtgctgtt tttagaagc caagggacaa 480 gctaagatag ggcccaagta atgctagtat ttacatttat ccacacaaaa cggacggggcc 540 tccgctgaac cagtgaggcc ccagacgtgc gcataaataa cccctgcgtg ctgcaccacc 600 tggggagagg ggggagacca cggtaaatgg agcgagcgca tagcaaaagg gacgcggggt 660 ccttttctct gccggtggca ctggggtagct gtggccaggt gtggtacttt gatggggccc 720 aggggctggag ctcaaggaag cgtcgcaggg tcacagatct gggggaaccc cggggaaaag 780 cactgaggca aaaccgccgc tcgtctccta caatatatgg gagggggagg ttgagtacgt 840 tctggattac tcataagacc tttttttttt ccttccggggc gcaaaaccgt gagctggatt 900 tataatcgcc ctataaagct ccagaggcgg tcaggcacct gcagaggagc cccgccgctc 960 cgccgactag ctgcccccgc gagcaacggc ctcgtgattt ccccgccgat ccggtccccg 1020 cctccccact ctgcccccgc ctaccccgga gccgtgcagc cgcctctccg aatctctctc 1080 ttctcctggc gctcgcgtgc gagagggaac tagcgagaac gaggaagcag ctggaggtga 1140 cgccggggcag attacgcctg tcaggggccga gccgagcgga tcgctgggcg ctgtgcagag 1200 gaaaggcggg agtgcccggc tcgctgtcgc agagccgagg tgggtaagct agcgaccacc 1260 tggacttccc agcgcccaac cgtggctttt cagccaggtc ctctcctccc gcggcttctc 1320 aaccaacccc atcccagcgc cggccaccca acctcccgaa atgagtgctt cctgccccag 1380 cagccgaagg cgctactagg aacggtaacc tgttactttt ccaggggccg tagtcgaccc 1440 gctgcccgag ttgctgtgcg actgcgcgcg cggggctaga gtgcaaggtg actgtggttc 1500 ttctctggcc aagtccgagg gagaacgtaa agatatggggc ctttttcccc ctctcacctt 1560 gtctcaccaa agtccctagt ccccggagca gttagcctct ttctttccag ggaattagcc 1620

- 124 046157

agacacaaca acgggaacca gacaccgaac cagacatgcc cgccccgtgc gccctccccg 1680 ctcgctgcct ttcctccctc ttgtctctcc agagccggat cttcaagggg agcctccgtg 1740 cccccggctg ctcagtccct ccggtgtgca ggaccccgga agtcctcccc gcacagctct 1800 cgcttctctt tgcagcctgt ttctgcgccg gaccagtcga ggactctgga cagtagaggc 1860 cccggggacga ccgagctg 1878

<210> 4 <211> 509 <212> DNA <213> Artificial sequence <220>

<223> Description of the artificial sequence: Synthetic polynucleotide <400> 4 gccctctagg ccacctgacc aggtcccctc agtccccccc ttcccacact cccacactca60 gcccccctcc cccccccccg acccctgcag gattatcctg tctgtgttcc tgactcagcc120 tgggagccac ctgggcagca ggggccaagg g tgtcctaga agggacctgg agtccacgct180 gggccaagcc tgccctttct ccctctgtct tccgtccctg cttgcggttc tgctgaatgt240 ggttatttct ctggctcctt ttacagagaa tgctgctgct aattttatgt ggagctctga300 ggcagtgtaa ttggaagcca g acaccctgt cagcagtggg ctcccgtcct gagctgccat360 gcttcctgct ctcctcccgt cccggctcct catttcatgc agccacctgt cccagggaga420 gaggagtcac ccaggcccct cagtccgccc cttaaataag aaagcctccg ttgctcggca480 cacataccaa gcagccgctg gtgcaatct509 <210> 5 <211> 1644 <212> DNA <2 13> Artificial sequence <220>

<223> Description of the artificial sequence: Synthetic polynucleotide <400> 5 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt60 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca120 atgggtggag tatttacggt aaactgccca cttgg cagta catcaagtgt atcatatgcc180 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta240 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac300 catgggtcga ggtgagcccc acgttctgct tcactctccc catctcc cccccctccccac360

- 125 046157

ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg 420 ggggggcgcg cgccaggcgg ggcggggcgg ggcgaggggc ggggcggggc gagggggaga 480 ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg 540 cggcggcggc ggccctataa aaagcgaagc gcgcggcggg cggggagtcgc tgcgttgcct 600 tcgccccgtg ccccgctccg cgccgcctcg cgccgcccgc cccggctctg actgaccgcg 660 ttactcccac aggtgagcgg gcgggacggc ccttctcctc cggggctgtaa ttagcgcttg 720 gtttaatgac ggctcgtttc ttttctgtgg ctgcgtgaaa gccttaaagg gctccggggag 780 ggccctttgt gcggggggga gcggctcggg gggtgcgtgc gtgtgtgtgt gcgtggggag 840 cgccgcgtgc ggcccgcgct gccccggcggc tgtgagcgct gcgggcgcgg cgcggggctt 900 tgtgcgctcc gcgtgtgcgc gagggagcg cggccggggg cggtgccccg cggtgcgggg 960 gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc gtggggggggt gagcagggggg 1020 tgtgggcgcg gcggtcgggc tgtaacccc ccctgcaccc ccctccccga gttgctgagc 1080 acggcccggc ttcgggtgcg gggctccgtg cggggcgtgg cgcggggctc gccgtgccgg 1140 gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg gccggggagg 1200 gctcggggga ggggcgcggc ggccccggag cgccggcggc tgtcgaggcg cggcgagccg 1260 cagccattgc cttttatggt aatcgtgcga gagggcgcag ggacttcctt tgtcccaaat 1320 ctggcggagc cgaaatctgg gagggcgccgc cgcaccccct ctagcggggcg cggggcgaagc 1380 ggtgcggcgc cggcaggaag gaaatggggcg ggggagggcct tcgtgcgtcg ccgcgccgcc 1440 gtccccttct ccatctccag cctcggggct gccgcagggg gacggctgcc ttcgggggggg 1500 acggggcagg gcggggttcg gcttctggcg tgtgaccggc ggctctagag cctctgctaa 1560 ccatgttcat gccttcttct ttttcctaca gctcctggggc aacgtgctgg ttgttgtgct 1620 gtctcatcat tttggcaaag aatt 1644

<210> 6 <211> 1335 <212> DNA <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic polynucleotide <400> 6

gagtaattca tacaaaagga ctcgcccctg ccttggggaa tccaggac cgtcgttaaa 60 ctccacctaa cgtagaaccc agagatcgct gcgttcccgc cccctcaccc gcccgctctc 120 gtcatcactg aggtggagaa gagcatgcgt gaggctccgg tgcccgtcag tgggcagagc 180 gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga accggtgcct 240

- 126 046157

agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc 300 ccgaggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca 360 acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg ttcccgcggg cctggcctct 420 ttacggggtta tggcccttgc gtgccttgaa ttacttccac gcccctggct gcagtacgtg 480 attcttgatc ccgagcttcg ggttggaagt gggtggggaga gttcgaggcc ttgcgcttaa 540 ggagcccctt cgcctcgtgc ttgagttgag gcctggcttg ggcgctgggg ccgccgcgtg 600 cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccattaa 660 aatttttgat gacctgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc 720 caagatctgc acactggtat ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg 780 tccagcgca catgttcggc gagggggggc ctgcgagcgc ggccaccgag aatcggacgg 840 gggtagtctc aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc 900 ccgccctggg cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg 960 cttcccggcc ctgctgcagg gagctcaaaa tggagaggacgc ggcgctcggg agagcggggcg 1020 ggtgagtcac ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac 1080 tccacggagt accggggcgcc gtccaggcac ctcgattagt tctcgagctt ttggagtacg 1140 tcgtctttag gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg 1200 gagactgaag ttaggccagc ttggcacttg atgtaattct ccttggaatt tgcccttttt 1260 gagtttggat tttcaggtgt cttggttcat cgtga tctcaagcct cagacagtgg ttcaaagttt ttttcttcca 1320 1335

<210> 7 <211> 214 <212> DNA <213> Artificial sequence <220>

<223> Description of the artificial sequence: Synthetic polynucleotide <400> 7 aaagagaccg gttcactgtg acagtaaaag agaccggttc actgtgagaa tgaaagagac60 cggttcactg tgatcggaaa agagaccggt tcactgtgag cggccttgaa acccagcaga120 caatgtagct cagtagaaac ccagcagaca atgtagctga atggaaaccc agcagacaat180 gtagcttcgg agaaacccag cagacaatgt agct214 <210> 8 <211> 21 <212> RNA <213> Homo sapiens

- 127 046157 <400> 8 ucacagugaa ccggucucuu u <210> 9 <211> 21 <212> DNA <213> Artificial sequence <220>

<223> Description of the artificial sequence: Synthetic oligonucleotide <400> 9 aaagagaccg gttcactgtg a 21 <210> 10 <211> 23 <212> RNA <213> Homo sapiens <400> 10 agcuacauug ucugcugggu uuc 23 <210> 11 <211> 23 <212> DNA <213> Artificial sequence <220>

<223> Description of the artificial sequence: Synthetic oligonucleotide <400> 11 gaaacccagc agacaatgta gct 23 <210> 12 <211> 23 <212> RNA <213> Homo sapiens <400> 12 agcuacaucu ggcuacuggg ucu <210> 13 <211> 23 < 212> DNA <213> Artificial sequence <220>

<223> Description of the artificial sequence: Synthetic oligonucleotide <400> 13 agaccagta gccagatgta gct 23 <210> 14 <211> 67

- 128 046157 <212> DNA <213> Artificial sequence <220>

<223> Description of the artificial sequence: Synthetic oligonucleotide <400> 14 gaaacccagc agacaatgta gctagaccca gtagccagat gtagctaaag agaccggttc 60 actgtga 67 <210> 15 <211> 134 <212> DNA <213> Artificial sequence <220>

<223> Description of the artificial sequence: Synthetic polynucleotide <400> 15 gaaacccagc agacaatgta gctagaccca gtagccagat gtagctaaag agaccggttc60 actgtgagaa acccagcaga caatgtagct agacccagta gccagatgta gctaaagaga120 ccggttcact gtga134 <210> 16 <211 > 62 <212> DNA <213> Artificial sequence <220>

<223> Description of the artificial sequence: Synthetic oligonucleotide <400> 16 aataaaagat ctttattttc attagatctg tgtgttggtt ttttgtgtgc ggaccgcacg 60 tg 62 <210> 17 <211> 477 <212> DNA <213> Artificial sequence <220>

<223> Description of the artificial sequence: Synthetic polynucleotide <400> 17 gggtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca60 gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc120 ttctataata ttatggggtg gagg ggggtg gtatggagca aggggcaagt tgggaagaca180 acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg240

- 129 046157

ctcactgcaa tctccgcctc ctggggttcaa gcgattctcc tgcctcagcc tcccgagttg 300 ttggggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg 360 ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat ctaccacct 420 tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc tgtcctt 477

<210> 18 <211> 18 <212> DNA <213> Homo sapiens <400> 18 ctaggtcaag tgtaggag <210> 19 <211> 18 <212> DNA <213> Homo sapiens <400> 19 acttgaccta gacagcct <210> 20 <211> 18 <212> DNA <213> Homo sapiens <400> 20 tgaataactc attagtga <210> 21 <211> 18 <212> DNA <213> Homo sapiens <400> 21 aaagtacatt aggctaat <210> 22 <211> 18 <212> DNA <213> Homo sapiens <400> 22 ccagcactgg tgcttcgt <210> 23 <211> 18 <212> DNA <213> Homo sapiens <400> 23 aaggctgtct aggtcaag

- 130 046157 <210> 24 <211> 24 <212> DNA <213> Homo sapiens <400> 24 ctaggtcaag tgtaggagac acac <210> 25 <211> 18 <212> DNA <213> Homo sapiens <400> 25 ggtcaagtgt aggagaca <210> 26 <211> 18 <212> DNA <213> Homo sapiens <400> 26 caagtgtagg agacacac <210> 27 <211> 24 <212> DNA <213> Homo sapiens <400> 27 agtgtaggag acacactgct ggcc <210> 28 <211> 18 <212> DNA <213> Homo sapiens <400> 28 agtgtaggag acacactg <210> 29 <211> 21 <212> DNA <213> Homo sapiens <400> 29 aggagacaca ctgctggcct g

<210> thirty <211> 18 <212> DNA <213> Homo

sapiens <400> 30 taggtaccat agagtgag

- 131 046157

<210> 31 <211> 18 <212> DNA <213> Homo sapiens <400> 31

gaggatactg cagaggtc <210> 32 <211> 27 <212> DNA <213> Homo sapiens <400> 32 taggtaccat agagtgaggc gaggatg <210> 33 <211> 27 <212> DNA <213> Homo sapiens <400> 33 atagagtgag gcgaggatga agccga g <210> 34 <211> 27 <212> DNA <213> Homo sapiens <400> 34 tgaagccgag aggatactgc agaggtc <210> 35 <211> 21 <212> DNA <213> Homo sapiens <400> 35 aaggctgtct aggtcaagtg t <210 > 36 <211> 21 <212> DNA <213> Homo sapiens <400> 36 tgttcctcca gattaacact t <210> 37 <211> 21 <212> DNA <213> Homo sapiens <400> 37 gatgaagccg agaggatact g

- 132 046157 <210> 38 <211> 21 <212> DNA <213> Homo sapiens <400> 38 gctgatttgt attaggtacc a <210> 39 <211> 21 <212> DNA <213> Homo sapiens <400> 39 agaaagctga tacagataca a <210> 40 <211> 21 <212> DNA <213> Homo sapiens <400> 40 ggtacgggca aagatttctt g <210> 41 <211> 21 <212> DNA <213> Homo sapiens <400> 41 agaaagctga tacagataca a < 210> 42 <211> 21 <212> DNA <213> Homo sapiens <400> 42 acacaatgag ccacctacaa g <210> 43 <211> 21 <212> DNA <213> Homo sapiens <400> 43 gtggctcatt gtgtgtgtgc c <210> 44 <211> 21 <212> DNA <213> Homo sapiens <400> 44 catatccctg caggttcaga a

- 133 046157 <210> 45 <211> 21 <212> DNA <213> Homo sapiens <400> 45 agagagagag agagagagag a <210> 46 <211> 21 <212> DNA <213> Homo sapiens <400> 46 ttctcagttt tgaaattaaa a <210> 47 <211> 21 <212> DNA <213> Homo sapiens <400> 47 tggattctct tctgaacctg c <210> 48 <211> 21 <212> DNA <213> Homo sapiens <400> 48 tgctgaggca ggacacagtg t < 210> 49 <211> 21 <212> DNA <213> Homo sapiens <400> 49 atcatctgta accatcaagg a <210> 50 <211> 21 <212> DNA <213> Homo sapiens <400> 50 tcctgcctac ttagtttcaa g <210> 51 <211> 21 <212> DNA <213> Homo sapiens <400> 51 attacagttc tgtcagcatg c

- 134 046157 <210> 52 <211> 21 <212> DNA <213> Homo sapiens <400> 52 tggtctcatt ctttttgtgg g <210> 53 <211> 21 <212> DNA <213> Homo sapiens <400> 53 cgatattttc atggattcct t <210> 54 <211> 21 <212> DNA <213> Homo sapiens <400> 54 ctgacactta ctttgtctaa a <210> 55 <211> 21 <212> DNA <213> Homo sapiens <400> 55 aaaactggaa ccgcattccc a < 210> 56 <211> 21 <212> DNA <213> Homo sapiens <400> 56 acaaagtaag tgtcagtgtg g <210> 57 <211> 21 <212> DNA <213> Homo sapiens <400> 57 aatagttg tgtcttttata a <210> 58 <211> 21 <212> DNA <213> Homo sapiens <400> 58 tgtacaagca gggctgcaaa g

- 135 046157 <210> 59 <211> 21 <212> DNA <213> Homo sapiens <400> 59 gttaacaaat acactaaaca c <210> 60 <211> 21 <212> DNA <213> Homo sapiens <400> 60 ttcaacaagc tcccaagaag t <210> 61 <211> 21 <212> DNA <213> Homo sapiens <400> 61 atgttcaagg tgcagaagga a <210> 62 <211> 21 <212> DNA <213> Homo sapiens <400> 62 tgtttgctca aacgtgcacc a < 210> 63 <211> 21 <212> DNA <213> Homo sapiens <400> 63 aaataagaca tgaaaacaag a <210> 64 <211> 21 <212> DNA <213> Homo sapiens <400> 64 aaatatgtac cacaagaaat g <210> 65 <211> 21 <212> DNA <213> Homo sapiens <400> 65 tatctggttt ctctcactgc t

- 136 046157 <210> 66 <211> 21 <212> DNA <213> Homo sapiens <400> 66 attgcaaagc aatttgga t <210> 67 <211> 3585 <212> DNA <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic polynucleotide

<400> 67 ggaggaagcc atcaactaaa ctacaatgac tgtaagatac aaaattggga atggtaacat 60 attttgaagt tctgttgaca taaagaatca tgatattaat gcccatggaa atgaaagggc 120 gatcaacact atggtttgaa aagggggaaa ttgtagagca cagatgtgtt cgtgtggcag 180 tgtgctgtct ctagcaatac tcagagaaga gagagaacaa tgaaattctg attggcccca 240 gtgtgagccc agatgaggtt cagctgccaa ctttctcttt cacatcttat gaaagtcatt 300 taagcacaac taactttttt tttttttttt tttttttgag acagagtctt gctctgttgc 360 ccaggacaga gtgcagtagt gactcaatct cggctcactg cagcctccac ctcctaggct 420 caaacggtcc tcctgcatca gcctcccaag tagctggaat tacaggagtg gccccacatg 480 cccagctaat ttttgtattt ttaatagata cggggggtttc accatatcac ccaggctggt 540 ctcgaactcc tggcctcaag tgatccacct gcctcggcct cccaaagtgc tggggattata 600 ggcgtcagcc actatgccca acccgaccaa ccttttttaa aataaatatt taaaaaattg 660 gtatttcaca tatatactag tatttacatt tatccacaca aaacggacgg gcctccgctg 720 aaccagtgag gccccagacg tgcgcataaa taacccctgc gtgctgcacc acctggggag 780 agggggga ccacggtaaa tggagcgagc gcatagcaaa agggacgcgg ggtccttttc 840 tctgccggtg gcactgggta gctgtggcca ggtgtggtac tttgatgggg cccagggctg 900 gagctcaagg aagcgtcgca gggtcacaga tctgggggaa ccccggggaa aagcactgag 960 gcaaaaccgc cgctcgtctc ctacaatata tgggggggg aggttgagta cgttctggat 1020 tactcataag accttttttt tttccttccg ggcgcaaaac cgtgagctgg atttataatc 1080 gccctataaa gctccagagg cggtcaggca cctgcagagg agccccgccg ctccgccgac 1140 tagctgcccc cgcgagcaac ggcctcgtga tttccccgcc gatccggtcc ccgcctcccc 1200 actctgcccc cgcctacccc ggagccgtgc agccgcctct ccgaatctct ctcttctcct 1260 ggcgctcgcg tgcgagagg aactagcgag aacgaggaag cagctggagg tgacgccggg 1320

- 137 046157

cagattacgc ctgtcagggc cgagccgagc ggatcgctgg gcgctgtgca gaggaaaggc 1380 ggggagtgccc ggctcgctgt cgcagagccg aggtgggtaa gctagcgacc acctggactt 1440 cccagcgccc aaccgtggct tttcagccag gtcctctcct cccgcggctt ctcaaccaac 1500 cccaccag cgccggccac ccaacctccc gaaatgagtg cttcctgccc cagcagccga 1560 aggcgctact aggaacggta acctgttact tttccagggg ccgtagtcga cccgctgccc 1620 gagttgctgt gcgactgcgc gcgcggggct agagtgcaag gtgactgtgg ttcttctctg 1680 gccaagtccg aggggagaacg taaagatatg ggcctttttc cccctctcac cttgtctcac 1740 caaagtccct agtccccgga gcagttagcc tctttctttc cagggaatta gccagacaca 1800 acaacgggaa ccagacaccg aaccagacat gcccgccccg tgcgccctcc ccgctcgctg 1860 cctttcctcc ctcttgtctc tccagagccg gatcttcaag gggagcctcc gtgcccccgg 1920 ctgctcagtc cctccggtgt gcaggacccc ggaagtcctc cccgcacagc tctcgcttct 1980 ctttgcagcc tgtttctgcg ccggaccagt cgaggactct ggacagtaga ggccccggga 2040 cgaccgagct ggaattcgcc accatggccc caaagaagaa gcggaaggtc ggtatccacg 2100 gagtcccagc agccctcgaa ccaggtgaaa aaccttacaa atgtcctgaa tgtgggaaat 2160 cattcagtcg cagcgacaac ctggtgagac atcaacgcac ccacagga gaaaaacctt 2220 aaaatgtcc agaatgtgga aagtccttct cacgagga taacttgcac actcatcaac 2280 gaacacatac tggtgaaaaa catacaagt gtcccgaatg tggtaaaagt tttagccgga 2340 gcgatgaact tgtccgacac caacgaaccc atacaggcga gaagccttac aaatgtcccg 2400 agtgtggcaa gagcttctca caatcaggga atctgactga gcatcaacga actcataccg 2460 gggaaaaacc ttacaagtgt ccagagtgtg ggaagagctt ttccacaagt ggacatctgg 2520 tacgccacca gaggacacat acaggggaga agccctacaa atgccccgaa tgcggtaaaa 2580 gtttctctca gaatagtacc ctgaccgaac accagcgaac acacactggg aaaaaaacga 2640 gtaaaaggcc ggcggccacg aaaaaggccg gccaggcaaa aaagaaaaag ggatcctacc 2700 catacgacgt accagattac gctctcgagg acgcgctgga cgatttcgat ctcgacatgc 2760 tggggttctga tgccctcgat gactttgacc tggatatgtt gggaagcgac gcattggatg 2820 actttgatct ggacatgctc ggctccgatg ctctggacga tttcgatctc gatatgttat 2880 aaactagtaa agagaccggt tcactgtgac agtaaaagag accggttcac tgtgagaatg 2940 aaagagaccg gttcactgtg atcggaaaag agaccggttc actgtgagcg gccttgaaac 3000 ccagcagaca atgtagctca gtagaaaccc agcagacaat gtagctgaat ggaaacccag 3060 cagacaatgt agcttcggag aaacccagca gacaatgtag ctaagcttgg gtggcatccc 3120 tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt gccccaccagc 3180 cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataattatt 3240

- 138 046157

atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc 3300 tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc 3360 tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag 3420 gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata 3480 ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa 3540 ttgctgggat tacaggcgtg aaccactgct cccttccctg tcctt 3585

<210> 68 <211> 3371 <212> DNA <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic polynucleotide

<400> 68 ggaggaagcc atcaactaaa ctacaatgac tgtaagatac aaaattggga atggtaacat 60 attttgaagt tctgttgaca taaagaatca tgatattaat gcccatggaa atgaaagggc 120 gatcaacact atggtttgaa aagggggaaa ttgtagagca cagatgtgtt cgtgtggcag 180 tgtgctgtct ctagcaatac tcagagaaga gagagaacaa tgaaattctg attggcccca 240 gtgtgagccc agatgaggtt cagctgccaa ctttctcttt cacatcttat gaaagtcatt 300 taagcacaac taactttttt tttttttttt tttttttgag acagagtctt gctctgttgc 360 ccaggacaga gtgcagtagt gactcaatct cggctcactg cagcctccac ctcctaggct 420 caaacggtcc tcctgcatca gcctcccaag tagctggaat tacaggagtg gccccacatg 480 cccagctaat ttttgtattt ttaatagata cggggggtttc accatatcac ccaggctggt 540 ctcgaactcc tggcctcaag tgatccacct gcctcggcct cccaaagtgc tggggattata 600 ggcgtcagcc actatgccca acccgaccaa ccttttttaa aataaatatt taaaaaattg 660 gtatttcaca tatatactag tatttacatt tatccacaca aaacggacgg gcctccgctg 720 aaccagtgag gccccagacg tgcgcataaa taacccctgc gtgctgcacc acctggggag 780 agggggga ccacggtaaa tggagcgagc gcatagcaaa agggacgcgg ggtccttttc 840 tctgccggtg gcactgggta gctgtggcca ggtgtggtac tttgatgggg cccagggctg 900 gagctcaagg aagcgtcgca gggtcacaga tctgggggaa ccccggggaa aagcactgag 960 gcaaaaccgc cgctcgtctc ctacaatata tgggggggg aggttgagta cgttctggat 1020 tactcataag accttttttt tttccttccg ggcgcaaaac cgtgagctgg atttataatc 1080 gccctataaa gctccagagg cggtcaggca cctgcagagg agccccgccg ctccgccgac 1140 tagctgcccc cgcgagcaac ggcctcgtga tttccccgcc gatccggtcc ccgcctcccc 1200

- 139 046157 actctgcccc cgcctacccc ggagccgtgc agccgcctct ccgaatctct ctcttctcct 1260 ggcgctcgcg tgcgagaggg aactagcgag aacgaggaag cagctggagg tgacccggg1320 cagattacgc ctgtcagggc cgagccgagc gg atcgctgg gcgctgtgca gaggaaaggc1380 gggagtgccc ggctcgctgt cgcagagccg aggtgggtaa gctagcgacc acctggactt1440 cccagcgccc aaccgtggct tttcagccag gtcctctcct cccgcggctt ctcaaccaac1500 cccacccag cgccggccac ccaacctccc gaaatgagtg cttcctgccc cagcagccga1560 aggcgctact aggaacggta acctgttact tttccagggg ccgtagtcga cccgctgccc1620 gagttgctgt gcgactgcgc gcgcggggct agagtgcaag gtgactgtgg ttcttctctg1680 gccaagtccg agggagaacg taaagatatg ggcctttttc cccctctcac cttgtctcac1740 caaagtccct agtccccgga gcagttagcc tctttctttc cagggaatta gccagacaca1800 acaacgggaa cgacacaccg aaccagacat gccc gccccg tgcgccctcc ccgctcgctg1860 cctttcctcc ctcttgtctc tccagagccg gatcttcaag gggagcctcc gtgcccccgg1920 ctgctcagtc cctccggtgt gcaggacccc ggaagtcctc cccgcacagc tctcgcttct1980 ctttgcagcc tgttt ctgcg ccggaccagt cgaggactct ggacagtaga ggccccggga2040 cgaccgagct ggaattcgcc accatggccc caaagaagaa gcggaaggtc ggtatccacg2100 gagtcccagc agccctcgaa ccaggtgaaa aaccttacaa atgtcctgaa tgtgggaaat2160 cattcagtcg cagcgacaac ctggtgagac atcaacgcac ccatacagga gaaaaacctt2220 ataaatgtcc agaatgtgga aagtccttct cacgagagga taacttgcac actcatcaac2280 gaacacatac tggtgaaaaa ccatacaagt gtcccgaatg tggtaaaagt tttagccgg a2340 gcgatgaact tgtccgacac caacgaaccc atacaggcga gaagccttac aaatgtcccg2400 agtgtggcaa gagcttctca caatcaggga atctgactga gcatcaacga actcataccg2460 gggaaaaacc ttacaagtgt ccagagtgtg ggaagagctt ttccacaagt ggacatctgg2 520 tacgccacca gaggacacat acaggggaga agccctacaa atgccccgaa tgcggtaaaa2580 gtttctctca gaatagtacc ctgaccgaac accagcgaac acacactggg aaaaaaacga2640 gtaaaaggcc ggcggccacg aaaaaggccg gccaggcaaa aaagaaaaag ggatcctacc2700 catacgacgt accagattac gctctcgagg acgcgctgga cgatttcgat ctcgacatgc2760 tgggttctga tgccctcgat gactttgacc tggatatgtt gggaagcgac gcattggatg2820 actttgatct g gacatgctc ggctccgatg ctctggacga tttcgatctc gatatgttat2880 aaactagtaa gcttgggtgg catccctgtg acccctcccc agtgcctctc ctggccctgg2940 aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc atcattttgt3000 ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg agcaaggggc3060

- 140 046157

aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc tggagtgcag 3120 tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc 3180 agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa tttttgtttt 3240 tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc taatctcagg 3300 tgatctacc accttggcct cccaaattgc tgggattaca ggcgtgaacc actngctccct 3360 tccctgtcct t 3371

<210> 69 <211> 4380 <212> DNA <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic polynucleotide

<400> 69 ggaggaagcc atcaactaaa ctacaatgac tgtaagatac aaaattggga atggtaacat 60 attttgaagt tctgttgaca taaagaatca tgatattaat gcccatggaa atgaaagggc 120 gatcaacact atggtttgaa aagggggaaa ttgtagagca cagatgtgtt cgtgtggcag 180 tgtgctgtct ctagcaatac tcagagaaga gagagaacaa tgaaattctg attggcccca 240 gtgtgagccc agatgaggtt cagctgccaa ctttctcttt cacatcttat gaaagtcatt 300 taagcacaac taactttttt tttttttttt tttttttgag acagagtctt gctctgttgc 360 ccaggacaga gtgcagtagt gactcaatct cggctcactg cagcctccac ctcctaggct 420 caaacggtcc tcctgcatca gcctcccaag tagctggaat tacaggagtg gccccacatg 480 cccagctaat ttttgtattt ttaatagata cggggggtttc accatatcac ccaggctggt 540 ctcgaactcc tggcctcaag tgatccacct gcctcggcct cccaaagtgc tggggattata 600 ggcgtcagcc actatgccca acccgaccaa ccttttttaa aataaatatt taaaaaattg 660 gtatttcaca tatatactag tatttacatt tatccacaca aaacggacgg gcctccgctg 720 aaccagtgag gccccagacg tgcgcataaa taacccctgc gtgctgcacc acctggggag 780 agggggga ccacggtaaa tggagcgagc gcatagcaaa agggacgcgg ggtccttttc 840 tctgccggtg gcactgggta gctgtggcca ggtgtggtac tttgatgggg cccagggctg 900 gagctcaagg aagcgtcgca gggtcacaga tctgggggaa ccccggggaa aagcactgag 960 gcaaaaccgc cgctcgtctc ctacaatata tgggggggg aggttgagta cgttctggat 1020 tactcataag accttttttt tttccttccg ggcgcaaaac cgtgagctgg atttataatc 1080 gccctataaa gctccagagg cggtcaggca cctgcagagg agccccgccg ctccgccgac 1140 tagctgcccc cgcgagcaac ggcctcgtga tttccccgcc gatccggtcc ccgcctcccc 1200

- 141 046157

actctgcccc cgcctacccc ggagccgtgc agccgcctct ccgaatctct ctcttctcct 1260 ggcgctcgcg tgcgagagg aactagcgag aacgaggaag cagctggagg tgacgccggg 1320 cagattacgc ctgtcagggc cgagccgagc ggatcgctgg gcgctgtgca gaggaaaggc 1380 ggggagtgccc ggctcgctgt cgcagagccg aggtgggtaa gctagcgacc acctggactt 1440 cccagcgccc aaccgtggct tttcagccag gtcctctcct cccgcggctt ctcaaccaac 1500 cccaccag cgccggccac ccaacctccc gaaatgagtg cttcctgccc cagcagccga 1560 aggcgctact aggaacggta acctgttact tttccagggg ccgtagtcga cccgctgccc 1620 gagttgctgt gcgactgcgc gcgcggggct agagtgcaag gtgactgtgg ttcttctctg 1680 gccaagtccg aggggagaacg taaagatatg ggcctttttc cccctctcac cttgtctcac 1740 caaagtccct agtccccgga gcagttagcc tctttctttc cagggaatta gccagacaca 1800 acaacgggaa ccagacaccg aaccagacat gcccgccccg tgcgccctcc ccgctcgctg 1860 cctttcctcc ctcttgtctc tccagagccg gatcttcaag gggagcctcc gtgcccccgg 1920 ctgctcagtc cctccggtgt gcaggacccc ggaagtcctc cccgcacagc tctcgcttct 1980 ctttgcagcc tgtttctgcg ccggaccagt cgaggactct ggacagtaga ggccccggga 2040 cgaccgagct ggaattcgcc accatggccc caaagaagaa gcggaaggtc ggtatccacg 2100 gagtcccagc agccctcgaa ccaggtgaaa aaccttacaa atgtcctgaa tgtgggaaat 2160 cattcagtcg cagcgacaac ctggtgagac atcaacgcac ccacagga gaaaaacctt 2220 aaaatgtcc agaatgtgga aagtccttct cacgagga taacttgcac actcatcaac 2280 gaacacatac tggtgaaaaa catacaagt gtcccgaatg tggtaaaagt tttagccgga 2340 gcgatgaact tgtccgacac caacgaaccc atacaggcga gaagccttac aaatgtcccg 2400 agtgtggcaa gagcttctca caatcaggga atctgactga gcatcaacga actcataccg 2460 gggaaaaacc ttacaagtgt ccagagtgtg ggaagagctt ttccacaagt ggacatctgg 2520 tacgccacca gaggacacat acaggggaga agccctacaa atgccccgaa tgcggtaaaa 2580 gtttctctca gaatagtacc ctgaccgaac accagcgaac acacactggg aaaaaaacga 2640 gtaaaaggcc ggcggccacg aaaaaggccg gccaggcaaa aaagaaaaag ggatcctacc 2700 catacgacgt accagattac gctctcgagg aggccagcgg ttccggacgg gctgacgcat 2760 tggacgattt tgatctggat atgctgggaa gtgacgccct cgatgatttt gaccttgaca 2820 tgcttggttc ggatgccctt gatgactttg acctcgacat gctcggcagt gacgcccttg 2880 atgatttcga cctggacatg ctgattaact ctagaagttc cggatctccg aaaaagaaac 2940 gcaaagttgg tagccagtac ctgcccgaca ccgacgaccg gcaccggatc gaggaaaagc 3000 ggaagcggac ctacgagaca ttcaagagca tcatgaagaa gtcccccttc agcggcccca 3060 ccgacctag acctccacct agaagaatcg ccgtgcccag cagatccagc gccagcgtgc 3120

- 142 046157

caaaacctgc cccccagcct taccccttca ccagcagcct gagcaccatc aactacgacg 3180 agttccctac catggtgttc cccagcggcc agatctctca ggcctctgct ctggctccag 3240 cccctcctca ggtgctgcct caggctcctg ctcctgcacc agctccagcc atggtgtctg 3300 cactggctca ggcaccagca cccgtgcctg tgctggctcc tggacctcca caggctgtgg 3360 ctccaccagc ccctaaacct acacaggccg gcgaggggcac actgtctgaa gctctgctgc 3420 agctgcagtt cgacgacgag gatctggggag ccctgctggg aaacagcacc gatcctgccg 3480 tgttcaccga cctggccagc gtggacaaca gcgagttcca gcagctgctg aaccagggca 3540 tccctgtggc ccctcaccacc accgagccca tgctgatgga ataccccgag gccatcaccc 3600 ggctcgtgac aggcgctcag aggcctcctg atccagctcc tgcccctctg ggagcaccag 3660 gcctgcctaa tggactgctg tctggcgacg aggacttcag ctctatcgcc gatatggatt 3720 tctcagcctt gctgggctct ggcagcggca gccggggattc cagggaaggg atgtttttgc 3780 cgaagcctga ggccggctcc gctattagtg acgtgtttga gggccgcgag gtgtgccagc 3840 caaaacgaat ccggccattt catcctccag gaagtccatg ggccaaccgc ccactccccg 3900 ccagcctcgc accaacacca accggtccag tacatgagcc agtcgggtca ctgaccccgg 3960 caccagtccc tcagccactg gatccagcgc ccgcagtgac tcccgaggcc agtcacctgt 4020 tggaggatcc cgatgaagag acgagccagg ctgtcaaagc ccttcggggag atggccgata 4080 ctgtgattcc ccagaaggaa gaggctgcaa tctgtggcca aatggacctt tcccatccgc 4140 ccccaagggg ccatctggat gagctgacaa cccacacttga gtccatgacc gaggatctga 4200 acctggactc acccctgacc ccggaattga acgagattct ggataccttc ctgaacgacg 4260 agtgcctctt gcatgccatg catatcagca caggactgtc catcttcgac acatctctgt 4320 tttaaactag taataaaaga tctttatttt cattagatct gtgtgttggt tttttgtgtg 4380

<210> 70 <211> 3332 <212> DNA <213> Artificial sequence <220>

<223> Description of the artificial sequence: Synthetic polynucleotide <400> 70 ggaggaagcc atcaactaaa ctacaatgac tgtaagatac aaaattggga atggtaacat60 attttgaagt tctgttgaca taaagaatca tgatattaat gcccatggaa atgaaagggc120 gatcaacact atggtttgaa aagggggaaa ttg tagagca cagatgtgtt cgtgtggcag180 tgtgctgtct ctagcaatac tcagagaaga gagagaacaa tgaaattctg attggcccca240 gtgtgagccc agatgaggtt cagctgccaa ctttctcttt cacatcttat gaaagtcatt300

- 143 046157

taagcacaac taactttttt tttttttttt tttttttgag acagagtctt gctctgttgc 360 ccaggacaga gtgcagtagt gactcaatct cggctcactg cagcctccac ctcctaggct 420 caaacggtcc tcctgcatca gcctcccaag tagctggaat tacaggagtg gccccacatg 480 cccagctaat ttttgtattt ttaatagata cggggggtttc accatatcac ccaggctggt 540 ctcgaactcc tggcctcaag tgatccacct gcctcggcct cccaaagtgc tggggattata 600 ggcgtcagcc actatgccca acccgaccaa ccttttttaa aataaatatt taaaaaattg 660 gtatttcaca tatatactag tatttacatt tatccacaca aaacggacgg gcctccgctg 720 aaccagtgag gccccagacg tgcgcataaa taacccctgc gtgctgcacc acctggggag 780 agggggga ccacggtaaa tggagcgagc gcatagcaaa agggacgcgg ggtccttttc 840 tctgccggtg gcactgggta gctgtggcca ggtgtggtac tttgatgggg cccagggctg 900 gagctcaagg aagcgtcgca gggtcacaga tctgggggaa ccccggggaa aagcactgag 960 gcaaaaccgc cgctcgtctc ctacaatata tgggggggg aggttgagta cgttctggat 1020 tactcataag accttttttt tttccttccg ggcgcaaaac cgtgagctgg atttataatc 1080 gccctataaa gctccagagg cggtcaggca cctgcagagg agccccgccg ctccgccgac 1140 tagctgcccc cgcgagcaac ggcctcgtga tttccccgcc gatccggtcc ccgcctcccc 1200 actctgcccc cgcctacccc ggagccgtgc agccgcctct ccgaatctct ctcttctcct 1260 ggcgctcgcg tgcgagagg aactagcgag aacgaggaag cagctggagg tgacgccggg 1320 cagattacgc ctgtcagggc cgagccgagc ggatcgctgg gcgctgtgca gaggaaaggc 1380 ggggagtgccc ggctcgctgt cgcagagccg aggtgggtaa gctagcgacc acctggactt 1440 cccagcgccc aaccgtggct tttcagccag gtcctctcct cccgcggctt ctcaaccaac 1500 cccaccag cgccggccac ccaacctccc gaaatgagtg cttcctgccc cagcagccga 1560 aggcgctact aggaacggta acctgttact tttccagggg ccgtagtcga cccgctgccc 1620 gagttgctgt gcgactgcgc gcgcggggct agagtgcaag gtgactgtgg ttcttctctg 1680 gccaagtccg aggggagaacg taaagatatg ggcctttttc cccctctcac cttgtctcac 1740 caaagtccct agtccccgga gcagttagcc tctttctttc cagggaatta gccagacaca 1800 acaacgggaa ccagacaccg aaccagacat gcccgccccg tgcgccctcc ccgctcgctg 1860 cctttcctcc ctcttgtctc tccagagccg gatcttcaag gggagcctcc gtgcccccgg 1920 ctgctcagtc cctccggtgt gcaggacccc ggaagtcctc cccgcacagc tctcgcttct 1980 ctttgcagcc tgtttctgcg ccggaccagt cgaggactct ggacagtaga ggccccggga 2040 cgaccgagct ggaattcgcc accatggccc caaagaagaa gcggaaggtc ggtatccacg 2100 gagtcccagc agccctcgaa ccaggtgaaa aaccttacaa atgtcctgaa tgtgggaaat 2160

- 144 046157

cattcagtcg cagcgacaac ctggtgagac atcaacgcac ccacagga gaaaaacctt 2220 aaaatgtcc agaatgtgga aagtccttct cacgagga taacttgcac actcatcaac 2280 gaacacatac tggtgaaaaa catacaagt gtcccgaatg tggtaaaagt tttagccgga 2340 gcgatgaact tgtccgacac caacgaaccc atacaggcga gaagccttac aaatgtcccg 2400 agtgtggcaa gagcttctca caatcaggga atctgactga gcatcaacga actcataccg 2460 gggaaaaacc ttacaagtgt ccagagtgtg ggaagagctt ttccacaagt ggacatctgg 2520 tacgccacca gaggacacat acaggggaga agccctacaa atgccccgaa tgcggtaaaa 2580 gtttctctca gaatagtacc ctgaccgaac accagcgaac acacactggg aaaaaaacga 2640 gtaaaaggcc ggcggccacg aaaaaggccg gccaggcaaa aaagaaaaag ggatccgacg 2700 cgctggacga tttcgatctc gacatgctgg gttctgatgc cctcgatgac tttgacctgg 2760 atatgttggg aagcgacgca ttggatgact ttgatctgga catgctcggc tccgatgctc 2820 tggacgattt cgatctcgat atgttataaa agcttgggtg gcatccctgt gacccctccc 2880 cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt gtcctaataa 2940 aattaagttg catcattttg tctgactagg tgtccttcta taatattatg gggtggaggg 3000 gggtggtatg gagcaagggg caagttggga agacaacctg tagggcctgc ggggtctatt 3060 gggaaccaag ctggagtgca gtggcacaat cttggctcac tgcaatctcc gcctcctggg 3120 ttcaagcgat tctcctgcct cagcctcccg agttgttggg attccaggca tgcatgacca 3180 ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg gccaggctgg 3240 tctccaactc ctaatctcag gtgatctacc caccttggcc tccaaattg ctggggattac 3300 aggcgtgaac cactgctccc ttccctgtcc tt 3332

<210> 71 <211> 3546 <212> DNA <213> Artificial sequence <220>

<223> Description of the artificial sequence: Synthetic polynucleotide <400> 71 ggaggaagcc atcaactaaa ctacaatgac tgtaagatac aaaattggga atggtaacat60 attttgaagt tctgttgaca taaagaatca tgatattaat gcccatggaa atgaaagggc120 gatcaacact atggtttgaa aagggggaaa ttg tagagca cagatgtgtt cgtgtggcag180 tgtgctgtct ctagcaatac tcagagaaga gagagaacaa tgaaattctg attggcccca240 gtgtgagccc agatgaggtt cagctgccaa ctttctcttt cacatcttat gaaagtcatt300 taagcacaac taactttttt tttttttt ttt tttttttgag acaagtctt gctctgttgc360

- 145 046157 ccaggacaga gtgcagtagt gactcaatct cggctcactg cagcctccac ctcctaggct 420 caaacggtcc tcctgcatca gcctcccaag tagctggaat tacaggagtg gcccaccatg480 cccagctaat ttttgtattt ttaatagata cgggggtttc accatatcac cca ggctggt540 ctcgaactcc tggcctcaag tgatccacct gcctcggcct cccaaagtgc tgggattata600 ggcgtcagcc actatgccca acccgaccaa ccttttttaa aataaatatt taaaaaattg660 gtatttcaca tatatactag tatttacatt tatccacaca aaacggacgg gcctccgctg72 0 aaccagtgag gccccagacg tgcgcataaa taacccctgc gtgctgcacc acctggggag780 agggggagga ccacggtaaa tggagcgagc gcatagcaaa agggacgcgg ggtccttttc840 tctgccggtg gcactggggta gctgtggcca ggtgtggtac tttgatgggg cccaggctg900 gagctcaagg aagcgtcgca gggtcacaga tctgggggaa ccccggggaa aagcactgag960 gcaaaaccgc cgctcgtctc ctacaatata tgggagggg aggttgagta cgttctggat1020 tactcataag accttttttt tttccttccg ggcgcaaaac cgtgagctgg atttataatc1080 gccctataaa gctccagagg cggtcaggca cctgcagagg agccccgccg ctccgccgac1140 tagctgcccc cgcgagcaac ggcctcgtga tttccccgcc gatccggtcc ccgcctcccc1200 actctgcccc cgcctacccc ggagccgtgc agccgcctct ccgaatctct ctcttctcct1260 ggcgctcgcg tgcgagaggg aactagcgag aacgaggaag cagctggagg tgacgccggg1320 cagattacgc ctgtcagggc cgagccgagc ggatcgctgg gcgctgtgca gaggaaaggc1380 gggagtgccc ggctcgctgt cgcagagccg aggtgggtaa gctagcgacc acctggactt1440 cccagcgccc aaccgtggct tttcagccag gtcctctcct cccgcgg ctt ctcaaccaac1500 cccacccag cgccggccac ccaacctccc gaaatgagtg cttcctgccc cagcagccga1560 aggcgctact aggaacggta acctgttact tttccagggg ccgtagtcga cccgctgccc1620 gagttgctgt gcgactgcgc gcgcggggct agag tgcaag gtgactgtgg ttcttctctg1680 gccaagtccg agggagaacg taaagatatg ggcctttttc cccctctcac cttgtctcac1740 caaagtccct agtccccgga gcagttagcc tctttctttc cagggaatta gccagacaca1800 acaacgggaa cgacacaccg aaccagacat gcccgccccg tgcgccctcc ccgctcgctg1860 cctttcctcc ctcttgtctc tccagagccg gatcttcaag gggagcctcc gtgcccccgg1920 ctgctcagtc cctccggtgt gcaggacccc ggaagtcctc cccgcacagc tctcgcttct1980 ctttg cagcc tgtttctgcg ccggaccagt cgaggactct ggacagtaga ggccccggga2040 cgaccgagct ggaattcgcc accatggccc caaagaagaa gcggaaggtc ggtatccacg2100 gagtcccagc agccctcgaa ccaggtgaaa aaccttacaa atgtcctgaa tgtgggaaat2160 cattcagtcg cagcgacaac ctggtgagac atcaacgcac ccatacagga gaaaaacctt2220 ataaatgtcc agaatgtgga aagtccttct cacgagagga taacttgcac actcatcaac2280

- 146 046157

gaacacatac tggtgaaaaa catacaagt gtcccgaatg tggtaaaagt tttagccgga 2340 gcgatgaact tgtccgacac caacgaaccc atacaggcga gaagccttac aaatgtcccg 2400 agtgtggcaa gagcttctca caatcaggga atctgactga gcatcaacga actcataccg 2460 gggaaaaacc ttacaagtgt ccagagtgtg ggaagagctt ttccacaagt ggacatctgg 2520 tacgccacca gaggacacat acaggggaga agccctacaa atgccccgaa tgcggtaaaa 2580 gtttctctca gaatagtacc ctgaccgaac accagcgaac acacactggg aaaaaaacga 2640 gtaaaaggcc ggcggccacg aaaaaggccg gccaggcaaa aaagaaaaag ggatccgacg 2700 cgctggacga tttcgatctc gacatgctgg gttctgatgc cctcgatgac tttgacctgg 2760 atatgttggg aagcgacgca ttggatgact ttgatctgga catgctcggc tccgatgctc 2820 tggacgattt cgatctcgat atgttataaa aagagaccgg ttcactgtga cagtaaaaga 2880 gaccggttca ctgtgagaat gaaagagacc ggttcactgt gatcggaaaa gagaccggtt 2940 cactgtgagc ggccttgaaa cccagcagac aatgtagctc agtagaaacc cagcagacaa 3000 tgtagctgaa tggaaaccca gcagacaatg tagcttcgga gaaacccagc agacaatgta 3060 gctaagcttg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc cctggaagtt 3120 gccactccag tgccccaccag ccttgtccta aaaaattaa gttgcatcat tttgtctgac 3180 taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa ggggcaagtt 3240 gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag tgcagtggca 3300 caatcttggc tcactgcaat ctccgcctcc tggggttcaag cgattctcct gcctcagcct 3360 cccgagttgt tggggattcca ggcatgcatg accaggctca gctaattttt gtttttttgg 3420 tagagacgggg gtttcaccat attggccagg ctggtctcca actcctaatc tcaggtgatc 3480 taccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc tcccttccct 3540 gtcctt 3546

<210> 72 <211> 1707 <212> DNA <213> Artificial sequence <220>

<223> Description of the artificial sequence: Synthetic polynucleotide <400> 72 atggccgcag atcacctgat gctggctgaa ggctacagac tggtgcagcg gcctccatct60 gccgctgccg cccacggccc ccacgccctg agaacactgc ccccctacgc cggccctggt120 cttgatagcg gactcagacc tagaggcgcc cctctgggcc ctccacctcc aagacagcct180 ggagccctgg cctacggcgc cttcggccct ccttctagct tccagccctt ccccgccgtg240

- 147 046157

cctcctccag ccgctggcat cgccacctg cagcctgtgg ccacccctta ccccggagaaga 300 gccgccgccc ctccaaacgc ccctggcgga cctcctggcc cccagcctgc tccaagcgcc 360 gctgcccctc cacctcctgc tcatgccctg ggcggcatgg acgccgagct gatcgacgag 420 gaagccctga ccagcctgga actggaactg ggcctgcaca gagtgcggga actgcctgag 480 ctgttcctgg gacagagcga gttcgactgc ttcagcgacc tggggcagcgc ccctcctgcc 540 ggctctgtgt cctgcgccga ccacctgatg ctcgccgagg gctaccgcct ggtgcagagg 600 ccgccgtccg ccgccgccgc ccatggccct catgcgctcc ggactctgcc gccgtacgcg 660 ggcccgggcc tggacagtgg gctgaggccg cggggggctc cgctggggcc gccgccgccc 720 cgccaacccg gggccctggc gtacggggcc ttcggggccgc cgtcctcctt ccagcccttt 780 ccggccgtgc ctccgccggc cgcgggcatc gcgcacctgc agcctgtggc gacgccgtac 840 cccggccgcg ccgccgcgcc ccccaacgct ccgggaggcc ccccggggccc gcagccggcc 900 ccaagcgccg cagccccgcc gccgcccgcg cacgccctgg gcggcatgga cgccgaactc 960 atcgacgagg aggcgctgac gtcgctggag ctggagctgg ggctgcaccg cgtgcgcgag 1020 ctgcccgagc tgttcctggg ccagagcgag ttcgactgct tctcggactt ggggtccgcg 1080 ccgcccgccg gctccgtgag ctgccagtcc cagctcatca aacccagccg catgcgcaag 1140 taccccaacc ggcccagcaa gacgcccccc cacgaacgcc cttacgcttg cccagtggag 1200 tcctgtgatc gccgcttctc ccgcagcgac aacctggtga gacacatccg catccacaca 1260 ggccagaagc ccttccagtg ccgcatctgc atgagaaact tcagccgaga ggataacttg 1320 cacactcaca tccgcaccca cacaggcgaa aagcccttcg cctgcgacat ctgtggaaga 1380 aagtttgccc ggagcgatga acttgtccga cataccaaga tccacttgcg gcagaaggac 1440 cgcccttacg cttgcccagt ggagtcctgt gatcgccgct tctcccaatc agggaatctg 1500 actgagcaca tccgcatcca cacaggccag aagcccttcc agtgccgcat ctgcatgaga 1560 aacttcagca caagtggaca tctggtacgc cacatccgca cccacacagg cgaaaagccc 1620 ttcgcctgcg acatctgtgg aagaaagttt gccagaata gtaccctgac cgaacatacc 1680 aagatccact tgcggcagaa ggacaag 1707

<210> 73 <211> 1755 <212> DNA <213> Artificial sequence <220>

<223> Description of the artificial sequence: Synthetic polynucleotide <400> 73 atggccgcag atcacctgat gctggctgaa ggctacagac tggtgcagcg gcctccatct 60

- 148 046157 gccgctgccg cccacggccc ccacgccctg agaacactgc ccccctacgc cggccctggt 120 cttgatagcg gactcagacc tagaggcgcc cctctgggcc ctccacctcc aagacagcct 180 ggagccctgg cctacggcgc cttcggccct ccttctagct tccag ccctt ccccgccgtg 240 cctcctccag ctgctggcat cgcccacctg cagcctgtgg ccacccctta ccccggaaga 300 gccgccgccc ctccaaacgc ccctggcgga cctcctggcc cccagcctgc tccaagcgcc 360 gctgcccctc cacctcctgc tcatgccctg ggcgg catgg acgccgagct gatcgacgag 420 gaagccctga ccagcctgga actggaactg ggcctgcaca gagtgcggga actgcctgag 480 ctgttcctgg gacagagcga gttcgactgc ttcagcgacc tgggcagcgc ccctcctgcc 540 ggctctgtgt cctgcggcgg cagcggcggc ggaagcggcg ccgaccacct gatgctcgcc 600 gagggctacc gcctggtgca gagccgccg t ccgccgccg ccgcccatgg ccctcatgcg 660 ctccggactc tgccgccgta cgcgggcccg ggcctggaca gtgggctgag gccgcggggg 720 gctccgctgg ggccgccgcc gccccgccaa cccggggccc tggcgtacgg ggccttcggg 780 ccgccgtcct ccttcca gcc ctttccggcc gtgcctccgc cggccgcggg catcgcgcac 840 ctgcagcctg tggcgacgcc gtaccccggc cgcgcggccg cgccccccaa cgctccggga 900 ggccccccgg gcccgcagcc ggccccaagc gccgcagccc cgccgccgcc cgcgcacgcc 960 ctgggcggca tggacgccga actcatcgac gaggaggcgc tgacgtcgct ggagctggag 1020 ctggggctgc accgcgtgcg cgagctgccc gagctgttcc t gggccagag cgagttcgac 1080 tgcttctcgg acttggggtc cgcgccgccc gccggctccg tgagctgcgg tggttctggt 1140 ggtggttctg gtcagtccca gctcatcaaa cccagccgca tgcgcaagta ccccaaccgg 1200 cccagcaaga cgccccccca cgaacgccct tacgcttgcc cagtggagtc ctgtgatcgc 1260 cgcttctccc gcagcgacaa cctggtgaga cacatccgca tccacacagg ccagaagccc 1320 ttccagtgcc gcatctgcat gagaaacttc agccgagagg ataacttgca cactcacatc 1380 cgcacccaca caggcgaaaa gcccttcgcc tgcgacatct gtggaagaaa gtttgcccgg 1440 agcgatgaac ttgtccgaca taccaagatc cacttgcggc agaaggaccg cccttac gct 1500 tgcccagtgg agtcctgtga tcgccgcttc tcccaatcag ggaatctgac tgagcacatc 1560 cgcatccaca caggccagaa gcccttccag tgccgcatct gcatgagaaa cttcagcaca 1620 agtggacatc tggtacgcca catccgcacc cacacaggcg aaaagccctt c gcctgcgac 1680 atctgtggaa gaaagtttgc ccagaatagt accctgaccg aacataccaa gatccacttg 1740 cggcagaagg acaag 1755 <210> 74 <211> 3438 <212> DNA

- 149 046157 <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic polynucleotide

<400> 74 ggaggaagcc atcaactaaa ctacaatgac tgtaagatac aaaattggga atggtaacat 60 attttgaagt tctgttgaca taaagaatca tgatattaat gcccatggaa atgaaagggc 120 gatcaacact atggtttgaa aagggggaaa ttgtagagca cagatgtgtt cgtgtggcag 180 tgtgctgtct ctagcaatac tcagagaaga gagagaacaa tgaaattctg attggcccca 240 gtgtgagccc agatgaggtt cagctgccaa ctttctcttt cacatcttat gaaagtcatt 300 taagcacaac taactttttt tttttttttt tttttttgag acagagtctt gctctgttgc 360 ccaggacaga gtgcagtagt gactcaatct cggctcactg cagcctccac ctcctaggct 420 caaacggtcc tcctgcatca gcctcccaag tagctggaat tacaggagtg gccccacatg 480 cccagctaat ttttgtattt ttaatagata cggggggtttc accatatcac ccaggctggt 540 ctcgaactcc tggcctcaag tgatccacct gcctcggcct cccaaagtgc tggggattata 600 ggcgtcagcc actatgccca acccgaccaa ccttttttaa aataaatatt taaaaaattg 660 gtatttcaca tatatactag tatttacatt tatccacaca aaacggacgg gcctccgctg 720 aaccagtgag gccccagacg tgcgcataaa taacccctgc gtgctgcacc acctggggag 780 agggggga ccacggtaaa tggagcgagc gcatagcaaa agggacgcgg ggtccttttc 840 tctgccggtg gcactgggta gctgtggcca ggtgtggtac tttgatgggg cccagggctg 900 gagctcaagg aagcgtcgca gggtcacaga tctgggggaa ccccggggaa aagcactgag 960 gcaaaaccgc cgctcgtctc ctacaatata tgggggggg aggttgagta cgttctggat 1020 tactcataag accttttttt tttccttccg ggcgcaaaac cgtgagctgg atttataatc 1080 gccctataaa gctccagagg cggtcaggca cctgcagagg agccccgccg ctccgccgac 1140 tagctgcccc cgcgagcaac ggcctcgtga tttccccgcc gatccggtcc ccgcctcccc 1200 actctgcccc cgcctacccc ggagccgtgc agccgcctct ccgaatctct ctcttctcct 1260 ggcgctcgcg tgcgagagg aactagcgag aacgaggaag cagctggagg tgacgccggg 1320 cagattacgc ctgtcagggc cgagccgagc ggatcgctgg gcgctgtgca gaggaaaggc 1380 ggggagtgccc ggctcgctgt cgcagagccg aggtgggtaa gctagcgacc acctggactt 1440 cccagcgccc aaccgtggct tttcagccag gtcctctcct cccgcggctt ctcaaccaac 1500 cccaccag cgccggccac ccaacctccc gaaatgagtg cttcctgccc cagcagccga 1560 aggcgctact aggaacggta acctgttact tttccagggg ccgtagtcga cccgctgccc 1620 gagttgctgt gcgactgcgc gcgcggggct agagtgcaag gtgactgtgg ttcttctctg 1680

- 150 046157 gccaagtccg agggagaacg taaagatatg ggcctttttc cccctctcac cttgtctcac 1740 caaagtccct agtccccgga gcagttagcc tctttctttc cagggaatta gccagacaca1800 acaacgggaa cgagacaccg aaccagacat gcccgccccg t gcgccctcc ccgctcgctg1860 cctttcctcc ctcttgtctc tccagagccg gatcttcaag gggagcctcc gtgcccccgg1920 ctgctcagtc cctccggtgt gcaggacccc ggaagtcctc cccgcacagc tctcgcttct1980 ctttgcagcc tgtttctgcg ccggaccagt cgaggactct ggacagtaga ggccccggga2040 cgaccgagct ggaattcgcc accatggccc caaagaagaa gcggaaggtc ggtatccacg2100 gagtcccagc agccctcgaa ccaggtgaaa aaccttacaa atgtcctgaa tgtgggaaat2160 cattcagtcg cagcgacaac ctggtgagac atcaacgcac ccatacagga gaaaaacctt2220 ataaatgtcc agaatgtgga aagtccttct cacgagagga taacttgcac actcatcaac2280 gaacacatac tggtgaaaaa ccatacaagt g tcccgaatg tggtaaaagt tttagccgga2340 gcgatgaact tgtccgacac caacgaaccc atacaggcga gaagccttac aaatgtcccg2400 agtgtggcaa gagcttctca caatcaggga atctgactga gcatcaacga actcataccg2460 gggaaaaacc ttacaagtgt cccagtgtg ggaagagctt ttccacaagt ggacatctgg2520 tacgccacca gaggacacat acaggggaga agccctacaa atgccccgaa tgcggtaaaa2580 gtttctctca gaatagtacc ctgaccgaac accagcgaac acacactggg aaaaaaacga2640 gtaaaaggcc ggcggccacg aaaaaggccg gccaggcaaa aaagaaaaag ggatcctacc2700 catacgacgt accagattac gctctcgagg acgcgctgga cgatttcgat ctcgacatgc2760 tgggttctga tgccctcgat gactttgacc tggatatgtt gggaagcgac gcatt ggatg2820 actttgatct ggacatgctc ggctccgatg ctctggacga tttcgatctc gatatgttat2880 aaactagtga aacccagcag acaatgtagc tagacccagt agccagatgt agctaaagag2940 accggttcac tgtgaaagct tgggtggcat ccctgtgacc cctccccagt gcctctcctg3000 gccctggaag ttgccactcc agtgcccacc agccttgtcc taataaaatt aagttgcatc3060 attttgtctg actaggtgtc cttctataat attatggggt ggaggggggt ggtatggagc3120 aaggggcaag ttgggaagac aacctgtagg gcctgcgggg tctattggga accaagctgg3180 agtgcagtgg cacaatcttg gctcactgca atctccgcct cctgggttca agcgattctc3240 ctgcctcagc ctcccgagtt gttgggattc caggcatgca tgaccaggct cagctaattt3300 ttgttttttt ggtagagacg gggtttcacc atattggcca ggctggtctc caactcctaa3360 tctcaggtga tctacccacc ttggcctccc aaattgctgg gattacaggc gtgaaccact 3420 gctcccttcc ctgtcctt3438 <210> 75 <211> 3505

- 151 046157 <212> DNA <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic polynucleotide

<400> 75 ggaggaagcc atcaactaaa ctacaatgac tgtaagatac aaaattggga atggtaacat 60 attttgaagt tctgttgaca taaagaatca tgatattaat gcccatggaa atgaaagggc 120 gatcaacact atggtttgaa aagggggaaa ttgtagagca cagatgtgtt cgtgtggcag 180 tgtgctgtct ctagcaatac tcagagaaga gagagaacaa tgaaattctg attggcccca 240 gtgtgagccc agatgaggtt cagctgccaa ctttctcttt cacatcttat gaaagtcatt 300 taagcacaac taactttttt tttttttttt tttttttgag acagagtctt gctctgttgc 360 ccaggacaga gtgcagtagt gactcaatct cggctcactg cagcctccac ctcctaggct 420 caaacggtcc tcctgcatca gcctcccaag tagctggaat tacaggagtg gccccacatg 480 cccagctaat ttttgtattt ttaatagata cggggggtttc accatatcac ccaggctggt 540 ctcgaactcc tggcctcaag tgatccacct gcctcggcct cccaaagtgc tggggattata 600 ggcgtcagcc actatgccca acccgaccaa ccttttttaa aataaatatt taaaaaattg 660 gtatttcaca tatatactag tatttacatt tatccacaca aaacggacgg gcctccgctg 720 aaccagtgag gccccagacg tgcgcataaa taacccctgc gtgctgcacc acctggggag 780 agggggga ccacggtaaa tggagcgagc gcatagcaaa agggacgcgg ggtccttttc 840 tctgccggtg gcactgggta gctgtggcca ggtgtggtac tttgatgggg cccagggctg 900 gagctcaagg aagcgtcgca gggtcacaga tctgggggaa ccccggggaa aagcactgag 960 gcaaaaccgc cgctcgtctc ctacaatata tgggggggg aggttgagta cgttctggat 1020 tactcataag accttttttt tttccttccg ggcgcaaaac cgtgagctgg atttataatc 1080 gccctataaa gctccagagg cggtcaggca cctgcagagg agccccgccg ctccgccgac 1140 tagctgcccc cgcgagcaac ggcctcgtga tttccccgcc gatccggtcc ccgcctcccc 1200 actctgcccc cgcctacccc ggagccgtgc agccgcctct ccgaatctct ctcttctcct 1260 ggcgctcgcg tgcgagagg aactagcgag aacgaggaag cagctggagg tgacgccggg 1320 cagattacgc ctgtcagggc cgagccgagc ggatcgctgg gcgctgtgca gaggaaaggc 1380 ggggagtgccc ggctcgctgt cgcagagccg aggtgggtaa gctagcgacc acctggactt 1440 cccagcgccc aaccgtggct tttcagccag gtcctctcct cccgcggctt ctcaaccaac 1500 cccaccag cgccggccac ccaacctccc gaaatgagtg cttcctgccc cagcagccga 1560 aggcgctact aggaacggta acctgttact tttccagggg ccgtagtcga cccgctgccc 1620 gagttgctgt gcgactgcgc gcgcggggct agagtgcaag gtgactgtgg ttcttctctg 1680

- 152 046157 gccaagtccg agggagaacg taaagatatg ggcctttttc cccctctcac cttgtctcac 1740 caaagtccct agtccccgga gcagttagcc tctttctttc cagggaatta gccagacaca1800 acaacgggaa cgagacaccg aaccagacat gcccgccccg t gcgccctcc ccgctcgctg1860 cctttcctcc ctcttgtctc tccagagccg gatcttcaag gggagcctcc gtgcccccgg1920 ctgctcagtc cctccggtgt gcaggacccc ggaagtcctc cccgcacagc tctcgcttct1980 ctttgcagcc tgtttctgcg ccggaccagt cgaggactct ggacagtaga ggccccggga2040 cgaccgagct ggaattcgcc accatggccc caaagaagaa gcggaaggtc ggtatccacg2100 gagtcccagc agccctcgaa ccaggtgaaa aaccttacaa atgtcctgaa tgtgggaaat2160 cattcagtcg cagcgacaac ctggtgagac atcaacgcac ccatacagga gaaaaacctt2220 ataaatgtcc agaatgtgga aagtccttct cacgagagga taacttgcac actcatcaac2280 gaacacatac tggtgaaaaa ccatacaagt g tcccgaatg tggtaaaagt tttagccgga2340 gcgatgaact tgtccgacac caacgaaccc atacaggcga gaagccttac aaatgtcccg2400 agtgtggcaa gagcttctca caatcaggga atctgactga gcatcaacga actcataccg2460 gggaaaaacc ttacaagtgt ccagagtgtg ggaagagctt ttccacaagt ggacatctgg2520 tacgccacca gaggacacat acaggggaga agccctacaa atgccccgaa tgcggtaaaa2580 gtttctctca gaatagtacc ctgaccgaac accagcgaac acacactggg aaaaaaacga2640 gtaaaaggcc ggcggccacg aaaaaggccg gccaggcaaa aaagaaaaag ggatcctacc2700 catacgacgt accagattac gctctcgagg acgcgctgga cgatttcgat ctcgacatgc2760 tgggttctga tgccctcgat gactttgacc tggatatgtt gggaagcgac gcatt ggatg2820 actttgatct ggacatgctc ggctccgatg ctctggacga tttcgatctc gatatgttat2880 aaactagtga aacccagcag acaatgtagc tagacccagt agccagatgt agctaaagag2940 accggttcac tgtgagaaac ccagcagaca atgtagctag acccagtagc cagat gtagc3000 taaagagacc ggttcactgt gaaagcttgg gtggcatccc tgtgacccct ccccagtgcc3060 tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag3120 ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt3180 atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc3240 aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc3300 gattctcctg cctcagcct c ccgagttgtt gggattccag gcatgcatga ccaggctcag3360 ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa3420 ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg3480 aaccactgct cc cttccctg tcctt3505

- 153 046157 <210> 76 <211> 3804 <212> DNA <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic polynucleotide

<400> 76 ggaggaagcc atcaactaaa ctacaatgac tgtaagatac aaaattggga atggtaacat 60 attttgaagt tctgttgaca taaagaatca tgatattaat gcccatggaa atgaaagggc 120 gatcaacact atggtttgaa aagggggaaa ttgtagagca cagatgtgtt cgtgtggcag 180 tgtgctgtct ctagcaatac tcagagaaga gagagaacaa tgaaattctg attggcccca 240 gtgtgagccc agatgaggtt cagctgccaa ctttctcttt cacatcttat gaaagtcatt 300 taagcacaac taactttttt tttttttttt tttttttgag acagagtctt gctctgttgc 360 ccaggacaga gtgcagtagt gactcaatct cggctcactg cagcctccac ctcctaggct 420 caaacggtcc tcctgcatca gcctcccaag tagctggaat tacaggagtg gccccacatg 480 cccagctaat ttttgtattt ttaatagata cggggggtttc accatatcac ccaggctggt 540 ctcgaactcc tggcctcaag tgatccacct gcctcggcct cccaaagtgc tggggattata 600 ggcgtcagcc actatgccca acccgaccaa ccttttttaa aataaatatt taaaaaattg 660 gtatttcaca tatatactag tatttacatt tatccacaca aaacggacgg gcctccgctg 720 aaccagtgag gccccagacg tgcgcataaa taacccctgc gtgctgcacc acctggggag 780 agggggga ccacggtaaa tggagcgagc gcatagcaaa agggacgcgg ggtccttttc 840 tctgccggtg gcactgggta gctgtggcca ggtgtggtac tttgatgggg cccagggctg 900 gagctcaagg aagcgtcgca gggtcacaga tctgggggaa ccccggggaa aagcactgag 960 gcaaaaccgc cgctcgtctc ctacaatata tgggggggg aggttgagta cgttctggat 1020 tactcataag accttttttt tttccttccg ggcgcaaaac cgtgagctgg atttataatc 1080 gccctataaa gctccagagg cggtcaggca cctgcagagg agccccgccg ctccgccgac 1140 tagctgcccc cgcgagcaac ggcctcgtga tttccccgcc gatccggtcc ccgcctcccc 1200 actctgcccc cgcctacccc ggagccgtgc agccgcctct ccgaatctct ctcttctcct 1260 ggcgctcgcg tgcgagagg aactagcgag aacgaggaag cagctggagg tgacgccggg 1320 cagattacgc ctgtcagggc cgagccgagc ggatcgctgg gcgctgtgca gaggaaaggc 1380 ggggagtgccc ggctcgctgt cgcagagccg aggtgggtaa gctagcgacc acctggactt 1440 cccagcgccc aaccgtggct tttcagccag gtcctctcct cccgcggctt ctcaaccaac 1500 cccaccag cgccggccac ccaacctccc gaaatgagtg cttcctgccc cagcagccga 1560

- 154 046157

aggcgctact aggaacggta acctgttact tttccagggg ccgtagtcga cccgctgccc 1620 gagttgctgt gcgactgcgc gcgcggggct agagtgcaag gtgactgtgg ttcttctctg 1680 gccaagtccg aggggagaacg taaagatatg ggcctttttc cccctctcac cttgtctcac 1740 caaagtccct agtccccgga gcagttagcc tctttctttc cagggaatta gccagacaca 1800 acaacgggaa ccagacaccg aaccagacat gcccgccccg tgcgccctcc ccgctcgctg 1860 cctttcctcc ctcttgtctc tccagagccg gatcttcaag gggagcctcc gtgcccccgg 1920 ctgctcagtc cctccggtgt gcaggacccc ggaagtcctc cccgcacagc tctcgcttct 1980 ctttgcagcc tgtttctgcg ccggaccagt cgaggactct ggacagtaga ggccccggga 2040 cgaccgagct ggaattcgcc accatggccg ccgaccacct gatgctcgcc gagggctacc 2100 gcctggtgca gaggccgccg tccgccgccg ccgcccatgg ccctcatgcg ctccggactc 2160 tgccgccgta cgcgggcccg ggcctggac gtggggctgag gccgcgggggg gctccgctgg 2220 ggccgccgcc gccccgccaa cccggggccc tggcgtacgg ggccttcggg ccgccgtcct 2280 ccttccagcc ctttccggcc gtgcctccgc cggccgcggg catcgcgcac ctgcagcctg 2340 tggcgacgcc gtaccccggc cgcgcggccg cgccccccaa cgctccggga ggccccccgg 2400 gcccgcagcc ggccccaagc gccgcagccc cgccgccgcc cgcgcacgcc ctggggcggca 2460 tggacgccga actcatcgac gaggaggcgc tgacgtcgct ggagctggag ctggggctgc 2520 accgcgtgcg cgagctgccc gagctgttcc tggggccagag cgagttcgac tgcttctcgg 2580 acttggggtc cgcgccgccc gccggctccg tgagctgcgg tggttctggt ggtggttctg 2640 gtcagtccca gctcatcaaa cccagccgca tgcgcaagta ccccaaccgg cccagcaaga 2700 cgcccccccca cgaacgccct tacgcttgcc cagtggagtc ctgtgatcgc cgcttctccc 2760 gcagcgacaa cctggtgaga cacatccgca tccacacagg ccagaagccc ttccagtgcc 2820 gcatctgcat gagaaacttc agccgagagg ataacttgca cactcacatc cgcaccaca 2880 caggcgaaaa gcccttcgcc tgcgacatct gtggaagaaa gtttgcccgg agcgatgaac 2940 ttgtccgaca taccaagatc cacttgcggc agaaggaccg cccttacgct tgccagtgg 3000 agtcctgtga tcgccgcttc tccaatcag ggaatctgac tgagcacatc cgcatccaca 3060 caggccagaa gcccttccag tgccgcatct gcatgagaaa cttcagcaca agtggacatc 3120 tggtacgcca catccgcacc cacacaggcg aaaagccctt cgcctgcgac atctgtggaa 3180 gaaagtttgc ccagaatagt accctgaccg aacataccaa gatccacttg cggcagaagg 3240 acaagtaact cgaggaaacc cagcagacaa tgtagctaga cccagtagcc agatgtagct 3300 aaagagaccg gttcactgtg aaagcttgggg tggcatccct gtgacccctc cccagtgcct 3360 ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat aaaattaagt 3420 tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag gggggtggta 3480

- 155 046157

tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta ttgggaacca 3540 agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg ggttcaagcg 3600 attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac caggctcagc 3660 taatttttgt ttttttggta gagacggggt ttcaccatat tggccaggct ggtctccaac 3720 tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctggggatt acaggcgtga 3780 accactgctc ccttccctgt cctt 3804

<210> 77 <211> 176 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic polypeptide <400> 77

Leu Glu Pro Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser 1 5 1015

Phe Ser Arg Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly 20 2530

Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Glu 35 4045

Asp Asn Leu His Thr His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr 50 5560

Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu Val 65 70 7580

Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu 85 9095

Cys Gly Lys Ser Phe Ser Gln Ser Gly Asn Leu Thr Glu His Gln Arg

100 105110

Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser

115 120125

Phe Ser Thr Ser Gly His Leu Val Arg His Gln Arg Thr His Thr Gly

130 135140

Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Asn

145 150 155160

- 156 046157

Ser Thr Leu Thr Glu His Gln Arg Thr His Thr Gly Lys Lys Thr Ser

165 170 175 <210> 78 <211> 176 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic polypeptide <400> 78

Leu Glu Pro Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser

5 1015

Phe Ser Thr Lys Asn Ser Leu Thr Glu His Gln Arg Thr His ThrGly

2530

Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ala 35 4045

Asp Asn Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr 50 5560

Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Leu Ala His LeuArg

70 7580

Ala His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys ProGlu

9095

Cys Gly Lys Ser Phe Ser Thr Lys Asn Ser Leu Thr Glu His Gln Arg 100 105110

Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser 115 120125

Phe Ser Gln Ala Gly His Leu Ala Ser His Gln Arg Thr His Thr Gly 130 135140

Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser ThrHis

145 150 155160

Leu Asp Leu Ile Arg His Gln Arg Thr His Thr Gly Lys Lys ThrSer

165 170175 <210> 79 <211> 176 <212> Protein <213> Artificial sequence

- 157 046157 <220>

<223> Description of the artificial sequence polypeptide <400> 79

Leu Glu Pro Gly Glu Lys Pro Tyr Lys Cys Pro Glu

5 10

Synthetic

Cys Gly Lys Ser 15

Phe Ser Gln Ala Gly His Leu Ala Ser His Gln Arg 20 25

Thr His Thr Gly 30

Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser 35 40

Phe Ser Arg Glu 45

Asp Asn Leu His Thr His Gln Arg Thr His Thr Gly

55 60

Glu Lys Pro Tyr

Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser 65 70 75

Gly Asn Leu Thr

Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr

90

Lys Cys Pro Glu

Cys Gly Lys Ser Phe Ser Thr His Leu Asp Leu Ile

100 105

Arg His Gln Arg 110

Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu

115 120

Cys Gly Lys Ser 125

Phe Ser Gln Lys Ser Ser Leu Ile Ala His Gln Arg

130 135 140

Thr His Thr Gly

Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser

145 150 155

Phe Ser Gln Ala

160

Gly His Leu Ala Ser His Gln Arg Thr His Thr Gly

165 170

Lys Lys Thr Ser 175 <210> 80 <211> 176 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial sequence polypeptide <400> 80

Leu Glu Pro Gly Glu Lys Pro Tyr Lys Cys Pro Glu

5 10

Synthetic

Cys Gly Lys Ser 15

- 158 046157

Phe Ser Thr Thr Gly Asn Leu Thr

Val His Gln Arg

Thr His Thr Gly 30

Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser 35 40

Phe Ser Thr Ser 45

Gly Glu Leu Val Arg His Gln Arg Thr His Thr Gly

55 60

Glu Lys Pro Tyr

Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Glu 65 70 75

Asp Asn Leu His 80

Thr His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr

90

Lys Cys Pro Glu

Cys Gly Lys Ser Phe Ser Thr Ser Gly Asn Leu Thr

100 105

Glu His Gln Arg 110

Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu

115 120

Cys Gly Lys Ser 125

Phe Ser Gln Ser Ser Ser Leu Val Arg His Gln Arg

130 135 140

Thr His Thr Gly

Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser

145 150 155

Phe Ser Gln Arg

160

Ala Asn Leu Arg Ala His Gln Arg Thr His Thr Gly

165 170

Lys Lys Thr Ser

175 <210> 81 <211> 176 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial sequence

Synthetic polypeptide <400> 81

Leu Glu Pro Gly Glu Lys

5

Phe Ser Ser Arg Arg Thr

Glu Lys Pro Tyr Lys Cys 35

Pro Tyr Lys Cys Pro Glu 10

Cys Arg Ala His Gln Arg 25

Pro Glu Cys Gly Lys Ser 40

Cys Gly Lys Ser 15

Thr His Thr Gly 30

Phe Ser Thr Thr 45

- 159 046157

Gly Ala Leu Thr Glu His Gln Arg

55

Thr His Thr Gly

Glu Lys Pro Tyr

Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser 65 70 75

Asp Glu Leu Val 80

Arg His Gln Arg Thr His 85

Thr Gly Glu Lys Pro Tyr

Lys Cys Pro Glu

Cys Gly Lys

Ser Phe

100

Ser Arg Asn Asp Ala Leu Thr

105

Glu His Gln Arg 110

Thr His

Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu 115 120

Cys Gly Lys Ser 125

Phe Ser Gln Ser Gly Asp Leu Arg Arg His Gln Arg

130 135 140

Thr His Thr Gly

Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser

145 150 155

Phe Ser Thr Ser

160

His Ser Leu Thr Glu His Gln Arg Thr His Thr Gly

165 170

Lys Lys Thr Ser 175 <210> 82 <211> 176 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial sequence

Synthetic polypeptide <400> 82

Leu Glu Pro Gly 1

Glu Lys Pro Tyr 5

Lys Cys Pro Glu 10

Cys Gly Lys Ser 15

Phe Ser Arg Lys

Asp Asn Leu Lys

Asn His Gln Arg 25

Thr His Thr Gly 30

Glu Lys Pro Tyr 35

Lys Cys Pro Glu 40

Cys Gly Lys Ser

Phe Ser Asp Pro 45

Gly Ala Leu Val 50

Arg His Gln Arg 55

Thr His Thr Gly

Glu Lys Pro Tyr

Lys Cys Pro Glu 65

Cys Gly Lys Ser 70

Phe Ser Arg Glu 75

Asp Asn Leu His 80

Thr His Gln Arg

Thr His Thr Gly

Glu Lys Pro Tyr

Lys Cys Pro Glu

- 160 046157

90

Cys Gly Lys Ser Phe Ser Asp Pro Gly Ala Leu Val

100 105

Arg His Gln Arg 110

Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu

115 120

Cys Gly Lys Ser 125

Phe Ser Thr Ser Gly Glu Leu Val Arg His Gln Arg

130 135 140

Thr His Thr Gly

Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser

145 150 155

Phe Ser Arg Lys

160

Asp Asn Leu Lys Asn His Gln Arg Thr His Thr Gly

165 170

Lys Lys Thr Ser

175 <210> 83 <211> 232 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial sequence

Synthetic polypeptide <400> 83

Leu Glu Pro Gly 1

Glu Lys Pro Tyr 5

Lys Cys Pro Glu 10

Cys Gly Lys Ser 15

Phe Ser Ser Lys 20

Lys Ala Leu Thr

Glu His Gln Arg 25

Thr His Thr Gly 30

Glu Lys Pro Tyr 35

Lys Cys Pro Glu 40

Cys Gly Lys Ser

Phe Ser Ser Pro 45

Ala Asp Leu Thr 50

Arg His Gln Arg 55

Thr His Thr Gly

Glu Lys Pro Tyr

Lys Cys Pro Glu 65

Cys Gly Lys Ser 70

Phe Ser Arg Ser 75

Asp Asn Leu Val 80

Arg His Gln Arg

Thr His Thr Gly 85

Glu Lys Pro Tyr 90

Lys Cys Pro Glu

Cys Gly Lys Ser

100

Phe Ser Arg Glu

Asp Asn Leu His 105

Thr His Gln Arg

110

Thr His Thr Gly 115

Glu Lys Pro Tyr

120

Lys Cys Pro Glu

Cys Gly Lys Ser

125

- 161 046157

Phe Ser Arg Ser Asp Glu Leu Val Arg His Gln Arg Thr His Thr Gly

130 135140

Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser

145 150 155160

Gly Asn Leu Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr

165 170175

Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly His Leu Val

180 185190

Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu

195 200205

Cys Gly Lys Ser Phe Ser Gln Asn Ser Thr Leu Thr Glu His Gln Arg

210 215220

Thr His Thr Gly Lys Lys Thr Ser

225230 <210> 84 <211> 176 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic polypeptide <400> 84

Leu Glu Pro Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser

5 1015

Phe Ser Ser Pro Ala Asp Leu Thr Arg His Gln Arg Thr His ThrGly

2530

Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser 35 4045

Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr 50 5560

Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Glu Asp Asn LeuHis

70 7580

Thr His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys ProGlu

9095

- 162 046157

Cys Gly Lys Ser Phe Ser Arg Ser

100

Asp Glu Leu Val

105

Arg His Gln Arg 110

Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu

115 120

Cys Gly Lys Ser 125

Phe Ser Gln Ser Gly Asn Leu Thr Glu His Gln Arg

130 135 140

Thr His Thr Gly

Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser

145 150 155

Phe Ser Thr Ser

160

Gly His Leu Val Arg His Gln Arg Thr His Thr Gly

165 170

Lys Lys Thr Ser

175 <210> 85 <211> 176 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial sequence polypeptide <400> 85

Leu Glu Pro Gly Glu Lys Pro Tyr Lys Cys Pro Glu

5 10

Synthetic

Cys Gly Lys Ser 15

Phe Ser Ser Lys Lys Ala Leu Thr Glu His Gln Arg 20 25

Thr His Thr Gly 30

Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser 35 40

Phe Ser Ser Pro 45

Ala Asp Leu Thr Arg His Gln Arg Thr His Thr Gly

55 60

Glu Lys Pro Tyr

Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser 65 70 75

Asp Asn Leu Val 80

Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr

90

Lys Cys Pro Glu

Cys Gly Lys Ser Phe Ser Arg Glu Asp Asn Leu His

100 105

Thr His Gln Arg 110

Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu

115 120

Cys Gly Lys Ser

125

- 163 046157

Phe Ser Arg Ser Asp Glu Leu Val Arg His Gln Arg

130 135 140

Thr His Thr Gly

Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser

145 150 155

Phe Ser Gln Ser

160

Gly Asn Leu Thr Glu His Gln Arg Thr His Thr Gly

165 170

Lys Lys Thr Ser

175 <210> 86 <211> 232 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial sequence polypeptide <400> 86

Leu Glu Pro Gly Glu Lys Pro Tyr Lys Cys Pro Glu

5 10

Synthetic

Cys Gly Lys Ser 15

Phe Ser Asp Cys Arg Asp Leu Ala Arg His Gln Arg 20 25

Thr His Thr Gly 30

Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser 35 40

Phe Ser Arg Asn 45

Asp Ala Leu Thr Glu His Gln Arg Thr His Thr Gly

55 60

Glu Lys Pro Tyr

Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Asn 65 70 75

Asp Ala Leu Thr

Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr

90

Lys Cys Pro Glu

Cys Gly Lys Ser Phe Ser Ser Pro Ala Asp Leu Thr

100 105

Arg His Gln Arg 110

Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu

115 120

Cys Gly Lys Ser 125

Phe Ser Asp Pro Gly Asn Leu Val Arg His Gln Arg

130 135 140

Thr His Thr Gly

Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser

145 150 155

Phe Ser Gln Arg

160

Ala His Leu Glu Arg His Gln Arg Thr His Thr Gly

Glu Lys Pro Tyr

- 164 046157

165

170

175

Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser

180 185

Ser Ser Leu Val

190

Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr

195 200

Lys Cys Pro Glu 205

Cys Gly Lys Ser Phe Ser His Arg Thr Thr Leu Thr

210 215 220

Asn His Gln Arg

Thr His Thr Gly Lys Lys Thr Ser

225 230 <210> 87 <211> 176 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial sequence polypeptide <400> 87

Leu Glu Pro Gly Glu Lys Pro Tyr Lys Cys Pro Glu

5 10

Synthetic

Cys Gly Lys Ser 15

Phe Ser Arg Asn Asp Ala Leu Thr Glu His Gln Arg 20 25

Thr His Thr Gly 30

Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser 35 40

Phe Ser Ser Pro 45

Ala Asp Leu Thr Arg His Gln Arg Thr His Thr Gly

55 60

Glu Lys Pro Tyr

Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro 65 70 75

Gly Asn Leu Val

Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr

90

Lys Cys Pro Glu

Cys Gly Lys Ser Phe Ser Gln Arg Ala His Leu Glu

100 105

Arg His Gln Arg 110

Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu

115 120

Cys Gly Lys Ser 125

Phe Ser Gln Ser Ser Ser Leu Val Arg His Gln Arg

130 135 140

Thr His Thr Gly

- 165 046157

Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser His Arg

145 150 155 160

Thr Thr Leu Thr Asn His Gln Arg Thr His

165 170

Thr Gly Lys Lys Thr Ser

175 <210> 88 <211> 204 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic polypeptide <400> 88

Leu Glu Pro Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser

5 1015

Phe Ser Arg Asn Asp Ala Leu Thr Glu His Gln Arg Thr His ThrGly

2530

Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro 35 4045

Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr 50 5560

Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser Gly Glu LeuVal

70 7580

Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys ProGlu

9095

Cys Gly Lys Ser Phe Ser Thr His Leu Asp Leu Ile Arg His Gln Arg 100 105110

Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser 115 120125

Phe Ser Ser Lys Lys Ala Leu Thr Glu His Gln Arg Thr His Thr Gly 130 135140

Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser GlnLeu

145 150 155160

Ala His Leu Arg Ala His Gln Arg Thr His Thr Gly Glu Lys ProTyr

165 170175

- 166 046157

Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser

180 185

Asp His Leu Thr 190

Asn His Gln Arg Thr His Thr Gly Lys Lys Thr Ser

195 200 <210> 89 <211> 176 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial sequence polypeptide <400> 89

Leu Glu Pro Gly Glu Lys Pro Tyr Lys Cys Pro Glu

5 10

Synthetic

Cys Gly Lys Ser 15

Phe Ser Arg Ser Asp Asn Leu Val Arg His Gln Arg 20 25

Thr His Thr Gly 30

Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser 35 40

Phe Ser His Arg 45

Thr Thr Leu Thr Asn His Gln Arg Thr His Thr Gly

55 60

Glu Lys Pro Tyr

Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Glu 65 70 75

Asp Asn Leu His 80

Thr His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr

90

Lys Cys Pro Glu

Cys Gly Lys Ser Phe Ser Thr Ser His Ser Leu Thr

100 105

Glu His Gln Arg 110

Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu

115 120

Cys Gly Lys Ser 125

Phe Ser Gln Ser Ser Ser Leu Val Arg His Gln Arg

130 135 140

Thr His Thr Gly

Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser

145 150 155

Phe Ser Arg Glu

160

Asp Asn Leu His Thr His Gln Arg Thr His Thr Gly

165 170

Lys Lys Thr Ser

175

- 167 046157 <210> 90 <211> 176 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial sequence polypeptide <400> 90

Leu Glu Pro Gly Glu Lys Pro Tyr Lys Cys Pro Glu

5 10

Phe Ser Asp Pro Gly Ala Leu Val Arg His Gln Arg

25

Synthetic

Cys Gly Lys Ser 15

Thr His Thr Gly 30

Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser 35 40

Phe Ser Arg Ser 45

Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly

55 60

Glu Lys Pro Tyr

Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Gln Ser 65 70 75

Gly Asp Leu Arg 80

Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr

90

Lys Cys Pro Glu

Cys Gly Lys Ser Phe Ser Thr His Leu Asp Leu Ile

100 105

Arg His Gln Arg 110

Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu

115 120

Cys Gly Lys Ser 125

Phe Ser Thr Ser Gly Asn Leu Val Arg His Gln Arg

130 135 140

Thr His Thr Gly

Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser

145 150 155

Phe Ser Arg Ser

160

Asp Asn Leu Val Arg His Gln Arg Thr His Thr Gly

165 170

Lys Lys Thr Ser 175 <210> 91 <211> 260 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial polypeptide sequence

Synthetic

- 168 046157 <400> 91

Leu Glu Pro Gly Glu Lys Pro Tyr Lys

5

Phe Ser Arg Arg Asp Glu Leu Asn Val

25

Glu Lys Pro Tyr Lys Cys Pro Glu Cys

40

Asp His Leu Thr Asn His Gln Arg Thr

55

Lys Cys Pro Glu Cys Gly Lys Ser Phe 65 70

Arg His Gln Arg Thr His Thr Gly Glu 85

Cys Gly Lys Ser Phe Ser Arg Ser Asp

100 105

Thr His Thr Gly Glu Lys Pro Tyr Lys

115 120

Phe Ser His Arg Thr Thr Leu Thr Asn

130 135

Glu Lys Pro Tyr Lys Cys Pro Glu Cys 145 150

Asp Asn Leu His Thr His Gln Arg Thr

165

Lys Cys Pro Glu Cys Gly Lys Ser Phe

180 185

Glu His Gln Arg Thr His Thr Gly Glu

195 200

Cys Gly Lys Ser Phe Ser Gln Ser Ser

210 215

Thr His Thr Gly Glu Lys Pro Tyr Lys

225 230

Phe Ser Arg Glu Asp Asn Leu His Thr

245

Pro Glu Cys Gly Lys Ser 15

Gln Arg Thr His Thr Gly 30

Lys Ser Phe Ser Arg Ser 45

Thr Gly Glu Lys Pro Tyr 60

Arg Ser Asp Asp Leu Val 75 80

Pro Tyr Lys Cys Pro Glu 95

Leu Val Arg His Gln Arg

110

Pro Glu Cys Gly Lys Ser

125

Gln Arg Thr His Thr Gly 140

Lys Ser Phe Ser Arg Glu

155 160

Thr Gly Glu Lys Pro Tyr

175

Thr Ser His Ser Leu Thr

190

Pro Tyr Lys Cys Pro Glu

205

Leu Val Arg His Gln Arg

220

Pro Glu Cys Gly Lys Ser

235 240

Gln Arg Thr His Thr Gly

255

- 169 046157

Lys Lys Thr Ser

260 <210> 92 <211> 177 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial sequence polypeptide <400> 92

Arg Pro Tyr Ala Cys Pro Val Glu Ser Cys Asp Arg

5 10

Synthetic

Arg Phe Ser Arg 15

Ser Asp Asn Leu Val Arg His Ile Arg Ile His Thr 20 25

Gly Gln Lys Pro 30

Phe Gln Cys Arg Ile Cys Met Arg Asn Phe Ser Arg 35 40

Glu Asp Asn Leu 45

His Thr His Ile Arg Thr His Thr Gly Glu Lys Pro

55 60

Phe Ala Cys Asp

Ile Cys Gly Arg Lys Phe Ala Arg Ser Asp Glu Leu

70 75

Val Arg His Thr

Lys Ile His Leu Arg Gln Lys Asp Arg Pro Tyr Ala

90

Cys Pro Val Glu

Ser Cys Asp Arg Arg Phe Ser Gln Ser Gly Asn Leu

100 105

Thr Glu His Ile

110

Arg Ile His Thr Gly Gln Lys Pro Phe Gln Cys Arg

115 120

Ile Cys Met Arg 125

Asn Phe Ser Thr Ser Gly His Leu Val Arg His Ile

130 135 140

Arg Thr His Thr

Gly Glu Lys Pro Phe Ala Cys Asp Ile Cys Gly Arg

145 150 155

Lys Phe Ala Gln

160

Asn Ser Thr Leu Thr Glu His Thr Lys Ile His Leu

165 170

Arg Gln Lys Asp

175

Lys

- 170 046157 <210> 93 <211> 177 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial sequence polypeptide <400> 93

Arg Pro Tyr Ala Cys Pro Val Glu Ser Cys Asp Arg

5 10

Synthetic

Arg Phe Ser Arg 15

Ser Asp Asn Leu Val Arg His Ile Arg Ile His Thr 20 25

Gly Gln Lys Pro 30

Phe Gln Cys Arg Ile Cys Met Arg Asn Phe Ser His 35 40

Arg Thr Thr Leu 45

Thr Asn His Ile Arg Thr His Thr Gly Glu Lys Pro

55 60

Phe Ala Cys Asp

Ile Cys Gly Arg Lys Phe Ala Arg Glu Asp Asn Leu 65 70 75

His Thr His Thr

Lys Ile His Leu Arg Gln Lys Asp Arg Pro Tyr Ala 85 90

Cys Pro Val Glu

Ser Cys Asp Arg Arg Phe Ser Thr Ser His Ser Leu

100 105

Thr Glu His Ile

110

Arg Ile His Thr Gly Gln Lys Pro Phe Gln Cys Arg

115 120

Ile Cys Met Arg 125

Asn Phe Ser Gln Ser Ser Ser Leu Val Arg His Ile

130 135 140

Arg Thr His Thr

Gly Glu Lys Pro Phe Ala Cys Asp Ile Cys Gly Arg

145 150 155

Lys Phe Ala Arg

160

Glu Asp Asn Leu His Thr His Thr Lys Ile His Leu

165 170

Arg Gln Lys Asp

175

Lys <210> 94 <211> 264 <212> Protein <213> Artificial sequence

- 171 046157 <220>

<223> Description of the artificial sequence

Synthetic polypeptide <400> 94

Arg Pro Tyr Ala 1

Cys Pro Val Glu Ser 5

Arg Asp Glu Leu

Asn Val His Ile Arg

Phe Gln Cys Arg

Ile Cys Met Arg Asn

Thr Asn His Ile

Arg Thr His Thr Gly 55

Cys Asp Arg 10

Ile His Thr

Phe Ser Arg

Glu Lys Pro 60

Arg Phe Ser Arg 15

Gly Gln Lys Pro 30

Ser Asp His Leu 45

Phe Ala Cys Asp

Ile Cys 65

Lys Ile

Ser Cys

Arg Ile

Asn Phe

130

Gly Glu 145

Glu Asp

Arg Pro

Ser His

Phe Gln

210

Val Arg

Gly Arg

His Leu

Asp Arg

100

His Thr 115

Ser His

Lys Pro

Asn Leu

Tyr Ala

180

Ser Leu

195

CysArg

His Ile

Lys Phe

Arg Gln 85

Arg Phe

Gly Gln

Arg Thr

Phe Ala

150

His Thr 165

Cys Pro

Thr Glu

Ile Cys

Arg Thr

Ala Arg

Lys Asp

Ser Arg

Lys Pro

120

Thr Leu 135

Cys Asp

His Thr

Val Glu

His Ile

200

Met Arg 215

His Thr

Ser Asp

Arg Pro

Ser Asp 105

Phe Gln

Thr Asn

Ile Cys

Lys Ile

170

Ser Cys 185

Arg Ile

Asn Phe

Gly Glu

Asp Leu 75

Tyr Ala

Asn Leu

CysArg

His Ile

140

Gly Arg 155

His Leu

Asp Arg

His Thr

Ser Gln

220

Lys Pro

Val Arg His Thr

Cys Pro Val Glu

Val Arg His Ile

110

Ile Cys Met Arg 125

Arg Thr His Thr

Lys Phe Ala Arg

160

Arg Gln Lys Asp

175

Arg Phe Ser Thr 190

Gly Gln Lys Pro 205

Ser Ser Ser Leu

Phe Ala Cys Asp

- 172 046157

225 230 235

240

Ile Cys Gly Arg Lys Phe Ala Arg Glu Asp Asn Leu

245 250

His Thr His Thr

255

Lys Ile His Leu Arg Gln Lys Asp

260 <210> 95 <211> 264 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial sequence polypeptide <400> 95

Arg Pro Tyr Ala Cys Pro Val Glu Ser Cys Asp Arg

5 10

Synthetic

Arg Phe Ser Asp

Pro Gly Ala Leu Val Arg

His Ile Arg Ile His Thr 25

Gly Gln Lys Pro 30

Phe Gln Cys Arg Ile Cys 35

Met Arg Asn Phe Ser Arg 40

Ser Asp Asn Leu 45

Val Arg His Ile Arg Thr 50

His Thr Gly Glu Lys Pro 55 60

Phe Ala Cys Asp

Ile Cys Gly Arg Lys Phe 65 70

Ala Gln Ser Gly Asp Leu 75

Arg Arg His Thr

Lys Ile His Leu Arg Gln 85

Lys Asp Arg Pro Tyr Ala 90

Cys Pro Val Glu

Ser Cys Asp Arg Arg Phe

100

Ser Thr His Leu Asp Leu

105

Ile Arg His Ile 110

Arg Ile His Thr Gly Gln 115

Lys Pro Phe Gln Cys Arg

120

Ile Cys Met Arg 125

Asn Phe Ser Thr Ser Gly

130

Asn Leu Val Arg His Ile

135 140

Arg Thr His Thr

Gly Glu Lys Pro Phe Ala

145 150

Cys Asp Ile Cys Gly Arg

155

Lys Phe Ala Arg

160

Ser Asp Asn Leu Val Arg

165

His Thr Lys Ile His Leu 170

Arg Gln Lys Asp

175

- 173 046157

Arg Pro Tyr Ala Cys Pro

180

Val Glu Ser Cys Asp Arg

185

Arg Phe Ser Gln

190

Ser Gly His Leu Thr Glu

195

His Ile Arg Ile His Thr

200

Gly Gln Lys Pro 205

Phe Gln Cys Arg Ile Cys

210

Met Arg Asn Phe Ser Glu

215 220

Arg Ser His Leu

Arg Glu His Ile Arg Thr

225 230

His Thr Gly Glu Lys Pro

235

Phe Ala Cys Asp

240

Ile Cys Gly Arg Lys Phe

245

Ala Gln Ala Gly His Leu

250

Ala Ser His Thr

255

Lys Ile His Leu Arg Gln

260

Lys Asp <210> 96 <211> 175 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial sequence polypeptide <400> 96

Arg Pro His Ala Cys Pro Ala Glu Gly Cys Asp Arg

5 10

Synthetic

Arg Phe Ser Arg 15

Ser Asp Asn Leu Val Arg His Leu Arg Ile His Thr 20 25

Gly His Lys Pro 30

Phe Gln Cys Arg Ile Cys Met Arg Ser Phe Ser Arg 35 40

Glu Asp Asn Leu 45

His Thr His Ile Arg Thr His Thr Gly Glu Lys Pro

55 60

Phe Ala Cys Glu

Phe Cys Gly Arg Lys Phe Ala Arg Ser Asp Glu Leu 65 70 75

Val Arg His Ala

Lys Ile His Leu Lys Gln Lys Glu His Ala Cys Pro

90

Ala Glu Gly Cys

Asp Arg Arg Phe Ser Gln Ser Gly Asn Leu Thr Glu

100 105

His Leu Arg Ile 110

- 174 046157

His Thr Gly His Lys Pro 115

Phe Gln Cys Arg Ile Cys

120

Met Arg Ser Phe 125

Ser Thr Ser Gly His Leu

130

Val Arg His Ile Arg Thr

135 140

His Thr Gly Glu

Lys Pro Phe Ala Cys Glu

145 150

Phe Cys Gly Arg Lys Phe

155

Ala Gln Asn Ser

160

Thr Leu Thr Glu His Ala

165

Lys Ile His Leu Lys Gln

170

Lys Glu Lys

175 <210> 97 <211> 175 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial sequence polypeptide <400> 97

Arg Pro His Ala Cys Pro Ala Glu Gly Cys Asp Arg

5 10

Synthetic

Arg Phe Ser Arg 15

Ser Asp Asn Leu Val Arg His Leu Arg Ile His Thr 20 25

Gly His Lys Pro 30

Phe Gln Cys Arg Ile Cys Met Arg Ser Phe Ser His 35 40

Arg Thr Thr Leu 45

Thr Asn His Ile Arg Thr His Thr Gly Glu Lys Pro

55 60

Phe Ala Cys Glu

Phe Cys Gly Arg Lys Phe Ala Arg Glu Asp Asn Leu 65 70 75

His Thr His Ala

Lys Ile His Leu Lys Gln Lys Glu His Ala Cys Pro

90

Ala Glu Gly Cys

Asp Arg Arg Phe Ser Thr Ser His Ser Leu Thr Glu

100 105

His Leu Arg Ile 110

His Thr Gly His Lys Pro Phe Gln Cys Arg Ile Cys

115 120

Met Arg Ser Phe 125

Ser Gln Ser Ser Ser Leu Val Arg His Ile Arg Thr

130 135 140

His Thr Gly Glu

- 175 046157

Lys Pro Phe Ala Cys Glu Phe Cys Gly Arg Lys Phe

145 150 155

Ala Arg Glu Asp

160

Asn Leu His Thr His Ala Lys Ile His Leu Lys Gln

165 170

Lys Glu Lys

175 <210> 98 <211> 261 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial sequence polypeptide <400> 98

Arg Pro His Ala Cys Pro Ala Glu Gly Cys Asp Arg

5 10

Synthetic

Arg Phe Ser Arg 15

Arg Asp Glu Leu Asn Val His Leu Arg Ile His Thr 20 25

Gly His Lys Pro 30

Phe Gln Cys Arg Ile Cys Met Arg Ser Phe Ser Arg 35 40

Ser Asp His Leu 45

Thr Asn His Ile Arg Thr His Thr Gly Glu Lys Pro

55 60

Phe Ala Cys Glu

Phe Cys Gly Arg Lys Phe Ala Arg Ser Asp Asp Leu 65 70 75

Val Arg His Ala

Lys Ile His Leu Lys Gln Lys Glu His Ala Cys Pro

90

Ala Glu Gly Cys

Asp Arg Arg Phe Ser Arg Ser Asp Asn Leu Val Arg

100 105

His Leu Arg Ile 110

His Thr Gly His Lys Pro Phe Gln Cys Arg Ile Cys

115 120

Met Arg Ser Phe 125

Ser His Arg Thr Thr Leu Thr Asn His Ile Arg Thr

130 135 140

His Thr Gly Glu

Lys Pro Phe Ala Cys Glu Phe Cys Gly Arg Lys Phe

145 150 155

Ala Arg Glu Asp

160

Asn Leu His Thr His Ala Lys Ile His Leu Lys Gln

165 170

Lys Glu His Ala 175

Cys Pro Ala Glu Gly Cys Asp Arg Arg Phe Ser Thr

Ser His Ser Leu

- 176 046157

180 185

190

Thr Glu His Leu Arg Ile His Thr Gly His Lys Pro

195 200

Phe Gln Cys Arg 205

Ile Cys Met Arg Ser Phe Ser Gln Ser Ser Ser Leu

210 215 220

Val Arg His Ile

Arg Thr His Thr Gly Glu Lys Pro Phe Ala Cys Glu

225 230 235

Phe Cys Gly Arg

240

Lys Phe Ala Arg Glu Asp Asn Leu His Thr His Ala

245 250

Lys Ile His Leu

255

Lys Gln Lys Glu Lys

260 <210> 99 <211> 753 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial sequence

Synthetic polypeptide <400> 99

Met Ala Pro Lys 1

Lys Lys Arg Lys 5

Val Gly Ile His 10

Gly Val Pro Ala 15

Ala Leu Glu Pro

Gly Glu Lys Pro

Tyr Lys Cys Pro 25

Glu Cys Gly Lys 30

Ser Phe Ser Arg

Ser Asp Asn Leu 40

Val Arg His Gln

Arg Thr His Thr 45

Gly Glu Lys Pro 50

Tyr Lys Cys Pro 55

Glu Cys Gly Lys 60

Ser Phe Ser His

Arg Thr Thr Leu 65

Thr Asn His Gln

Arg Thr His Thr 75

Gly Glu Lys Pro 80

Tyr Lys Cys Pro

Glu Cys Gly Lys 85

Ser Phe Ser Arg 90

Glu Asp Asn Leu

His Thr His Gln

100

Arg Thr His Thr

Gly Glu Lys Pro 105

Tyr Lys Cys Pro 110

Glu Cys Gly Lys

115

Ser Phe Ser Thr

120

Ser His Ser Leu

Thr Glu His Gln 125

- 177 046157

Arg Thr His Thr Gly Glu Lys Pro

130 135

Tyr Lys Cys Pro Glu Cys Gly

140

Ser Phe Ser Gln

145

Ser Ser Ser Leu Val Arg His Gln Arg Thr His

150 155

Gly Glu Lys Pro

Tyr Lys Cys Pro Glu Cys Gly Lys

165 170

Ser Phe Ser

175

Glu Asp Asn Leu His

180

Thr His Gln Arg Thr His Thr Gly Lys Lys

185 190

Ser Lys Arg Pro Ala Ala Thr Lys Lys Ala Gly Gln Ala Lys Lys

195 200 205

Lys Gly Ser Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Leu Glu Glu

210 215 220

Ser Gly Ser Gly Arg Ala Asp Ala Leu Asp Asp Phe Asp Leu Asp

225 230 235

Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly

245 250 255

Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala

260 265 270

Asp Asp Phe Asp Leu Asp Met Leu Ile Asn Ser Arg Ser Ser Gly

275 280 285

Pro Lys Lys Lys Arg Lys Val Gly Ser Gln Tyr Leu Pro Asp Thr

290 295 300

Asp Arg His Arg Ile Glu Glu Lys Arg Lys Arg Thr Tyr Glu Thr

305 310 315

Lys Ser Ile Met Lys Lys Ser Pro Phe Ser Gly Pro Thr Asp Pro

325 330 335

Pro Pro Pro Arg Arg Ile Ala Val Pro Ser Arg Ser Ser Ala Ser

340 345 350

Pro Lys Pro Ala Pro Gln Pro Tyr Pro Phe Thr Ser Ser Leu Ser

355 360 365

Ile Asn Tyr Asp Glu Phe Pro Thr Met Val Phe Pro Ser Gly Gln

Lys

Thr 160

Arg

Thr

Lys

Ala

Met 240

Ser

Leu

Ser

Asp

Phe 320

Arg

Val

Thr

Ile

- 178 046157

370

375

380

Ser Gln Ala Ser Ala Leu Ala Pro Ala Pro Pro Gln Val Leu Pro

385 390 395

Ala Pro Ala Pro Ala

405

Pro Ala Pro Ala Met

410

Val Ser Ala Leu Ala

415

Ala Pro Ala Pro Val

420

Pro Val Leu Ala Pro

425

Gly Pro Pro Gln Ala

430

Ala Pro Pro Ala Pro

435

Lys Pro Thr Gln Ala

440

Gly Glu Gly Thr Leu 445

Glu Ala Leu Leu Gln

450

Leu Gln Phe Asp Asp

455

Glu Asp Leu Gly Ala

460

Leu Gly Asn Ser Thr 465

Asp Pro Ala Val Phe 470

Thr Asp Leu Ala Ser 475

Asp Asn Ser Glu Phe

485

Gln Gln Leu Leu Asn

490

Gln Gly Ile Pro Val

495

Pro His Thr Thr Glu

500

Pro Met Leu Met Glu

505

Tyr Pro Glu Ala Ile

510

Arg Leu Val Thr Gly 515

Ala Gln Arg Pro Pro 520

Asp Pro Ala Pro Ala 525

Leu Gly Ala Pro Gly

530

Leu Pro Asn Gly Leu

535

Leu Ser Gly Asp Glu

540

Phe Ser Ser Ile Ala 545

Asp Met Asp Phe Ser 550

Ala Leu Leu Gly Ser 555

Ser Gly Ser Arg Asp

565

Ser Arg Glu Gly Met

570

Phe Leu Pro Lys Pro

575

Ala Gly Ser Ala Ile

580

Ser Asp Val Phe Glu

585

Gly Arg Glu Val Cys

590

Pro Lys Arg Ile Arg 595

Pro Phe His Pro Pro

600

Gly Ser Pro Trp Ala 605

Arg Pro Leu Pro Ala

610

Ser Leu Ala Pro Thr

615

Pro Thr Gly Pro Val

620

Gln 400

Gln

Val

Ser

Leu

Val 480

Ala

Thr

Pro

Asp

Gly 560

Glu

Gln

Asn

His

- 179 046157

Glu Pro Val 625

Pro Ala Pro

Asp Glu Glu

Thr Val Ile 675

Leu Ser His 690

Leu Glu Ser 705

Glu Leu Asn

His Ala Met

Phe

Gly Ser Leu

630

Ala Val Thr

645

Thr Ser Gln 660

Pro Gln Lys

Pro Pro Pro

Met Thr Glu

710

Glu Ile Leu

725

His Ile Ser 740

Thr Pro Ala

Pro Glu Ala

Ala Val Lys 665

Glu Glu Ala 680

Arg Gly His 695

Asp Leu Asn

Asp Thr Phe

Thr Gly Leu

745

Pro Val Pro

635

Ser His Leu 650

Ala Leu Arg

Ala Ile Cys

Leu Asp Glu

700

Leu Asp Ser

715

Leu Asn Asp 730

Ser Ile Phe

Gln Pro Leu Asp

640

Leu Glu Asp Pro

655

Glu Met Ala Asp

670

Gly Gln Met Asp 685

Leu Thr Thr Thr

Pro Leu Thr Pro

720

Glu Cys Leu Leu

735

Asp Thr Ser Leu 750 <210> 100 <211> 753 <212> Protein <213> Artificial Sequence <220>

<223> Description of the artificial sequence polypeptide <400> 100

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Ile His

510

Ala Leu Glu Pro Gly Glu Lys Pro Tyr Lys Cys Pro 2025

Ser Phe Ser Arg Ser Asp Asn Leu Val Arg His Gln 3540

Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys 50 5560

Glu Asp Asn Leu His Thr His Gln Arg Thr His Thr

Synthetic

Gly Val Pro Ala 15

Glu Cys Gly Lys 30

Arg Thr His Thr 45

Ser Phe Ser Arg

Gly Glu Lys Pro

- 180 046157

70 75

Tyr Lys Cys Pro Glu 85

Cys Gly Lys Ser Phe

Ser Arg Ser Asp Glu

Val Arg His Gln Arg

100

Thr His Thr Gly Glu

105

Lys Pro Tyr Lys Cys

110

Glu Cys Gly Lys Ser 115

Phe Ser Gln Ser Gly 120

Asn Leu Thr Glu His

125

Arg Thr His Thr Gly 130

Glu Lys Pro Tyr Lys 135

Cys Pro Glu Cys Gly

140

Ser Phe Ser Thr Ser 145

Gly His Leu Val Arg 150

His Gln Arg Thr His 155

Gly Glu Lys Pro Tyr

165

Lys Cys Pro Glu Cys

170

Gly Lys Ser Phe Ser

175

Asn Ser Thr Leu Thr

180

Glu His Gln Arg Thr

185

His Thr Gly Lys Lys

190

Ser Lys Arg Pro Ala

195

Ala Thr Lys Lys Ala 200

Gly Gln Ala Lys Lys 205

Lys Gly Ser Tyr Pro

210

Tyr Asp Val Pro Asp

215

Tyr Ala Leu Glu Glu

220

Ser Gly Ser Gly Arg 225

Ala Asp Ala Leu Asp 230

Asp Phe Asp Leu Asp 235

Leu Gly Ser Asp Ala

245

Leu Asp Asp Phe Asp

250

Leu Asp Met Leu Gly

255

Asp Ala Leu Asp Asp

260

Phe Asp Leu Asp Met

265

Leu Gly Ser Asp Ala

270

Asp Asp Phe Asp Leu 275

Asp Met Leu Ile Asn 280

Ser Arg Ser Ser Gly

285

Pro Lys Lys Lys Arg 290

Lys Val Gly Ser Gln

295

Tyr Leu Pro Asp Thr

300

Asp Arg His Arg Ile 305

Glu Glu Lys Arg Lys 310

Arg Thr Tyr Glu Thr 315

Leu

Pro

Gln

Lys

Thr 160

Gln

Thr

Lys

Ala

Met 240

Ser

Leu

Ser

Asp

Phe 320

- 181 046157

Lys

Ser Ile Met Lys Lys Ser Pro

325

Phe Ser Gly Pro Thr Asp Pro

330 335

Pro Pro Pro Arg Arg Ile Ala Val Pro

340 345

Ser Arg Ser

Ser Ala Ser

350

Pro Lys Pro Ala Pro Gln Pro Tyr Pro Phe Thr Ser Ser Leu Ser

355 360 365

Ile Asn Tyr Asp Glu

370

Phe Pro Thr Met Val

375

Phe Pro Ser Gly Gln 380

Ser Gln Ala Ser Ala 385

Leu Ala Pro Ala Pro 390

Pro Gln Val Leu Pro 395

Ala Pro Ala Pro Ala

405

Pro Ala Pro Ala Met

410

Val Ser Ala Leu Ala

415

Ala Pro Ala Pro Val

420

Pro Val Leu Ala Pro

425

Gly Pro Pro Gln Ala

430

Ala Pro Pro Ala Pro

435

Lys Pro Thr Gln Ala

440

Gly Glu Gly Thr Leu 445

Glu Ala Leu Leu Gln

450

Leu Gln Phe Asp Asp 455

Glu Asp Leu Gly Ala

460

Leu Gly Asn Ser Thr 465

Asp Pro Ala Val Phe 470

Thr Asp Leu Ala Ser 475

Asp Asn Ser Glu Phe

485

Gln Gln Leu Leu Asn

490

Gln Gly Ile Pro Val

495

Pro His Thr Thr Glu

500

Pro Met Leu Met Glu

505

Tyr Pro Glu Ala Ile

510

Arg Leu Val Thr Gly 515

Ala Gln Arg Pro Pro 520

Asp Pro Ala Pro Ala 525

Leu Gly Ala Pro Gly

530

Leu Pro Asn Gly Leu

535

Leu Ser Gly Asp Glu 540

Phe Ser Ser Ile Ala 545

Asp Met Asp Phe Ser 550

Ala Leu Leu Gly Ser 555

Ser Gly Ser Arg Asp

565

Ser Arg Glu Gly Met

570

Phe Leu Pro Lys Pro

575

Arg

Val

Thr

Ile

Gln 400

Gln

Val

Ser

Leu

Val 480

Ala

Thr

Pro

Asp

Gly 560

Glu

- 182 046157

Ala Gly Ser

Ala Ile Ser 580

Asp Val Phe

585

Glu Gly Arg

Glu Val Cys Gln 590

Pro Lys Arg 595

Arg Pro Leu 610

Ile Arg Pro

Phe His Pro

600

Pro Gly Ser

Pro Trp Ala Asn 605

Pro Ala Ser

Leu Ala Pro 615

Thr Pro Thr

620

Gly Pro Val His

Glu Pro Val 625

Gly Ser Leu

630

Thr Pro Ala

Pro Val Pro

635

Gln Pro Leu Asp

640

Pro Ala Pro

Asp Glu Glu

Thr Val Ile 675

Leu Ser His 690

Leu Glu Ser 705

Glu Leu Asn

Ala Val Thr

645

Pro Glu Ala

Ser His Leu 650

Leu Glu Asp Pro

655

His Ala Met

Phe

Thr Ser Gln 660

Pro Gln Lys

Pro Pro Pro

Met Thr Glu

710

Glu Ile Leu

725

His Ile Ser 740

Ala Val Lys

665

Ala Leu Arg

Glu Met Ala Asp

670

Glu Glu Ala

680

Ala Ile Cys

Gly Gln Met Asp 685

Arg Gly His 695

Asp Leu Asn

Asp Thr Phe

Thr Gly Leu

745

Leu Asp Glu

700

Leu Asp Ser

715

Leu Asn Asp 730

Ser Ile Phe <210> 101 <211> 272 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial sequence polypeptide <400> 101

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Ile His

5 10

Leu Thr Thr Thr

Pro Leu Thr Pro

720

Glu Cys Leu Leu

735

Asp Thr Ser Leu 750

Synthetic

Gly Val Pro Ala 15

- 183 046157

Ala Leu Glu Pro Gly

Glu Lys Pro Tyr Lys 25

Cys Pro Glu Cys Gly 30

Ser Phe Ser Arg Ser 35

Asp Asn Leu Val Arg 40

His Gln Arg Thr His 45

Gly Glu Lys Pro Tyr 50

Lys Cys Pro Glu Cys 55

Gly Lys Ser Phe Ser 60

Arg Thr Thr Leu Thr 65

Asn His Gln Arg Thr 70

His Thr Gly Glu Lys 75

Tyr Lys Cys Pro Glu 85

Cys Gly Lys Ser Phe

Ser Arg Glu Asp Asn

His Thr His Gln Arg

100

Thr His Thr Gly Glu

105

Lys Pro Tyr Lys Cys

110

Glu Cys Gly Lys Ser 115

Phe Ser Thr Ser His

120

Ser Leu Thr Glu His

125

Arg Thr His Thr Gly 130

Glu Lys Pro Tyr Lys 135

Cys Pro Glu Cys Gly

140

Ser Phe Ser Gln Ser 145

Ser Ser Leu Val Arg 150

His Gln Arg Thr His 155

Gly Glu Lys Pro Tyr

165

Lys Cys Pro Glu Cys

170

Gly Lys Ser Phe Ser

175

Glu Asp Asn Leu His

180

Thr His Gln Arg Thr

185

His Thr Gly Lys Lys

190

Ser Lys Arg Pro Ala

195

Ala Thr Lys Lys Ala

200

Gly Gln Ala Lys Lys 205

Lys Gly Ser Tyr Pro

210

Tyr Asp Val Pro Asp

215

Tyr Ala Leu Glu Asp

220

Leu Asp Asp Phe Asp 225

Leu Asp Met Leu Gly 230

Ser Asp Ala Leu Asp 235

Phe Asp Leu Asp Met

245

Leu Gly Ser Asp Ala

250

Leu Asp Asp Phe Asp

255

Asp Met Leu Gly Ser

260

Asp Ala Leu Asp Asp

265

Phe Asp Leu Asp Met

270

Lys

Thr

His

Pro 80

Leu

Pro

Gln

Lys

Thr 160

Arg

Thr

Lys

Ala

Asp 240

Leu

Leu

- 184 046157 <210> 102 <211> 272 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic polypeptide <400> 102

Met Ala Pro Lys Lys Lys Arg Lys 1 5

Val Gly Ile His Gly Val Pro Ala

15

Ala Leu Glu Pro Gly Glu Lys Pro 20

Tyr Lys Cys Pro Glu Cys Gly Lys 25 30

Ser Phe Ser Arg Ser Asp Asn Leu

40

Val Arg His Gln Arg Thr His Thr 45

Gly Glu Lys Pro Tyr Lys Cys Pro

55

Glu Cys Gly Lys Ser Phe Ser Arg 60

Glu Asp Asn Leu His Thr His Gln 65 70

Arg Thr His Thr Gly Glu Lys Pro

80

Tyr Lys Cys Pro Glu Cys Gly Lys 85

Ser Phe Ser Arg Ser Asp Glu Leu

95

Val Arg His Gln Arg Thr His Thr

100

Gly Glu Lys Pro Tyr Lys Cys Pro

105 110

Glu Cys Gly Lys Ser Phe Ser Gln

115 120

Ser Gly Asn Leu Thr Glu His Gln

125

Arg Thr His Thr Gly Glu Lys Pro

130 135

Tyr Lys Cys Pro Glu Cys Gly Lys

140

Ser Phe Ser Thr Ser Gly His Leu

145 150

Val Arg His Gln Arg Thr His Thr

155 160

Gly Glu Lys Pro Tyr Lys Cys Pro

165

Glu Cys Gly Lys Ser Phe Ser Gln

170 175

Asn Ser Thr Leu Thr Glu His Gln

180

Arg Thr His Thr Gly Lys Lys Thr 185 190

Ser Lys Arg Pro Ala Ala Thr Lys

195 200

Lys Ala Gly Gln Ala Lys Lys Lys

205

- 185 046157

Lys Gly Ser Tyr Pro Tyr

210

Asp Val Pro Asp Tyr Ala

215 220

Leu Glu Asp Ala

Leu Asp Asp Phe Asp Leu

225 230

Asp Met Leu Gly Ser Asp

235

Ala Leu Asp Asp

240

Phe Asp Leu Asp Met Leu

245

Gly Ser Asp Ala Leu Asp

250

Asp Phe Asp Leu

255

Asp Met Leu Gly Ser Asp

260

Ala Leu Asp Asp Phe Asp

265

Leu Asp Met Leu 270 <210>

<211>

<212>

103

580 Protein <213> Artificial sequence <220>

<223> Description of the artificial sequence polypeptide <400> 103

Synthetic

Met Ala Ala 1

Arg Pro Pro

Leu Pro Pro

Gly Ala Pro 50

Tyr Gly Ala 65

Pro Pro Pro

Tyr Pro Gly

Gly Pro Gln

115

Ala Leu Gly

130

Ser Leu Glu

Asp His Leu 5

Ser Ala Ala 20

Tyr Ala Gly

Leu Gly Pro

Phe Gly Pro 70

Ala Ala Gly 85

Arg Ala Ala 100

Pro Ala Pro

Gly Met Asp

Leu Glu Leu

Met Leu Ala

Ala Ala His

Pro Gly Leu 40

Pro Pro Pro 55

Pro Ser Ser

Ile Ala His

Ala Pro Pro

105

Ser Ala Ala

120

Ala Glu Leu 135

Gly Leu His

Glu Gly Tyr 10

Gly Pro His

Asp Ser Gly

Arg Gln Pro 60

Phe Gln Pro 75

Leu Gln Pro 90

Asn Ala Pro

Ala Pro Pro

Ile Asp Glu

140

Arg Val Arg

Arg Leu Val Gln 15

Ala Leu Arg Thr 30

Leu Arg Pro Arg 45

Gly Ala Leu Ala

Phe Pro Ala Val

Val Ala Thr Pro

Gly Gly Pro Pro 110

Pro Pro Ala His 125

Glu Ala Leu Thr

Glu Leu Pro Glu

- 186 046157

145

150

155

Leu Phe Leu Gly Gln

165

Ser Glu Phe Asp Cys

170

Phe Ser Asp Leu Gly

175

Ala Pro Pro Ala Gly

180

Ser Val Ser Cys Gly

185

Gly Ser Gly Gly Gly

190

Gly Gln Ser Gln Leu

195

Ile Lys Pro Ser Arg 200

Met Arg Lys Tyr Pro 205

Arg Pro Ser Lys Thr 210

Pro Pro His Glu Arg 215

Pro Tyr Ala Cys Pro 220

Glu Ser Cys Asp Arg 225

Arg Phe Ser Arg Ser 230

Asp Asn Leu Val Arg 235

Ile Arg Ile His Thr

245

Gly Gln Lys Pro Phe

250

Gln Cys Arg Ile Cys

255

Arg Asn Phe Ser His

260

Arg Thr Thr Leu Thr

265

Asn His Ile Arg Thr

270

Thr Gly Glu Lys Pro

275

Phe Ala Cys Asp Ile 280

Cys Gly Arg Lys Phe

285

Arg Glu Asp Asn Leu

290

His Thr His Thr Lys

295

Ile His Leu Arg Gln

300

Asp Arg Pro Tyr Ala 305

Cys Pro Val Glu Ser 310

Cys Asp Arg Arg Phe 315

Thr Ser His Ser Leu

325

Thr Glu His Ile Arg

330

Ile His Thr Gly Gln

335

Pro Phe Gln Cys Arg

340

Ile Cys Met Arg Asn

345

Phe Ser Gln Ser Ser

350

Leu Val Arg His Ile 355

Arg Thr His Thr Gly 360

Glu Lys Pro Phe Ala 365

Asp Ile Cys Gly Arg

370

Lys Phe Ala Arg Glu

375

Asp Asn Leu His Thr 380

Thr Lys Ile His Leu 385

Arg Gln Lys Asp Lys 390

Leu Glu Met Ala Asp 395

160

Ser

Ser

Asn

Val

His 240

Met

His

Ala

Lys

Ser 320

Lys

Ser

Cys

His

His 400

- 187 046157

Leu Met Leu

Ala Glu Gly

405

Tyr Arg Leu

Val Gln Arg 410

Pro Pro Ser Ala

415

Ala Ala Ala

His Gly Pro 420

His Ala Leu

425

Arg Thr Leu

Pro Pro Tyr Ala

430

Gly Pro Gly

435

Leu Asp Ser

Gly Leu Arg

440

Pro Arg Gly

Ala Pro Leu Gly 445

Pro Pro Pro

450

Pro Arg Gln

Pro Gly Ala 455

Leu Ala Tyr

460

Gly Ala Phe Gly

Pro Pro Ser 465

Ser Phe Gln

470

Pro Phe Pro

Ala Val Pro

475

Pro Pro Ala Ala

480

Gly Ile Ala

His Leu Gln

485

Pro Val Ala

Thr Pro Tyr 490

Pro Gly Arg Ala

495

Ala Ala Pro

Pro Asn Ala 500

Pro Gly Gly

505

Pro Pro Gly

Pro Gln Pro Ala

510

Pro Ser Ala

515

Ala Ala Pro

Pro Pro Pro

520

Ala His Ala

Leu Gly Gly Met 525

Asp Ala Glu

530

Leu Ile Asp

Glu Glu Ala 535

Leu Thr Ser

540

Leu Glu Leu Glu

Leu Gly Leu 545

His Arg Val

550

Arg Glu Leu

Pro Glu Leu

555

Phe Leu Gly Gln

560

Ser Glu Phe

Asp Cys Phe

565

Ser Asp Leu

Gly Ser Ala 570

Pro Pro Ala Gly

575

Ser Val Ser Cys

580 <210> 104 <211> 394 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial polypeptide sequence

Synthetic <400> 104

Met Ala Ala Asp His Leu Met Leu Ala Glu Gly Tyr 1 5 10

Arg Leu Val Gln 15

Arg Pro Pro Ser Ala Ala Ala Ala His Gly Pro His

Ala Leu Arg Thr

- 188 046157

Leu Pro Pro Tyr Ala 35

Gly Pro Gly Leu Asp 40

Ser Gly Leu Arg Pro 45

Gly Ala Pro Leu Gly 50

Pro Pro Pro Pro Arg 55

Gln Pro Gly Ala Leu 60

Tyr Gly Ala Phe Gly 65

Pro Pro Ser Ser Phe 70

Gln Pro Phe Pro Ala 75

Pro Pro Pro Ala Ala

Gly Ile Ala His Leu

Gln Pro Val Ala Thr

Tyr Pro Gly Arg Ala

100

Ala Ala Pro Pro Asn

105

Ala Pro Gly Gly Pro

110

Gly Pro Gln Pro Ala

115

Pro Ser Ala Ala Ala

120

Pro Pro Pro Pro Ala

125

Ala Leu Gly Gly Met

130

Asp Ala Glu Leu Ile 135

Asp Glu Glu Ala Leu 140

Ser Leu Glu Leu Glu 145

Leu Gly Leu His Arg 150

Val Arg Glu Leu Pro 155

Leu Phe Leu Gly Gln

165

Ser Glu Phe Asp Cys

170

Phe Ser Asp Leu Gly

175

Ala Pro Pro Ala Gly

180

Ser Val Ser Cys Gly

185

Gly Ser Gly Gly Gly

190

Gly Gln Ser Gln Leu 195

Ile Lys Pro Ser Arg 200

Met Arg Lys Tyr Pro 205

Arg Pro Ser Lys Thr 210

Pro Pro His Glu Arg 215

Pro Tyr Ala Cys Pro 220

Glu Ser Cys Asp Arg 225

Arg Phe Ser Arg Ser 230

Asp Asn Leu Val Arg 235

Ile Arg Ile His Thr

245

Gly Gln Lys Pro Phe

250

Gln Cys Arg Ile Cys

255

Arg Asn Phe Ser His

260

Arg Thr Thr Leu Thr

265

Asn His Ile Arg Thr

270

Arg

Ala

Val 80

Pro

Pro

His

Thr

Glu 160

Ser

Ser

Asn

Val

His 240

Met

His

- 189 046157

Thr Gly Glu

275

Lys Pro Phe

Ala Cys Asp

280

Arg Glu Asp

290

Asn Leu His

Thr His Thr 295

Asp Arg Pro 305

Thr Ser His

Pro Phe Gln

Leu Val Arg

355

Asp Ile Cys

370

Thr Lys Ile 385

Tyr Ala Cys 310

Ser Leu Thr 325

Cys Arg Ile 340

His Ile Arg

Gly Arg Lys

His Leu Arg 390

Pro Val Glu

Glu His Ile

Cys Met Arg

345

Thr His Thr

360

Phe Ala Arg 375

Gln Lys Asp

Ile Cys Gly

Lys Ile His 300

Ser Cys Asp 315

Arg Ile His 330

Asn Phe Ser

Gly Glu Lys

Glu Asp Asn

380

Lys

Arg Lys Phe Ala 285

Leu Arg Gln Lys

Arg Arg Phe Ser

320

Thr Gly Gln Lys

335

Gln Ser Ser Ser

350

Pro Phe Ala Cys 365

Leu His Thr His <210> 105 <211> 580 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial sequence

Synthetic polypeptide <400> 105

Met Ala Ala Asp 1

His Leu Met Leu 5

Ala Glu Gly Tyr 10

Arg Leu Val Gln 15

Arg Pro Pro Ser

Ala Ala Ala Ala

His Gly Pro His 25

Ala Leu Arg Thr 30

Leu Pro Pro Tyr 35

Ala Gly Pro Gly 40

Leu Asp Ser Gly

Leu Arg Pro Arg 45

Gly Ala Pro Leu 50

Gly Pro Pro Pro 55

Pro Arg Gln Pro

Gly Ala Leu Ala

Tyr Gly Ala Phe 65

Gly Pro Pro Ser 70

Ser Phe Gln Pro

Phe Pro Ala Val

Pro Pro Pro Ala

Ala Gly Ile Ala

His Leu Gln Pro

Val Ala Thr Pro

- 190 046157

Tyr Pro Gly Arg Ala

100

Ala Ala Pro Pro Asn

105

Ala Pro Gly Gly Pro

110

Gly Pro Gln Pro Ala

115

Pro Ser Ala Ala Ala

120

Pro Pro Pro Pro Ala

125

Ala Leu Gly Gly Met

130

Asp Ala Glu Leu Ile 135

Asp Glu Glu Ala Leu 140

Ser Leu Glu Leu Glu 145

Leu Gly Leu His Arg 150

Val Arg Glu Leu Pro 155

Leu Phe Leu Gly Gln

165

Ser Glu Phe Asp Cys

170

Phe Ser Asp Leu Gly

175

Ala Pro Pro Ala Gly

180

Ser Val Ser Cys Gly

185

Gly Ser Gly Gly Gly

190

Gly Gln Ser Gln Leu 195

Ile Lys Pro Ser Arg 200

Met Arg Lys Tyr Pro 205

Arg Pro Ser Lys Thr

210

Pro Pro His Glu Arg 215

Pro Tyr Ala Cys Pro 220

Glu Ser Cys Asp Arg 225

Arg Phe Ser Arg Ser 230

Asp Asn Leu Val Arg 235

Ile Arg Ile His Thr

245

Gly Gln Lys Pro Phe

250

Gln Cys Arg Ile Cys

255

Arg Asn Phe Ser Arg

260

Glu Asp Asn Leu His

265

Thr His Ile Arg Thr

270

Thr Gly Glu Lys Pro

275

Phe Ala Cys Asp Ile 280

Cys Gly Arg Lys Phe

285

Arg Ser Asp Glu Leu

290

Val Arg His Thr Lys

295

Ile His Leu Arg Gln

300

Asp Arg Pro Tyr Ala 305

Cys Pro Val Glu Ser 310

Cys Asp Arg Arg Phe 315

Gln Ser Gly Asn Leu

325

Thr Glu His Ile Arg

330

Ile His Thr Gly Gln

335

Pro

His

Thr

Glu 160

Ser

Ser

Asn

Val

His 240

Met

His

Ala

Lys

Ser 320

Lys

- 191 046157

Pro Phe Gln Cys Arg Ile Cys Met Arg

340 345

Leu Val Arg His Ile Arg Thr His Thr

355 360

Asp Ile Cys Gly Arg Lys Phe Ala Gln

370 375

Thr Lys Ile His Leu Arg Gln Lys Asp

385 390

Leu Met Leu Ala Glu Gly Tyr Arg Leu

405

Ala Ala Ala His Gly Pro His Ala Leu

420 425

Gly Pro Gly Leu Asp Ser Gly Leu Arg

435 440

Pro Pro Pro Pro Arg Gln Pro Gly Ala

450 455

Pro Pro Ser Ser Phe Gln Pro Phe Pro

465 470

Gly Ile Ala His Leu Gln Pro Val Ala 485

Ala Ala Pro Pro Asn Ala Pro Gly Gly

500 505

Pro Ser Ala Ala Ala Pro Pro Pro Pro

515 520

Asp Ala Glu Leu Ile Asp Glu Glu Ala

530 535

Leu Gly Leu His Arg Val Arg Glu Leu

545 550

Ser Glu Phe Asp Cys Phe Ser Asp Leu

565

Phe Ser Thr Ser Gly His

350

Glu Lys Pro Phe Ala Cys 365

Ser Thr Leu Thr Glu His

380

Leu Glu Met Ala Asp His

395 400

Gln Arg Pro Pro Ser Ala

415

Thr Leu Pro Pro Tyr Ala

430

Arg Gly Ala Pro Leu Gly

445

Ala Tyr Gly Ala Phe Gly

460

Val Pro Pro Pro Ala Ala

475 480

Pro Tyr Pro Gly Arg Ala

495

Pro Gly Pro Gln Pro Ala

510

His Ala Leu Gly Gly Met

525

Thr Ser Leu Glu Leu Glu

540

Glu Leu Phe Leu Gly Gln

555 560

Ser Ala Pro Pro Ala Gly

575

Ser Val Ser Cys

580

- 192 046157 <210> 106 <211> 394 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic polypeptide <400> 106

Met Ala Ala Asp His Leu Met Leu 1 5

Ala Glu Gly Tyr Arg Leu Val Gln

15

Arg Pro Pro Ser Ala Ala Ala Ala 20

His Gly Pro His Ala Leu Arg Thr 25 30

Leu Pro Pro Tyr Ala Gly Pro Gly

40

Leu Asp Ser Gly Leu Arg Pro Arg 45

Gly Ala Pro Leu Gly Pro Pro Pro

55

Pro Arg Gln Pro Gly Ala Leu Ala 60

Tyr Gly Ala Phe Gly Pro Pro Ser 65 70

Ser Phe Gln Pro Phe Pro Ala Val

80

Pro Pro Pro Ala Ala Gly Ile Ala 85

His Leu Gln Pro Val Ala Thr Pro

95

Tyr Pro Gly Arg Ala Ala Ala Pro

100

Pro Asn Ala Pro Gly Gly Pro Pro

105 110

Gly Pro Gln Pro Ala Pro Ser Ala

115 120

Ala Ala Pro Pro Pro Pro Ala His

125

Ala Leu Gly Gly Met Asp Ala Glu

130 135

Leu Ile Asp Glu Glu Ala Leu Thr

140

Ser Leu Glu Leu Glu Leu Gly Leu

145 150

His Arg Val Arg Glu Leu Pro Glu

155 160

Leu Phe Leu Gly Gln Ser Glu Phe

165

Asp Cys Phe Ser Asp Leu Gly Ser

170 175

Ala Pro Pro Ala Gly Ser Val Ser

180

Cys Gly Gly Ser Gly Gly Gly Ser

185 190

Gly Gln Ser Gln Leu Ile Lys Pro

195 200

Ser Arg Met Arg Lys Tyr Pro Asn

205

- 193 046157

Arg Pro

210

Ser Lys

Thr Pro

Glu Ser 225

Cys Asp

Arg Arg

230

Ile Arg

Ile His

Thr Gly 245

Arg Asn

Phe Ser

260

Arg Glu

Pro His 215

Phe Ser

Gln Lys

Asp Asn

Glu Arg

Arg Ser

Pro Phe

250

Leu His 265

Thr Gly

Glu Lys 275

Pro Phe

Ala Cys

280

Asp Ile

Pro Tyr 220

Asp Asn 235

Gln Cys

Thr His

Cys Gly

Ala Cys Pro Val

Leu Val Arg His

240

Arg Ile Cys Met

255

Ile Arg Thr His

270

Arg Lys Phe Ala 285

Arg Ser

290

Asp Glu

Leu Val

Arg His 295

Thr Lys

Ile His

300

Leu Arg Gln Lys

Asp Arg 305

Pro Tyr

Ala Cys

310

Pro Val

Glu Ser

Cys Asp 315

Arg Arg Phe Ser

320

Gln Ser

Gly Asn

Leu Thr 325

Glu His

Ile Arg

330

Ile His

Thr Gly Gln Lys

335

Pro Phe

Gln Cys

340

Arg Ile

Cys Met

Arg Asn 345

Phe Ser

Thr Ser Gly His 350

Leu Val

Arg His 355

Ile Arg

Thr His

360

Thr Gly

Glu Lys

Pro Phe Ala Cys 365

Asp Ile

370

Cys Gly

Arg Lys

Phe Ala 375

Gln Asn

Ser Thr

380

Leu Thr Glu His

Thr Lys 385

Ile His

Leu Arg

390

Gln Lys

Asp Lys <210> 107 <211> 388 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial polypeptide sequence

Synthetic <400> 107

Met Gln Ser Gln Leu Ile Lys Pro Ser Arg Met Arg 1 5 10

Lys Tyr Pro Asn

Arg Pro Ser Lys Thr Pro Pro His Glu Arg Pro Tyr

Ala Cys Pro Val

- 194 046157

Glu Ser Cys Asp Arg 35

Arg Phe Ser Arg Ser 40

Asp Asn Leu Val Arg 45

Ile Arg Ile His Thr 50

Gly Gln Lys Pro Phe 55

Gln Cys Arg Ile Cys 60

Arg Asn Phe Ser Arg 65

Glu Asp Asn Leu His 70

Thr His Ile Arg Thr 75

Thr Gly Glu Lys Pro

Phe Ala Cys Asp Ile

Cys Gly Arg Lys Phe

Arg Ser Asp Glu Leu

100

Val Arg His Thr Lys

105

Ile His Leu Arg Gln

110

Asp Arg Pro Tyr Ala 115

Cys Pro Val Glu Ser 120

Cys Asp Arg Arg Phe 125

Gln Ser Gly Asn Leu

130

Thr Glu His Ile Arg

135

Ile His Thr Gly Gln

140

Pro Phe Gln Cys Arg 145

Ile Cys Met Arg Asn 150

Phe Ser Thr Ser Gly 155

Leu Val Arg His Ile

165

Arg Thr His Thr Gly

170

Glu Lys Pro Phe Ala

175

Asp Ile Cys Gly Arg

180

Lys Phe Ala Gln Asn

185

Ser Thr Leu Thr Glu

190

Thr Lys Ile His Leu 195

Arg Gln Lys Asp Lys

200

Leu Glu Met Ala Asp

205

Leu Met Leu Ala Glu

210

Gly Tyr Arg Leu Val

215

Gln Arg Pro Pro Ser

220

Ala Ala Ala His Gly 225

Pro His Ala Leu Arg 230

Thr Leu Pro Pro Tyr 235

Gly Pro Gly Leu Asp

245

Ser Gly Leu Arg Pro

250

Arg Gly Ala Pro Leu

255

Pro Pro Pro Pro Arg

260

Gln Pro Gly Ala Leu

265

Ala Tyr Gly Ala Phe

270

His

Met

His 80

Ala

Lys

Ser

Lys

His 160

Cys

His

His

Ala

Ala 240

Gly

Gly

- 195 046157

Pro Pro Ser Ser Phe Gln Pro Phe Pro Ala Val Pro

275 280

Pro Pro Ala Ala 285

Gly Ile Ala His Leu Gln Pro Val Ala Thr Pro Tyr

290 295 300

Pro Gly Arg Ala

Ala Ala Pro Pro Asn Ala Pro Gly Gly Pro Pro Gly

305 310 315

Pro Gln Pro Ala

320

Pro Ser Ala Ala Ala Pro Pro Pro Pro Ala His Ala

325 330

Leu Gly Gly Met

335

Asp Ala Glu Leu Ile Asp Glu Glu Ala Leu Thr Ser

340 345

Leu Glu Leu Glu

350

Leu Gly Leu His Arg Val Arg Glu Leu Pro Glu Leu

355 360

Phe Leu Gly Gln 365

Ser Glu Phe Asp Cys Phe Ser Asp Leu Gly Ser Ala

370 375 380

Pro Pro Ala Gly

Ser Val Ser Cys 385 <210> 108 <211> 479 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial sequence

Synthetic polypeptide <400> 108

Met Gln Ser Gln 1

Leu Ile Lys Pro 5

Ser Arg Met Arg 10

Lys Tyr Pro Asn 15

Arg Pro Ser Lys

Thr Pro Pro His

Glu Arg Pro Tyr 25

Ala Cys Pro Val 30

Glu Ser Cys Asp 35

Arg Arg Phe Ser 40

Arg Ser Asp Asn

Leu Val Arg His 45

Ile Arg Ile His 50

Thr Gly Gln Lys 55

Pro Phe Gln Cys 60

Arg Ile Cys Met

Arg Asn Phe Ser 65

Arg Glu Asp Asn 70

Leu His Thr His

Ile Arg Thr His

Thr Gly Glu Lys

Pro Phe Ala Cys

Asp Ile Cys Gly

Arg Lys Phe Ala

- 196 046157

90 95

Arg

Asp

Gln

Pro 145

Leu

Asp

Thr

Glu

Gly 225

Gln

Phe

Asn

Gly

Ser 305

Ala

Ser Asp Glu Leu Val

100

Arg His Thr Lys Ile 105

His Leu Arg Gln Lys 110

Arg Pro Tyr Ala Cys 115

Pro Val Glu Ser Cys

120

Asp Arg Arg Phe Ser 125

Ser Gly Asn Leu Thr 130

Glu His Ile Arg Ile 135

His Thr Gly Gln Lys 140

Phe Gln Cys Arg Ile

150

Cys Met Arg Asn Phe

155

Ser Thr Ser Gly His

160

Val Arg His Ile Arg

165

Thr His Thr Gly Glu

170

Lys Pro Phe Ala Cys

175

Ile Cys Gly Arg Lys

180

Phe Ala Gln Asn Ser

185

Thr Leu Thr Glu His

190

Lys Ile His Leu Arg

195

Gln Lys Asp Lys Leu

200

Glu Met Ser Gly Leu 205

Met Ala Asp His Met 210

Met Ala Met Asn His 215

Gly Arg Phe Pro Asp 220

Thr Asn Gly Leu His

230

His His Pro Ala His

235

Arg Met Gly Met Gly

240

Phe Pro Ser Pro His

245

His His Gln Gln Gln

250

Gln Pro Gln His Ala

255

Asn Ala Leu Met Gly 260

Ala Thr Ser Gly Ile 275

Gly His Pro Pro Ser 290

Gln Phe Met Gly Pro

310

Ser Met Gln Leu Gln

325

Glu His Ile His Tyr 265

Gly Ala Gly Asn Met 270

Arg His Ala Met Gly 280

Ala Leu Ala Pro Ala 295

Pro Val Ala Ser Gln

315

Lys Leu Asn Asn Gln

330

Pro Gly Thr Val Asn

285

Ala Arg Phe Asn Asn 300

Gly Gly Ser Leu Pro

320

Tyr Phe Asn His His

335

- 197 046157

Pro Tyr Pro His Asn His Tyr Met Pro Asp Leu His

340 345

Pro Ala Ala Gly

350

His Gln Met Asn Gly Thr Asn Gln His Phe Arg Asp

355 360

Cys Asn Pro Lys 365

His Ser Gly Gly Ser Ser Thr Pro Gly Gly Ser Gly

370 375 380

Gly Ser Ser Thr

Pro Gly Gly Ser Gly Ser Ser Ser Gly Gly Gly Ala

385 390 395

Gly Ser Ser Asn

400

Ser Gly Gly Gly

Ser Gly Ser Gly Asn Met Pro Ala

405 410

Ser Val Ala His

415

Val Pro Ala Ala Met Leu Pro Pro Asn Val Ile Asp

420 425

Thr Asp Phe Ile 430

Asp Glu Glu Val Leu Met Ser Leu Val Ile Glu Met

435 440

Gly Leu Asp Arg 445

Ile Lys Glu Leu Pro Glu Leu Trp Leu Gly Gln Asn

450 455 460

Glu Phe Asp Phe

Met Thr Asp Phe Val Cys Lys Gln Gln Pro Ser Arg

465 470 475

Val Ser Cys <210> 109 <211> 476 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial sequence

Synthetic polypeptide <400> 109

Met Ser Gly Leu 1

Glu Met Ala Asp 5

His Met Met Ala

Met Asn His Gly 15

Arg Phe Pro Asp

Gly Thr Asn Gly

Leu His His His 25

Pro Ala His Arg 30

Met Gly Met Gly 35

Gln Phe Pro Ser

Pro His His His

Gln Gln Gln Gln 45

Pro Gln His Ala 50

Phe Asn Ala Leu

Met Gly Glu His

Ile His Tyr Gly

Ala Gly Asn Met

Asn Ala Thr Ser

Gly Val Arg His

Ala Met Gly Pro

- 198 046157

Gly

Arg

Gly

Phe

Pro 145

Cys

Gly

Gly

Ser

Thr 225

Gly

Glu

Val

Pro

Pro 305

75 80

Thr Val Asn Gly Gly

Phe Asn Asn Ser Gln

100

Ser Leu Pro Ala Ser

115

Asn His His Pro Tyr 130

Ala Ala Gly His Gln

150

Asn Pro Lys His Ser

165

Ser Ser Thr Pro Gly 180

Ser Ser Asn Ser Gly

195

Val Ala His Val Pro 210

Asp Phe Ile Asp Glu

230

Leu Asp Arg Ile Lys

245

Phe Asp Phe Met Thr 260

Ser Cys Gln Ser Gln

275

Asn Arg Pro Ser Lys 290

Val Glu Ser Cys Asp

310

His Pro Pro Ser Ala

Phe Met Gly Pro Pro 105

Met Gln Leu Gln Lys

120

Pro His Asn His Tyr 135

Met Asn Gly Thr Asn

155

Gly Gly Ser Ser Thr

170

Gly Ser Gly Ser Ser 185

Gly Gly Ser Gly Ser 200

Ala Ala Met Leu Pro 215

Glu Val Leu Met Ser

235

Glu Leu Pro Glu Leu

250

Asp Phe Val Cys Lys 265

Leu Ile Lys Pro Ser 280

Thr Pro Pro His Glu 295

Arg Arg Phe Ser Arg

315

Leu Ala Pro Ala Ala

Val Ala Ser Gln Gly

110

Leu Asn Asn Gln Tyr

125

Met Pro Asp Leu His 140

Gln His Phe Arg Asp

160

Pro Gly Gly Ser Gly

175

Ser Gly Gly Gly Ala

190

Gly Asn Met Pro Ala 205

Pro Asn Val Ile Asp 220

Leu Val Ile Glu Met

240

Trp Leu Gly Gln Asn

255

Gln Gln Pro Ser Arg 270

Arg Met Arg Lys Tyr

285

Arg Pro Tyr Ala Cys 300

Ser Asp Asn Leu Val

320

- 199 046157

Arg His Ile Arg Ile His Thr Gly Gln Lys Pro

325 330

Phe

Gln Cys Arg Ile

335

Cys Met Arg Asn Phe Ser Arg Glu Asp Asn Leu His

340 345

Thr His Ile Arg

350

Thr His Thr Gly Glu Lys Pro Phe Ala Cys Asp Ile

355 360

Cys Gly Arg Lys 365

Phe Ala Arg Ser Asp Glu Leu Val Arg His

370 375

Thr Lys

380

Ile His Leu Arg

Gln Lys Asp 385

Arg Pro Tyr

390

Phe Ser Gln

Ser Gly Asn

405

Gln Lys Pro

Phe Gln Cys 420

Gly His Leu

435

Val Arg His

Ala Cys Asp

450

Ile Cys Gly

Glu His Thr 465

Lys Ile His

470

Ala Cys Pro

Leu Thr Glu

Arg Ile Cys 425

Ile Arg Thr 440

Arg Lys Phe 455

Leu Arg Gln

Val Glu Ser

395

His Ile Arg 410

Met Arg Asn

His Thr Gly

Ala Gln Asn

460

Lys Asp Lys

475 <210> 110 <211> 554 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial sequence polypeptide <400> 110

Met Ala Ala Asp His Leu Met Leu Ala Glu Gly Tyr

5 10

Arg Pro Pro Ser Ala Ala Ala Ala His Gly Pro His 20 25

Leu Pro Pro Tyr Ala Gly Pro Gly Leu Asp Ser Gly 35 40

Cys Asp Arg Arg

400

Ile His Thr Gly 415

Phe Ser Thr Ser

430

Glu Lys Pro Phe 445

Ser Thr Leu Thr

Synthetic

Arg Leu Val Gln 15

Ala Leu Arg Thr 30

Leu Arg Pro Arg 45

Gly Ala Pro Leu Gly Pro Pro Pro Pro Arg Gln Pro

Gly Ala Leu Ala

- 200 046157

55 60

Tyr 65

Pro

Tyr

Gly

Ala

Ser 145

Leu

Ala

Gly

Arg

Pro 225

Leu

Glu

Ala

Cys

Gly Ala Phe Gly Pro Pro 70

Ser Ser Phe Gln Pro Phe Pro Ala Val

80

Pro Pro Ala Ala Gly Ile Ala His 85

Leu Gln Pro Val Ala Thr Pro

95

Pro Gly Arg Ala Ala

100

Pro Gln Pro Ala Pro

115

Leu Gly Gly Met Asp 130

Leu Glu Leu Glu Leu

150

Phe Leu Gly Gln Ser

165

Pro Pro Ala Gly Ser

180

Arg Pro His Ala Cys 195

Ser Asp Asn Leu Val 210

Phe Gln Cys Arg Ile

230

His Thr His Ile Arg

245

Phe Cys Gly Arg Lys 260

Lys Ile His Leu Lys 275

Asp Arg Arg Phe Ser 290

Ala Pro Pro Asn Ala

105

Pro Gly Gly Pro Pro

110

Ser Ala Ala Ala Pro

120

Pro Pro Pro Ala His

125

Ala Glu Leu Ile Asp 135

Glu Glu Ala Leu Thr 140

Gly Leu His Arg Val

155

Glu Phe Asp Cys Phe

170

Val Ser Cys Gly Gly

185

Pro Ala Glu Gly Cys 200

Arg His Leu Arg Ile 215

Cys Met Arg Ser Phe

235

Thr His Thr Gly Glu

250

Phe Ala Arg Ser Asp

265

Gln Lys Glu His Ala

280

Gln Ser Gly Asn Leu 295

Arg Glu Leu Pro Glu

160

Ser Asp Leu Gly Ser

175

Ser Gly Gly Gly Ser

190

Asp Arg Arg Phe Ser 205

His Thr Gly His Lys 220

Ser Arg Glu Asp Asn

240

Lys Pro Phe Ala Cys

255

Glu Leu Val Arg His 270

Cys Pro Ala Glu Gly

285

Thr Glu His Leu Arg 300

- 201 046157

Ile His Thr Gly His 305

Lys Pro Phe Gln Cys 310

Arg Ile Cys Met Arg 315

Phe Ser Thr Ser Gly

325

His Leu Val Arg His

330

Ile Arg Thr His Thr

335

Glu Lys Pro Phe Ala

340

Cys Glu Phe Cys Gly

345

Arg Lys Phe Ala Gln

350

Ser Thr Leu Thr Glu

355

His Ala Lys Ile His 360

Leu Lys Gln Lys Glu 365

Leu Glu Met Ala Asp

370

His Leu Met Leu Ala

375

Glu Gly Tyr Arg Leu 380

Gln Arg Pro Pro Ser 385

Ala Ala Ala Ala His 390

Gly Pro His Ala Leu 395

Thr Leu Pro Pro Tyr

405

Ala Gly Pro Gly Leu

410

Asp Ser Gly Leu Arg

415

Arg Gly Ala Pro Leu

420

Gly Pro Pro Pro Pro

425

Arg Gln Pro Gly Ala

430

Ala Tyr Gly Ala Phe 435

Gly Pro Pro Ser Ser 440

Phe Gln Pro Phe Pro

445

Val Pro Pro Pro Ala

450

Ala Gly Ile Ala His 455

Leu Gln Pro Val Ala

460

Pro Tyr Pro Gly Arg 465

Ala Ala Ala Pro Pro 470

Asn Ala Pro Gly Gly 475

Pro Gly Pro Gln Pro

485

Ala Pro Ser Ala Ala

490

Ala Pro Pro Pro Pro

495

His Ala Leu Gly Gly

500

Met Asp Ala Glu Leu

505

Ile Asp Glu Glu Ala

510

Thr Ser Leu Glu Leu

515

Glu Leu Gly Leu His 520

Arg Val Arg Glu Leu 525

Glu Leu Phe Leu Gly

530

Gln Ser Glu Phe Asp 535

Cys Phe Ser Asp Leu

540

Ser 320

Gly

Asn

Lys

Val

Arg 400

Pro

Leu

Ala

Thr

Pro 480

Ala

Leu

Pro

Gly

Ser Ala Pro Pro Ala 545

Gly Ser Val Ser Cys 550

- 202 046157 <210> 111 <211> 362 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial sequence

Synthetic polypeptide <400> 111

Met Arg Pro His Ala Cys 1 5

Pro Ala Glu Gly Cys Asp 10

Arg Arg Phe Ser 15

Arg Ser Asp Asn Leu Val 20

Arg His Leu Arg Ile His 25

Thr Gly His Lys 30

Pro Phe Gln Cys Arg Ile 35

Cys Met Arg Ser Phe Ser 40

Arg Glu Asp Asn 45

Leu His Thr His Ile Arg 50

Thr His Thr Gly Glu Lys 55 60

Pro Phe Ala Cys

Glu Phe Cys Gly Arg Lys 65 70

Phe Ala Arg Ser Asp Glu 75

Leu Val Arg His

Ala Lys Ile His Leu Lys 85

Gln Lys Glu His Ala Cys 90

Pro Ala Glu Gly

Cys Asp Arg Arg Phe Ser

100

Gln Ser Gly Asn Leu Thr

105

Glu His Leu Arg 110

Ile His Thr Gly His Lys 115

Pro Phe Gln Cys Arg Ile

120

Cys Met Arg Ser 125

Phe Ser Thr Ser Gly His

130

Leu Val Arg His Ile Arg

135 140

Thr His Thr Gly

Glu Lys Pro Phe Ala Cys

145 150

Glu Phe Cys Gly Arg Lys

155

Phe Ala Gln Asn

160

Ser Thr Leu Thr Glu His

165

Ala Lys Ile His Leu Lys

170

Gln Lys Glu Lys 175

Leu Glu Met Ala Asp His

180

Leu Met Leu Ala Glu Gly

185

Tyr Arg Leu Val 190

Gln Arg Pro Pro Ser Ala

195

Ala Ala Ala His Gly Pro 200

His Ala Leu Arg 205

- 203 046157

Thr Leu Pro Pro Tyr Ala Gly Pro Gly Leu Asp Ser

210 215 220

Gly Leu Arg Pro

Arg Gly Ala Pro Leu Gly Pro Pro Pro Pro Arg Gln

225 230 235

Pro Gly Ala Leu

240

Ala Tyr Gly Ala Phe Gly Pro Pro Ser Ser Phe Gln

245 250

Pro Phe Pro Ala

255

Val Pro Pro Pro Ala Ala Gly Ile Ala His Leu Gln

260 265

Pro Val Ala Thr

270

Pro Tyr Pro Gly Arg Ala Ala Ala Pro Pro Asn Ala

275 280

Pro Gly Gly Pro 285

Pro Gly Pro Gln Pro Ala Pro Ser Ala Ala Ala Pro

290 295 300

Pro Pro Pro Ala

His Ala Leu Gly Gly Met Asp Ala Glu Leu Ile Asp

305 310 315

Glu Glu Ala Leu

320

Thr Ser Leu Glu Leu Glu Leu Gly Leu His Arg Val

325 330

Arg Glu Leu Pro

335

Glu Leu Phe Leu Gly Gln Ser Glu Phe Asp Cys Phe

340 345

Ser Asp Leu Gly

350

Ser Ala Pro Pro Ala Gly Ser Val Ser Cys

355 360 <210> 112 <211> 479 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial sequence polypeptide <400> 112

Met Arg Pro His Ala Cys Pro Ala Glu Gly Cys Asp

510

Arg Ser Asp Asn Leu Val Arg His Leu Arg Ile His 2025

Pro Phe Gln Cys Arg Ile Cys Met Arg Ser Phe Ser 3540

Leu His Thr His Ile Arg Thr His Thr Gly Glu Lys

Synthetic

Arg Arg Phe Ser 15

Thr Gly His Lys 30

Arg Glu Asp Asn 45

Pro Phe Ala Cys

- 204 046157

55 60

Glu Phe Cys Gly Arg Lys Phe Ala Arg Ser Asp Glu Leu Val Arg 65 70 75

Ala Lys Ile His Leu

Lys Gln Lys Glu His

Ala Cys Pro Ala Glu

Cys Asp Arg Arg Phe

100

Ser Gln Ser Gly Asn

105

Leu Thr Glu His Leu

110

Ile His Thr Gly His 115

Lys Pro Phe Gln Cys

120

Arg Ile Cys Met Arg 125

Phe Ser Thr Ser Gly

130

His Leu Val Arg His

135

Ile Arg Thr His Thr 140

Glu Lys Pro Phe Ala 145

Cys Glu Phe Cys Gly 150

Arg Lys Phe Ala Gln 155

Ser Thr Leu Thr Glu

165

His Ala Lys Ile His

170

Leu Lys Gln Lys Glu

175

Lys Ala Glu Lys Gly

180

Gly Ala Pro Ser Ala

185

Ser Ser Ala Pro Pro

190

Ser Leu Ala Pro Val

195

Val Thr Thr Cys Ala

200

Leu Glu Met Ser Gly

205

Glu Met Ala Asp His 210

Met Met Ala Met Asn

215

His Gly Arg Phe Pro

220

Gly Thr Asn Gly Leu 225

His His His Pro Ala 230

His Arg Met Gly Met 235

Gln Phe Pro Ser Pro

245

His His His Gln Gln

250

Gln Gln Pro Gln His

255

Phe Asn Ala Leu Met

260

Gly Glu His Ile His

265

Tyr Gly Ala Gly Asn

270

Asn Ala Thr Ser Gly

275

Ile Arg His Ala Met 280

Gly Pro Gly Thr Val 285

Gly Gly His Pro Pro

290

Ser Ala Leu Ala Pro

295

Ala Ala Arg Phe Asn

300

His 80

Gly

Arg

Ser

Gly

Asn 160

Lys

Val

Leu

Asp

Gly 240

Ala

Met

Asn

Asn

- 205 046157

Ser Gln Phe Met Gly Pro Pro Val Ala Ser Gln Gly

305 310 315

Gly Ser Leu Pro

320

Ala Ser Met Gln Leu Gln Lys Leu Asn Asn Gln Tyr

325 330

Phe Asn His His

335

Pro Tyr Pro His Asn His Tyr Met Pro Asp Leu His

340 345

Pro Ala Ala Gly

350

His Gln Met Asn Gly Thr Asn Gln His Phe Arg Asp

355 360

Cys Asn Pro Lys 365

His Ser Gly Gly Ser Ser Thr Pro Gly Gly Ser Gly

370 375 380

Gly Ser Ser Thr

Pro Gly Gly Ser Gly Ser Ser Ser Gly Gly Gly Ala

385 390 395

Gly Ser Ser Asn

400

Ser Gly Gly Gly Ser Gly Ser Gly Asn Met Pro Ala

405 410

Ser Val Ala His

415

Val Pro Ala Ala Met Leu Pro Pro Asn Val Ile Asp

420 425

Thr Asp Phe Ile

430

Asp Glu Glu Val Leu Met Ser Leu Val Ile Glu Met

435 440

Gly Leu Asp Arg 445

Ile Lys Glu Leu Pro Glu Leu Trp Leu Gly Gln Asn

450 455 460

Glu Phe Asp Phe

Met Thr Asp Phe Val Cys Lys Gln Gln Pro Ser Arg

465 470 475

Val Ser Cys <210> 113 <211> 476 <212> Protein <213>

Artificial sequence <220>

<223> Description of the artificial sequence

Synthetic <400>

Met 1

Arg polypeptide

113

Ser

Phe

Gly

Pro

Leu

Glu 5

Met

Ala

Asp

His

Met 10

Met

Ala

Met Asn His Gly 15

Asp 20

Gly

Thr

Asn

Gly

Leu 25

His

His

His

Pro Ala His Arg 30

Met

Gly

Met

Gly

Gln

Phe

Pro

Ser

Pro

His

His

His

Gln Gln Gln Gln

- 206 046157

40 45

Pro

Ala 65

Gly

Arg

Gly

Phe

Pro 145

Cys

Gly

Gly

Ser

Thr 225

Gly

Glu

Val

Gln His Ala Phe Asn 50

Gly Asn Met Asn Ala

Thr Val Asn Gly Gly

Phe Asn Asn Ser Gln

100

Ser Leu Pro Ala Ser

115

Asn His His Pro Tyr 130

Ala Ala Gly His Gln

150

Asn Pro Lys His Ser

165

Ser Ser Thr Pro Gly 180

Ser Ser Asn Ser Gly

195

Val Ala His Val Pro 210

Asp Phe Ile Asp Glu

230

Leu Asp Arg Ile Lys

245

Phe Asp Phe Met Thr 260

Ser Cys Arg Pro His

275

Ala Leu Met Gly Glu 55

Thr Ser Gly Val Arg 75

His Pro Pro Ser Ala

Phe Met Gly Pro Pro 105

Met Gln Leu Gln Lys

120

Pro His Asn His Tyr 135

Met Asn Gly Thr Asn

155

Gly Gly Ser Ser Thr

170

Gly Ser Gly Ser Ser 185

Gly Gly Ser Gly Ser

200

Ala Ala Met Leu Pro 215

Glu Val Leu Met Ser

235

Glu Leu Pro Glu Leu

250

Asp Phe Val Cys Lys 265

Ala Cys Pro Ala Glu

280

His Ile His Tyr Gly 60

His Ala Met Gly Pro

Leu Ala Pro Ala Ala

Val Ala Ser Gln Gly

110

Leu Asn Asn Gln Tyr

125

Met Pro Asp Leu His 140

Gln His Phe Arg Asp

160

Pro Gly Gly Ser Gly

175

Ser Gly Gly Gly Ala

190

Gly Asn Met Pro Ala

205

Pro Asn Val Ile Asp 220

Leu Val Ile Glu Met

240

Trp Leu Gly Gln Asn

255

Gln Gln Pro Ser Arg 270

Gly Cys Asp Arg Arg

285

- 207 046157

Phe Ser Arg

290

Ser Asp Asn

Leu Val Arg 295

His Leu Arg

300

Ile His Thr Gly

His Lys Pro 305

Phe Gln Cys

310

Arg Ile Cys

Met Arg Ser

315

Phe Ser Arg Glu

320

Asp Asn Leu

His Thr His

325

Ile Arg Thr

His Thr Gly 330

Glu Lys Pro Phe

335

Ala Cys Glu

Phe Cys Gly 340

Arg Lys Phe

345

Ala Arg Ser

Asp Glu Leu Val

350

Arg His Ala

355

Lys Ile His

Leu Lys Gln

360

Lys Glu His

Ala Cys Pro Ala 365

Glu Gly Cys

370

Asp Arg Arg

Phe Ser Gln 375

Ser Gly Asn

380

Leu Thr Glu His

Leu Arg Ile 385

His Thr Gly

390

His Lys Pro

Phe Gln Cys

395

Arg Ile Cys Met

400

Arg Ser Phe

Ser Thr Ser

405

Gly His Leu

Val Arg His 410

Ile Arg Thr His 415

Thr Gly Glu

Lys Pro Phe 420

Ala Cys Glu

425

Phe Cys Gly

Arg Lys Phe Ala

430

Gln Asn Ser

435

Thr Leu Thr

Glu His Ala

440

Lys Ile His

Leu Lys Gln Lys 445

Glu Lys Lys

450

Ala Glu Lys

Gly Gly Ala 455

Pro Ser Ala

460

Ser Ser Ala Pro

Pro Val Ser 465

Leu Ala Pro

470

Val Val Thr

Thr Cys Ala

475 <210> 114 <211> 559 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial polypeptide sequence

Synthetic <400> 114

Met Thr Gly Lys Leu Ala Glu Lys Leu Pro Val Thr 1 5 10

Met Ser Ser Leu

Leu Asn Gln Leu Pro Asp Asn Leu Tyr Pro Glu Glu

Ile Pro Ser Ala

- 208 046157

Leu Asn Leu Phe Ser

Gly Ser Ser Asp Ser 40

Val Val His Tyr Asn 45

Met Ala Thr Glu Asn

Val Met Asp Ile Gly 55

Leu Thr Asn Glu Lys 60

Asn Pro Glu Leu Ser 65

Tyr Ser Gly Ser Phe 70

Gln Pro Ala Pro Gly 75

Lys Thr Val Thr Tyr

Leu Gly Lys Phe Ala

Phe Asp Ser Pro Ser

Trp Cys Gln Asp Asn

100

Ile Ile Ser Leu Met

105

Ser Ala Gly Ile Leu

110

Val Pro Pro Ala Ser

115

Gly Ala Leu Ser Thr 120

Gln Thr Ser Thr Ala

125

Met Val Gln Pro

130

Gln Gly Asp Val Glu

135

Ala Met Tyr Pro Ala 140

Pro Pro Tyr Ser Asn 145

Cys Gly Asp Leu Tyr 150

Ser Glu Pro Val Ser 155

His Asp Pro Gln Gly

165

Asn Pro Gly Leu Ala

170

Tyr Ser Pro Gln Asp

175

Gln Ser Ala Lys Pro

180

Ala Leu Asp Ser Asn

185

Leu Phe Pro Met Ile

190

Asp Tyr Asn Leu Tyr 195

His His Pro Asn Asp

200

Met Gly Ser Ile Pro 205

His Lys Pro Phe Gln

210

Gly Met Asp Pro Ile 215

Arg Val Asn Pro Pro 220

Ile Thr Pro Leu Glu 225

Thr Ile Lys Ala Phe 230

Lys Asp Lys Gln Ile 235

Pro Gly Phe Gly Ser

245

Leu Pro Gln Pro Pro

250

Leu Thr Leu Lys Pro

255

Arg Pro Arg Lys Tyr

260

Pro Asn Arg Pro Ser

265

Lys Thr Pro Leu His

270

Gln

Pro

Asn 80

Asn

Gly

Ser

Leu

Phe 160

Tyr

Pro

Glu

Pro

His 240

Ile

Glu

- 209 046157

Arg Pro His Ala Cys 275

Pro Ala Glu Gly Cys 280

Asp Arg Arg Phe Ser 285

Arg Asp Glu Leu Asn

290

Val His Leu Arg Ile

295

His Thr Gly His Lys 300

Phe Gln Cys Arg Ile 305

Cys Met Arg Ser Phe 310

Ser Arg Ser Asp His 315

Thr Asn His Ile Arg

325

Thr His Thr Gly Glu

330

Lys Pro Phe Ala Cys

335

Phe Cys Gly Arg Lys

340

Phe Ala Arg Ser Asp

345

Asp Leu Val Arg His

350

Lys Ile His Leu Lys 355

Gln Lys Glu His Ala 360

Cys Pro Ala Glu Gly

365

Asp Arg Arg Phe Ser

370

Arg Ser Asp Asn Leu

375

Val Arg His Leu Arg

380

His Thr Gly His Lys 385

Pro Phe Gln Cys Arg 390

Ile Cys Met Arg Ser 395

Ser His Arg Thr Thr

405

Leu Thr Asn His Ile

410

Arg Thr His Thr Gly

415

Lys Pro Phe Ala Cys

420

Glu Phe Cys Gly Arg

425

Lys Phe Ala Arg Glu

430

Asn Leu His Thr His

435

Ala Lys Ile His Leu 440

Lys Gln Lys Glu His 445

Cys Pro Ala Glu Gly

450

Cys Asp Arg Arg Phe

455

Ser Thr Ser His Ser

460

Thr Glu His Leu Arg 465

Ile His Thr Gly His 470

Lys Pro Phe Gln Cys 475

Ile Cys Met Arg Ser

485

Phe Ser Gln Ser Ser

490

Ser Leu Val Arg His

495

Arg Thr His Thr Gly

500

Glu Lys Pro Phe Ala

505

Cys Glu Phe Cys Gly

510

Lys Phe Ala Arg Glu

515

Asp Asn Leu His Thr 520

His Ala Lys Ile His 525

Arg

Pro

Leu 320

Glu

Ala

Cys

Ile

Phe 400

Glu

Asp

Ala

Leu

Arg 480

Ile

Arg

Leu

- 210 046157

Lys Gln Lys Glu Lys Lys Ala Glu

Lys Gly Gly Ala

Pro Ser Ala Ser

530

535

540

Ser Ala Pro Pro Val

Ser Leu Ala Pro Val Val Thr

Thr Cys Ala

545 550 555 <210> 115 <211> 631 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial sequence polypeptide <400> 115

Met Ala Ala Ala Lys Ala Glu Met Gln Leu Met Ser

5 10

Synthetic

Pro Leu Gln Ile

Ser Asp Pro Phe Gly Ser Phe Pro His Ser Pro Thr 20 25

Met Asp Asn Tyr 30

Pro Lys Leu Glu Glu Met Met Leu Leu Ser Asn Gly 35 40

Ala Pro Gln Phe 45

Leu Gly Ala Ala Gly Ala Pro Glu Gly Ser Gly Ser

55 60

Asn Ser Ser Ser

Ser Ser Ser Gly Gly Gly Gly Gly Gly Gly Gly Gly 65 70 75

Ser Asn Ser Ser

Ser Ser Ser Ser Thr Phe Asn Pro Gln Ala Asp Thr

90

Gly Glu Gln Pro

Tyr Glu His Leu Thr Ala Glu Ser Phe Pro Asp Ile

100 105

Ser Leu Asn Asn

110

Glu Lys Val Leu Val Glu Thr Ser Tyr Pro Ser Gln

115 120

Thr Thr Arg Leu 125

Pro Pro Ile Thr Tyr Thr Gly Arg Phe Ser Leu Glu

130 135 140

Pro Ala Pro Asn

Ser Gly Asn Thr Leu Trp Pro Glu Pro Leu Phe Ser

145 150 155

Leu Val Ser Gly

160

Leu Val Ser Met Thr Asn Pro Pro Ala Ser Ser Ser

165 170

Ser Ala Pro Ser

175

- 211 046157

Pro Ala Ala Ser Ser Ala Ser Ala

Ser Gln Ser Pro Pro Leu Ser

180 185 190

Ala Val Pro Ser Asn Asp Ser Ser Pro Ile Tyr Ser Ala Ala Pro

195 200 205

Phe Pro

210

Thr Pro Asn

Thr Asp Ile Phe Pro Glu Pro Gln Ser Gln

215 220

Phe Pro Gly Ser Ala Gly Thr Ala Leu Gln Tyr Pro Pro Pro Ala

225 230 235

Pro Ala Ala Lys Gly Gly Phe Gln Val Pro Met Ile Pro Asp Tyr

245 250 255

Phe Pro Gln Gln Gln Gly Asp Leu Gly Leu Gly Thr Pro Asp Gln

260 265 270

Pro Phe Gln Gly Leu Glu Ser Arg Thr Gln Gln Pro Ser Leu Thr

275 280 285

Leu Ser Thr Ile Lys Ala Phe Ala Thr Gln Ser Gly Ser Gln Asp

290 295 300

Lys Ala Leu Asn Thr Ser Tyr Gln Ser Gln Leu Ile Lys Pro Ser

305 310 315

Met Arg Lys Tyr Pro Asn Arg Pro Ser Lys Thr Pro Pro His Glu

325 330 335

Pro Tyr Ala Cys Pro Val Glu Ser Cys Asp Arg Arg Phe Ser Arg

340 345 350

Asp Asn Leu Val Arg His Ile Arg Ile His Thr Gly Gln Lys Pro

355 360 365

Gln Cys Arg Ile Cys Met Arg Asn Phe Ser His Arg Thr Thr Leu

370 375 380

Asn His Ile Arg Thr His Thr Gly Glu Lys Pro Phe Ala Cys Asp

385 390 395

Cys Gly Arg Lys Phe Ala Arg Glu Asp Asn Leu His Thr His Thr

405 410 415

Ile His Leu Arg Gln Lys Asp Arg Pro Tyr Ala Cys Pro Val Glu

420 425 430

Cys

Thr

Ala

Tyr 240

Leu

Lys

Pro

Leu

Arg 320

Arg

Ser

Phe

Thr

Ile 400

Lys

Ser

- 212 046157

Cys Asp

Ile His

450

Phe Ser 465

Glu Lys

Arg Arg 435

Thr Gly

Gln Ser

Pro Phe

Phe Ser

Gln Lys

Ser Ser

470

Ala Cys 485

Asp Asn

Leu His

500

Thr His

Thr Ser 440

Pro Phe 455

Leu Val

Asp Ile

Thr Lys

His Ser

Gln Cys

Arg His

Cys Gly

490

Ile His

505

Leu Thr

Arg Ile

460

Ile Arg 475

Arg Lys

Leu Arg

Lys Ala

Asp Lys 515

Ser Val

Val Ala

520

Ser Ser

Ala Thr

Glu His Ile Arg 445

Cys Met Arg Asn

Thr His Thr Gly

480

Phe Ala Arg Glu

495

Gln Lys Asp Lys 510

Ser Ser Leu Ser 525

Ser Tyr

530

Pro Ser

Pro Val

Ala Thr 535

Ser Tyr

Pro Ser

540

Pro Val Thr Thr

Ser Tyr 545

Pro Ser

Pro Ala

550

Thr Thr

Ser Tyr

Pro Ser 555

Pro Val Pro Thr

560

Ser Phe

Ser Ser

Pro Gly 565

Ser Ser

Thr Tyr

570

Pro Ser

Pro Val His Ser

575

Gly Phe

Pro Ser

580

Pro Ser

Val Ala

Thr Thr

585

Tyr Ser

Ser Val Pro Pro

590

Ala Phe

Pro Ala 595

Gln Val

Ser Ser

600

Phe Pro

Ser Ser

Ala Val Thr Asn 605

Ser Phe

610

Ser Ala

Ser Thr

Gly Leu 615

Ser Asp

Met Thr

620

Ala Thr Phe Ser

Pro Arg 625

Thr Ile

Glu Ile

630

Cys <210> 116 <211> 719 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial sequence polypeptide <400> 116

Synthetic

- 213 046157

Met Ala 1

Ala Ala

Lys Ala 5

GluMet

Gln Leu

Met Ser

ProLeu

Ser Asp

Pro Phe

Gly Ser

Phe Pro

His Ser 25

Pro Thr

Met Asp

Pro Lys

Leu Glu 35

GluMet

Met Leu

Leu Ser

Asn Gly

Ala Pro 45

Gln Ile 15

Asn Tyr

Gln Phe

Leu Gly Ala Ala Gly Ala Pro Glu Gly Ser Gly Ser Asn Ser Ser Ser 50 5560

Ser Ser Ser Gly Gly Gly Gly Gly Gly Gly Gly Gly Ser Asn Ser Ser 65 70 7580

Ser Ser Ser Ser Thr Phe Asn Pro Gln Ala Asp Thr Gly Glu Gln Pro 85 9095

Tyr Glu His Leu Thr Ala Glu Ser Phe Pro Asp Ile Ser Leu Asn Asn

100 105110

Glu Lys Val Leu Val Glu Thr Ser Tyr Pro Ser Gln Thr Thr Arg Leu

115 120125

Pro Pro Ile Thr Tyr Thr Gly Arg Phe Ser Leu Glu Pro Ala Pro Asn

130 135140

Ser Gly Asn Thr Leu Trp Pro Glu Pro Leu Phe Ser Leu Val Ser Gly

145 150 155160

Leu Val Ser Met Thr Asn Pro Pro Ala Ser Ser Ser Ser Ala Pro Ser

165 170175

Pro Ala Ala Ser Ser Ala Ser Ala Ser Gln Ser Pro Pro Leu Ser Cys

180 185190

Ala Val Pro Ser Asn Asp Ser Ser Pro Ile Tyr Ser Ala Ala Pro Thr

195 200205

Phe Pro Thr Pro Asn Thr Asp Ile Phe Pro Glu Pro Gln Ser Gln Ala

210 215220

Phe Pro Gly Ser Ala Gly Thr Ala Leu Gln Tyr Pro Pro Pro Ala Tyr

225 230 235240

Pro Ala Ala Lys Gly Gly Phe Gln Val Pro Met Ile Pro Asp Tyr Leu

245 250255

- 214 046157

Phe

Pro

Leu

Lys 305

Met

Pro

Asp

Gln

Asn 385

Cys

Ile

Cys

Ile

Phe 465

Glu

Asp

Pro Gln Gln Gln Gly 260

Phe Gln Gly Leu Glu

275

Ser Thr Ile Lys Ala 290

Ala Leu Asn Thr Ser

310

Arg Lys Tyr Pro Asn

325

Tyr Ala Cys Pro Val

340

Glu Leu Asn Val His

355

Cys Arg Ile Cys Met 370

His Ile Arg Thr His

390

Gly Arg Lys Phe Ala

405

His Leu Arg Gln Lys 420

Asp Arg Arg Phe Ser

435

His Thr Gly Gln Lys 450

Ser His Arg Thr Thr

470

Lys Pro Phe Ala Cys

485

Asn Leu His Thr His

500

Asp Leu Gly Leu Gly 265

Ser Arg Thr Gln Gln

280

Phe Ala Thr Gln Ser 295

Tyr Gln Ser Gln Leu

315

Arg Pro Ser Lys Thr

330

Glu Ser Cys Asp Arg 345

Ile Arg Ile His Thr

360

Arg Asn Phe Ser Arg 375

Thr Gly Glu Lys Pro

395

Arg Ser Asp Asp Leu

410

Asp Arg Pro Tyr Ala 425

Arg Ser Asp Asn Leu

440

Pro Phe Gln Cys Arg 455

Leu Thr Asn His Ile

475

Asp Ile Cys Gly Arg

490

Thr Lys Ile His Leu 505

Thr Pro Asp Gln Lys

270

Pro Ser Leu Thr Pro

285

Gly Ser Gln Asp Leu 300

Ile Lys Pro Ser Arg

320

Pro Pro His Glu Arg

335

Arg Phe Ser Arg Arg

350

Gly Gln Lys Pro Phe

365

Ser Asp His Leu Thr 380

Phe Ala Cys Asp Ile

400

Val Arg His Thr Lys 415

Cys Pro Val Glu Ser

430

Val Arg His Ile Arg 445

Ile Cys Met Arg Asn 460

Arg Thr His Thr Gly

480

Lys Phe Ala Arg Glu

495

Arg Gln Lys Asp Arg 510

- 215 046157

Pro Tyr

His Ser

530

Gln Cys 545

Arg His

Ala Cys 515

Leu Thr

Arg Ile

Ile Arg

Pro Val

Glu His

Cys Met

550

Thr His 565

Cys Gly

Arg Lys

580

Phe Ala

Ile His

Leu Arg 595

Gln Lys

Ser Ser

610

Ala Thr

Ser Ser

Ser Tyr 625

Pro Ser

Pro Val

630

Ser Tyr

Pro Ser

Pro Val 645

Thr Tyr

Pro Ser

660

Pro Val

Thr Thr

Tyr Ser 675

Ser Val

Glu Ser

520

Ile Arg 535

Arg Asn

Thr Gly

Arg Glu

Asp Lys

600

Leu Ser 615

Thr Thr

Pro Thr

His Ser

Pro Pro

680

Cys Asp

Ile His

Phe Ser

Glu Lys

570

Asp Asn 585

Lys Ala

Ser Tyr

Ser Tyr

Ser Phe

650

Gly Phe 665

Ala Phe

Arg Arg

Thr Gly

540

Gln Ser 555

Pro Phe

Leu His

Asp Lys

Pro Ser

620

Pro Ser 635

Ser Ser

Pro Ser

Pro Ala

Phe Ser Thr Ser 525

Gln Lys Pro Phe

Ser Ser Leu Val

560

Ala Cys Asp Ile

575

Thr His Thr Lys

590

Ser Val Val Ala 605

Pro Val Ala Thr

Pro Ala Thr Thr

640

Pro Gly Ser Ser

655

Pro Ser Val Ala

670

Gln Val Ser Ser 685

Phe Pro

690

Ser Ser

Ala Val

Thr Asn 695

Ser Phe

Ser Ala

700

Ser Thr Gly Leu

Ser Asp 705

Met Thr

Ala Thr

710

Phe Ser

Pro Arg

Thr Ile 715

Glu Ile Cys <210> 117 <211> 627 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial polypeptide sequence

Synthetic

- 216 046157 <400> 117 Met Ala Ala 1

Ala Lys Ala Glu Met Gln Leu Met Ser Pro Leu Gln

10 15

Ser Asp Pro

Phe Gly Ser Phe Pro His Ser Pro Thr Met Asp Asn

25 30

Pro Lys Leu 35

Glu Glu Met Met Leu Leu Ser Asn Gly Ala Pro Gln

45

Leu Gly Ala 50

Ala Gly Ala Pro Glu Gly Ser Gly Ser Asn Ser Ser 55 60

Ser Ser Ser 65

Gly Gly Gly Gly Gly Gly Gly Gly Gly Ser Asn Ser 70 75

Ser Ser Ser

Ser Thr Phe Asn Pro Gln Ala Asp Thr Gly Glu Gln

90 95

Tyr Glu His

Leu Thr Ala Glu Ser Phe Pro Asp Ile Ser Leu Asn

100 105 110

Glu Lys Val

115

Leu Val Glu Thr Ser Tyr Pro Ser Gln Thr Thr Arg

120 125

Pro Pro Ile

130

Thr Tyr Thr Gly Arg Phe Ser Leu Glu Pro Ala Pro

135 140

Ser Gly Asn 145

Thr Leu Trp Pro Glu Pro Leu Phe Ser Leu Val Ser

150 155

Leu Val Ser

Met Thr Asn Pro Pro Ala Ser Ser Ser Ser Ala Pro

165 170 175

Pro Ala Ala

Ser Ser Ala Ser Ala Ser Gln Ser Pro Pro Leu Ser

180 185 190

Ala Val Pro

195

Ser Asn Asp Ser Ser Pro Ile Tyr Ser Ala Ala Pro

200 205

Phe Pro Thr

210

Pro Asn Thr Asp Ile Phe Pro Glu Pro Gln Ser Gln

215 220

Phe Pro Gly 225

Ser Ala Gly Thr Ala Leu Gln Tyr Pro Pro Pro Ala

230 235

Pro Ala Ala

Lys Gly Gly Phe Gln Val Pro Met Ile Pro Asp Tyr

245 250 255

Ile

Tyr

Phe

Ser

Ser 80

Pro

Asn

Leu

Asn

Gly 160

Ser

Cys

Thr

Ala

Tyr 240

Leu

- 217 046157

Phe Pro Gln

Gln Gln Gly 260

Asp Leu Gly

265

Leu Gly Thr

Pro Asp Gln

270

Pro Phe Gln

275

Gly Leu Glu

Ser Arg Thr

280

Gln Gln Pro

Ser Leu Thr 285

Leu Ser Thr

290

Ile Lys Ala

Phe Ala Thr 295

Gln Ser Gly

300

Ser Gln Asp

Lys Ala Leu 305

Asn Thr Ser

310

Tyr Gln Ser

Gln Leu Ile

315

Lys Pro Ser

Met Arg Lys

Tyr Pro Asn

325

Arg Pro Ser

Lys Thr Pro 330

Pro His Glu

335

Pro Tyr Ala

Cys Pro Val 340

Glu Ser Cys

345

Asp Arg Arg

Phe Ser Arg

350

Asp Asn Leu

355

Val Arg His

Ile Arg Ile

360

His Thr Gly

Gln Lys Pro 365

Gln Cys Arg

370

Ile Cys Met

Arg Asn Phe 375

Ser His Arg

380

Thr Thr Leu

Asn His Ile 385

Arg Thr His

390

Thr Gly Glu

Lys Pro Phe

395

Ala Cys Asp

Cys Gly Arg

Lys Phe Ala

405

Arg Glu Asp

Asn Leu His 410

Thr His Ile

415

Thr His Thr

Gly Glu Lys 420

Pro Phe Ala

425

Cys Asp Ile

Cys Gly Arg

430

Phe Ser Thr

435

Ser His Ser

Leu Thr Glu

440

His Ile Arg

Ile His Thr 445

Gln Lys Pro

450

Phe Gln Cys

Arg Ile Cys 455

Met Arg Asn

460

Phe Ser Gln

Ser Ser Leu 465

Val Arg His

470

Ile Arg Thr

His Thr Gly 475

Glu Lys Pro

Ala Cys Asp

Ile Cys Gly

485

Arg Lys Phe

Ala Arg Glu 490

Asp Asn Leu

495

Thr His Thr

Lys Ile His

Leu Arg Gln

Lys Asp Lys

Lys Ala Asp

Lys

Pro

Leu

Arg 320

Arg

Ser

Phe

Thr

Ile 400

Arg

Lys

Gly

Ser

Phe 480

His

Lys

- 218 046157

Ser Val

Pro Val

530

Pro Ala 545

Pro Gly

Pro Ser

Gln Val

Ser Thr

610

500

Val Ala 515

Ala Thr

Thr Thr

Ser Ser

Val Ala

580

Ser Ser 595

Gly Leu

Ser Ser

Ser Tyr

Ser Tyr

550

Thr Tyr 565

Thr Thr

Phe Pro

Ser Asp

Ala Thr

520

Pro Ser 535

Pro Ser

Pro Ser

Tyr Ser

Ser Ser

600

Met Thr 615

505

Ser Ser

Pro Val

Pro Val

Pro Val

570

Ser Val

585

Ala Val

Ala Thr

510

Leu Ser

Thr Thr

540

Pro Thr 555

His Ser

Pro Pro

Thr Asn

Phe Ser

620

Ser Tyr Pro Ser 525

Ser Tyr Pro Ser

Ser Phe Ser Ser

560

Gly Phe Pro Ser

575

Ala Phe Pro Ala

590

Ser Phe Ser Ala 605

Pro Arg Thr Ile

Glu Ile Cys 625 <210> 118 <211> 631 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial sequence

Synthetic polypeptide <400> 118

Met Ala Ala Ala 1

Lys Ala Glu Met 5

Gln Leu Met Ser

Pro Leu Gln Ile

Ser Asp Pro Phe

Gly Ser Phe Pro

His Ser Pro Thr 25

Met Asp Asn Tyr 30

Pro Lys Leu Glu 35

Glu Met Met Leu

Leu Ser Asn Gly

Ala Pro Gln Phe 45

Leu Gly Ala Ala 50

Gly Ala Pro Glu 55

Gly Ser Gly Ser 60

Asn Ser Ser Ser

Ser Ser Ser Gly 65

Gly Gly Gly Gly 70

Gly Gly Gly Gly 75

Ser Asn Ser Ser

- 219 046157

Ser Ser Ser Ser Thr Phe Asn Pro

Gln Ala Asp Thr Gly Glu Gln

95

Tyr Glu His

Leu Thr Ala Glu Ser Phe Pro Asp Ile Ser Leu Asn

100 105 110

Glu Lys Val

115

Leu Val Glu Thr

Ser Tyr Pro Ser Gln Thr Thr Arg

120 125

Pro Pro

130

Ile Thr

Tyr Thr Gly Arg Phe Ser Leu Glu Pro Ala Pro

135 140

Ser Gly Asn Thr Leu Trp Pro Glu Pro Leu Phe Ser Leu Val Ser

145 150 155

Leu Val Ser Met Thr Asn Pro Pro Ala Ser Ser Ser Ser Ala Pro

165 170 175

Pro Ala Ala Ser Ser Ala Ser Ala Ser Gln Ser Pro Pro Leu Ser

180 185 190

Ala Val Pro Ser Asn Asp Ser Ser Pro Ile Tyr Ser Ala Ala Pro

195 200 205

Phe Pro Thr Pro Asn Thr Asp Ile Phe Pro Glu Pro Gln Ser Gln

210 215 220

Phe Pro Gly Ser Ala Gly Thr Ala Leu Gln Tyr Pro Pro Pro Ala

225 230 235

Pro Ala Ala Lys Gly Gly Phe Gln Val Pro Met Ile Pro Asp Tyr

245 250 255

Phe Pro Gln Gln Gln Gly Asp Leu Gly Leu Gly Thr Pro Asp Gln

260 265 270

Pro Phe Gln Gly Leu Glu Ser Arg Thr Gln Gln Pro Ser Leu Thr

275 280 285

Leu Ser Thr Ile Lys Ala Phe Ala Thr Gln Ser Gly Ser Gln Asp

290 295 300

Lys Ala Leu Asn Thr Ser Tyr Gln Ser Gln Leu Ile Lys Pro Ser

305 310 315

Pro

Asn

Leu

Asn

Gly 160

Ser

Cys

Thr

Ala

Tyr 240

Leu

Lys

Pro

Leu

Arg 320

Arg

Met Arg Lys Tyr Pro Asn Arg Pro Ser Lys Thr Pro Pro His Glu

- 220 046157

325

330 335

Pro

Asp

Gln

Thr 385

Cys

Ile

Cys

Ile

Phe 465

Glu

Ser

Lys

Ser

Ser 545

Ser

Tyr Ala Cys Pro Val Glu Ser 340

Cys Asp Arg Arg Phe Ser Arg

345 350

Asn Leu Val Arg His Ile Arg

355 360

Cys Arg Ile Cys Met Arg Asn

370 375

His Ile Arg Thr His Thr Gly

390

Gly Arg Lys Phe Ala Arg Ser 405

His Leu Arg Gln Lys Asp Arg 420

Asp Arg Arg Phe Ser Gln Ser

435 440

His Thr Gly Gln Lys Pro Phe

450 455

Ser Thr Ser Gly His Leu Val 470

Lys Pro Phe Ala Cys Asp Ile

485

Thr Leu Thr Glu His Thr Lys

500

Ala Asp Lys Ser Val Val Ala

515 520

Tyr Pro Ser Pro Val Ala Thr

530 535

Tyr Pro Ser Pro Ala Thr Thr

550

Phe Ser Ser Pro Gly Ser Ser 565

Ile His Thr Gly Gln Lys Pro

365

Phe Ser Arg Glu Asp Asn Leu

380

Glu Lys Pro Phe Ala Cys Asp 395

Asp Glu Leu Val Arg His Thr

410 415

Pro Tyr Ala Cys Pro Val Glu

425 430

Gly Asn Leu Thr Glu His Ile 445

Gln Cys Arg Ile Cys Met Arg 460

Arg His Ile Arg Thr His Thr 475

Cys Gly Arg Lys Phe Ala Gln

490 495

Ile His Leu Arg Gln Lys Asp

505 510

Ser Ser Ala Thr Ser Ser Leu

525

Ser Tyr Pro Ser Pro Val Thr 540

Ser Tyr Pro Ser Pro Val Pro 555

Thr Tyr Pro Ser Pro Val His

570 575

Ser

Phe

His

Ile 400

Lys

Ser

Arg

Asn

Gly 480

Asn

Lys

Ser

Thr

Thr 560

Ser

- 221 046157

Gly Phe

Pro Ser

580

Pro Ser

Val Ala

Ala Phe

Pro Ala 595

Gln Val

Ser Ser

600

Ser Phe

610

Ser Ala

Ser Thr

Gly Leu 615

Thr Thr 585

Phe Pro

Ser Asp

Tyr Ser

Ser Ser

Met Thr

620

Ser Val Pro Pro

590

Ala Val Thr Asn 605

Ala Thr Phe Ser

Pro Arg Thr Ile Glu Ile Cys

625 630 <210> 119 <211> 473 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial sequence polypeptide <400> 119

Synthetic

Met Thr Gly 1

Leu Asn Gln

Leu Asn Leu

Met Ala Thr

Asn Pro Glu 65

Lys Thr Val

Trp Cys Gln

Val Pro Pro

115

Met Val Gln

130

Pro Pro Tyr

Lys Leu Ala 5

Leu Pro Asp 20

Phe Ser Gly

Glu Asn Val

Leu Ser Tyr 70

Thr Tyr Leu 85

Asp Asn Ile 100

Ala Ser Gly

Pro Pro Gln

Ser Asn Cys

Glu Lys Leu

Asn Leu Tyr

Ser Ser Asp 40

Met Asp Ile 55

Ser Gly Ser

Gly Lys Phe

Ile Ser Leu

105

Ala Leu Ser

120

Gly Asp Val 135

Gly Asp Leu

Pro Val Thr 10

Pro Glu Glu

Ser Val Val

Gly Leu Thr 60

Phe Gln Pro 75

Ala Phe Asp 90

Met Ser Ala

Thr Gln Thr

Glu Ala Met

140

Tyr Ser Glu

Met Ser Ser Leu

Ile Pro Ser Ala

His Tyr Asn Gln 45

Asn Glu Lys Pro

Ala Pro Gly Asn 80

Ser Pro Ser Asn

Gly Ile Leu Gly 110

Ser Thr Ala Ser 125

Tyr Pro Ala Leu

Pro Val Ser Phe

- 222 046157

145

150

155

His Asp Pro Gln Gly

165

Asn Pro Gly Leu Ala

170

Tyr Ser Pro Gln Asp

175

Gln Ser Ala Lys Pro

180

Ala Leu Asp Ser Asn

185

Leu Phe Pro Met Ile

190

Asp Tyr Asn Leu Tyr 195

His His Pro Asn Asp

200

Met Gly Ser Ile Pro 205

His Lys Pro Phe Gln 210

Gly Met Asp Pro Ile 215

Arg Val Asn Pro Pro 220

Ile Thr Pro Leu Glu 225

Thr Ile Lys Ala Phe 230

Lys Asp Lys Gln Ile 235

Pro Gly Phe Gly Ser

245

Leu Pro Gln Pro Pro

250

Leu Thr Leu Lys Pro

255

Arg Pro Arg Lys Tyr

260

Pro Asn Arg Pro Ser

265

Lys Thr Pro Leu His

270

Arg Pro His Ala Cys 275

Pro Ala Glu Gly Cys 280

Asp Arg Arg Phe Ser 285

Ser Asp Asn Leu Val

290

Arg His Leu Arg Ile 295

His Thr Gly His Lys

300

Phe Gln Cys Arg Ile 305

Cys Met Arg Ser Phe 310

Ser His Arg Thr Thr 315

Thr Asn His Ile Arg

325

Thr His Thr Gly Glu

330

Lys Pro Phe Ala Cys

335

Phe Cys Gly Arg Lys

340

Phe Ala Arg Glu Asp

345

Asn Leu His Thr His

350

Lys Ile His Leu Lys 355

Gln Lys Glu His Ala 360

Cys Pro Ala Glu Gly

365

Asp Arg Arg Phe Ser

370

Thr Ser His Ser Leu

375

Thr Glu His Leu Arg

380

His Thr Gly His Lys 385

Pro Phe Gln Cys Arg 390

Ile Cys Met Arg Ser 395

160

Tyr

Pro

Glu

Pro

His 240

Ile

Glu

Arg

Pro

Leu 320

Glu

Ala

Cys

Ile

Phe 400

- 223 046157

Ser Gln

Ser Ser

Lys Pro

Phe Ala

420

Asn Leu

His Thr 435

Ala Glu

450

Lys Gly

Leu Ala 465

Pro Val

Ser Leu 405

Cys Glu

His Ala

Gly Ala

Val Thr

470

Val Arg

Phe Cys

Lys Ile 440

Pro Ser 455

Thr Cys

His Ile

410

Gly Arg 425

His Leu

Ala Ser

Ala

Arg Thr

Lys Phe

Lys Gln

Ser Ala

460

His Thr Gly Glu 415

Ala Arg Glu Asp

430

Lys Glu Lys Lys 445

Pro Pro Val Ser <210> 120 <211> 719 <212> Protein <213> Artificial Sequence <220>

<223> Description of the artificial sequence

Synthetic <400>

Met

Ser

Pro

Leu

Ser 65

Ser

Tyr

Glu

Pro polypeptide

120

Ala

Asp

Lys

Gly 50

Ser

Ser

Glu

Lys

Pro

Ala

Ala

Lys 5

Ala

Glu

Met

Gln

Leu

Met

Ser

Pro Leu Gln Ile

Pro

Phe 20

Gly

Ser

Phe

Pro

His 25

Ser

Pro

Thr

Met Asp Asn Tyr 30

Leu 35

Glu

Glu

Met

Met

Leu 40

Leu

Ser

Asn

Gly

Ala Pro Gln Phe 45

Ala

Ala

Gly

Ala

Pro 55

Glu

Gly

Ser

Gly

Ser 60

Asn Ser Ser Ser

Ser

Ser

His

Val

115

Ile

Gly

Gly

Gly 70

Gly

Gly

Gly

Gly

Gly 75

Gly

Ser Asn Ser Ser

Ser

Thr 85

Phe

Asn

Pro

Gln

Ala 90

Asp

Thr

Gly Glu Gln Pro 95

Leu

100

Leu

Thr

Thr

Val

Tyr

Ala

Glu

Thr

Glu

Thr

Gly

Ser

Ser

120

Arg

Phe

105

Pro

Asp

Ile

Ser Leu Asn Asn

110

Tyr

Phe

Pro

Ser

Ser

Leu

Gln

Glu

Thr Thr Arg Leu 125

Pro Ala Pro Asn

- 224 046157

130

135

140

Ser Gly Asn Thr Leu Trp Pro Glu Pro Leu Phe Ser Leu Val Ser

145 150 155

Leu Val Ser Met Thr Asn Pro Pro Ala Ser Ser Ser Ser Ala Pro

165 170 175

Pro Ala Ala Ser Ser Ala Ser Ala Ser Gln Ser Pro Pro Leu Ser

180 185 190

Ala Val Pro Ser Asn Asp Ser Ser Pro Ile Tyr Ser Ala Ala Pro

195 200 205

Phe Pro Thr Pro Asn Thr Asp Ile Phe Pro Glu Pro Gln Ser Gln

210 215 220

Phe Pro Gly Ser Ala Gly Thr Ala Leu Gln Tyr Pro Pro Pro Ala

225 230 235

Pro Ala Ala Lys Gly Gly Phe Gln Val Pro Met Ile Pro Asp Tyr

245 250 255

Phe Pro Gln Gln Gln Gly Asp Leu Gly Leu Gly Thr Pro Asp Gln

260 265 270

Pro Phe Gln Gly Leu Glu Ser Arg Thr Gln Gln Pro Ser Leu Thr

275 280 285

Leu Ser Thr Ile Lys Ala Phe Ala Thr Gln Ser Gly Ser Gln Asp

290 295 300

Lys Ala Leu Asn Thr Ser Tyr Gln Ser Gln Leu Ile Lys Pro Ser

305 310 315

Met Arg Lys Tyr Pro Asn Arg Pro Ser Lys Thr Pro Pro His Glu

325 330 335

Pro Tyr Ala Cys Pro Val Glu Ser Cys Asp Arg Arg Phe Ser Asp

340 345 350

Gly Ala Leu Val Arg His Ile Arg Ile His Thr Gly Gln Lys Pro

355 360 365

Gln Cys Arg Ile Cys Met Arg Asn Phe Ser Arg Ser Asp Asn Leu

370 375 380

Gly 160

Ser

Cys

Thr

Ala

Tyr 240

Leu

Lys

Pro

Leu

Arg 320

Arg

Pro

Phe

Val

- 225 046157

Arg 385

Cys

Ile

Cys

Ile

Phe 465

Glu

Asp

Pro

Gly

Gln 545

Glu

Cys

Ile

Ser

Ser 625

His Ile Arg Thr His

390

Gly Arg Lys Phe Ala

405

His Leu Arg Gln Lys 420

Asp Arg Arg Phe Ser

435

His Thr Gly Gln Lys 450

Ser Thr Ser Gly Asn

470

Lys Pro Phe Ala Cys

485

Asn Leu Val Arg His 500

Tyr Ala Cys Pro Val

515

His Leu Thr Glu His 530

Cys Arg Ile Cys Met

550

His Ile Arg Thr His

565

Gly Arg Lys Phe Ala

580

His Leu Arg Gln Lys

595

Ser Ala Thr Ser Ser 610

Tyr Pro Ser Pro Val

630

Thr Gly Glu Lys Pro

395

Gln Ser Gly Asp Leu

410

Asp Arg Pro Tyr Ala 425

Thr His Leu Asp Leu

440

Pro Phe Gln Cys Arg 455

Leu Val Arg His Ile

475

Asp Ile Cys Gly Arg

490

Thr Lys Ile His Leu 505

Glu Ser Cys Asp Arg 520

Ile Arg Ile His Thr 535

Arg Asn Phe Ser Glu

555

Thr Gly Glu Lys Pro

570

Gln Ala Gly His Leu 585

Asp Lys Lys Ala Asp 600

Leu Ser Ser Tyr Pro 615

Thr Thr Ser Tyr Pro

635

Phe Ala Cys Asp Ile

400

Arg Arg His Thr Lys 415

Cys Pro Val Glu Ser

430

Ile Arg His Ile Arg 445

Ile Cys Met Arg Asn 460

Arg Thr His Thr Gly

480

Lys Phe Ala Arg Ser

495

Arg Gln Lys Asp Arg 510

Arg Phe Ser Gln Ser

525

Gly Gln Lys Pro Phe 540

Arg Ser His Leu Arg

560

Phe Ala Cys Asp Ile

575

Ala Ser His Thr Lys 590

Lys Ser Val Val Ala

605

Ser Pro Val Ala Thr 620

Ser Pro Ala Thr Thr

640

- 226 046157

Ser Tyr

Pro Ser

Thr Tyr

Pro Ser

660

Thr Thr

Tyr Ser 675

Phe Pro

690

Ser Ser

Pro Val 645

Pro Val

Ser Val

Ala Val

Pro Thr

His Ser

Pro Pro

680

Thr Asn 695

Ser Phe

650

Gly Phe 665

Ala Phe

Ser Phe

Ser Asp Met Thr Ala Thr Phe Ser Pro Arg

705 710 <210> 121 <211> 627 <212> Protein <213> Artificial sequence <220>

Ser Ser

Pro Ser

Pro Ala

Ser Ala

700

Thr Ile 715 <223> Description of the artificial sequence polypeptide <400> 121 Met Ala Ala 1

Ser Asp Pro

Pro Lys Leu 35

Leu Gly Ala 50

Ala Lys Ala 5

Phe Gly Ser 20

Glu Glu Met

Ala Gly Ala

Glu Met Gln

Phe Pro His

Met Leu Leu

Pro Glu Gly 55

Leu Met Ser 10

Ser Pro Thr

Ser Asn Gly

Ser Gly Ser 60

Ser Ser Ser Gly Gly Gly Gly Gly Gly Gly Gly Gly 65 70 75

Ser

Ser Ser Ser Thr Phe Asn Pro Gln Ala Asp Thr

90

Tyr Glu His Leu Thr Ala Glu Ser Phe Pro Asp Ile

100 105

Glu Lys Val Leu Val Glu Thr Ser

115 120

Tyr Pro Ser Gln

Pro Gly Ser Ser

655

Pro Ser Val Ala

670

Gln Val Ser Ser 685

Ser Thr Gly Leu

Glu Ile Cys

Synthetic

Pro Leu Gln Ile

Met Asp Asn Tyr 30

Ala Pro Gln Phe 45

Asn Ser Ser Ser

Ser Asn Ser Ser

Gly Glu Gln Pro

Ser Leu Asn Asn

110

Thr Thr Arg Leu 125

- 227 046157

Pro Pro Ile Thr Tyr Thr Gly Arg Phe

130 135

Ser Gly Asn Thr Leu Trp Pro Glu Pro

145 150

Leu Val Ser Met Thr Asn Pro Pro Ala

165

Pro Ala Ala Ser Ser Ala Ser Ala Ser

180 185

Ala Val Pro Ser Asn Asp Ser Ser Pro

195 200

Phe Pro Thr Pro Asn Thr Asp Ile Phe

210 215

Phe Pro Gly Ser Ala Gly Thr Ala Leu 225 230

Pro Ala Ala Lys Gly Gly Phe Gln Val

245

Phe Pro Gln Gln Gln Gly Asp Leu Gly

260 265

Pro Phe Gln Gly Leu Glu Ser Arg Thr

275 280

Leu Ser Thr Ile Lys Ala Phe Ala Thr

290 295

Lys Ala Leu Asn Thr Ser Tyr Gln Ser

305 310

Met Arg Lys Tyr Pro Asn Arg Pro Ser

325

Pro Tyr Ala Cys Pro Val Glu Ser Cys

340 345

Asp Asn Leu Thr Arg His Ile Arg Ile

355 360

Gln Cys Arg Ile Cys Met Arg Asn Phe

370 375

Leu Glu Pro Ala Pro Asn

140

Phe Ser Leu Val Ser Gly

155 160

Ser Ser Ser Ala Pro Ser

175

Ser Pro Pro Leu Ser Cys

190

Tyr Ser Ala Ala Pro Thr

205

Glu Pro Gln Ser Gln Ala

220

Tyr Pro Pro Pro Ala Tyr

235 240

Met Ile Pro Asp Tyr Leu

255

Gly Thr Pro Asp Gln Lys

270

Gln Pro Ser Leu Thr Pro

285

Ser Gly Ser Gln Asp Leu

300

Leu Ile Lys Pro Ser Arg

315 320

Thr Pro Pro His Glu Arg

335

Arg Arg Phe Ser Arg Ser

350

Thr Gly Gln Lys Pro Phe

365

His Ser Thr Thr Leu Thr

380

- 228 046157

Asn 385

Cys

Thr

Phe

Gln

Ser 465

Ala

Thr

Ser

Pro

Pro 545

Pro

Pro

Gln

Ser

Glu 625

His Ile

Gly Arg

His Thr

Ser Thr

435

Lys Pro 450

Ser Leu

Cys Asp

His Thr

Val Val

515

Val Ala 530

Ala Thr

Gly Ser

Ser Val

Val Ser

595

Thr Gly 610

Ile Cys

Arg Thr

Lys Phe

405

Gly Glu 420

Ser His

Phe Gln

Thr Arg

Ile Cys

485

Lys Ile 500

Ala Ser

Thr Ser

Thr Ser

Ser Thr

565

Ala Thr 580

Ser Phe

Leu Ser

His Thr 390

Ala Arg

Lys Pro

Ser Leu

CysArg

455

His Ile 470

Gly Arg

His Leu

Ser Ala

Tyr Pro

535

Tyr Pro 550

Tyr Pro

Thr Tyr

Pro Ser

Asp Met

615

Gly Glu

Ser Asp

Phe Ala

425

Thr Glu 440

Ile Cys

Arg Thr

Lys Phe

Arg Gln

505

Thr Ser 520

Ser Pro

Ser Pro

Ser Pro

Ser Ser

585

Ser Ala 600

Thr Ala

Lys Pro

395

Phe Ala

Cys Asp

Asn Arg 410

Cys Asp

His Ile

Met Arg

His Thr

475

Ala Arg 490

Lys Asp

Ser Leu

Val Thr

Val Pro

555

Val His 570

Val Pro

Val Thr

Thr Phe

Lys Thr

Ile Cys

Arg Ile

445

Asn Phe 460

Gly Glu

Ser Asp

Lys Lys

Ser Ser

525

Thr Ser 540

Thr Ser

Ser Gly

Pro Ala

Asn Ser

605

Ser Pro 620

His Ile

415

Gly Arg 430

His Thr

Ser Gln

Lys Pro

Asn Arg

495

Ala Asp 510

Tyr Pro

Tyr Pro

Phe Ser

Phe Pro

575

Phe Pro 590

Phe Ser

Arg Thr

Ile 400

Arg

Lys

Gly

Ser

Phe 480

Lys

Lys

Ser

Ser

Ser 560

Ser

Ala

Ala

Ile

- 229 046157 <210> 122 <211> 473 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial sequence polypeptide <400> 122

Met Thr Gly Lys Leu Ala Glu Lys Leu Pro Val Thr

5 10

Synthetic

Met Ser Ser Leu

Leu Asn Gln Leu Pro Asp 20

Asn Leu Tyr Pro Glu Glu 25

Ile Pro Ser Ala

Leu Asn Leu Phe Ser Gly 35

Ser Ser Asp Ser Val Val 40

His Tyr Asn Gln 45

Met Ala Thr Glu Asn Val

Met Asp Ile Gly Leu Thr 55 60

Asn Glu Lys Pro

Asn Pro Glu Leu Ser Tyr 65 70

Ser Gly Ser Phe Gln Pro 75

Ala Pro Gly Asn 80

Lys Thr Val Thr Tyr Leu 85

Gly Lys Phe Ala Phe Asp 90

Ser Pro Ser Asn

Trp Cys Gln Asp Asn Ile

100

Ile Ser Leu Met Ser Ala

105

Gly Ile Leu Gly 110

Val Pro Pro Ala Ser Gly

115

Ala Leu Ser Thr Gln Thr

120

Ser Thr Ala Ser 125

Met Val Gln Pro Pro Gln

130

Gly Asp Val Glu Ala Met

135 140

Tyr Pro Ala Leu

Pro Pro Tyr Ser Asn Cys

145 150

Gly Asp Leu Tyr Ser Glu

155

Pro Val Ser Phe

160

His Asp Pro Gln Gly Asn

165

Pro Gly Leu Ala Tyr Ser

170

Pro Gln Asp Tyr

175

Gln Ser Ala Lys Pro Ala

180

Leu Asp Ser Asn Leu Phe

185

Pro Met Ile Pro

190

Asp Tyr Asn Leu Tyr His

195

His Pro Asn Asp Met Gly

200

Ser Ile Pro Glu 205

- 230 046157

His Lys Pro Phe Gln 210

Gly Met Asp Pro Ile 215

Arg Val Asn Pro Pro 220

Ile Thr Pro Leu Glu 225

Thr Ile Lys Ala Phe 230

Lys Asp Lys Gln Ile 235

Pro Gly Phe Gly Ser

245

Leu Pro Gln Pro Pro

250

Leu Thr Leu Lys Pro

255

Arg Pro Arg Lys Tyr

260

Pro Asn Arg Pro Ser

265

Lys Thr Pro Leu His

270

Arg Pro His Ala Cys 275

Pro Ala Glu Gly Cys 280

Asp Arg Arg Phe Ser 285

Ser Asp Asn Leu Val

290

Arg His Leu Arg Ile 295

His Thr Gly His Lys

300

Phe Gln Cys Arg Ile 305

Cys Met Arg Ser Phe 310

Ser Arg Glu Asp Asn 315

His Thr His Ile Arg

325

Thr His Thr Gly Glu

330

Lys Pro Phe Ala Cys

335

Phe Cys Gly Arg Lys

340

Phe Ala Arg Ser Asp

345

Glu Leu Val Arg His

350

Lys Ile His Leu Lys 355

Gln Lys Glu His Ala 360

Cys Pro Ala Glu Gly 365

Asp Arg Arg Phe Ser

370

Gln Ser Gly Asn Leu 375

Thr Glu His Leu Arg

380

His Thr Gly His Lys 385

Pro Phe Gln Cys Arg 390

Ile Cys Met Arg Ser 395

Ser Thr Ser Gly His

405

Leu Val Arg His Ile

410

Arg Thr His Thr Gly 415

Lys Pro Phe Ala Cys

420

Glu Phe Cys Gly Arg

425

Lys Phe Ala Gln Asn

430

Thr Leu Thr Glu His

435

Ala Lys Ile His Leu 440

Lys Gln Lys Glu Lys 445

Ala Glu Lys Gly Gly 450

Ala Pro Ser Ala Ser

455

Ser Ala Pro Pro Val

460

Pro

His 240

Ile

Glu

Arg

Pro

Leu 320

Glu

Ala

Cys

Ile

Phe 400

Glu

Ser

Lys

Ser

- 231 046157

Leu Ala Pro Val Val Thr Thr Cys Ala

465

470 <210> 123 <211> 627 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic polypeptide <400> 123

Met Ala Ala Ala Lys Ala Glu Met 1 5

Gln Leu Met Ser Pro Leu Gln Ile

15

Ser Asp Pro Phe Gly Ser Phe Pro 20

His Ser Pro Thr Met Asp Asn Tyr 25 30

Pro Lys Leu Glu Glu Met Met Leu

40

Leu Ser Asn Gly Ala Pro Gln Phe 45

Leu Gly Ala Ala Gly Ala Pro Glu

55

Gly Ser Gly Ser Asn Ser Ser Ser 60

Ser Ser Ser Gly Gly Gly Gly Gly 65 70

Gly Gly Gly Gly Ser Asn Ser Ser

80

Ser Ser Ser Ser Thr Phe Asn Pro

Gln Ala Asp Thr Gly Glu Gln Pro

95

Tyr Glu His Leu Thr Ala Glu Ser

100

Phe Pro Asp Ile Ser Leu Asn Asn

105 110

Glu Lys Val Leu Val Glu Thr Ser

115 120

Tyr Pro Ser Gln Thr Thr Arg Leu

125

Pro Pro Ile Thr Tyr Thr Gly Arg

130 135

Phe Ser Leu Glu Pro Ala Pro Asn

140

Ser Gly Asn Thr Leu Trp Pro Glu

145 150

Pro Leu Phe Ser Leu Val Ser Gly

155 160

Leu Val Ser Met Thr Asn Pro Pro

165

Ala Ser Ser Ser Ser Ala Pro Ser

170 175

Pro Ala Ala Ser Ser Ala Ser Ala

180

Ser Gln Ser Pro Pro Leu Ser Cys

185 190

- 232 046157

Ala Val Pro Ser Asn Asp Ser Ser Pro Ile Tyr Ser Ala Ala Pro

195 200 205

Phe Pro Thr Pro Asn Thr Asp Ile Phe Pro Glu Pro Gln Ser Gln

210 215 220

Phe Pro Gly Ser Ala Gly Thr Ala Leu Gln Tyr Pro Pro Pro Ala 225 230 235

Pro Ala Ala Lys Gly Gly Phe Gln Val Pro Met Ile Pro Asp Tyr

245 250 255

Phe Pro Gln Gln Gln Gly Asp Leu Gly Leu Gly Thr Pro Asp Gln

260 265 270

Pro Phe Gln Gly Leu Glu Ser Arg Thr Gln Gln Pro Ser Leu Thr

275 280 285

Leu Ser Thr Ile Lys Ala Phe Ala Thr Gln Ser Gly Ser Gln Asp

290 295 300

Lys Ala Leu Asn Thr Ser Tyr Gln Ser Gln Leu Ile Lys Pro Ser

305 310 315

Met Arg Lys Tyr Pro Asn Arg Pro Ser Lys Thr Pro Pro His Glu

325 330 335

Pro Tyr Ala Cys Pro Val Glu Ser Cys Asp Arg Arg Phe Ser Arg

340 345 350

Asp Asn Leu Thr Arg His Ile Arg Ile His Thr Gly Gln Lys Pro

355 360 365

Gln Cys Arg Ile Cys Met Arg Asn Phe Ser Arg Ser Asp Asn Leu

370 375 380

Thr His Ile Arg Thr His Thr Gly Glu Lys Pro Phe Ala Cys Asp

385 390 395

Cys Gly Arg Lys Phe Ala Arg Ser Asp Glu Arg Lys Arg His Ile

405 410 415

Thr His Thr Gly Glu Lys Pro Phe Ala Cys Asp Ile Cys Gly Arg

420 425 430

Phe Ser Gln Ser Gly Asn Leu Thr Glu His Ile Arg Ile His Thr

435 440 445

Thr

Ala

Tyr 240

Leu

Lys

Pro

Leu

Arg 320

Arg

Ser

Phe

Thr

Ile 400

Arg

Lys

Gly

- 233 046157

Gln Lys Pro Phe Gln Cys Arg Ile Cys Met Arg Asn

450 455 460

Phe Ser Thr Ser

Gly His Leu Thr Arg His Ile Arg Thr His Thr Gly

465 470 475

Glu Lys Pro Phe

480

Ala Cys Asp Ile Cys Gly Arg Lys Phe Ala Gln Ser

485 490

Ser Thr Arg Lys

495

Glu His Thr Lys Ile His Leu Arg Gln Lys Asp Lys

500 505

Lys Ala Asp Lys

510

Ser Val Val Ala Ser Ser Ala Thr Ser Ser Leu Ser

515 520

Ser Tyr Pro Ser 525

Pro Val Ala Thr Ser Tyr Pro Ser Pro Val Thr Thr

530 535 540

Ser Tyr Pro Ser

Pro Ala Thr Thr Ser Tyr Pro Ser Pro Val Pro Thr

545 550 555

Ser Phe Ser Ser

560

Pro Gly Ser Ser Thr Tyr Pro Ser Pro Val His Ser

565 570

Gly Phe Pro Ser

575

Pro Ser Val Ala Thr Thr Tyr Ser Ser Val Pro Pro

580 585

Ala Phe Pro Ala

590

Gln Val Ser Ser Phe Pro Ser Ser Ala Val Thr Asn

595 600

Ser Phe Ser Ala 605

Ser Thr Gly Leu Ser Asp Met Thr Ala Thr Phe Ser

610 615 620

Pro Arg Thr Ile

Glu Ile Cys 625 <210> 124 <211> 627 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial sequence polypeptide <400> 124

Met Ala Ala Ala Lys Ala Glu Met Gln Leu Met Ser

5 10

Synthetic

Pro Leu Gln Ile

- 234 046157

Ser Asp Pro Phe Gly Ser Phe Pro His Ser Pro

25

Thr Met Asp Asn

Pro Lys

Leu Glu Glu Met Met 35

Leu Leu Ser Asn Gly Ala Pro Gln 40 45

Leu Gly Ala Ala

Gly Ala Pro Glu Gly

Ser Gly Ser Asn Ser

Ser

Ser Ser Ser Gly Gly Gly Gly Gly Gly Gly Gly Gly Ser Asn Ser 65 70 75

Ser Ser Ser Ser Thr Phe Asn Pro Gln Ala Asp Thr Gly Glu Gln 85 90 95

Tyr Glu His Leu Thr Ala Glu Ser Phe Pro Asp Ile Ser Leu Asn

100 105 110

Glu Lys Val Leu Val Glu Thr Ser Tyr Pro Ser Gln Thr Thr Arg

115 120 125

Pro Pro Ile Thr Tyr Thr Gly Arg Phe Ser Leu Glu Pro Ala Pro

130 135 140

Ser Gly Asn Thr Leu Trp Pro Glu Pro Leu Phe Ser Leu Val Ser

145 150 155

Leu Val Ser Met Thr Asn Pro Pro Ala Ser Ser Ser Ser Ala Pro

165 170 175

Pro Ala Ala Ser Ser Ala Ser Ala Ser Gln Ser Pro Pro Leu Ser

180 185 190

Ala Val Pro Ser Asn Asp Ser Ser Pro Ile Tyr Ser Ala Ala Pro

195 200 205

Phe Pro Thr Pro Asn Thr Asp Ile Phe Pro Glu Pro Gln Ser Gln

210 215 220

Phe Pro Gly Ser Ala Gly Thr Ala Leu Gln Tyr Pro Pro Pro Ala

225 230 235

Pro Ala Ala Lys

Gly Gly Phe Gln Val

245

Pro Met 250

Ile Pro Asp Tyr

255

Phe Pro Gln Gln Gln Gly Asp Leu

260

Gly Leu Gly Thr Pro Asp Gln

265 270

Tyr

Phe

Ser

Ser 80

Pro

Asn

Leu

Asn

Gly 160

Ser

Cys

Thr

Ala

Tyr 240

Leu

Lys

- 235 046157

Pro Phe Gln Gly Leu

275

Glu Ser Arg Thr Gln 280

Gln Pro Ser Leu Thr

285

Leu Ser Thr Ile Lys 290

Ala Phe Ala Thr Gln

295

Ser Gly Ser Gln Asp

300

Lys Ala Leu Asn Thr 305

Ser Tyr Gln Ser Gln 310

Leu Ile Lys Pro Ser 315

Met Arg Lys Tyr Pro

325

Asn Arg Pro Ser Lys

330

Thr Pro Pro His Glu

335

Pro Tyr Ala Cys Pro

340

Val Glu Ser Cys Asp

345

Arg Arg Phe Ser Arg

350

Asp Asn Leu Val Arg 355

His Ile Arg Ile His 360

Thr Gly Gln Lys Pro 365

Gln Cys Arg Ile Cys

370

Met Arg Asn Phe Ser 375

Arg Glu Asp Asn Leu 380

Thr His Ile Arg Thr 385

His Thr Gly Glu Lys 390

Pro Phe Ala Cys Asp 395

Cys Gly Arg Lys Phe

405

Ala Arg Ser Asp Glu

410

Leu Val Arg His Ile

415

Thr His Thr Gly Glu

420

Lys Pro Phe Ala Cys

425

Asp Ile Cys Gly Arg

430

Phe Ser Gln Ser Gly

435

Asn Leu Thr Glu His

440

Ile Arg Ile His Thr 445

Gln Lys Pro Phe Gln 450

Cys Arg Ile Cys Met

455

Arg Asn Phe Ser Thr

460

Gly His Leu Val Arg 465

His Ile Arg Thr His 470

Thr Gly Glu Lys Pro 475

Ala Cys Asp Ile Cys

485

Gly Arg Lys Phe Ala

490

Gln Asn Ser Thr Leu

495

Glu His Thr Lys Ile

500

His Leu Arg Gln Lys

505

Asp Lys Lys Ala Asp

510

Ser Val Val Ala Ser

Ser Ala Thr Ser Ser

Leu Ser Ser Tyr Pro

Pro

Leu

Arg 320

Arg

Ser

Phe

His

Ile 400

Arg

Lys

Gly

Ser

Phe 480

Thr

Lys

Ser

- 236 046157

Pro Val

530

Pro Ala 545

Pro Gly

Pro Ser

Gln Val

Ser Thr

610

515

Ala Thr

Thr Thr

Ser Ser

Val Ala

580

Ser Ser 595

Gly Leu

Ser Tyr

Ser Tyr

550

Thr Tyr 565

Thr Thr

Phe Pro

Ser Asp

520

Pro Ser 535

Pro Ser

Pro Ser

Tyr Ser

Ser Ser

600

Met Thr 615

525

Pro Val

Pro Val

Pro Val

570

Ser Val

585

Ala Val

Ala Thr

Glu Ile Cys 625

Thr Thr

540

Pro Thr 555

His Ser

Pro Pro

Thr Asn

Phe Ser

620 <210> 125 <211> 272 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial sequence polypeptide <400> 125

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Ile His

5 10

Ala Leu Glu Pro Gly Glu Lys Pro Tyr Lys Cys Pro 20 25

Ser Phe Ser Ser Pro Ala Asp Leu Thr Arg His Gln

40

Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys

55 60

Ser Asp Asn Leu Val Arg His Gln Arg Thr His Thr

70 75

Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg

90

Ser Tyr Pro Ser

Ser Phe Ser Ser

560

Gly Phe Pro Ser

575

Ala Phe Pro Ala

590

Ser Phe Ser Ala 605

Pro Arg Thr Ile

Synthetic

Gly Val Pro Ala 15

Glu Cys Gly Lys 30

Arg Thr His Thr 45

Ser Phe Ser Arg

Gly Glu Lys Pro 80

Glu Asp Asn Leu

- 237 046157

His Thr His Gln Arg Thr His Thr Gly Glu Lys

100 105

Pro Tyr Lys Cys Pro

110

Glu Cys Gly Lys Ser Phe Ser Arg Ser Asp Glu Leu Val Arg His Gln

115 120 125

Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys

130 135 140

Ser Phe Ser Gln Ser Gly Asn Leu Thr Glu His Gln Arg Thr His Thr

145 150 155 160

Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr

165 170 175

Ser Gly His Leu Val Arg His Gln Arg Thr His Thr Gly Lys Lys Thr

180 185 190

Ser Lys Arg Pro Ala Ala Thr Lys Lys Ala Gly Gln Ala Lys Lys Lys

195 200 205

Lys Gly Ser Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Leu Glu Asp Ala

210 215 220

Leu Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp

225 230 235 240

Phe Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu

245 250 255

Asp Met Leu Gly Ser Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu

260 265 270 <210> 126 <211> 272 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic polypeptide <400> 126

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Ile His Gly Val Pro Ala

5 10 15

Ala Leu Glu Pro Gly Glu Lys Pro Tyr

25

Lys Cys Pro Glu Cys Gly Lys 30

- 238 046157

Ser Phe Ser Asp Pro 35

Gly Ala Leu Val Arg 40

His Gln Arg Thr His 45

Gly Glu Lys Pro Tyr 50

Lys Cys Pro Glu Cys 55

Gly Lys Ser Phe Ser 60

Ser Asp Asn Leu Val 65

Arg His Gln Arg Thr 70

His Thr Gly Glu Lys 75

Tyr Lys Cys Pro Glu 85

Cys Gly Lys Ser Phe

Ser Gln Ser Gly Asp

Arg Arg His Gln Arg

100

Thr His Thr Gly Glu

105

Lys Pro Tyr Lys Cys

110

Glu Cys Gly Lys Ser 115

Phe Ser Thr His Leu

120

Asp Leu Ile Arg His 125

Arg Thr His Thr Gly 130

Glu Lys Pro Tyr Lys 135

Cys Pro Glu Cys Gly

140

Ser Phe Ser Thr Ser 145

Gly Asn Leu Val Arg 150

His Gln Arg Thr His 155

Gly Glu Lys Pro Tyr

165

Lys Cys Pro Glu Cys

170

Gly Lys Ser Phe Ser

175

Ser Asp Asn Leu Val

180

Arg His Gln Arg Thr

185

His Thr Gly Lys Lys

190

Ser Lys Arg Pro Ala

195

Ala Thr Lys Lys Ala 200

Gly Gln Ala Lys Lys 205

Lys Gly Ser Tyr Pro

210

Tyr Asp Val Pro Asp

215

Tyr Ala Leu Glu Asp

220

Leu Asp Asp Phe Asp 225

Leu Asp Met Leu Gly 230

Ser Asp Ala Leu Asp 235

Phe Asp Leu Asp Met

245

Leu Gly Ser Asp Ala

250

Leu Asp Asp Phe Asp

255

Asp Met Leu Gly Ser

260

Asp Ala Leu Asp Asp

265

Phe Asp Leu Asp Met

270

Thr

Arg

Pro 80

Leu

Pro

Gln

Lys

Thr 160

Arg

Thr

Lys

Ala

Asp 240

Leu

Leu <210> 127 <211> 261

- 239 046157 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial sequence polypeptide <400> 127

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Ile His

5 10

Ala Leu Glu Pro Gly Glu Lys Pro Tyr Lys Cys Pro 20 25

Synthetic

Gly Val Pro Ala 15

Glu Cys Gly Lys 30

Ser Phe Ser Arg Ser Asp 35

Asn Leu Val Arg His Gln 40

Arg Thr His Thr 45

Gly Glu Lys Pro Tyr Lys 50

Cys Pro Glu Cys Gly Lys 55 60

Ser Phe Ser Arg

Glu Asp Asn Leu His Thr 65 70

His Gln Arg Thr His Thr 75

Gly Glu Lys Pro 80

Tyr Lys Cys Pro Glu Cys 85

Gly Lys Ser Phe Ser Arg 90

Ser Asp Glu Leu 95

Val Arg His Gln Arg Thr

100

His Thr Gly Glu Lys Pro

105

Tyr Lys Cys Pro 110

Glu Cys Gly Lys Ser Phe 115

Ser Gln Ser Gly Asn Leu

120

Thr Glu His Gln 125

Arg Thr His Thr Gly Glu 130

Lys Pro Tyr Lys Cys Pro

135 140

Glu Cys Gly Lys

Ser Phe Ser Thr Ser Gly

145 150

His Leu Val Arg His Gln 155

Arg Thr His Thr

160

Gly Glu Lys Pro Tyr Lys

165

Cys Pro Glu Cys Gly Lys

170

Ser Phe Ser Gln

175

Asn Ser Thr Leu Thr Glu

180

His Gln Arg Thr His Thr

185

Gly Lys Lys Thr 190

Ser Lys Arg Pro Ala Ala

195

Thr Lys Lys Ala Gly Gln

200

Ala Lys Lys Lys 205

Lys Gly Ser Asp Ala Leu

210

Asp Asp Phe Asp Leu Asp

215 220

Met Leu Gly Ser

- 240 046157

Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly

225 230 235

Ser Asp Ala Leu

240

Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala

245 250

Leu Asp Asp Phe

255

Asp Leu Asp Met Leu

260 <210> 128 <211> 386 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial sequence polypeptide <400> 128

Met Ala Ala Asp His Leu Met Leu Ala Glu Gly Tyr

5 10

Synthetic

Arg Leu Val Gln 15

Arg Pro Pro Ser Ala Ala Ala Ala His Gly Pro His 20 25

Ala Leu Arg Thr 30

Leu Pro Pro Tyr Ala Gly Pro Gly Leu Asp Ser Gly

40

Leu Arg Pro Arg 45

Gly Ala Pro Leu Gly Pro Pro Pro Pro Arg Gln Pro

55 60

Gly Ala Leu Ala

Tyr Gly Ala Phe Gly Pro Pro Ser Ser Phe Gln Pro 65 70 75

Phe Pro Ala Val

Pro Pro Pro Ala Ala Gly Ile Ala His Leu Gln Pro 85 90

Val Ala Thr Pro

Tyr Pro Gly Arg Ala Ala Ala Pro Pro Asn Ala Pro

100 105

Gly Gly Pro Pro 110

Gly Pro Gln Pro Ala Pro Ser Ala Ala Ala Pro Pro

115 120

Pro Pro Ala His 125

Ala Leu Gly Gly Met Asp Ala Glu Leu Ile Asp Glu

130 135 140

Glu Ala Leu Thr

Ser Leu Glu Leu Glu Leu Gly Leu His Arg Val Arg

145 150 155

Glu Leu Pro Glu

160

- 241 046157

Leu Phe Leu Gly Gln

165

Ser Glu Phe Asp Cys

170

Phe Ser Asp Leu Gly

175

Ala Pro Pro Ala Gly

180

Ser Val Ser Cys Gln

185

Ser Gln Leu Ile Lys

190

Ser Arg Met Arg Lys

195

Tyr Pro Asn Arg Pro

200

Ser Lys Thr Pro Pro

205

Glu Arg Pro Tyr Ala

210

Cys Pro Val Glu Ser

215

Cys Asp Arg Arg Phe 220

Arg Ser Asp Asn Leu 225

Val Arg His Ile Arg 230

Ile His Thr Gly Gln 235

Pro Phe Gln Cys Arg

245

Ile Cys Met Arg Asn

250

Phe Ser Arg Glu Asp

255

Leu His Thr His Ile

260

Arg Thr His Thr Gly

265

Glu Lys Pro Phe Ala

270

Asp Ile Cys Gly Arg 275

Lys Phe Ala Arg Ser 280

Asp Glu Leu Val Arg 285

Thr Lys Ile His Leu 290

Arg Gln Lys Asp Arg 295

Pro Tyr Ala Cys Pro 300

Glu Ser Cys Asp Arg 305

Arg Phe Ser Gln Ser 310

Gly Asn Leu Thr Glu 315

Ile Arg Ile His Thr

325

Gly Gln Lys Pro Phe

330

Gln Cys Arg Ile Cys

335

Arg Asn Phe Ser Thr

340

Ser Gly His Leu Val

345

Arg His Ile Arg Thr

350

Thr Gly Glu Lys Pro

355

Phe Ala Cys Asp Ile 360

Cys Gly Arg Lys Phe 365

Gln Asn Ser Thr Leu

370

Thr Glu His Thr Lys

375

Ile His Leu Arg Gln

380

Ser

Pro

His

Ser

Lys 240

Asn

Cys

His

Val

His 320

Met

His

Ala

Lys

Asp Lys 385 <210> 129 <211> 402 <212> Protein

- 242 046157 <213> Artificial sequence <220>

<223> Description of the artificial sequence polypeptide <400> 129

Synthetic

Met Ala Ala 1

Asp His Leu 5

Met Leu Ala

Glu Gly Tyr 10

Arg Leu Val Gln 15

Arg Pro Pro

Ser Ala Ala 20

Ala Ala His

Gly Pro His

Ala Leu Arg Thr 30

Leu Pro Pro

Tyr Ala Gly

Pro Gly Leu 40

Asp Ser Gly

Leu Arg Pro Arg 45

Gly Ala Pro 50

Leu Gly Pro

Pro Pro Pro 55

Arg Gln Pro

Gly Ala Leu Ala

Tyr Gly Ala 65

Phe Gly Pro

Pro Ser Ser

Phe Gln Pro

Phe Pro Ala Val

Pro Pro Pro

Ala Ala Gly 85

Ile Ala His

Leu Gln Pro 90

Val Ala Thr Pro

Tyr Pro Gly

Arg Ala Ala 100

Ala Pro Pro

105

Asn Ala Pro

Gly Gly Pro Pro 110

Gly Pro Gln

115

Pro Ala Pro

Ser Ala Ala

120

Ala Pro Pro

Pro Pro Ala His 125

Ala Leu Gly

130

Gly Met Asp

Ala Glu Leu 135

Ile Asp Glu

140

Glu Ala Leu Thr

Ser Leu Glu 145

Leu Glu Leu

150

Gly Leu His

Arg Val Arg

155

Glu Leu Pro Glu

160

Leu Phe Leu

Gly Gln Ser

165

Glu Phe Asp

Cys Phe Ser 170

Asp Leu Gly Ser

175

Ala Pro Pro

Ala Gly Ser 180

Val Ser Cys

185

Gly Gly Ser

Gly Gly Gly Ser

190

Gly Gly Gly

195

Ser Gly Gly

Gly Ser Gly

200

Gln Ser Gln

Leu Ile Lys Pro 205

Ser Arg Met

210

Arg Lys Tyr

Pro Asn Arg 215

Pro Ser Lys

220

Thr Pro Pro His

- 243 046157

Glu Arg Pro 225

Arg Ser Asp

Pro Phe Gln

Leu His Thr

275

Tyr Ala Cys

230

Pro Val Glu

Ser Cys Asp

235

Arg Arg Phe Ser

240

Asp Ile

290

Thr Lys 305

Glu Ser

Ile Arg

Arg Asn

Thr Gly

370

Gln Asn 385

Asp Lys <210>

<211>

<212>

Asn Leu Val

245

Arg His Ile

Arg Ile His 250

Thr Gly Gln Lys

255

Cys Arg Ile 260

His Ile Arg

Cys Met Arg

265

Thr His Thr

280

Asn Phe Ser

Gly Glu Lys

Cys Gly

Arg Lys

Ile His

Leu Arg

310

Cys Asp

Arg Arg 325

Ile His

340

Thr Gly

Phe Ser 355

Glu Lys

Ser Thr

130

569 Protein

Thr Ser

Pro Phe

Leu Thr

390

Phe Ala 295

Gln Lys

Phe Ser

Gln Lys

Gly His 360

Ala Cys 375

Glu His

Arg Ser

Asp Glu

300

Asp Arg

Pro Tyr 315

Arg Glu Asp Asn 270

Pro Phe Ala Cys 285

Leu Val Arg His

Ala Cys Pro Val

320

Gln Ser

330

Gly Asn

Leu Thr Glu His

335

Pro Phe 345

Gln Cys

Arg Ile Cys Met

350

Leu Val

Arg His

Ile Arg Thr His 365

Asp Ile

Cys Gly

380

Arg Lys Phe Ala

Thr Lys

Ile His

395

Leu Arg Gln Lys

400 <213> Artificial sequence <220>

<223> Description of the artificial polypeptide sequence

Synthetic <400> 130

Met Ala Ala Asp His Leu Met Leu Ala Glu Gly Tyr

5 10

Arg Leu Val Gln 15

Arg Pro Pro Ser Ala Ala Ala Ala His Gly Pro His

Ala Leu Arg Thr

- 244 046157

Leu Pro Pro Tyr Ala 35

Gly Pro Gly Leu Asp 40

Ser Gly Leu Arg Pro 45

Gly Ala Pro Leu Gly 50

Pro Pro Pro Pro Arg 55

Gln Pro Gly Ala Leu 60

Tyr Gly Ala Phe Gly 65

Pro Pro Ser Ser Phe 70

Gln Pro Phe Pro Ala 75

Pro Pro Pro Ala Ala

Gly Ile Ala His Leu

Gln Pro Val Ala Thr

Tyr Pro Gly Arg Ala

100

Ala Ala Pro Pro Asn

105

Ala Pro Gly Gly Pro

110

Gly Pro Gln Pro Ala

115

Pro Ser Ala Ala Ala

120

Pro Pro Pro Pro Ala

125

Ala Leu Gly Gly Met

130

Asp Ala Glu Leu Ile 135

Asp Glu Glu Ala Leu 140

Ser Leu Glu Leu Glu 145

Leu Gly Leu His Arg 150

Val Arg Glu Leu Pro 155

Leu Phe Leu Gly Gln

165

Ser Glu Phe Asp Cys

170

Phe Ser Asp Leu Gly

175

Ala Pro Pro Ala Gly

180

Ser Val Ser Cys Ala

185

Asp His Leu Met Leu

190

Glu Gly Tyr Arg Leu 195

Val Gln Arg Pro Pro

200

Ser Ala Ala Ala Ala

205

Gly Pro His Ala Leu

210

Arg Thr Leu Pro Pro 215

Tyr Ala Gly Pro Gly

220

Asp Ser Gly Leu Arg 225

Pro Arg Gly Ala Pro 230

Leu Gly Pro Pro Pro 235

Arg Gln Pro Gly Ala

245

Leu Ala Tyr Gly Ala

250

Phe Gly Pro Pro Ser

255

Phe Gln Pro Phe Pro

260

Ala Val Pro Pro Pro

265

Ala Ala Gly Ile Ala

270

Arg

Ala

Val 80

Pro

Pro

His

Thr

Glu 160

Ser

Ala

His

Leu

Pro 240

Ser

His

- 245 046157

Leu Gln Pro Val Ala

275

Thr Pro Tyr Pro Gly

280

Arg Ala Ala Ala Pro 285

Asn Ala Pro Gly Gly

290

Pro Pro Gly Pro Gln

295

Pro Ala Pro Ser Ala

300

Ala Pro Pro Pro Pro 305

Ala His Ala Leu Gly 310

Gly Met Asp Ala Glu 315

Ile Asp Glu Glu Ala

325

Leu Thr Ser Leu Glu

330

Leu Glu Leu Gly Leu

335

Arg Val Arg Glu Leu

340

Pro Glu Leu Phe Leu

345

Gly Gln Ser Glu Phe

350

Cys Phe Ser Asp Leu

355

Gly Ser Ala Pro Pro 360

Ala Gly Ser Val Ser 365

Gln Ser Gln Leu Ile

370

Lys Pro Ser Arg Met

375

Arg Lys Tyr Pro Asn

380

Pro Ser Lys Thr Pro 385

Pro His Glu Arg Pro 390

Tyr Ala Cys Pro Val 395

Ser Cys Asp Arg Arg

405

Phe Ser Arg Ser Asp

410

Asn Leu Val Arg His

415

Arg Ile His Thr Gly

420

Gln Lys Pro Phe Gln

425

Cys Arg Ile Cys Met

430

Asn Phe Ser Arg Glu 435

Asp Asn Leu His Thr 440

His Ile Arg Thr His 445

Gly Glu Lys Pro Phe

450

Ala Cys Asp Ile Cys 455

Gly Arg Lys Phe Ala

460

Ser Asp Glu Leu Val 465

Arg His Thr Lys Ile 470

His Leu Arg Gln Lys 475

Arg Pro Tyr Ala Cys

485

Pro Val Glu Ser Cys

490

Asp Arg Arg Phe Ser

495

Ser Gly Asn Leu Thr

500

Glu His Ile Arg Ile

505

His Thr Gly Gln Lys

510

Phe Gln Cys Arg Ile 515

Cys Met Arg Asn Phe

520

Ser Thr Ser Gly His

525

Pro

Ala

Leu 320

His

Asp

Cys

Arg

Glu 400

Ile

Arg

Thr

Arg

Asp 480

Gln

Pro

Leu

- 246 046157

Val Arg His Ile Arg Thr His Thr

530 535

Gly Glu Lys

Pro Phe Ala Cys Asp 540

Ile Cys Gly Arg Lys Phe Ala Gln Asn Ser Thr Leu Thr Glu His Thr

545 550 555 560

Lys Ile His Leu Arg Gln Lys Asp Lys

565 <210> 131 <211> 585 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic polypeptide <400> 131

Met Ala Ala Asp His Leu Met Leu Ala Glu Gly Tyr Arg Leu Val Gln

5 1015

Arg Pro Pro Ser Ala Ala Ala Ala His Gly Pro His Ala Leu Arg Thr 20 2530

Leu Pro Pro Tyr Ala Gly Pro Gly Leu Asp Ser Gly Leu Arg Pro Arg 35 4045

Gly Ala Pro Leu Gly Pro Pro Pro Pro Arg Gln Pro Gly Ala Leu Ala 50 5560

Tyr Gly Ala Phe Gly Pro Pro Ser Ser Phe Gln Pro Phe Pro Ala Val 65 70 7580

Pro Pro Pro Ala Ala Gly Ile Ala His Leu Gln Pro Val Ala Thr Pro 85 9095

Tyr Pro Gly Arg Ala Ala Ala Pro Pro Asn Ala Pro Gly Gly Pro Pro

100 105110

Gly Pro Gln Pro Ala Pro Ser Ala Ala Ala Pro Pro Pro Pro Ala His

115 120125

Ala Leu Gly Gly Met Asp Ala Glu Leu Ile Asp Glu Glu Ala Leu Thr

130 135140

Ser Leu Glu Leu Glu Leu Gly Leu His Arg Val Arg Glu Leu Pro Glu

145 150 155160

- 247 046157

Leu Phe Leu Gly Gln

165

Ser Glu Phe Asp Cys

170

Phe Ser Asp Leu Gly

175

Ala Pro Pro Ala Gly

180

Ser Val Ser Cys Gly

185

Gly Ser Gly Gly Gly

190

Gly Ala Asp His Leu

195

Met Leu Ala Glu Gly 200

Tyr Arg Leu Val Gln 205

Pro Pro Ser Ala Ala

210

Ala Ala His Gly Pro 215

His Ala Leu Arg Thr

220

Pro Pro Tyr Ala Gly 225

Pro Gly Leu Asp Ser 230

Gly Leu Arg Pro Arg 235

Ala Pro Leu Gly Pro

245

Pro Pro Pro Arg Gln

250

Pro Gly Ala Leu Ala

255

Gly Ala Phe Gly Pro

260

Pro Ser Ser Phe Gln

265

Pro Phe Pro Ala Val

270

Pro Pro Ala Ala Gly

275

Ile Ala His Leu Gln

280

Pro Val Ala Thr Pro

285

Pro Gly Arg Ala Ala 290

Ala Pro Pro Asn Ala

295

Pro Gly Gly Pro Pro 300

Pro Gln Pro Ala Pro 305

Ser Ala Ala Ala Pro 310

Pro Pro Pro Ala His 315

Leu Gly Gly Met Asp Ala Glu Leu Ile Asp Glu Glu Ala Leu Thr

325 330 335

Leu Glu Leu Glu Leu Gly Leu His Arg Val Arg Glu Leu Pro Glu

340 345 350

Phe Leu Gly Gln Ser Glu Phe Asp Cys Phe Ser Asp Leu Gly Ser

355 360 365

Pro Pro Ala Gly Ser Val Ser Cys Gly Gly Ser Gly Gly Gly Ser

370 375 380

Gln Ser Gln Leu Ile Lys Pro Ser Arg Met Arg Lys Tyr Pro Asn

385 390 395

Pro Ser Lys Thr Pro Pro His Glu Arg Pro Tyr Ala Cys Pro Val

405 410 415

Ser

Ser

Arg

Leu

Gly 240

Tyr

Pro

Tyr

Gly

Ala 320

Ser

Leu

Ala

Gly

Arg 400

Glu

- 248 046157

Ser Cys Asp

Arg Arg Phe 420

Ser Arg Ser

425

Asp Asn Leu

Val Arg His Ile

430

Arg Ile His

435

Thr Gly Gln

Lys Pro Phe

440

Gln Cys Arg

Ile Cys Met Arg 445

Asn Phe Ser

450

Arg Glu Asp

Asn Leu His 455

Thr His Ile

460

Arg Thr His Thr

Gly Glu Lys 465

Pro Phe Ala

470

Cys Asp Ile

Cys Gly Arg

475

Lys Phe Ala Arg

480

Ser Asp Glu

Leu Val Arg

485

His Thr Lys

Ile His Leu 490

Arg Gln Lys Asp

495

Arg Pro Tyr

Ala Cys Pro 500

Val Glu Ser

505

Cys Asp Arg

Arg Phe Ser Gln

510

Ser Gly Asn

515

Leu Thr Glu

His Ile Arg

520

Ile His Thr

Gly Gln Lys Pro 525

Phe Gln Cys

530

Arg Ile Cys

Met Arg Asn 535

Phe Ser Thr

540

Ser Gly His Leu

Val Arg His 545

Ile Arg Thr

550

His Thr Gly

Glu Lys Pro 555

Phe Ala Cys Asp

560

Ile Cys Gly

Arg Lys Phe

565

Ala Gln Asn

Ser Thr Leu 570

Thr Glu His Thr

575

Lys Ile His

Leu Arg Gln 580

Lys Asp Lys

585 <210> 132 <211> 523 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial polypeptide sequence

Synthetic <400> 132

Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly

5 10

Ser Asp Ala Leu 15

Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala 20 25

Leu Asp Asp Phe 30

- 249 046157

Asp

Met

Val 65

Glu

Ser

Ala

Pro

Pro 145

Ala

Ala

Val

Pro

Gln 225

Pro

Gln

Met

Leu Asp Met

Leu Gly Ser Asp

Ala Leu Asp Asp Phe Asp Leu Asp 45

Leu Ile Asn Ser Arg Ser Ser Gly Ser Pro Lys Lys Lys Arg Lys 50 55 60

Gly Ser Gln

Tyr

Leu Pro Asp Thr Asp Asp Arg His Arg Ile Glu 70 75 80

Lys Arg Lys Arg Thr 85

Pro Phe Ser Gly Pro

100

Val Pro Ser Arg Ser 115

Tyr Pro Phe Thr Ser 130

Thr Met Val Phe Pro

150

Pro Ala Pro Pro Gln

165

Pro Ala Met Val Ser

180

Leu Ala Pro Gly Pro

195

Thr Gln Ala Gly Glu 210

Phe Asp Asp Glu Asp

230

Ala Val Phe Thr Asp

245

Leu Leu Asn Gln Gly 260

Leu Met Glu Tyr Pro

275

Tyr Glu Thr Phe Lys

Thr Asp Pro Arg Pro

105

Ser Ala Ser Val Pro

120

Ser Leu Ser Thr Ile 135

Ser Gly Gln Ile Ser

155

Val Leu Pro Gln Ala

170

Ala Leu Ala Gln Ala

185

Pro Gln Ala Val Ala

200

Gly Thr Leu Ser Glu 215

Leu Gly Ala Leu Leu

235

Leu Ala Ser Val Asp

250

Ile Pro Val Ala Pro

265

Glu Ala Ile Thr Arg

280

Ser Ile Met Lys Lys

Pro Pro Arg Arg Ile

110

Lys Pro Ala Pro Gln

125

Asn Tyr Asp Glu Phe 140

Gln Ala Ser Ala Leu

160

Pro Ala Pro Ala Pro

175

Pro Ala Pro Val Pro

190

Pro Pro Ala Pro Lys 205

Ala Leu Leu Gln Leu 220

Gly Asn Ser Thr Asp

240

Asn Ser Glu Phe Gln

255

His Thr Thr Glu Pro

270

Leu Val Thr Gly Ala

285

- 250 046157

Gln Arg Pro Pro Asp

290

Pro Ala Pro Ala Pro

295

Leu Gly Ala Pro Gly 300

Pro Asn Gly Leu Leu 305

Ser Gly Asp Glu Asp 310

Phe Ser Ser Ile Ala 315

Met Asp Phe Ser Ala

325

Leu Leu Gly Ser Gly

330

Ser Gly Ser Arg Asp

335

Arg Glu Gly Met Phe

340

Leu Pro Lys Pro Glu

345

Ala Gly Ser Ala Ile

350

Asp Val Phe Glu Gly 355

Arg Glu Val Cys Gln 360

Pro Lys Arg Ile Arg 365

Phe His Pro Pro Gly

370

Ser Pro Trp Ala Asn

375

Arg Pro Leu Pro Ala 380

Leu Ala Pro Thr Pro 385

Thr Gly Pro Val His 390

Glu Pro Val Gly Ser 395

Thr Pro Ala Pro Val

405

Pro Gln Pro Leu Asp

410

Pro Ala Pro Ala Val

415

Pro Glu Ala Ser His

420

Leu Leu Glu Asp Pro

425

Asp Glu Glu Thr Ser

430

Ala Val Lys Ala Leu 435

Arg Glu Met Ala Asp 440

Thr Val Ile Pro Gln

445

Glu Glu Ala Ala Ile

450

Cys Gly Gln Met Asp

455

Leu Ser His Pro Pro

460

Arg Gly His Leu Asp 465

Glu Leu Thr Thr Thr 470

Leu Glu Ser Met Thr 475

Asp Leu Asn Leu Asp

485

Ser Pro Leu Thr Pro

490

Glu Leu Asn Glu Ile

495

Asp Thr Phe Leu Asn

500

Asp Glu Cys Leu Leu

505

His Ala Met His Ile

510

Leu

Asp 320

Ser

Ser

Pro

Ser

Leu 400

Thr

Gln

Lys

Pro

Glu 480

Leu

Ser

Thr Gly Leu Ser Ile 515

Phe Asp Thr Ser Leu 520

Phe <210> 133 <211> 50

- 251 046157 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial sequence polypeptide <400> 133

Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu Gly

510

Asp Asp Phe Asp Leu Asp Met Leu Gly Ser Asp Ala 2025

Asp Leu Asp Met Leu Gly Ser Asp Ala Leu Asp Asp 3540

Synthetic

Ser Asp Ala Leu 15

Leu Asp Asp Phe 30

Phe Asp Leu Asp 45

Met Leu <210> 134 <211> 275 <212> Protein <213> Artificial Sequence <220>

<223> Description of the artificial sequence polypeptide <400> 134

Synthetic

Met Ser Gly 1

Arg Phe Pro

Met Gly Met 35

Pro Gln His 50

Ala Gly Asn 65

Gly Thr Val

Leu Glu Met

Asp Gly Thr 20

Gly Gln Phe

Ala Phe Asn

Met Asn Ala

Asn Gly Gly 85

Arg Phe Asn

Asn Ser Gln 100

Gly Ser Leu

Pro Ala Ser

Ala Asp His

Asn Gly Leu 25

Pro Ser Pro 40

Ala Leu Met 55

Thr Ser Gly

His Pro Pro

Phe Met Gly

105

Met Gln Leu

Met Met Ala 10

His His His

His His His

Gly Glu His 60

Ile Arg His 75

Ser Ala Leu 90

Pro Pro Val

Gln Lys Leu

Met Asn His Gly 15

Pro Ala His Arg 30

Gln Gln Gln Gln 45

Ile His Tyr Gly

Ala Met Gly Pro 80

Ala Pro Ala Ala

Ala Ser Gln Gly 110

Asn Asn Gln Tyr

- 252 046157

115 120125

Phe Asn His Pro Tyr Pro His Asn His Tyr Met Pro Asp Leu His

130 135140

Pro Ala Ala Gly His Gln Met Asn Gly Thr Asn Gln His Phe Arg Asp

145 150 155160

Cys Asn Pro Lys His Ser Gly Gly Ser Ser Thr Pro Gly Gly Ser Gly

165 170175

Gly Ser Ser Thr Pro Gly Gly Ser Gly Ser Ser Ser Gly Gly Gly Ala

180 185190

Gly Ser Ser Asn Ser Gly Gly Gly Ser Gly Ser Gly Asn Met Pro Ala

195 200205

Ser Val Ala His Val Pro Ala Ala Met Leu Pro Pro Asn Val Ile Asp

210 215220

Thr Asp Phe Ile Asp Glu Glu Val Leu Met Ser Leu Val Ile Glu Met

225 230 235240

Gly Leu Asp Arg Ile Lys Glu Leu Pro Glu Leu Trp Leu Gly Gln Asn

245 250255

Glu Phe Asp Phe Met Thr Asp Phe Val Cys Lys Gln Gln Pro Ser Arg

260 265270

Val Ser Cys

275 <210> 135 <211> 183 <212> Protein <213>

Artificial sequence <220>

<223> Description of artificial sequence: Synthetic polypeptide <400> 135

Ala Asp His Leu Met Leu Ala Glu Gly Tyr Arg Leu Val Gln Arg Pro

5 1015

Pro Ser Ala Ala Ala Ala His Gly Pro His Ala Leu Arg Thr LeuPro

2530

Pro Tyr Ala Gly Pro Gly Leu Asp Ser Gly Leu Arg Pro Arg Gly Ala 35 4045

- 253 046157

ProLeu

Gly Pro

Pro Pro

Ala Phe 65

Gly Pro

Pro Ser

Pro Ala

Ala Gly

Ile Ala 85

Gly Arg

Ala Ala

100

Ala Pro

Glin Pro

Ala Pro 115

Ser Ala

Gly Gly

130

Met Asp

Ala Glu

Glu Leu 145

Glu Leu

Gly Leu

150

Leu Gly

Gln Ser

Glu Phe 165

Pro Ala

Gly Ser

180

Val Ser

Pro Arg 55

Ser Phe

His Leu

Pro Asn

Ala Ala

120

Leu Ile 135

His Arg

Asp Cys

Cys

Glin Pro

Glin Pro

Glin Pro

Ala Pro 105

Pro Pro

Asp Glu

Val Arg

Phe Ser

170

Gly Ala 60

Phe Pro 75

Val Ala

Gly Gly

Pro Pro

Glu Ala

140

Glu Leu 155

Asp Leu

Leu Ala Tyr Gly

Ala Val Pro Pro

Thr Pro Tyr Pro 95

Pro Pro Gly Pro

110

Ala His Ala Leu 125

Leu Thr Ser Leu

Pro Glu Leu Phe

160

Gly Ser Ala Pro

175 <210> 136 <211> 7 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial sequence

Synthetic peptide <220>

<221> MOD_RES <222> (2)..(2) <223> Any amino acid <220>

<221> MOD_RES <222> (4)..(4) <223> Any amino acid <220>

<221> MOD_RES <222> (6)..(6) <223> Any amino acid

- 254 046157 <400> 136

Cys Xaa Cys Xaa His Xaa His 1 5 <210> 137 <211> 17 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic peptide <220>

<221> MOD_RES <222> (2)..(2) <223> Any amino acid <220>

<221> MOD_RES <222> (4)..(4) <223> Any amino acid <220>

<221> MOD_RES <222> (6)..(6) <223> Any amino acid <220>

<221> MOD_RES <222> (8)..(10) <223> Any amino acid <220>

<221> MOD_RES <222> (12)..(12) <223> Any amino acid <220>

<221> MOD_RES <222> (14)..(14) <223> Any amino acid <220>

<221> MOD_RES <222> (16)..(16) <223> Any amino acid <400> 137

Cys Xaa Cys Xaa Cys Xaa His Xaa Xaa Xaa Cys Xaa Cys Xaa Cys Xaa

5 10 15

Cys <210> 138 <211> 17 <212> Protein <213> Artificial sequence

- 255 046157 <220>

<223> Description of artificial sequence: Synthetic peptide <220>

<221> MOD_RES <222> (2)..(2) <223> Any amino acid <220>

<221> MOD_RES <222> (4)..(4) <223> Any amino acid <220>

<221> MOD_RES <222> (6)..(6) <223> Any amino acid <220>

<221> MOD_RES <222> (8)..(10) <223> Any amino acid <220>

<221> MOD_RES <222> (12)..(12) <223> Any amino acid <220>

<221> MOD_RES <222> (14)..(14) <223> Any amino acid <220>

<221> MOD_RES <222> (16)..(16) <223> Any amino acid <400> 138

Cys Xaa Cys Xaa Cys Xaa Cys Xaa Xaa Xaa His Xaa Cys Xaa Cys Xaa

5 10 15

Cys <210> 139 <211> 15 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic peptide <220>

<221> MOD_RES <222> (2)..(2)

- 256 046157 <223> Any amino acid <220>

<221> MOD_RES <222> (4)..(4) <223> Any amino acid <220>

<221> MOD_RES <222> (6)..(6) <223> Any amino acid <220>

<221> MOD_RES <222> (8)..(8) <223> Any amino acid <220>

<221> MOD_RES <222> (10)..(10) <223> Any amino acid <220>

<221> MOD_RES <222> (12)..(12) <223> Any amino acid <220>

<221> MOD_RES <222> (14)..(14) <223> Any amino acid <220>

<221> MOD_RES <222> (15) . . (15) <223> Cys, His, or Asp <400> 139

Cys Xaa Cys Xaa His Xaa Cys Xaa Cys Xaa Cys Xaa Cys Xaa Xaa

5 10 15 <210> 140 <211> 17 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic peptide <220>

<221> MOD_RES <222> (2)..(2) <223> Any amino acid <220>

<221> MOD_RES <222> (4)..(4) <223> Any amino acid <220>

- 257 046157 <221> MOD_RES <222> (6)..(6) <223> Any amino acid <220>

<221> MOD_RES <222> (8)..(10) <223> Any amino acid <220>

<221> MOD_RES <222> (12)..(12) <223> Any amino acid <220>

<221> MOD_RES <222> (14) .. (14) <223> Any amino acid <220>

<221> MOD_RES <222> (16)..(16) <223> Any amino acid <400> 140

Cys Xaa Cys Xaa Cys Xaa Cys Xaa Xaa Xaa Cys Xaa Cys Xaa Cys Xaa

5 10 15

Cys <210> 141 <211> 7 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic peptide <220>

<221> MOD_RES <222> (2)..(2) <223> Any amino acid <220>

<221> MOD_RES <222> (4)..(4) <223> Any amino acid <220>

<221> MOD_RES <222> (6)..(6) <223> Any amino acid <400> 141

Cys Xaa Cys Xaa Cys Xaa His 1 5

- 258 046157 <210> 142 <211> 7 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic peptide <220>

<221> MOD_RES <222> (2)..(2) <223> Any amino acid <220>

<221> MOD_RES <222> (4)..(4) <223> Any amino acid <220>

<221> MOD_RES <222> (6)..(6) <223> Any amino acid <400> 142

Cys Xaa Cys Xaa His Xaa Cys 1 5 <210> 143 <211> 17 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic peptide <220>

<221> MOD_RES <222> (2)..(2) <223> Any amino acid <220>

<221> MOD_RES <222> (4)..(4) <223> Any amino acid <220>

<221> MOD_RES <222> (6)..(6) <223> Any amino acid <220>

<221> MOD_RES <222> (8)..(10) <223> Any amino acid <220>

<221> MOD_RES <222> (12)..(12)

- 259 046157 <223> Any amino acid <220>

<221> MOD_RES <222> (14) .. (14) <223> Any amino acid <220>

<221> MOD_RES <222> (16)..(16) <223> Any amino acid <400> 143

Cys Xaa Cys Xaa His Xaa Cys Xaa Xaa Xaa Cys Xaa Cys Xaa His Xaa

5 10 15

Cys <210> 144 <211> 17 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic peptide <220>

<221> MOD_RES <222> (2)..(2) <223> Any amino acid <220>

<221> MOD_RES <222> (4)..(4) <223> Any amino acid <220>

<221> MOD_RES <222> (6)..(6) <223> Any amino acid <220>

<221> MOD_RES <222> (8)..(10) <223> Any amino acid <220>

<221> MOD_RES <222> (12)..(12) <223> Any amino acid <220>

<221> MOD_RES <222> (14)..(14) <223> Any amino acid <220>

<221> MOD_RES

- 260 046157 <222> (16)..(16) <223> Any amino acid <400> 144

Cys Xaa Cys Xaa Cys Xaa Cys Xaa Xaa Xaa Cys Xaa Cys Xaa His Xaa

5 10 15

Cys <210> 145 <211> 7 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic peptide <220>

<221> MOD_RES <222> (2)..(2) <223> Any amino acid <220>

<221> MOD_RES <222> (4)..(4) <223> Any amino acid <220>

<221> MOD_RES <222> (6)..(6) <223> Any amino acid <400> 145

Cys Xaa Cys Xaa Cys Xaa Cys 1 5 <210> 146 <211> 17 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic peptide <220>

<221> MOD_RES <222> (2)..(2) <223> Any amino acid <220>

<221> MOD_RES <222> (4)..(4) <223> Any amino acid <220>

- 261 046157 <221> MOD_RES <222> (6)..(6) <223> Any amino acid <220>

<221> MOD_RES <222> (8)..(10) <223> Any amino acid <220>

<221> MOD_RES <222> (12)..(12) <223> Any amino acid <220>

<221> MOD_RES <222> (14) .. (14) <223> Any amino acid <220>

<221> MOD_RES <222> (16)..(16) <223> Any amino acid <400> 146

Cys Xaa Cys Xaa Cys Xaa Cys Xaa Xaa Xaa Cys Xaa His Xaa His Xaa

5 10 15

Cys <210> 147 <211> 344 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic polypeptide <220>

<221> MISC_FEATURE <222> (8)..(322) <223> This region may contain 1-15 repeat units Tyr Lys Cys Pro Glu Cys

Gly Lys Ser Phe Ser His Gln Arg Thr His Thr Gly Glu Lys Pro <220>

<221> NON_CONS <222> (18)..(19) <220>

<221> NON_CONS <222> (39)..(40) <220>

<221> NON_CONS <222> (60)..(61)

- 262 046157

NON_CONS (81) .. (82)

NON_CONS (102) .. (103)

NON_CONS (123)..(124)

NON_CONS (144) .. (145)

NON_CONS (165)..(166)

NON_CONS (186)..(187)

NON_CONS (207)..(208)

NON_CONS (228)..(229)

NON_CONS (249)..(250)

NON_CONS (270)..(271)

NON_CONS (291)..(292)

NON_CONS (312)..(313)

NON_CONS (333)..(334)

See the original application description for a detailed description of substitutions and preferred embodiments <400>

147

Leu Glu Pro 1

Gly

Glu Lys Pro Tyr Lys 5

Cys Pro Glu Cys Gly 10

Lys Ser 15

Phe Ser His

Gln 20

Arg Thr His Thr Gly

Glu Lys Pro Tyr Lys

Cys Pro

- 263 046157

Glu Cys Gly Lys Ser 35

Phe Ser His Gln Arg 40

Thr His Thr Gly Glu Lys 45

Pro Tyr Lys Cys Pro 50

Glu Cys Gly Lys Ser 55

Phe Ser His Gln Arg Thr 60

His Thr Gly Glu Lys 65

Pro Tyr Lys Cys Pro 70

Glu Cys Gly Lys Ser Phe 75 80

Ser His Gln Arg Thr

His Thr Gly Glu Lys

Pro Tyr Lys Cys Pro Glu 95

Cys Gly Lys Ser Phe

100

Ser His Gln Arg Thr

105

His Thr Gly Glu Lys Pro

110

Tyr Lys Cys Pro Glu 115

Cys Gly Lys Ser Phe 120

Ser His Gln Arg Thr His

125

Thr Gly Glu Lys Pro

130

Tyr Lys Cys Pro Glu

135

Cys Gly Lys Ser Phe Ser 140

His Gln Arg Thr His 145

Thr Gly Glu Lys Pro 150

Tyr Lys Cys Pro Glu Cys

155 160

Gly Lys Ser Phe Ser

165

His Gln Arg Thr His

170

Thr Gly Glu Lys Pro Tyr

175

Lys Cys Pro Glu Cys

180

Gly Lys Ser Phe Ser

185

His Gln Arg Thr His Thr

190

Gly Glu Lys Pro Tyr

195

Lys Cys Pro Glu Cys 200

Gly Lys Ser Phe Ser His

205

Gln Arg Thr His Thr

210

Gly Glu Lys Pro Tyr

215

Lys Cys Pro Glu Cys Gly

220

Lys Ser Phe Ser His 225

Gln Arg Thr His Thr 230

Gly Glu Lys Pro Tyr Lys

235 240

Cys Pro Glu Cys Gly

245

Lys Ser Phe Ser His

250

Gln Arg Thr His Thr Gly

255

Glu Lys Pro Tyr Lys

260

Cys Pro Glu Cys Gly

265

Lys Ser Phe Ser His Gln

270

Arg Thr His Thr Gly

Glu Lys Pro Tyr Lys

Cys Pro Glu Cys Gly Lys

- 264 046157

275

280

Ser Phe Ser His Gln Arg Thr His Thr Gly Glu Lys

290 295300

Pro Glu Cys Gly Lys Ser Phe Ser His Gln Arg Thr

305 310315

Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe

325330

285

Pro Tyr Lys Cys

His Thr Gly Glu

320

Ser His Gln Arg

335

Thr His Thr Gly Lys Lys Thr Ser

340 <210> 148 <211> 292 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial polypeptide sequence

Synthetic <220>

<221> MOD_RES <222> (8) .. (57) <223> Any amino acid <220>

< 221> MISC_FEATURE < 222> (8)..(57) < 223> This region can contain 1-50 residues <220>

< 221> MOD_RES < 222> (65).. (114) < 223> Any amino acid <220>

< 221> MISC_FEATURE < 222> (65)..(114) < 223> This region can contain 1-50 residues <220>

< 221> MOD_RES < 222> (122)..(171) < 223> Any amino acid <220>

< 221> MISC_FEATURE < 222> (122)..(171) < 223> This region can contain 1-50 residues <220>

< 221> MOD_RES < 222> (179)..(228) < 223> Any amino acid

- 265 046157 <220>

<221> MISC_FEATURE <222> (179)..(228) <223> This area can contain 1<220>

<221> MOD_RES <222> (236)..(285) <223> Any amino acid <220>

<221> MISC_FEATURE <222> (236)..(285) <223> This area can contain 1<400> 148

Arg Ser Asp Asn Leu Val Arg Xaa Xaa 1 5

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

25

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

40

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

55

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 65 70

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 85

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

100 105

Xaa Xaa Arg Ser Asp Glu Leu Val Arg

115 120

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

130 135

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

145 150

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 165

Thr Glu Xaa Xaa Xaa Xaa Xaa Xaa Xaa

180 185 statok statok

Xaa Xaa Xaa Xaa Xaa Xaa

Xaa Xaa Xaa Xaa Xaa Xaa 30

Xaa Xaa Xaa Xaa Xaa Xaa 45

Glu Asp Asn Leu His Thr 60

Xaa Xaa Xaa Xaa Xaa Xaa

80

Xaa Xaa Xaa Xaa Xaa Xaa 95

Xaa Xaa Xaa Xaa Xaa Xaa

110

Xaa Xaa Xaa Xaa Xaa Xaa

125

Xaa Xaa Xaa Xaa Xaa Xaa

140

Xaa Xaa Xaa Xaa Xaa Xaa

155 160

Xaa Gln Ser Gly Asn Leu 175

Xaa Xaa Xaa Xaa Xaa Xaa

190

- 266 046157

Xaa Xaa

Xaa Xaa

210

Xaa Xaa 225

Xaa Xaa

Xaa Xaa

Xaa Xaa

Xaa Xaa 195

Xaa Xaa

Xaa Xaa

Xaa Xaa

Xaa Xaa 260

Xaa Xaa 275

Xaa Xaa

Xaa Xaa

Thr Ser

230

Xaa Xaa 245

Xaa Xaa

Xaa Xaa

Xaa Xaa

200

Xaa Xaa 215

Gly His

Xaa Xaa

Xaa Xaa

Xaa Xaa

280

Xaa Xaa

Xaa Xaa

Xaa Xaa

Xaa Xaa

220

Leu Val Arg Xaa

235

Xaa Xaa

250

Xaa Xaa

Xaa Xaa 265

Xaa Xaa

Xaa Xaa 205

Xaa Xaa

Xaa Xaa

Xaa Xaa

Xaa Xaa

270

Xaa Xaa

Xaa Xaa

Xaa Xaa

240

Xaa Xaa 255

Xaa Xaa

Xaa Xaa

Xaa Xaa

Xaa Gln Asn Ser

285

Thr Leu Thr Glu

290 <210> 149 <211> 292 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic polypeptide <220>

<221> MOD_RES <222> (8) .. (57) <223> Any amino acid <220>

< 221> MISC_FEATURE < 222> (8)..(57) < 223> This region can contain 1-50 residues <220>

< 221> MOD_RES < 222> (65)..(114) < 223> Any amino acid <220>

< 221> MISC_FEATURE < 222> (65)..(114) <223> This region can contain 1-50 residues <220>

<221> MOD_RES <222> (122)..(171)

- 267 046157 <223> Any amino acid <220>

<221> MISC_FEATURE <222> (122)..(171) <223> This region can contain 1-50 residues <220>

<221> MOD_RES <222> (179)..(228) <223> Any amino acid <220>

<221> MISC_FEATURE <222> (179)..(228) <223> This region can contain 1-50 residues <220>

<221> MOD_RES <222> (236)..(285) <223> Any amino acid <220>

<221> MISC_FEATURE <222> (236)..(285) <223> This region can contain 1-50 residues <400> 149 Arg Ser Asp 1

Asn Leu Val

Arg Xaa Xaa

Xaa Xaa Xaa 10

Xaa Xaa Xaa

Xaa Xaa Xaa

Xaa Xaa Xaa 20

Xaa Xaa Xaa

Xaa Xaa Xaa

Xaa Xaa Xaa

Xaa Xaa Xaa

Xaa Xaa Xaa

Xaa Xaa Xaa

Xaa Xaa Xaa

Xaa Xaa Xaa 45

Xaa Xaa Xaa

Xaa Xaa Xaa

Xaa Xaa Xaa 55

His Arg Thr

Thr Leu Thr

Xaa Xaa Xaa 65

Xaa Xaa Xaa

Xaa Xaa Xaa

Xaa Xaa Xaa

Xaa Xaa Xaa

Xaa Xaa Xaa

Xaa Xaa Xaa

Xaa Xaa Xaa

Xaa Xaa Xaa 90

Xaa Xaa Xaa

Xaa Xaa Xaa

Xaa Xaa Xaa 100

Xaa Xaa Xaa

105

Xaa Xaa Xaa

Xaa Xaa Xaa

110

Xaa Xaa Arg

115

Glu Asp Asn

Leu His Thr

120

Xaa Xaa Xaa

Xaa Xaa Xaa 125

Xaa Xaa Xaa

130

Xaa Xaa Xaa

Xaa Xaa Xaa 135

Xaa Xaa Xaa

140

Xaa Xaa Xaa

Xaa

Xaa

Xaa

Asn

Xaa 80

Xaa

Xaa

Xaa

Xaa

- 268 046157

Xaa Xaa 145

Xaa Xaa

Thr Glu

Xaa Xaa

Xaa Xaa 210

Xaa Xaa 225

Xaa Xaa

Xaa Xaa

Xaa Xaa

Xaa Xaa

Xaa Xaa

Xaa Xaa

180

Xaa Xaa 195

Xaa Xaa

Xaa Xaa

Xaa Xaa

Xaa Xaa

260

Xaa Xaa 275

Asn Leu His Thr

290 <210> 150 <211> 292 <212> Protein

Xaa Xaa

150

Xaa Xaa 165

Xaa Xaa

Xaa Xaa

Xaa Xaa

Gln Ser

230

Xaa Xaa 245

Xaa Xaa

Xaa Xaa

Xaa Xaa

Xaa Xaa

Xaa Xaa

Xaa Xaa

200

Xaa Xaa 215

Ser Ser

Xaa Xaa

Xaa Xaa

Xaa Xaa

280

Xaa Xaa

Xaa Xaa

170

Xaa Xaa 185

Xaa Xaa

Xaa Xaa

Xaa Xaa 155

Xaa Thr

Xaa Xaa

Xaa Xaa

Xaa Xaa 220

Xaa Xaa

Ser His

Xaa Xaa

190

Xaa Xaa 205

Xaa Xaa

Xaa Xaa

160

Ser Leu 175

Xaa Xaa

Xaa Xaa

Xaa Xaa

Leu Val Arg Xaa

235

Xaa Xaa

Xaa Xaa

240

Xaa Xaa

250

Xaa Xaa 265

Xaa Xaa

Xaa Xaa

Xaa Xaa

Xaa Xaa

Xaa Xaa

Xaa Xaa

270 <213> Artificial sequence <220>

Xaa Xaa 255

Xaa Xaa

Xaa Arg Glu Asp 285 <223> Description of artificial sequence: Synthetic polypeptide <220>

<221> MOD_RES <222> (8) .. (57) <223> Any amino acid <220>

<221> MISC_FEATURE <222> (8)..(57) <223> This region can contain 1-50 residues <221> MOD_RES <220>

- 269 046157 <222> (65) .. (114) <223> Any amino acid <220>

<221> MISC_FEATURE <222> (65)..(114) <223> This region can contain 1-50 residues <220>

<221> MOD_RES <222> (122)..(171) <223> Any amino acid <220>

<221> MISC_FEATURE <222> (122)..(171) <223> This region can contain 1-50 residues <220>

<221> MOD_RES <222> (179)..(228) <223> Any amino acid <220>

<221> MISC_FEATURE <222> (179)..(228) <223> This region can contain 1-50 residues <220>

<221> MOD_RES <222> (236)..(285) <223> Any amino acid <220>

<221> MISC_FEATURE <222> (236)..(285) <223> This region can contain 1-50 residues <400> 150

Asp Pro Gly Ala Leu Val Arg Xaa 1 5

Xaa Xaa

Xaa Xaa

Xaa Xaa

Xaa Xaa

Xaa Xaa

Xaa Xaa

Xaa Xaa

Xaa Xaa 25

Xaa Xaa

Xaa Xaa

Xaa Xaa

Xaa Xaa 35

Xaa Xaa

Xaa Xaa

Xaa Xaa

Xaa Xaa

Xaa Xaa 45

Xaa Xaa 15

Xaa Xaa

Xaa Xaa

Xaa Xaa

Xaa Xaa

Xaa Xaa

Xaa Xaa 55

Xaa Arg Ser Asp Asn Leu Val Arg 60

Xaa Xaa 65

Xaa Xaa

Xaa Xaa

Xaa Xaa

Xaa Xaa

Xaa Xaa 75

Xaa Xaa

Xaa Xaa

Xaa Xaa

Xaa Xaa 85

Xaa Xaa

Xaa

Xaa Xaa Xaa 90

Xaa Xaa

Xaa Xaa 80

Xaa Xaa 95

- 270 046157

Xaa

Xaa

Xaa

Xaa

100

Xaa

Xaa

Xaa

Xaa

Xaa

105

Xaa

Xaa

Xaa

Xaa

Xaa

110

Xaa

Xaa

Xaa

Xaa

Gln

115

Ser

Gly

Asp

Leu

Arg 120

Arg

Xaa

Xaa

Xaa

Xaa

125

Xaa

Xaa

Xaa

Xaa

Xaa

130

Xaa

Xaa

Xaa

Xaa

Xaa

135

Xaa

Xaa

Xaa

Xaa

Xaa

140

Xaa

Xaa

Xaa

Xaa

Xaa

145

Xaa

Xaa

Xaa

Xaa

Xaa

150

Xaa

Xaa

Xaa

Xaa

Xaa

155

Xaa

Xaa

Xaa

Xaa

Xaa

160

Xaa

Xaa

Xaa

Xaa

Xaa

165

Xaa

Xaa

Xaa

Xaa

Xaa

170

Xaa

Thr

His

Leu

Asp 175

Leu

Ile

Arg

Xaa

Xaa

180

Xaa

Xaa

Xaa

Xaa

Xaa

185

Xaa

Xaa

Xaa

Xaa

Xaa

190

Xaa

Xaa

Xaa

Xaa

Xaa

195

Xaa

Xaa

Xaa

Xaa

Xaa

200

Xaa

Xaa

Xaa

Xaa

Xaa

205

Xaa

Xaa

Xaa

Xaa

Xaa

210

Xaa

Xaa

Xaa

Xaa

Xaa

215

Xaa

Xaa

Xaa

Xaa

Xaa

220

Xaa

Xaa

Xaa

Xaa

Xaa

225

Xaa

Xaa

Xaa

Thr

Ser

230

Gly

Asn

Leu

Val

Arg 235

Xaa

Xaa

Xaa

Xaa

Xaa

240

Xaa

Xaa

Xaa

Xaa

Xaa

245

Xaa

Xaa

Xaa

Xaa

Xaa

250

Xaa

Xaa

Xaa

Xaa

Xaa

255

Xaa

Xaa

Xaa

Xaa

Xaa

260

Xaa

Xaa

Xaa

Xaa

Xaa

265

Xaa

Xaa

Xaa

Xaa

Xaa

270

Xaa

Xaa

Xaa

Xaa

Xaa

275

Xaa

Xaa

Xaa

Xaa

Xaa

280

Xaa

Xaa

Xaa

Xaa

Xaa

285

Arg

Ser

Asp

Asn Leu Val Arg

290 <210> 151 <211> 463 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic polypeptide <220>

- 271 046157 <221> MOD_RES <222> (8) .. (57) <223> Any amino acid <220>

<221> MISC_FEATURE <222> (8)..(57) <223> This area can contain 1-5 <220>

<221> MOD_RES <222> (65).. (114) <223> Any amino acid <220>

<221> MISC_FEATURE <222> (65)..(114) <223> This area can contain 1-5 <220>

<221> MOD_RES <222> (122)..(171) <223> Any amino acid <220>

<221> MISC_FEATURE <222> (122)..(171) <223> This area can contain 1-5 <220>

<221> MOD_RES <222> (179)..(228) <223> Any amino acid <220>

<221> MISC_FEATURE <222> (179)..(228) <223> This area can contain 1-5 <220>

<221> MOD_RES <222> (236)..(285) <223> Any amino acid <220>

<221> MISC_FEATURE <222> (236)..(285) <223> This area can contain 1-5 <220>

<221> MOD_RES <222> (293)..(342) <223> Any amino acid <220>

<221> MISC_FEATURE <222> (293)..(342) <223> This area can contain 1-5 <220>

<221> MOD_RES <222> (350)..(399) <223> Any amino acid residues residues residues residues residues

- 272 046157 <220>

<221> MISC_FEATURE <222> (350)..(399) <223> This area can contain 1<220>

<221> MOD_RES <222> (407)..(456) <223> Any amino acid <220>

<221> MISC_FEATURE <222> (407)..(456) <223> This area can contain 1<400> 151

Arg Arg Asp Glu Leu Asn Val Xaa Xaa 1 5

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

25

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

40

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

55

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 65 70

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 85

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

100 105

Xaa Xaa Arg Ser Asp Asp Leu Val Arg

115 120

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

130 135

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

145 150

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 165

Val Arg Xaa Xaa Xaa Xaa Xaa Xaa Xaa

180 185 statok statok

Xaa Xaa Xaa Xaa Xaa Xaa

Xaa Xaa Xaa Xaa Xaa Xaa 30

Xaa Xaa Xaa Xaa Xaa Xaa 45

Ser Asp His Leu Thr Asn 60

Xaa Xaa Xaa Xaa Xaa Xaa

80

Xaa Xaa Xaa Xaa Xaa Xaa 95

Xaa Xaa Xaa Xaa Xaa Xaa

110

Xaa Xaa Xaa Xaa Xaa Xaa

125

Xaa Xaa Xaa Xaa Xaa Xaa

140

Xaa Xaa Xaa Xaa Xaa Xaa

155 160

Xaa Arg Ser Asp Asn Leu 175

Xaa Xaa Xaa Xaa Xaa Xaa

190

- 273 046157

Xaa Xaa Xaa

195

Xaa Xaa Xaa

Xaa Xaa Xaa

200

Xaa Xaa Xaa

Xaa Xaa Xaa 205

Xaa Xaa Xaa

210

Xaa Xaa Xaa

Xaa Xaa Xaa 215

Xaa Xaa Xaa

220

Xaa Xaa Xaa

Xaa Xaa Xaa 225

Xaa His Arg

230

Thr Thr Leu

Thr Asn Xaa

235

Xaa Xaa Xaa

Xaa Xaa Xaa

Xaa Xaa Xaa

245

Xaa Xaa Xaa

Xaa Xaa Xaa 250

Xaa Xaa Xaa

255

Xaa Xaa Xaa

Xaa Xaa Xaa 260

Xaa Xaa Xaa

265

Xaa Xaa Xaa

Xaa Xaa Xaa

270

Xaa Xaa Xaa

275

Xaa Xaa Xaa

Xaa Xaa Xaa

280

Xaa Xaa Xaa

Xaa Arg Glu 285

Asn Leu His

290

Thr Xaa Xaa

Xaa Xaa Xaa 295

Xaa Xaa Xaa

300

Xaa Xaa Xaa

Xaa Xaa Xaa 305

Xaa Xaa Xaa

310

Xaa Xaa Xaa

Xaa Xaa Xaa

315

Xaa Xaa Xaa

Xaa Xaa Xaa

Xaa Xaa Xaa

325

Xaa Xaa Xaa

Xaa Xaa Xaa 330

Xaa Xaa Xaa

335

Xaa Xaa Xaa

Xaa Xaa Xaa 340

Thr Ser His

345

Ser Leu Thr

Glu Xaa Xaa

350

Xaa Xaa Xaa

355

Xaa Xaa Xaa

Xaa Xaa Xaa

360

Xaa Xaa Xaa

Xaa Xaa Xaa 365

Xaa Xaa Xaa

370

Xaa Xaa Xaa

Xaa Xaa Xaa 375

Xaa Xaa Xaa

380

Xaa Xaa Xaa

Xaa Xaa Xaa 385

Xaa Xaa Xaa

390

Xaa Xaa Xaa

Xaa Xaa Xaa

395

Xaa Xaa Xaa

Ser Ser Ser

Leu Val Arg

405

Xaa Xaa Xaa

Xaa Xaa Xaa 410

Xaa Xaa Xaa

415

Xaa Xaa Xaa

Xaa Xaa Xaa 420

Xaa Xaa Xaa

425

Xaa Xaa Xaa

Xaa Xaa Xaa

430

Xaa Xaa Xaa

Xaa Xaa Xaa

Xaa Xaa Xaa

Xaa Xaa Xaa

Xaa Xaa Xaa

Xaa

Xaa

Xaa 240

Xaa

Xaa

Asp

Xaa

XAA 320

Xaa

Xaa

Xaa

Xaa

Gln 400

Xaa

Xaa

Xaa

- 274 046157

435

440

445

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Arg Glu Asp Asn Leu His Thr

450 455 460 <210> 152 <211> 7 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic peptide <400> 152

Arg Ser Asp Asn Leu Val Arg

5 <210> 153 <211> 7 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic peptide <400> 153

Arg Glu Asp Asn Leu His Thr

5 <210> 154 <211> 7 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial sequence: Synthetic peptide <400> 154 Arg Ser Asp Glu Leu Val Arg 1 5 <210> 155 <211> 7 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic peptide <400> 155 Gln Ser Gly Asn Leu Thr Glu 1 5

- 275 046157 <210> 156 <211> 7 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic peptide <400> 156

Thr Ser Gly His Leu Val Arg

5 <210> 157 <211> 7 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic peptide <400> 157

Gln Asn Ser Thr Leu Thr Glu

5 <210> 158 <211> 7 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic peptide <400> 158

Asp Pro Gly Ala Leu Val Arg

5 <210> 159 <211> 7 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic peptide <400> 159

His Arg Thr Thr Leu Thr Asn

5 <210> 160 <211> 7 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic

- 276 046157 peptide <400> 160

Gln Ser Gly Asp Leu Arg Arg <210> 161 <211> 7 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic peptide <400> 161

Thr Ser His Ser Leu Thr Glu

5 <210> 162 <211> 7 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic peptide <400> 162

Thr His Leu Asp Leu Ile Arg

5 <210> 163 <211> 7 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic peptide <400> 163

Gln Ser Ser Ser Leu Val Arg

5 <210> 164 <211> 7 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic peptide <400> 164

Thr Ser Gly Asn Leu Val Arg

5

- 277 046157 <210> 165 <211> 7 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic peptide <400> 165

Arg Arg Asp Glu Leu Asn Val

5 <210> 166 <211> 7 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic peptide <400> 166

Arg Ser Asp Asp Leu Val Arg

5 <210> 167 <211> 7 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic peptide <400> 167

Arg Ser Asp His Leu Thr Asn

5 <210> 168 <211> 16 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic peptide <400> 168

Leu Ile Lys Pro Ser Arg Met Arg Lys Tyr Pro Asn Arg Pro Ser Lys 1 5 10 15 <210> 169 <211> 7 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic

- 278 046157 peptide <400> 169

Pro Lys Lys Lys Arg Lys Val <210> 170 <211> 16 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic peptide <400> 170

Lys Arg Pro Ala Ala Thr Lys Lys Ala Gly Gln Ala Lys Lys Lys Lys 1 5 10 15 <210> 171 <211> 9 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic peptide <400> 171

Tyr Pro Tyr Asp Val Pro Asp Tyr Ala

5 <210> 172 <211> 2009 <212> Protein <213> Homo sapiens <400> 172

Met Glu Gln Thr Val Leu Val Pro Pro Gly Pro Asp Ser Phe Asn Phe

5 1015

Phe Thr Arg Glu Ser Leu Ala Ala Ile Glu Arg Arg Ile Ala Glu Glu 20 2530

Lys Ala Lys Asn Pro Lys Pro Asp Lys Lys Asp Asp Asp Glu Asn Gly 35 4045

Pro Lys Pro Asn Ser Asp Leu Glu Ala Gly Lys Asn Leu Pro Phe Ile 50 5560

Tyr Gly Asp Ile Pro Pro Glu Met Val Ser Glu Pro Leu Glu Asp Leu 65 70 7580

Asp Pro Tyr Tyr Ile Asn Lys Lys Thr Phe Ile Val Leu Asn Lys Gly 85 9095

- 279 046157

Lys Ala Ile Phe Arg

100

Phe Ser Ala Thr Ser

105

Ala Leu Tyr Ile Leu

110

Pro Phe Asn Pro Leu

115

Arg Lys Ile Ala Ile 120

Lys Ile Leu Val His 125

Leu Phe Ser Met Leu

130

Ile Met Cys Thr Ile

135

Leu Thr Asn Cys Val 140

Met Thr Met Ser Asn 145

Pro Pro Asp Trp Thr 150

Lys Asn Val Glu Tyr 155

Phe Thr Gly Ile Tyr

165

Thr Phe Glu Ser Leu

170

Ile Lys Ile Ile Ala

175

Gly Phe Cys Leu Glu

180

Asp Phe Thr Phe Leu

185

Arg Asp Pro Trp Asn

190

Leu Asp Phe Thr Val 195

Ile Thr Phe Ala Tyr

200

Val Thr Glu Phe Val

205

Leu Gly Asn Val Ser

210

Ala Leu Arg Thr Phe 215

Arg Val Leu Arg Ala

220

Lys Thr Ile Ser Val 225

Ile Pro Gly Leu Lys 230

Thr Ile Val Gly Ala 235

Ile Gln Ser Val Lys

245

Lys Leu Ser Asp Val

250

Met Ile Leu Thr Val

255

Cys Leu Ser Val Phe

260

Ala Leu Ile Gly Leu

265

Gln Leu Phe Met Gly

270

Leu Arg Asn Lys Cys

275

Ile Gln Trp Pro Pro

280

Thr Asn Ala Ser Leu

285

Glu His Ser Ile Glu

290

Lys Asn Ile Thr Val

295

Asn Tyr Asn Gly Thr

300

Ile Asn Glu Thr Val 305

Phe Glu Phe Asp Trp 310

Lys Ser Tyr Ile Gln 315

Ser Arg Tyr His Tyr

325

Phe Leu Glu Gly Phe

330

Leu Asp Ala Leu Leu

335

Gly Asn Ser Ser Asp

Ala Gly Gln Cys Pro

Glu Gly Tyr Met Cys

Thr

Ser

Phe

Thr 160

Arg

Trp

Asp

Leu

Leu 240

Phe

Asn

Glu

Leu

Asp 320

Cys

Val

- 280 046157

340

345

350

Lys Ala Gly Arg Asn

355

Pro Asn Tyr Gly Tyr 360

Thr Ser Phe Asp Thr 365

Ser Trp Ala Phe Leu

370

Ser Leu Phe Arg Leu

375

Met Thr Gln Asp Phe 380

Glu Asn Leu Tyr Gln 385

Leu Thr Leu Arg Ala 390

Ala Gly Lys Thr Tyr 395

Ile Phe Phe Val Leu

405

Val Ile Phe Leu Gly

410

Ser Phe Tyr Leu Ile

415

Leu Ile Leu Ala Val

420

Val Ala Met Ala Tyr

425

Glu Glu Gln Asn Gln

430

Thr Leu Glu Glu Ala

435

Glu Gln Lys Glu Ala 440

Glu Phe Gln Gln Met

445

Glu Gln Leu Lys Lys 450

Gln Gln Glu Ala Ala

455

Gln Gln Ala Ala Thr

460

Thr Ala Ser Glu His 465

Ser Arg Glu Pro Ser 470

Ala Ala Gly Arg Leu 475

Asp Ser Ser Ser Glu

485

Ala Ser Lys Leu Ser

490

Ser Lys Ser Ala Lys

495

Arg Arg Asn Arg Arg

500

Lys Lys Arg Lys Gln

505

Lys Glu Gln Ser Gly

510

Glu Glu Lys Asp Glu 515

Asp Glu Phe Gln Lys 520

Ser Glu Ser Glu Asp

525

Ile Arg Arg Lys Gly

530

Phe Arg Phe Ser Ile 535

Glu Gly Asn Arg Leu

540

Tyr Glu Lys Arg Tyr 545

Ser Ser Pro His Gln 550

Ser Leu Leu Ser Ile 555

Gly Ser Leu Phe Ser

565

Pro Arg Arg Asn Ser

570

Arg Thr Ser Leu Phe

575

Phe Arg Gly Arg Ala

580

Lys Asp Val Gly Ser

585

Glu Asn Asp Phe Ala

590

Phe

Trp

Met 400

Asn

Ala

Ile

Ala

Ser 480

Glu

Gly

Ser

Thr

Arg 560

Ser

Asp

- 281 046157

Asp Glu His Ser Thr Phe Glu Asp Asn

595 600

Phe Val Pro Arg Arg His Gly Glu Arg

610 615

Thr Ser Arg Ser Ser Arg Met Leu Ala

625 630

Met His Ser Thr Val Asp Cys Asn Gly 645

Pro Ser Val Pro Thr Ser Pro Val Gly

660 665

Ile Asp Lys Pro Ala Thr Asp Asp Asn

675 680

Met Arg Lys Arg Arg Ser Ser Ser Phe

690 695

Glu Asp Pro Ser Gln Arg Gln Arg Ala

705 710

Thr Asn Thr Val Glu Glu Leu Glu Glu

725

Cys Trp Tyr Lys Phe Ser Asn Ile Phe

740 745

Tyr Trp Leu Lys Val Lys His Val Val

755 760

Phe Val Asp Leu Ala Ile Thr Ile Cys

770 775

Met Ala Met Glu His Tyr Pro Met Thr 785 790

Thr Val Gly Asn Leu Val Phe Thr Gly

805

Leu Lys Ile Ile Ala Met Asp Pro Tyr

820 825

Asn Ile Phe Asp Gly Phe Ile Val Thr

835 840

Ser Arg Arg Asp Ser Leu

605

Asn Ser Asn Leu Ser Gln

620

Phe Pro Ala Asn Gly Lys

635 640

Val Ser Leu Val Gly Gly

655

Leu Leu Pro Glu Val Ile

670

Thr Thr Thr Glu Thr Glu

685

Val Ser Met Asp Phe Leu

700

Ser Ile Ala Ser Ile Leu

715 720

Arg Gln Lys Cys Pro Pro

735

Ile Trp Asp Cys Ser Pro

750

Leu Val Val Met Asp Pro 765

Val Leu Asn Thr Leu Phe

780

His Phe Asn Asn Val Leu

795 800

Phe Thr Ala Glu Met Phe

815

Tyr Phe Gln Glu Gly Trp

830

Ser Leu Val Glu Leu Gly 845

- 282 046157

Leu

Arg 865

Lys

Leu

Gly

Leu

Phe 945

Glu

Val

Ser

Met

Ala

Ile

Leu

Ile

Ser

Ala Asn Val Glu Gly 850

Val Phe Lys Leu Ala

870

Ile Ile Gly Asn Ser

885

Ala Ile Ile Val Phe

900

Lys Ser Tyr Lys Asp

915

Pro Arg Trp His Met 930

Arg Val Leu Cys Gly

950

Val Ala Gly Gln Ala

965

Ile Gly Asn Leu Val 980

Leu Ser Val Leu Arg 855

Lys Ser Trp Pro Thr

875

Val Gly Ala Leu Gly

890

Ile Phe Ala Val Val

905

Cys Val Cys Lys Ile 920

Asn Asp Phe Phe His 935

Glu Trp Ile Glu Thr

955

Met Cys Leu Thr Val

970

Val Leu Asn Leu Phe

985

Ser Phe Ser Ala Asp Asn

995

Ser Phe Arg Leu Leu 860

Leu Asn Met Leu Ile

880

Asn Leu Thr Leu Val

895

Gly Met Gln Leu Phe 910

Ala Ser Asp Cys Gln 925

Ser Phe Leu Ile Val 940

Met Trp Asp Cys Met

960

Phe Met Met Val Met

975

Leu Ala Leu Leu Leu

990

Leu Ala Ala Thr Asp Asp Asp Asn Glu 1000 1005

Asn 1010 Asn Leu Gln Ile Ala 1015 Val Asp Arg Met His 1020 Lys Gly Val Tyr 1025 Val Lys Arg Lys Ile 1030 Tyr Glu Phe Ile Gln 1035 Gln Ser Phe Arg 1040 Lys Gln Lys Ile Leu 1045 Asp Glu Ile Lys Pro 1050 Leu Asp Asp Asn 1055 Asn Lys Lys Asp Ser 1060 Cys Met Ser Asn His 1065 Thr Ala Glu Gly 1070 Lys Asp Leu Asp Tyr 1075 Leu Lys Asp Val Asn 1080 Gly Thr Thr Gly 1085 Ile Gly Thr Gly Ser 1090 Ser Val Glu Lys Tyr 1095 Ile Ile Asp

- 283 046157

Glu Ser 1100 Asp Tyr Met Ser Phe 1105 Ile Asn Asn Pro Ser 1110 Leu Thr Val Thr Val 1115 Pro Ile Ala Val Gly 1120 Glu Ser Asp Phe Glu 1125 Asn Leu Asn Thr Glu 1130 Asp Phe Ser Ser Glu 1135 Ser Asp Leu Glu Glu 1140 Ser Lys Glu Lys Leu 1145 Asn Glu Ser Ser Ser 1150 Ser Ser Glu Gly Ser 1155 Thr Val Asp Ile Gly 1160 Ala Pro Val Glu Glu 1165 Gln Pro Val Val Glu 1170 Pro Glu Glu Thr Leu 1175 Glu Pro Glu Ala Cys 1180 Phe Thr Glu Gly Cys 1185 Val Gln Arg Phe Lys 1190 Cys Cys Gln Ile Asn 1195 Val Glu Glu Gly Arg 1200 Gly Lys Gln Trp Trp 1205 Asn Leu Arg Arg Thr 1210 Cys Phe Arg Ile Val 1215 Glu His Asn Trp Phe 1220 Glu Thr Phe Ile Val 1225 Phe Met Ile Leu Leu 1230 Ser Ser Gly Ala Leu 1235 Ala Phe Glu Asp Ile 1240 Tyr Ile Asp Gln Arg 1245 Lys Thr Ile Lys Thr 1250 Met Leu Glu Tyr Ala 1255 Asp Lys Val Phe Thr 1260 Tyr Ile Phe Ile Leu 1265 Glu Met Leu Leu Lys 1270 Trp Val Ala Tyr Gly 1275 Tyr Gln Thr Tyr Phe 1280 Thr Asn Ala Trp Cys 1285 Trp Leu Asp Phe Leu 1290 Ile Val Asp Val Ser 1295 Leu Val Ser Leu Thr 1300 Ala Asn Ala Leu Gly 1305 Tyr Ser Glu Leu Gly 1310 Ala Ile Lys Ser Leu 1315 Arg Thr Leu Arg Ala 1320 Leu Arg Pro Leu Arg Ala Leu Ser Arg Phe Glu Gly Met Arg Val Val Val Asn

- 284 046157

1325 1330 1335

Ala Leu 1340 Leu Gly Ala Ile Pro 1345 Ser Ile Met Asn Val 1350 Leu Leu Val Cys Leu 1355 Ile Phe Trp Leu Ile 1360 Phe Ser Ile Met Gly 1365 Val Asn Leu Phe Ala 1370 Gly Lys Phe Tyr His 1375 Cys Ile Asn Thr Thr 1380 Thr Gly Asp Arg Phe 1385 Asp Ile Glu Asp Val 1390 Asn Asn His Thr Asp 1395 Cys Leu Lys Leu Ile 1400 Glu Arg Asn Glu Thr 1405 Ala Arg Trp Lys Asn 1410 Val Lys Val Asn Phe 1415 Asp Asn Val Gly Phe 1420 Gly Tyr Leu Ser Leu 1425 Leu Gln Val Ala Thr 1430 Phe Lys Gly Trp Met 1435 Asp Ile Met Tyr Ala 1440 Ala Val Asp Ser Arg 1445 Asn Val Glu Leu Gln 1450 Pro Lys Tyr Glu Glu 1455 Ser Leu Tyr Met Tyr 1460 Leu Tyr Phe Val Ile 1465 Phe Ile Ile Phe Gly 1470 Ser Phe Phe Thr Leu 1475 Asn Leu Phe Ile Gly 1480 Val Ile Ile Asp Asn 1485 Phe Asn Gln Gln Lys 1490 Lys Lys Phe Gly Gly 1495 Gln Asp Ile Phe Met 1500 Thr Glu Glu Gln Lys 1505 Lys Tyr Tyr Asn Ala 1510 Met Lys Lys Leu Gly 1515 Ser Lys Lys Pro Gln 1520 Lys Pro Ile Pro Arg 1525 Pro Gly Asn Lys Phe 1530 Gln Gly Met Val Phe 1535 Asp Phe Val Thr Arg 1540 Gln Val Phe Asp Ile 1545 Ser Ile Met Ile Leu Ile Cys Leu Asn Met Val Thr Met Met Val Glu Thr Asp

1550 1555 1560

- 285 046157

Asp Gln 1565 Ser Glu Tyr Val Thr 1570 Thr Ile Leu Ser Arg 1575 Ile Asn Leu Val Phe Ile Val Leu Phe Thr Gly Glu Cys Val Leu Lys Leu Ile 1580 1585 1590 Ser Leu Arg His Tyr Tyr Phe Thr Ile Gly Trp Asn Ile Phe Asp 1595 1600 1605 Phe Val Val Val Ile Leu Ser Ile Val Gly Met Phe Leu Ala Glu 1610 1615 1620 Leu Ile Glu Lys Tyr Phe Val Ser Pro Thr Leu Phe Arg Val Ile 1625 1630 1635

Arg Leu 1640 Ala Arg Ile Gly Arg 1645 Ile Leu Arg Leu Ile 1650 Lys Gly Ala Lys Gly 1655 Ile Arg Thr Leu Leu 1660 Phe Ala Leu Met Met 1665 Ser Leu Pro Ala Leu 1670 Phe Asn Ile Gly Leu 1675 Leu Leu Phe Leu Val 1680 Met Phe Ile Tyr Ala 1685 Ile Phe Gly Met Ser 1690 Asn Phe Ala Tyr Val 1695 Lys Arg Glu Val Gly 1700 Ile Asp Asp Met Phe 1705 Asn Phe Glu Thr Phe 1710 Gly Asn Ser Met Ile 1715 Cys Leu Phe Gln Ile 1720 Thr Thr Ser Ala Gly 1725 Trp Asp Gly Leu Leu 1730 Ala Pro Ile Leu Asn 1735 Ser Lys Pro Pro Asp 1740 Cys Asp Pro Asn Lys 1745 Val Asn Pro Gly Ser 1750 Ser Val Lys Gly Asp 1755 Cys Gly Asn Pro Ser 1760 Val Gly Ile Phe Phe 1765 Phe Val Ser Tyr Ile 1770 Ile Ile Ser Phe Leu 1775 Val Val Val Asn Met 1780 Tyr Ile Ala Val Ile 1785 Leu Glu Asn Phe Ser 1790 Val Ala Thr Glu Glu 1795 Ser Ala Glu Pro Leu 1800 Ser Glu Asp

- 286 046157

Asp Phe 1805 Glu Met Phe Tyr Glu 1810 Val Trp Glu Lys Phe 1815 Asp Pro Asp Ala Thr 1820 Gln Phe Met Glu Phe 1825 Glu Lys Leu Ser Gln 1830 Phe Ala Ala Ala Leu 1835 Glu Pro Pro Leu Asn 1840 Leu Pro Gln Pro Asn 1845 Lys Leu Gln Leu Ile 1850 Ala Met Asp Leu Pro 1855 Met Val Ser Gly Asp 1860 Arg Ile His Cys Leu 1865 Asp Ile Leu Phe Ala 1870 Phe Thr Lys Arg Val 1875 Leu Gly Glu Ser Gly 1880 Glu Met Asp Ala Leu 1885 Arg Ile Gln Met Glu 1890 Glu Arg Phe Met Ala 1895 Ser Asn Pro Ser Lys 1900 Val Ser Tyr Gln Pro 1905 Ile Thr Thr Thr Leu 1910 Lys Arg Lys Gln Glu 1915 Glu Val Ser Ala Val 1920 Ile Ile Gln Arg Ala 1925 Tyr Arg Arg His Leu 1930 Leu Lys Arg Thr Val 1935 Lys Gln Ala Ser Phe 1940 Thr Tyr Asn Lys Asn 1945 Lys Ile Lys Gly Gly 1950 Ala Asn Leu Leu Ile 1955 Lys Glu Asp Met Ile 1960 Ile Asp Arg Ile Asn 1965 Glu Asn Ser Ile Thr 1970 Glu Lys Thr Asp Leu 1975 Thr Met Ser Thr Ala 1980 Ala Cys Pro Pro Ser 1985 Tyr Asp Arg Val Thr 1990 Lys Pro Ile Val Glu 1995 Lys His Glu Gln Glu 2000 Gly Lys Asp Glu Lys 2005 Ala Lys Gly Lys

<210> <211> <212> <213> 173 1052 Protein Homo sapiens <400> 173

- 287 046157

Lys Arg Asn Tyr Ile Leu Gly Leu Ala 1 5

Tyr Gly Ile Ile Asp Tyr Glu Thr Arg

25

Arg Leu Phe Lys Glu Ala Asn Val Glu

40

Lys Arg Gly Ala Arg Arg Leu Lys Arg

55

Arg Val Lys Lys Leu Leu Phe Asp Tyr 65 70

Glu Leu Ser Gly Ile Asn Pro Tyr Glu 85

Gln Lys Leu Ser Glu Glu Glu Phe Ser

100 105

Lys Arg Arg Gly Val His Asn Val Asn

115 120

Asn Glu Leu Ser Thr Lys Glu Gln Ile

130 135

Glu Glu Lys Tyr Val Ala Glu Leu Gln

145 150

Gly Glu Val Arg Gly Ser Ile Asn Arg

165

Lys Glu Ala Lys Gln Leu Leu Lys Val

180 185

Asp Gln Ser Phe Ile Asp Thr Tyr Ile

195 200

Thr Tyr Tyr Glu Gly Pro Gly Glu Gly

210 215

Ile Lys Glu Trp Tyr Glu Met Leu Met

225 230

Glu Glu Leu Arg Ser Val Lys Tyr Ala

245

Gly Ile Thr Ser Val Gly 15

Val Ile Asp Ala Gly Val

Asn Glu Gly Arg Arg Ser 45

Arg Arg His Arg Ile Gln 60

Leu Leu Thr Asp His Ser 75 80

Arg Val Lys Gly Leu Ser 95

Ala Leu Leu His Leu Ala

110

Val Glu Glu Asp Thr Gly

125

Arg Asn Ser Lys Ala Leu

140

Glu Arg Leu Lys Lys Asp

155 160

Lys Thr Ser Asp Tyr Val

175

Lys Ala Tyr His Gln Leu

190

Leu Leu Glu Thr Arg Arg

205

Pro Phe Gly Trp Lys Asp

220

His Cys Thr Tyr Phe Pro

235 240

Asn Ala Asp Leu Tyr Asn

255

- 288 046157

Ala Leu Asn Asp Leu

260

Asn Asn Leu Val Ile

265

Thr Arg Asp Glu Asn

270

Lys Leu Glu Tyr Tyr 275

Glu Lys Phe Gln Ile

280

Ile Glu Asn Val Phe

285

Gln Lys Lys Lys Pro 290

Thr Leu Lys Gln Ile

295

Ala Lys Glu Ile Leu

300

Asn Glu Glu Asp Ile 305

Lys Gly Tyr Arg Val 310

Thr Ser Thr Gly Lys 315

Glu Phe Thr Asn Leu

325

Lys Val Tyr His Asp

330

Ile Lys Asp Ile Thr

335

Arg Lys Glu Ile Ile

340

Glu Asn Ala Glu Leu

345

Leu Asp Gln Ile Ala

350

Ile Leu Thr Ile Tyr

355

Gln Ser Ser Glu Asp 360

Ile Gln Glu Glu Leu

365

Asn Leu Asn Ser Glu

370

Leu Thr Gln Glu Glu

375

Ile Glu Gln Ile Ser

380

Leu Lys Gly Tyr Thr 385

Gly Thr His Asn Leu 390

Ser Leu Lys Ala Ile 395

Leu Ile Leu Asp Glu

405

Leu Trp His Thr Asn

410

Asp Asn Gln Ile Ala

415

Phe Asn Arg Leu Lys

420

Leu Val Pro Lys Lys

425

Val Asp Leu Ser Gln

430

Lys Glu Ile Pro Thr

435

Thr Leu Val Asp Asp

440

Phe Ile Leu Ser Pro

445

Val Lys Arg Ser Phe

450

Ile Gln Ser Ile Lys 455

Val Ile Asn Ala Ile

460

Lys Lys Tyr Gly Leu 465

Pro Asn Asp Ile Ile 470

Ile Glu Leu Ala Arg 475

Lys Asn Ser Lys Asp

485

Ala Gln Lys Met Ile

490

Asn Glu Met Gln Lys

495

Asn Arg Gln Thr Asn

500

Glu Arg Ile Glu Glu

505

Ile Ile Arg Thr Thr

510

Glu

Lys

Val

Pro 320

Ala

Lys

Thr

Asn

Asn 400

Ile

Gln

Val

Ile

Glu 480

Arg

Gly

- 289 046157

Lys

Gln

Leu 545

Ser

Glu

Ser

Asn

Leu 625

Ile

Asn

Lys

Phe

Leu 705

Asp

Ala

Phe

Glu Asn Ala Lys Tyr 515

Glu Gly Lys Cys Leu 530

Leu Asn Asn Pro Phe

550

Val Ser Phe Asp Asn

565

Glu Ala Ser Lys Lys

580

Ser Asp Ser Lys Ile

595

Leu Ala Lys Gly Lys 610

Leu Glu Glu Arg Asp

630

Asn Arg Asn Leu Val

645

Leu Leu Arg Ser Tyr 660

Ser Ile Asn Gly Gly

675

Lys Lys Glu Arg Asn 690

Ile Ile Ala Asn Ala

710

Lys Ala Lys Lys Val

725

Glu Ser Met Pro Glu

740

Ile Thr Pro His Gln

Leu Ile Glu Lys Ile 520

Tyr Ser Leu Glu Ala 535

Asn Tyr Glu Val Asp

555

Ser Phe Asn Asn Lys

570

Gly Asn Arg Thr Pro 585

Ser Tyr Glu Thr Phe

600

Gly Arg Ile Ser Lys 615

Ile Asn Arg Phe Ser

635

Asp Thr Arg Tyr Ala

650

Phe Arg Val Asn Asn 665

Phe Thr Ser Phe Leu

680

Lys Gly Tyr Lys His 695

Asp Phe Ile Phe Lys

715

Met Glu Asn Gln Met

730

Ile Glu Thr Glu Gln

745

Ile Lys His Ile Lys

Lys Leu His Asp Met

525

Ile Pro Leu Glu Asp 540

His Ile Ile Pro Arg

560

Val Leu Val Lys Gln

575

Phe Gln Tyr Leu Ser 590

Lys Lys His Ile Leu 605

Thr Lys Lys Glu Tyr 620

Val Gln Lys Asp Phe

640

Thr Arg Gly Leu Met

655

Leu Asp Val Lys Val

670

Arg Arg Lys Trp Lys

685

His Ala Glu Asp Ala 700

Glu Trp Lys Lys Leu

720

Phe Glu Glu Lys Gln

735

Glu Tyr Lys Glu Ile 750

Asp Phe Lys Asp Tyr

- 290 046157

755 760 765

Lys

Asp 785

Val

Lys

Pro

Glu

Thr 865

Tyr

Pro

Phe

Asn

Cys 945

Phe

Leu

Val

Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Glu Leu Ile Asn

770 775 780

Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu Ile

790 795 800

Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu Lys

805 810 815

Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His Asp

820 825 830

Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly Asp

835 840 845

Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr Leu

850 855 860

Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile Lys

870 875 880

Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp Tyr

885 890 895

Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr Arg

900 905 910

Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val Lys

915 920 925

Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser Lys

930 935 940

Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala Glu

950 955 960

Ile Ala Ser Phe Tyr Asn Asn Asp Leu Ile Lys Ile Asn Gly Glu

965 970 975

Tyr Arg Val Ile Gly Val Asn Asn Asp Leu Leu Asn Arg Ile Glu

980 985 990

Asn Met Ile Asp Ile Thr Tyr Arg Glu Tyr Leu Glu Asn Met Asn

995 1000 1005

- 291 046157

Asp

Lys 1010

Arg

Pro

Pro

Arg

Ile 1015

Ile

Lys

Thr

Ile

Ala

1020

Ser

Lys Thr

Gln

Ser

1025

Ile

Lys

Lys

Tyr

Ser

1030

Thr

Asp

Ile

Leu

Gly 1035

Asn

Leu Tyr

Glu

Val

1040

Lys

Ser

Lys

Lys

His

1045

Pro

Gln

Ile

Ile

Lys 1050

Lys

Gly <210> 174 <211> 1148 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic polypeptide <400> 174

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Ile His Gly Val Pro Ala

5 1015

Ala Lys Arg Asn Tyr Ile Leu Gly Leu Ala Ile Gly Ile Thr Ser Val 20 2530

Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 35 4045

Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 50 5560

Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 65 70 7580

Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 85 9095

Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu

100 105110

Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu

115 120125

Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr

130 135140

Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala

145 150 155160

Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys

- 292 046157

165

170

175

Asp Gly Glu Val Arg

180

Gly Ser Ile Asn Arg

185

Phe Lys Thr Ser Asp

190

Val Lys Glu Ala Lys

195

Gln Leu Leu Lys Val 200

Gln Lys Ala Tyr His 205

Leu Asp Gln Ser Phe

210

Ile Asp Thr Tyr Ile

215

Asp Leu Leu Glu Thr

220

Arg Thr Tyr Tyr Glu 225

Gly Pro Gly Glu Gly 230

Ser Pro Phe Gly Trp 235

Asp Ile Lys Glu Trp

245

Tyr Glu Met Leu Met

250

Gly His Cys Thr Tyr

255

Pro Glu Glu Leu Arg

260

Ser Val Lys Tyr Ala

265

Tyr Asn Ala Asp Leu

270

Asn Ala Leu Asn Asp 275

Leu Asn Asn Leu Val

280

Ile Thr Arg Asp Glu

285

Glu Lys Leu Glu Tyr

290

Tyr Glu Lys Phe Gln 295

Ile Ile Glu Asn Val

300

Lys Gln Lys Lys Lys 305

Pro Thr Leu Lys Gln 310

Ile Ala Lys Glu Ile 315

Val Asn Glu Glu Asp

325

Ile Lys Gly Tyr Arg

330

Val Thr Ser Thr Gly

335

Pro Glu Phe Thr Asn

340

Leu Lys Val Tyr His

345

Asp Ile Lys Asp Ile

350

Ala Arg Lys Glu Ile 355

Ile Glu Asn Ala Glu

360

Leu Leu Asp Gln Ile 365

Lys Ile Leu Thr Ile 370

Tyr Gln Ser Ser Glu 375

Asp Ile Gln Glu Glu 380

Thr Asn Leu Asn Ser 385

Glu Leu Thr Gln Glu 390

Glu Ile Glu Gln Ile 395

Asn Leu Lys Gly Tyr

405

Thr Gly Thr His Asn

410

Leu Ser Leu Lys Ala

415

Tyr

Gln

Arg

Lys 240

Phe

Tyr

Asn

Phe

Leu 320

Lys

Thr

Ala

Leu

Ser 400

Ile

- 293 046157

Asn Leu Ile Leu Asp

420

Glu Leu Trp His Thr

425

Asn Asp Asn Gln Ile

430

Ile Phe Asn Arg Leu

435

Lys Leu Val Pro Lys

440

Lys Val Asp Leu Ser

445

Gln Lys Glu Ile Pro 450

Thr Thr Leu Val Asp

455

Asp Phe Ile Leu Ser

460

Val Val Lys Arg Ser 465

Phe Ile Gln Ser Ile 470

Lys Val Ile Asn Ala 475

Ile Lys Lys Tyr Gly

485

Leu Pro Asn Asp Ile

490

Ile Ile Glu Leu Ala

495

Glu Lys Asn Ser Lys

500

Asp Ala Gln Lys Met

505

Ile Asn Glu Met Gln

510

Arg Asn Arg Gln Thr 515

Asn Glu Arg Ile Glu 520

Glu Ile Ile Arg Thr 525

Gly Lys Glu Asn Ala

530

Lys Tyr Leu Ile Glu 535

Lys Ile Lys Leu His 540

Met Gln Glu Gly Lys 545

Cys Leu Tyr Ser Leu 550

Glu Ala Ile Pro Leu 555

Asp Leu Leu Asn Asn

565

Pro Phe Asn Tyr Glu

570

Val Asp His Ile Ile

575

Arg Ser Val Ser Phe

580

Asp Asn Ser Phe Asn

585

Asn Lys Val Leu Val

590

Gln Glu Glu Ala Ser

595

Lys Lys Gly Asn Arg

600

Thr Pro Phe Gln Tyr

605

Ser Ser Ser Asp Ser 610

Lys Ile Ser Tyr Glu 615

Thr Phe Lys Lys His 620

Leu Asn Leu Ala Lys 625

Gly Lys Gly Arg Ile 630

Ser Lys Thr Lys Lys 635

Tyr Leu Leu Glu Glu

645

Arg Asp Ile Asn Arg

650

Phe Ser Val Gln Lys

655

Phe Ile Asn Arg Asn

660

Leu Val Asp Thr Arg

665

Tyr Ala Thr Arg Gly

670

Ala

Gln

Pro

Ile 480

Arg

Lys

Thr

Asp

Glu 560

Pro

Lys

Leu

Ile

Glu 640

Asp

Leu

- 294 046157

Met

Val

Lys 705

Ala

Leu

Gln

Ile

Tyr 785

Asn

Ile

Lys

Asp

Asp 865

Leu

Lys

Tyr

Asn Leu Leu Arg Ser 675

Lys Ser Ile Asn Gly 690

Phe Lys Lys Glu Arg

710

Leu Ile Ile Ala Asn

725

Asp Lys Ala Lys Lys 740

Ala Glu Ser Met Pro

755

Phe Ile Thr Pro His 770

Lys Tyr Ser His Arg

790

Asp Thr Leu Tyr Ser

805

Val Asn Asn Leu Asn

820

Lys Leu Ile Asn Lys

835

Pro Gln Thr Tyr Gln 850

Glu Lys Asn Pro Leu

870

Thr Lys Tyr Ser Lys

885

Tyr Tyr Gly Asn Lys 900

Pro Asn Ser Arg Asn 915

Tyr Phe Arg Val Asn 680

Gly Phe Thr Ser Phe 695

Asn Lys Gly Tyr Lys

715

Ala Asp Phe Ile Phe

730

Val Met Glu Asn Gln

745

Glu Ile Glu Thr Glu

760

Gln Ile Lys His Ile 775

Val Asp Lys Lys Pro

795

Thr Arg Lys Asp Asp

810

Gly Leu Tyr Asp Lys 825

Ser Pro Glu Lys Leu

840

Lys Leu Lys Leu Ile 855

Tyr Lys Tyr Tyr Glu

875

Lys Asp Asn Gly Pro

890

Leu Asn Ala His Leu

905

Lys Val Val Lys Leu 920

Asn Leu Asp Val Lys

685

Leu Arg Arg Lys Trp 700

His His Ala Glu Asp

720

Lys Glu Trp Lys Lys

735

Met Phe Glu Glu Lys 750

Gln Glu Tyr Lys Glu 765

Lys Asp Phe Lys Asp 780

Asn Arg Glu Leu Ile

800

Lys Gly Asn Thr Leu

815

Asp Asn Asp Lys Leu 830

Leu Met Tyr His His 845

Met Glu Gln Tyr Gly 860

Glu Thr Gly Asn Tyr

880

Val Ile Lys Lys Ile

895

Asp Ile Thr Asp Asp 910

Ser Leu Lys Pro Tyr

925

- 295 046157

Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val

930 935940

Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser

945 950 955960

Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala

965 970975

Glu Phe Ile Ala Ser Phe Tyr Asn Asn Asp Leu Ile Lys Ile Asn Gly

980 985990

Glu Leu Tyr Arg Val Ile

995

Gly Val Asn Asn Asp Leu Leu Asn Arg Ile

10001005

Glu Val Asn Met Ile Asp Ile 1015 Thr Tyr Arg Glu Tyr 1020 Leu Glu Asn 1010 Met Asn 1025 Asp Lys Arg Pro Pro 1030 Arg Ile Ile Lys Thr 1035 Ile Ala Ser Lys Thr 1040 Gln Ser Ile Lys Lys 1045 Tyr Ser Thr Asp Ile 1050 Leu Gly Asn Leu Tyr 1055 Glu Val Lys Ser Lys 1060 Lys His Pro Gln Ile 1065 Ile Lys Lys Gly Lys 1070 Arg Pro Ala Ala Thr 1075 Lys Lys Ala Gly Gln 1080 Ala Lys Lys Lys Lys 1085 Gly Ser Tyr Pro Tyr 1090 Asp Val Pro Asp Tyr 1095 Ala Leu Glu Asp Ala 1100 Leu Asp Asp Phe Asp 1105 Leu Asp Met Leu Gly 1110 Ser Asp Ala Leu Asp 1115 Asp Phe Asp Leu Asp 1120 Met Leu Gly Ser Asp 1125 Ala Leu Asp Asp Phe 1130 Asp Leu Asp Met Leu 1135 Gly Ser Asp Ala Leu 1140 Asp Asp Phe

Asp Leu Asp Met Leu

1145 <210> 175

- 296 046157 <211> 387 <212> Protein <213> Homo sapiens <400> 175

Met Thr Gly Lys Leu 1 5

Ala Glu Lys Leu Pro

Val Thr Met Ser Ser

Leu Asn Gln Leu Pro

Asp Asn Leu Tyr Pro

Glu Glu Ile Pro Ser

Leu Asn Leu Phe Ser

Gly Ser Ser Asp Ser 40

Val Val His Tyr Asn 45

Met Ala Thr Glu Asn 50

Val Met Asp Ile Gly 55

Leu Thr Asn Glu Lys 60

Asn Pro Glu Leu Ser 65

Tyr Ser Gly Ser Phe 70

Gln Pro Ala Pro Gly 75

Lys Thr Val Thr Tyr

Leu Gly Lys Phe Ala

Phe Asp Ser Pro Ser

Trp Cys Gln Asp Asn

100

Ile Ile Ser Leu Met

105

Ser Ala Gly Ile Leu

110

Val Pro Pro Ala Ser

115

Gly Ala Leu Ser Thr 120

Gln Thr Ser Thr Ala

125

Met Val Gln Pro

130

Gln Gly Asp Val Glu 135

Ala Met Tyr Pro Ala 140

Pro Pro Tyr Ser Asn 145

Cys Gly Asp Leu Tyr 150

Ser Glu Pro Val Ser 155

His Asp Pro Gln Gly

165

Asn Pro Gly Leu Ala

170

Tyr Ser Pro Gln Asp

175

Gln Ser Ala Lys Pro

180

Ala Leu Asp Ser Asn

185

Leu Phe Pro Met Ile

190

Asp Tyr Asn Leu Tyr 195

His His Pro Asn Asp

200

Met Gly Ser Ile Pro 205

His Lys Pro Phe Gln 210

Gly Met Asp Pro Ile 215

Arg Val Asn Pro Pro 220

Ile Thr Pro Leu Glu 225

Thr Ile Lys Ala Phe 230

Lys Asp Lys Gln Ile 235

Leu

Ala

Gln

Pro

Asn 80

Asn

Gly

Ser

Leu

Phe 160

Tyr

Pro

Glu

Pro

His 240

- 297 046157

Pro Gly Phe Gly Ser Leu Pro Gln Pro Pro Leu Thr Leu Lys Pro Ile

245 250255

Arg Pro Arg Lys Tyr Pro Asn Arg Pro Ser Lys Thr Pro Leu His Glu

260 265270

Arg Pro His Ala Cys Pro Ala Glu Gly Cys Asp Arg Arg Phe Ser Arg

275 280285

Ser Asp Glu Leu Thr Arg His Leu Arg Ile His Thr Gly His Lys Pro

290 295300

Phe Gln Cys Arg Ile Cys Met Arg Ser Phe Ser Arg Ser Asp His Leu

305 310 315320

Thr Thr His Ile Arg Thr His Thr Gly Glu Lys Pro Phe Ala Cys Glu

325 330335

Phe Cys Gly Arg Lys Phe Ala Arg Ser Asp Glu Arg Lys Arg His Ala

340 345350

Lys Ile His Leu Lys Gln Lys Glu Lys Lys Ala Glu Lys Gly Gly Ala

355 360365

Pro Ser Ala Ser Ser Ala Pro Pro Val Ser Leu Ala Pro Val Val Thr

370 375380

Thr Cys Ala 385 <210> 176 <211> 543 <212> Protein <213> Homo sapiens <400> 176

Met Ala Ala Ala Lys Ala Glu Met Gln Leu Met Ser Pro Leu Gln Ile

5 1015

Ser Asp Pro Phe Gly Ser Phe Pro His Ser Pro Thr Met Asp Asn Tyr 20 2530

Pro Lys Leu Glu Glu Met Met Leu Leu Ser Asn Gly Ala Pro Gln Phe 35 4045

Leu Gly Ala Ala Gly Ala Pro Glu Gly Ser Gly Ser Asn Ser Ser Ser 50 5560

- 298 046157

Ser Ser Ser Gly Gly Gly Gly Gly Gly Gly Gly Gly Ser Asn Ser 65 70 75

Ser Ser Ser Ser Thr Phe Asn Pro Gln Ala Asp Thr Gly Glu Gln 85 90 95

Tyr Glu His Leu Thr Ala Glu Ser Phe Pro Asp Ile Ser Leu Asn

100 105 110

Glu Lys Val Leu Val Glu Thr Ser Tyr Pro Ser Gln Thr Thr Arg

115 120 125

Pro Pro Ile Thr Tyr Thr Gly Arg Phe Ser Leu Glu Pro Ala Pro

130 135 140

Ser Gly Asn Thr Leu Trp Pro Glu Pro Leu Phe Ser Leu Val Ser

145 150 155

Leu Val Ser Met Thr Asn Pro Pro Ala Ser Ser Ser Ser Ala Pro

165 170 175

Pro Ala Ala Ser Ser Ala Ser Ala Ser Gln Ser Pro Pro Leu Ser

180 185 190

Ala Val Pro Ser Asn Asp Ser Ser Pro Ile Tyr Ser Ala Ala Pro

195 200 205

Phe Pro Thr Pro Asn Thr Asp Ile Phe Pro Glu Pro Gln Ser Gln

210 215 220

Phe Pro Gly Ser Ala Gly Thr Ala Leu Gln Tyr Pro Pro Pro Ala

225 230 235

Pro Ala Ala Lys Gly Gly Phe Gln Val Pro Met Ile Pro Asp Tyr

245 250 255

Phe Pro Gln Gln Gln Gly Asp Leu Gly Leu Gly Thr Pro Asp Gln

260 265 270

Pro Phe Gln Gly Leu Glu Ser Arg Thr Gln Gln Pro Ser Leu Thr

275 280 285

Leu Ser Thr Ile Lys Ala Phe Ala Thr Gln Ser Gly Ser Gln Asp

290 295 300

Lys Ala Leu Asn Thr Ser Tyr Gln Ser Gln Leu Ile Lys Pro Ser

305 310 315

Ser 80

Pro

Asn

Leu

Asn

Gly 160

Ser

Cys

Thr

Ala

Tyr 240

Leu

Lys

Pro

Leu

Arg 320

- 299 046157

Met Arg Lys Tyr Pro Asn Arg Pro Ser Lys Thr Pro Pro His Glu Arg

325 330335

Pro Tyr Ala Cys Pro Val Glu Ser Cys Asp Arg Arg Phe Ser Arg Ser

340 345350

Asp Glu Leu Thr Arg His Ile Arg Ile His Thr Gly Gln Lys Pro Phe

355 360365

Gln Cys Arg Ile Cys Met Arg Asn Phe Ser Arg Ser Asp His Leu Thr

370 375380

Thr His Ile Arg Thr His Thr Gly Glu Lys Pro Phe Ala Cys Asp Ile

385 390 395400

Cys Gly Arg Lys Phe Ala Arg Ser Asp Glu Arg Lys Arg His Thr Lys

405 410415

Ile His Leu Arg Gln Lys Asp Lys Lys Ala Asp Lys Ser Val Val Ala

420 425430

Ser Ser Ala Thr Ser Ser Leu Ser Ser Tyr Pro Ser Pro Val Ala Thr

435 440445

Ser Tyr Pro Ser Pro Val Thr Thr Ser Tyr Pro Ser Pro Ala Thr Thr

450 455460

Ser Tyr Pro Ser Pro Val Pro Thr Ser Phe Ser Ser Pro Gly Ser Ser

465 470 475480

Thr Tyr Pro Ser Pro Val His Ser Gly Phe Pro Ser Pro Ser Val Ala

485 490495

Thr Thr Tyr Ser Ser Val Pro Pro Ala Phe Pro Ala Gln Val Ser Ser

500 505510

Phe Pro Ser Ser Ala Val Thr Asn Ser Phe Ser Ala Ser Thr Gly Leu

515 520525

Ser Asp Met Thr Ala Thr Phe Ser Pro Arg Thr Ile Glu Ile Cys

530 535540 <210> 177 <211> 8 <212> Protein <213> Artificial sequence

- 300 046157 <220>

<223> Description of artificial sequence: Synthetic peptide <400> 177

Gly Gly Ser Gly Gly Gly Ser Gly

5 <210> 178 <211> 16 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic peptide <400> 178

Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly 1 5 10 15 <210> 179 <211> 4 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic peptide <220>

<223> See the original application description for a detailed description of substitutions and preferred embodiments <400> 179

Gly Gly Gly Ser <210> 180 <211> 5 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic peptide <220>

<223> See original application description for detailed description of substitutions and preferred embodiments <400> 180

Gly Gly Gly Gly Ser

5 <210> 181 <211> 4 <212> Protein <213> Artificial sequence

- 301 046157 <220>

<223> Description of artificial sequence: Synthetic peptide <220>

<223> See the original application description for a detailed description of substitutions and preferred embodiments <400> 181

Gly Gly Ser Gly <210> 182 <211> 9 <212> DNA <213> Homo sapiens <400> 182 gcgkgggcg 9 <210> 183 <211> 76 <212> DNA <213> Artificial sequence <220>

<223> Description of the artificial sequence: Synthetic oligonucleotide <400> 183 gttttagtac tctggaaaca gaatctacta aaacaaggca aaatgccgtg tttatctcgt 60 caacttgttg gcgaga 76 <210> 184 <211> 3871 <212> DNA <213> Artificial sequence <2 20>

<223> Description of artificial sequence: Synthetic polynucleotide

<400> 184 ggaggaagcc atcaactaaa ctacaatgac tgtaagatac aaaattggga atggtaacat 60 attttgaagt tctgttgaca taaagaatca tgatattaat gcccatggaa atgaaagggc 120 gatcaacact atggtttgaa aagggggaaa ttgtagagca cagatgtgtt cgtgtggcag 180 tgtgctgtct ctagcaatac tcagagaaga gagagaacaa tgaaattctg attggcccca 240 gtgtgagccc agatgaggtt cagctgccaa ctttctcttt cacatcttat gaaagtcatt 300 taagcacaac taactttttt tttttttttt tttttttgag acagagtctt gctctgttgc 360 ccaggacaga gtgcagtagt gactcaatct cggctcactg cagcctccac ctcctaggct 420 caaacggtcc tcctgcatca gcctcccaag tagctggaat tacaggagtg gccccacatg 480

- 302 046157 cccagctaat tttttattt ttaatagata cgggggtttc accatatcac ccaggctggt 540 ctcgaactcc tggcctcaag tgatccacct gcctcggcct cccaaagtgc tgggattata600 ggcgtcagcc actatgccca acccgaccaa ccttttttaa aataaat att taaaaaattg660 gtatttcaca tatatactag tatttacatt tatccacaca aaacggacgg gcctccgctg720 aaccagtgag gccccagacg tgcgcataaa taacccctgc gtgctgcacc acctggggag780 agggggagga ccacggtaaa tggagcgagc gcatagcaaa agggacgcgg ggtccttttc840 tctgccggtg gcactgggta gctgtggcca ggtgtggtac tttgatgggg cccagggctg900 gagctcaagg aagcgtcgca gggtcacaga tctgggggaa ccccggggaa aagcactgag960 gcaaaaccgc cgctcgtctc ctacaatata tgggagggggg aggttgagta cgttctggat1020 tactcataag accttttttt tttccttccg ggcgcaaaac cgtgagctgg atttataatc1080 gccctataaa gctccagagg cggtcaggca cctgcagagg agccccgccg ctccgccgac1140 tagctgcccc cgcgagcaac ggcctcgtga tttccccgcc gatccggtcc ccgcctcccc1200 actctgcccc cgcctacccc ggagccgtgc agccgcctct ccgaatctct ctcttctcct1260 ggcgctcgcg tgcgaga ggg aactagcgag aacgaggaag cagctggagg tgacgccggg1320 cagattacgc ctgtcagggc cgagccgagc ggatcgctgg gcgctgtgca gaggaaaggc1380 gggagtgccc ggctcgctgt cgcagagccg aggtgggtaa gctagcgacc acctggactt1440 cccagcgccc aaccgtggct tttcagccag gtcctctcct cccgcggctt ctcaaccaac1500 cccatcccag cgccggccac ccaacctccc gaaatgagtg cttcctgccc cagcagccga1560 aggcgctact aggaacggta acctgttact tttccagggg ccgtagtcga cccgct gccc1620 gagttgctgt gcgactgcgc gcgcggggct agagtgcaag gtgactgtgg ttcttctctg1680 gccaagtccg agggaacg taaagatatg ggcctttttc cccctctcac cttgtctcac1740 caaagtccct agtccccgga gcagttagcc tctttct ttc cagggaatta gccagacaca1800 acaacgggaa cgacacaccg aaccagacat gcccgccccg tgcgccctcc ccgctcgctg1860 cctttcctcc ctcttgtctc tccagagccg gatcttcaag gggagcctcc gtgcccccgg1920 ctgctcagtc cctccggtgt gcaggacccc ggaagtcctc cccgcacagc tctcgcttct1980 ctttgcagcc tgtttctgcg ccggaccagt cgaggactct ggacagtaga ggccccggga2040 cgaccgagct ggaattcgcc accatggccg ccgaccacct gatgctcgcc gagggctacc2100 gcctggtgca gaggcc gccg tccgccgccg ccgcccatgg ccctcatgcg ctccggactc2160 tgccgccgta cgcgggcccg ggcctggaca gtgggctgag gccgcggggg gctccgctgg2220 ggccgccgcc gccccgccaa cccggggccc tggcgtacgg ggccttcggg ccgccgtcct2280 ccttccagcc ctttccggcc gtgcctccgc cggccgcggg catcgcgcac ctgcagcctg2340

- 303 046157 tggcgacgcc gtaccccggc cgcgcggccg cgccccccaa cgctccggga ggccccccgg 2400 gcccgcagcc ggccccaagc gccgcagccc cgccgccgcc cgcgcacgcc ctgggcggca2460 tggacgccga actcatcgac gaggaggcgc tgacg tcgct ggagctggag ctggggctgc2520 accgcgtgcg cgagctgccc gagctgttcc tgggccagag cgagttcgac tgcttctcgg2580 acttggggtc cgcgccgccc gccggctccg tgagctgcgg tggttctggt ggtggttctg2640 gtcagtccca gctcat caaa cccagccgca tgcgcaagta ccccaaccgg cccagcaaga2700 cgccccccca cgaacgccct tacgcttgcc cagtggagtc ctgtgatcgc cgcttctccc2760 gcagcgacaa cctggtgaga cacatccgca tccacacagg ccagaagccc ttccagtgcc2820 gcatctgcat gagaaacttc agccgagagg ataacttgca cactcacatc cgcacccaca2880 caggcgaaaa gcccttcgcc tgcgacatct gtggaagaaa gtttgcccgg agcgatgaac2940 ttgtccgaca taccaagatc cacttgcggc agaa ggaccg cccttacgct tgcccagtgg3000 agtcctgtga tcgccgcttc tcccaatcag ggaatctgac tgagcacatc cgcatccaca3060 caggccagaa gcccttccag tgccgcatct gcatgagaaa cttcagcaca agtggacatc3120 tggtacgcca catccgcacc cacacaggcg aaa agccctt cgcctgcgac atctgtggaa3180 gaaagtttgc ccagaatagt accctgaccg aacataccaa gatccacttg cggcagaagg3240 acaagtaact cgaggaaacc cagcagacaa tgtagctaga cccagtagcc agatgtagct3300 aaagagaccg gttcactgtg agaaacccag cagacaatgt agctagaccc agtagccaga3360 tgtagctaaa gagaccggtt cactgtgaaa gcttgggtgg catccctgtg acccctcccc3420 agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataa a3480 attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg3540 ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg3600 ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctcc g cctcctgggt3660 tcaagcgatt ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag3720 gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt3780 ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca3840 ggcgtgaacc actgctccct tccctgtcct t3871 <210> 185 <211> 22 <212> DNA <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic primer <400> 185

- 304 046157 tgtctcggca ttgagaacat tc <210> 186 <211> 21 <212> DNA <213> Artificial sequence <220>

<223> Description of the artificial sequence: Synthetic primer <400> 186 attggtggga ggccattgta t 21 <210> 187 <211> 20 <212> DNA <213> Artificial sequence <220>

<223> Description of the artificial sequence: Synthetic primer <400> 187 accaccagtcc atgccatcac 20 <210> 188 <211> 20 <212> DNA <213> Artificial sequence <220>

<223> Description of the artificial sequence: Synthetic primer <400> 188 tccaccaccc tgttgctgta 20 <210> 189 <211> 20 <212> DNA <213> Artificial sequence <220>

<223> Description of the artificial sequence: Synthetic primer <400> 189 caaaaaagcc acaaaagcct 20 <210> 190 <211> 20 <212> DNA <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic primer

- 305 046157 <400> 190 ttagctccgc aagaaacatc <210> 191 <211> 23 <212> DNA <213> Artificial sequence <220>

<223> Description of the artificial sequence: Synthetic primer <400> 191 ccatggaact ggctcgattt cac 23 <210> 192 <211> 21 <212> DNA <213> Artificial sequence <220>

<223> Description of the artificial sequence: Synthetic primer <400> 192 attggtggga ggccactgta t 21 <210> 193 <211> 27 <212> DNA <213> Artificial sequence <220>

<223> Description of the artificial sequence: Synthetic probe <400> 193 aggcctgaaa accattgtgg gagccct 27 <210> 194 <211> 24 <212> DNA <213> Artificial sequence <220>

<223> Description of the artificial sequence: Synthetic primer <400> 194 gaatgtggga aatcattcag tcgc 24 <210> 195 <211> 24 <212> DNA <213> Artificial sequence <220>

- 306 046157 <223> Description of the artificial sequence: Synthetic primer <400> 195 gcaagttatc ctctcgtgag aagg <210> 196 <211> 29 <212> DNA <213> Artificial sequence <220>

<223> Description of the artificial sequence: Synthetic probe <400> 196 gcgacaacct ggtgagacat caacgcacc 29 <210> 197 <211> 22 <212> DNA <213> Artificial sequence <220>

<223> Description of the artificial sequence: Synthetic primer <400> 197 gctgttatct cttgtgggct gt 22 <210> 198 <211> 22 <212> DNA <213> Artificial sequence <220>

<223> Description of the artificial sequence: Synthetic primer <400> 198 aaactcatgg gagctgccgg tt 22 <210> 199 <211> 25 <212> DNA <213> Artificial sequence <220>

<223> Description of the artificial sequence: Synthetic probe <400> 199 ccacacaaat ctctccctgg cattg 25 <210> 200 <211> 24 <212> DNA <213> Artificial sequence

- 307 046157 <220>

<223> Description of the artificial sequence: Synthetic primer <400> 200 gaatgtggga aatcattcag tcgc 24 <210> 201 <211> 24 <212> DNA <213> Artificial sequence <220>

<223> Description of the artificial sequence: Synthetic primer <400> 201 gcaagttatc ctctcgtgag aagg 24 <210> 202 <211> 29 <212> DNA <213> Artificial sequence <220>

<223> Description of the artificial sequence: Synthetic probe <400> 202 gcgacaacct ggtgagacat caacgcacc 29 <210> 203 <211> 1182 <212> DNA <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic polynucleotide <400> 203

atggccgccg accacctgat gctcgccgag ggctaccgcc tggtgcagag gccgccgtcc 60 gccgccgccg cccatggccc tcatgcgctc cggactctgc cgccgtacgc gggcccggggc 120 ctggacagtg ggctgaggcc gcggggggct ccgctggggc cgccgccgcc ccgccaaccc 180 ggggccctgg cgtacggggc cttcggggccg ccgtcctcct tccagccctt tccggccgtg 240 cctccgccgg ccgcgggcat cgcgcacctg cagcctgtgg cgacgccgta ccccggccgc 300 gcggccgcgc cccccaacgc tccggggggc cccccggggcc cgcagccggc cccaagcgcc 360 gcagccccgc cgccgcccgc gcacgccctg ggcggcatgg acgccgaact catcgacgag 420 gaggcgctga cgtcgctgga gctggagctg gggctgcacc gcgtgcgcga gctgcccgag 480 ctgttcctgg gccagagcga gttcgactgc ttctcggact tggggtccgc gccgcccgcc 540

- 308 046157

ggctccgtga gctgcggtgg ttctggtggt ggttctggtc agtccagct catcaaaccc 600 agccgcatgc gcaagtaccc caaccggccc agcaagacgc ccccccacga acgcccttac 660 gcttgcccag tggagtcctg tgatcgccgc ttctcccgca gcgacaacct ggtgagacac 720 atccgcatcc acacaggcca gaagcccttc cagtgccgca tctgcatgag aaacttcagc 780 cgagaggata acttgcacac tcacatccgc acccacacag gcgaaaagcc cttcgcctgc 840 gacatctgtg gaagaaagtt tgccggggagc gatgaacttg tccgacatac caagatccac 900 ttgcggcaga aggaccgccc ttacgcttgc ccagtggagt cctgtgatcg ccgcttctcc 960 caatcaggga atctgactga gcacatccgc atccacacag gccagaagcc cttccagtgc 1020 cgcatctgca tgagaaactt cagcacaagt ggacatctgg tacgccacat ccgcaccac 1080 acaggcgaaa agcccttcgc ctgcgacatc tgtggaagaa agtttgccca gaatagtacc 1140 ctgaccgaac ataccaagat ccacttgcgg cagaaggaca ag 1182

<210> 204 <211> 1206 <212> DNA <213> Artificial sequence <220>

<223> Description of the artificial sequence: Synthetic polynucleotide <400> 204 atggccgccg accacctgat gctcgccgag ggctaccgcc tggtgcagag gccgccgtcc60 gccgccgccg cccatggccc tcatgcgctc cggactctgc cgccgtacgc gggcccgggc120 ctggacagtg ggctgaggcc g cggggggct ccgctggggc cgccgccgcc ccgccaaccc180 ggggccctgg cgtacggggc cttcgggccg ccgtcctcct tccagccctt tccggccgtg240 cctccgccgg ccgcgggcat cgcgcacctg cagcctgtgg cgacgccgta ccccggccgc300 gcggccgcg c cccccaacgc tccgggaggc cccccggggcc cgcagccggc cccaagcgcc360 gcagccccgc cgccgcccgc gcacgccctg ggcggcatgg acgccgaact catcgacgag420 gaggcgctga cgtcgctgga gctggagctg gggctgcacc gcgtgcgcga gctgcccgag480 ctgttcctgg gccagagcga gttcgactgc ttctcggact tggggtccgc gccgcccgcc5 40 ggctccgtga gctgcggtgg ttctggtggt ggttctggtg gtggcagcgg gggaggttct600 ggtcagtccc agctcatcaa acccagccgc atgcgcaagt accccaaccg gcccagcaag660 acgccccccc acgaacgccc ttacgcttgc ccagtggagt cctgtgatcg ccgcttctcc720 cgcagcgaca acctggtgag acacatccgc atccacacag gccagaagcc cttccagtgc780 cgcatctgca tgagaaactt cagccgagag gataacttgc acactcacat ccgcacccac840 acaggcgaaa agcccttcgc ctgcgacatc tgtggaagaa agtttgcccg gagcgatgaa900

- 309 046157

cttgtccgac ataccaagat ccacttgcgg cagaaggacc gcccttacgc ttgcccagtg 960 gagtcctgtg atcgccgctt ctcccaatca gggaatctga ctgagcacat ccgcatccac 1020 acaggccaga agcccttcca gtgccgcatc tgcatgagaa acttcagcac aagtggacat 1080 ctggtacgcc acatccgcac ccacacaggc gaaaagccct tcgcctgcga catctgtgga 1140 agaaagtttg cccagaatag taccctgacc gaacatacca agatccactt gcggcagaag 1200 gacaag 1206

<210> 205 <211> 402 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic polypeptide <400> 205

Met Ala Ala Asp His Leu Met Leu Ala Glu Gly Tyr Arg Leu Val Gln

5 1015

Arg Pro Pro Ser Ala Ala Ala Ala His Gly Pro His Ala Leu Arg Thr 20 2530

Leu Pro Pro Tyr Ala Gly Pro Gly Leu Asp Ser Gly Leu Arg Pro Arg 35 4045

Gly Ala Pro Leu Gly Pro Pro Pro Pro Arg Gln Pro Gly Ala Leu Ala 50 5560

Tyr Gly Ala Phe Gly Pro Pro Ser Ser Phe Gln Pro Phe Pro Ala Val 65 70 7580

Pro Pro Pro Ala Ala Gly Ile Ala His Leu Gln Pro Val Ala Thr Pro 85 9095

Tyr Pro Gly Arg Ala Ala Ala Pro Pro Asn Ala Pro Gly Gly Pro Pro

100 105110

Gly Pro Gln Pro Ala Pro Ser Ala Ala Ala Pro Pro Pro Pro Ala His

115 120125

Ala Leu Gly Gly Met Asp Ala Glu Leu Ile Asp Glu Glu Ala Leu Thr

130 135140

Ser Leu Glu Leu Glu Leu Gly Leu His Arg Val Arg Glu Leu Pro Glu

145 150 155160

- 310 046157

Leu Phe Leu Gly Gln

165

Ser Glu Phe Asp Cys

170

Phe Ser Asp Leu Gly

175

Ala Pro Pro Ala Gly

180

Ser Val Ser Cys Gly

185

Gly Ser Gly Gly Gly

190

Gly Gly Gly Ser Gly 195

Gly Gly Ser Gly Gln 200

Ser Gln Leu Ile Lys

205

Ser Arg Met Arg Lys

210

Tyr Pro Asn Arg Pro

215

Ser Lys Thr Pro Pro

220

Glu Arg Pro Tyr Ala 225

Cys Pro Val Glu Ser 230

Cys Asp Arg Arg Phe 235

Arg Ser Asp Asn Leu

245

Val Arg His Ile Arg

250

Ile His Thr Gly Gln

255

Pro Phe Gln Cys Arg

260

Ile Cys Met Arg Asn

265

Phe Ser Arg Glu Asp

270

Leu His Thr His Ile

275

Arg Thr His Thr Gly

280

Glu Lys Pro Phe Ala 285

Asp Ile Cys Gly Arg 290

Lys Phe Ala Arg Ser

295

Asp Glu Leu Val Arg 300

Thr Lys Ile His Leu 305

Arg Gln Lys Asp Arg 310

Pro Tyr Ala Cys Pro 315

Glu Ser Cys Asp Arg

325

Arg Phe Ser Gln Ser

330

Gly Asn Leu Thr Glu

335

Ile Arg Ile His Thr

340

Gly Gln Lys Pro Phe

345

Gln Cys Arg Ile Cys

350

Arg Asn Phe Ser Thr 355

Ser Gly His Leu Val 360

Arg His Ile Arg Thr 365

Thr Gly Glu Lys Pro

370

Phe Ala Cys Asp Ile 375

Cys Gly Arg Lys Phe 380

Gln Asn Ser Thr Leu 385

Thr Glu His Thr Lys 390

Ile His Leu Arg Gln 395

Ser

Ser

Pro

His

Ser 240

Lys

Asn

Cys

His

Val 320

His

Met

His

Ala

Lys 400

Asp Lys

- 311 046157 <210> 206 <211> 1707 <212> DNA <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic polynucleotide

<400> 206 atggccgcag atcacctgat gctggctgaa ggctacagac tggtgcagcg gcctccactct 60 gccgctgccg cccacggccc ccacgccctg agaacactgc ccccctacgc cggccctggt 120 cttgatagcg gactcagacc tagaggcgcc cctctggggcc ctccacctcc aagacagcct 180 ggagccctgg cctacggcgc cttcggccct ccttctagct tccagccctt ccccgccgtg 240 cctcctccag ccgctggcat cgccacctg cagcctgtgg ccacccctta ccccggagaaga 300 gccgccgccc ctccaaacgc ccctggcgga cctcctggcc cccagcctgc tccaagcgcc 360 gctgcccctc cacctcctgc tcatgccctg ggcggcatgg acgccgagct gatcgacgag 420 gaagccctga ccagcctgga actggaactg ggcctgcaca gagtgcggga actgcctgag 480 ctgttcctgg gacagagcga gttcgactgc ttcagcgacc tggggcagcgc ccctcctgcc 540 ggctctgtgt cctgcgccga ccacctgatg ctcgccgagg gctaccgcct ggtgcagagg 600 ccgccgtccg ccgccgccgc ccatggccct catgcgctcc ggactctgcc gccgtacgcg 660 ggcccgggcc tggacagtgg gctgaggccg cggggggctc cgctggggcc gccgccgccc 720 cgccaacccg gggccctggc gtacggggcc ttcggggccgc cgtcctcctt ccagcccttt 780 ccggccgtgc ctccgccggc cgcgggcatc gcgcacctgc agcctgtggc gacgccgtac 840 cccggccgcg ccgccgcgcc ccccaacgct ccgggaggcc ccccggggccc gcagccggcc 900 ccaagcgccg cagccccgcc gccgcccgcg cacgccctgg gcggcatgga cgccgaactc 960 atcgacgagg aggcgctgac gtcgctggag ctggagctgg ggctgcaccg cgtgcgcgag 1020 ctgcccgagc tgttcctggg ccagagcgag ttcgactgct tctcggactt ggggtccgcg 1080 ccgcccgccg gctccgtgag ctgccagtcc cagctcatca aacccagccg catgcgcaag 1140 taccccaacc ggcccagcaa gacgcccccc cacgaacgcc cttacgcttg cccagtggag 1200 tcctgtgatc gccgcttctc ccgcagcgac aacctggtga gacacatccg catccacaca 1260 ggccagaagc ccttccagtg ccgcatctgc atgagaaact tcagccgaga ggataacttg 1320 cacactcaca tccgcaccca cacaggcgaa aagcccttcg cctgcgacat ctgtggaaga 1380 aagtttgccc ggagcgatga acttgtccga cataccaaga tccacttgcg gcagaaggac 1440 cgcccttacg cttgcccagt ggagtcctgt gatcgccgct tctcccaatc agggaatctg 1500 actgagcaca tccgcatcca cacaggccag aagcccttcc agtgccgcat ctgcatgaga 1560

- 312 046157

aacttcagca caagtggaca tctggtacgc cacatccgca cccacacagg cgaaaagccc 1620 ttcgcctgcg acatctgtgg aagaaagttt gccagaata gtaccctgac cgaacatacc 1680 aagatccact tgcggcagaa ggacaag 1707

<210> 207 <211> 569 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic polypeptide <400> 207

Met Ala Ala Asp His Leu Met Leu Ala Glu Gly Tyr Arg Leu Val Gln

5 1015

Arg Pro Pro Ser Ala Ala Ala Ala His Gly Pro His Ala Leu Arg Thr 20 2530

Leu Pro Pro Tyr Ala Gly Pro Gly Leu Asp Ser Gly Leu Arg Pro Arg 35 4045

Gly Ala Pro Leu Gly Pro Pro Pro Pro Arg Gln Pro Gly Ala Leu Ala 50 5560

Tyr Gly Ala Phe Gly Pro Pro Ser Ser Phe Gln Pro Phe Pro Ala Val 65 70 7580

Pro Pro Pro Ala Ala Gly Ile Ala His Leu Gln Pro Val Ala Thr Pro 85 9095

Tyr Pro Gly Arg Ala Ala Ala Pro Pro Asn Ala Pro Gly Gly Pro Pro

100 105110

Gly Pro Gln Pro Ala Pro Ser Ala Ala Ala Pro Pro Pro Pro Ala His

115 120125

Ala Leu Gly Gly Met Asp Ala Glu Leu Ile Asp Glu Glu Ala Leu Thr

130 135140

Ser Leu Glu Leu Glu Leu Gly Leu His Arg Val Arg Glu Leu Pro Glu

145 150 155160

Leu Phe Leu Gly Gln Ser Glu Phe Asp Cys Phe Ser Asp Leu Gly Ser

165 170175

Ala Pro Pro Ala Gly Ser Val Ser Cys Ala Asp His Leu Met Leu Ala

180 185190

- 313 046157

Glu Gly Tyr Arg Leu

195

Val Gln Arg Pro Pro

200

Ser Ala Ala Ala Ala

205

Gly Pro His Ala Leu

210

Arg Thr Leu Pro Pro 215

Tyr Ala Gly Pro Gly

220

Asp Ser Gly Leu Arg 225

Pro Arg Gly Ala Pro 230

Leu Gly Pro Pro Pro 235

Arg Gln Pro Gly Ala

245

Leu Ala Tyr Gly Ala

250

Phe Gly Pro Pro Ser

255

Phe Gln Pro Phe Pro

260

Ala Val Pro Pro Pro

265

Ala Ala Gly Ile Ala

270

Leu Gln Pro Val Ala

275

Thr Pro Tyr Pro Gly

280

Arg Ala Ala Ala Pro 285

Asn Ala Pro Gly Gly 290

Pro Pro Gly Pro Gln 295

Pro Ala Pro Ser Ala

300

Ala Pro Pro Pro Pro 305

Ala His Ala Leu Gly 310

Gly Met Asp Ala Glu 315

Ile Asp Glu Glu Ala

325

Leu Thr Ser Leu Glu

330

Leu Glu Leu Gly Leu

335

Arg Val Arg Glu Leu

340

Pro Glu Leu Phe Leu

345

Gly Gln Ser Glu Phe

350

Cys Phe Ser Asp Leu

355

Gly Ser Ala Pro Pro 360

Ala Gly Ser Val Ser 365

Gln Ser Gln Leu Ile

370

Lys Pro Ser Arg Met

375

Arg Lys Tyr Pro Asn 380

Pro Ser Lys Thr Pro 385

Pro His Glu Arg Pro 390

Tyr Ala Cys Pro Val 395

Ser Cys Asp Arg Arg

405

Phe Ser Arg Ser Asp

410

Asn Leu Val Arg His

415

Arg Ile His Thr Gly

420

Gln Lys Pro Phe Gln

425

Cys Arg Ile Cys Met

430

Asn Phe Ser Arg Glu

Asp Asn Leu His Thr

His Ile Arg Thr His

His

Leu

Pro 240

Ser

His

Pro

Ala

Leu 320

His

Asp

Cys

Arg

Glu 400

Ile

Arg

Thr

- 314 046157

Gly Glu

450

Ser Asp 465

Arg Pro

435

Lys Pro

Glu Leu

Tyr Ala

Phe Ala

Val Arg

470

Cys Pro 485

Ser Gly

Asn Leu

500

Thr Glu

440

Cys Asp 455

His Thr

Val Glu

His Ile

Ile Cys

Lys Ile

Ser Cys

490

Arg Ile 505

Phe Gln

Cys Arg 515

Ile Cys

Met Arg

520

Asn Phe

Gly Arg

460

His Leu 475

Asp Arg

His Thr

Ser Thr

Val Arg

530

His Ile

Arg Thr

His Thr 535

Gly Glu

Lys Pro

540

Ile Cys 545

Gly Arg

Lys Phe

550

Ala Gln

Asn Ser

Thr Leu 555

445

Lys Phe

Arg Gln

Arg Phe

Gly Gln

510

Ser Gly 525

Phe Ala

Thr Glu

Ala Arg

Lys Asp

480

Ser Gln

495

Lys Pro

His Leu

Cys Asp

His Thr

560

Lys Ile His Leu Arg Gln Lys Asp Lys

565 <210> 208 <211> 1755 <212> DNA <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic polynucleotide

<400> 208 atggccgcag atcacctgat gctggctgaa ggctacagac tggtgcagcg gcctccactct gccgctgccg cccacggccc ccacgccctg agaacactgc ccccctacgc cggccctggt cttgatagcg gactcagacc tagaggcgcc cctctggggcc ctccacctcc aagacagcct ggagccctgg cctacggcgc cttcggccct ccttctagct tccagccctt ccccgccgtg cctcctccag ctgctggcat cgccacctg cagcctgtgg ccacccctta ccccggagaaga gccgccgccc ctccaaacgc ccctggcgga cctcctggcc cccagcctgc tccaagcgcc gctgcccctc cacctcctgc tcatgccctg ggcggcatgg acgccgagct gatcgacgag gaagccctga ccagcctgga actggaactg ggcctgcaca gagtgcggga actgcctgag ctgttcctgg gacagagcga gttcgactgc ttcagcgacc tggggcagcgc ccctcctgcc

120

180

240

300

360

420

480

540

- 315 046157

ggctctgtgt cctgcggcgg cagcggcggc ggaagcggcg ccgaccacct gatgctcgcc 600 gagggctacc gcctggtgca gaggccgccg tccgccgccg ccgcccatgg ccctcatgcg 660 ctccggactc tgccgccgta cgcgggcccg ggcctggac gtggggctgag gccgcgggggg 720 gctccgctgg ggccgccgcc gccccgccaa cccggggccc tggcgtacgg ggccttcggg 780 ccgccgtcct ccttccagcc ctttccggcc gtgcctccgc cggccgcggg catcgcgcac 840 ctgcagcctg tggcgacgcc gtaccccggc cgcgcggccg cgccccccaa cgctccggga 900 ggccccccgg gcccgcagcc ggccccaagc gccgcagccc cgccgccgcc cgcgcacgcc 960 ctggggcggca tggacgccga actcatcgac gaggaggcgc tgacgtcgct ggagctggag 1020 ctggggctgc accgcgtgcg cgagctgccc gagctgttcc tggggccagag cgagttcgac 1080 tgcttctcgg acttggggtc cgcgccgccc gccggctccg tgagctgcgg tggttctggt 1140 ggtggttctg gtcagtccca gctcatcaaa cccagccgca tgcgcaagta ccccaaccgg 1200 cccagcaaga cgcccccccca cgaacgccct tacgcttgcc cagtggagtc ctgtgatcgc 1260 cgcttctccc gcagcgacaa cctggtgaga cacatccgca tccacacagg ccagaagccc 1320 ttccagtgcc gcatctgcat gagaaacttc agccgagagg ataacttgca cactcacatc 1380 cgcaccaca caggcgaaaa gcccttcgcc tgcgacatct gtggaagaaa gtttgcccgg 1440 agcgatgaac ttgtccgaca taccaagatc cacttgcggc agaaggaccg cccttacgct 1500 tgccagtgg agtcctgtga tcgccgcttc tccaatcag ggaatctgac tgagcacatc 1560 cgcatccaca caggccagaa gcccttccag tgccgcatct gcatgagaaa cttcagcaca 1620 agtggacatc tggtacgcca catccgcacc cacacaggcg aaaagccctt cgcctgcgac 1680 atctgtggaa gaaagtttgc ccagaatagt accctgaccg aacataccaa gatccacttg 1740 cggcagaagg acaag 1755

<210> 209 <211> 585 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic polypeptide <400> 209

Met Ala Ala Asp His Leu Met Leu 1 5

Ala Glu Gly Tyr Arg Leu Val Gln

15

Arg Pro Pro Ser Ala Ala Ala Ala 20

His Gly Pro His Ala Leu Arg Thr 25 30

Leu Pro Pro Tyr Ala Gly Pro Gly

40

Leu Asp Ser Gly Leu Arg Pro Arg 45

- 316 046157

Gly Ala Pro Leu Gly 50

Pro Pro Pro Pro Arg 55

Gln Pro Gly Ala Leu 60

Tyr Gly Ala Phe Gly 65

Pro Pro Ser Ser Phe 70

Gln Pro Phe Pro Ala 75

Pro Pro Pro Ala Ala

Gly Ile Ala His Leu

Gln Pro Val Ala Thr

Tyr Pro Gly Arg Ala

100

Ala Ala Pro Pro Asn

105

Ala Pro Gly Gly Pro

110

Gly Pro Gln Pro Ala

115

Pro Ser Ala Ala Ala

120

Pro Pro Pro Pro Ala

125

Ala Leu Gly Gly Met

130

Asp Ala Glu Leu Ile 135

Asp Glu Glu Ala Leu 140

Ser Leu Glu Leu Glu 145

Leu Gly Leu His Arg 150

Val Arg Glu Leu Pro 155

Leu Phe Leu Gly Gln

165

Ser Glu Phe Asp Cys

170

Phe Ser Asp Leu Gly

175

Ala Pro Pro Ala Gly

180

Ser Val Ser Cys Gly

185

Gly Ser Gly Gly Gly

190

Gly Ala Asp His Leu

195

Met Leu Ala Glu Gly

200

Tyr Arg Leu Val Gln 205

Pro Pro Ser Ala Ala

210

Ala Ala His Gly Pro 215

His Ala Leu Arg Thr

220

Pro Pro Tyr Ala Gly 225

Pro Gly Leu Asp Ser 230

Gly Leu Arg Pro Arg 235

Ala Pro Leu Gly Pro

245

Pro Pro Pro Arg Gln

250

Pro Gly Ala Leu Ala

255

Gly Ala Phe Gly Pro

260

Pro Ser Ser Phe Gln

265

Pro Phe Pro Ala Val

270

Pro Pro Ala Ala Gly

275

Ile Ala His Leu Gln

280

Pro Val Ala Thr Pro

285

Pro Gly Arg Ala Ala

Ala Pro Pro Asn Ala

Pro Gly Gly Pro Pro

Ala

Val 80

Pro

Pro

His

Thr

Glu 160

Ser

Ser

Arg

Leu

Gly 240

Tyr

Pro

Tyr

Gly

- 317 046157

290

295

300

Pro Gln Pro Ala Pro Ser Ala Ala Ala Pro Pro Pro Pro Ala His

305 310 315

Leu Gly Gly Met Asp Ala Glu Leu

325

Ile Asp Glu Glu Ala Leu Thr

330 335

Leu Glu Leu Glu Leu Gly Leu His Arg Val Arg Glu Leu Pro Glu

340 345 350

Phe Leu Gly Gln Ser Glu Phe Asp Cys Phe Ser Asp Leu Gly Ser

355 360 365

Pro Pro Ala Gly Ser Val Ser Cys Gly Gly Ser Gly Gly Gly Ser

370 375 380

Gln Ser Gln Leu Ile Lys Pro Ser Arg Met Arg Lys Tyr Pro Asn

385 390 395

Pro Ser Lys Thr Pro Pro His Glu Arg Pro Tyr Ala Cys Pro Val

405 410 415

Ser Cys Asp Arg Arg Phe Ser Arg Ser Asp Asn Leu Val Arg His

420 425 430

Arg Ile His Thr Gly Gln Lys Pro Phe Gln Cys Arg Ile Cys Met

435 440 445

Asn Phe Ser Arg Glu Asp Asn Leu His Thr His Ile Arg Thr His

450 455 460

Gly Glu Lys Pro Phe Ala Cys Asp Ile Cys Gly Arg Lys Phe Ala

465 470 475

Ser Asp Glu Leu Val Arg His Thr Lys Ile His Leu Arg Gln Lys

485 490 495

Arg Pro Tyr Ala Cys Pro Val Glu

500

Ser Cys Asp Arg Arg Phe Ser

505 510

Ser Gly Asn Leu Thr Glu His 515

Ile Arg Ile His Thr Gly Gln Lys

520 525

Phe Gln Cys Arg Ile Cys Met Arg Asn Phe Ser

530 535

Thr Ser Gly His 540

Ala 320

Ser

Leu

Ala

Gly

Arg 400

Glu

Ile

Arg

Thr

Arg 480

Asp

Gln

Pro

Leu

- 318 046157

Val Arg His Ile Arg Thr His Thr Gly Glu Lys Pro Phe Ala Cys Asp

545

550

555

560

Ile Cys Gly Arg Lys Phe Ala Gln Asn Ser Thr Leu Thr Glu His Thr

565

570

575

Lys Ile His Leu Arg Gln Lys Asp Lys

580

585 <210> 210 <211> 1113 <212> DNA <213> Homo sapiens <400> 210 atggagctgg aattggatgc tggtgaccaa gacctgctgg ccttcctgct agaggaaagt60 ggagatttgg ggacggcacc cgatgaggcc gtgagggccc cactggactg ggcgctgcc g120 ctttctgagg tgccgagcga ctgggaagta gatgatttgc tgtgctccct gctgagtccc180 ccagcgtcgt tgaacattct cagctcctcc aacccctgcc ttgtccacca tgaccacacc240 tactccctcc cacgggaaac tgtctctatg gatctagaga gtgagagctg tagaaaagag300 gggacccaga tgactccaca gcatatggag gagctggcag agcaggagat tgctaggcta360 gtactgacag atgaggagaa gagtctattg gagaaggagg ggcttattct gcctgagaca420 cttcctctca ctaagacaga ggaacaaatt ctgaaacgtg tgcggaggaa g attcgaaat480 aaaagatctg ctcaagagag ccgcaggaaa aagaaggtgt acgttggggg tttagagagc540 cgggtcttga aatacacagc ccagaatatg gagcttcaga acaaagtaca gcttctggag600 gaacagaatt tgtcccttct agatcaactg aggaaactcc aggccatggt gattg agatc660 tcaaacaaaa ccagcagcag cagcacctgc atcttggtcc tgctagtctc cttctgcctc720 ctccttgtac ctgctatgta ctcctctgac acaaggggga gcctgccagc tgagcatgga780 gtgttgtccc gccagcttcg tgccctcccc agtgaggacc cttaccagct ggagctgcct840 gccctgcagt cagaagtgcc gaaagacagc acacaccagt ggttggacgg ctcagactgt900 gtactccagg cccctggcaa cacttcctgc ctgctgcatt acatgcctca ggctcccagt 960 gcagagcctc ccctggagtg gcccttccct gacctcttct cagagcctct ctgccgaggt1020 cccatcctcc ccctgcaggc aaatctcaca aggaagggag gatggcttcc tactggtagc1080 ccctctgtca ttttgcagga cagatactca ggc1113 <210> 211 <21 1> 371 <212> Protein <213> Homo sapiens <400> 211

- 319 046157

Met 1

Leu

Ala

Glu

Asn 65

Tyr

Cys

Ala

Leu

Lys 145

Lys

Gly

Gln

Gln

Ser 225

Leu

Glu Leu Glu Leu Asp 5

Glu Glu Ser Gly Asp 20

Pro Leu Asp Trp Ala 35

Val Asp Asp Leu Leu 50

Ile Leu Ser Ser Ser

Ser Leu Pro Arg Glu 85

Arg Lys Glu Gly Thr 100

Glu Gln Glu Ile Ala

115

Leu Glu Lys Glu Gly 130

Thr Glu Glu Gln Ile

150

Arg Ser Ala Gln Glu

165

Leu Glu Ser Arg Val

180

Asn Lys Val Gln Leu 195

Leu Arg Lys Leu Gln 210

Ser Ser Ser Thr Cys

230

Leu Val Pro Ala Met

245

Ala Gly Asp Gln Asp

Leu Gly Thr Ala Pro 25

Leu Pro Leu Ser Glu 40

Cys Ser Leu Leu Ser 55

Asn Pro Cys Leu Val 75

Thr Val Ser Met Asp

Gln Met Thr Pro Gln

105

Arg Leu Val Leu Thr 120

Leu Ile Leu Pro Glu 135

Leu Lys Arg Val Arg

155

Ser Arg Arg Lys Lys

170

Leu Lys Tyr Thr Ala

185

Leu Glu Glu Gln Asn

200

Ala Met Val Ile Glu 215

Ile Leu Val Leu Leu

235

Tyr Ser Ser Asp Thr

250

Leu Leu Ala Phe Leu

Asp Glu Ala Val Arg 30

Val Pro Ser Asp Trp 45

Pro Pro Ala Ser Leu 60

His His Asp His Thr

Leu Glu Ser Glu Ser

His Met Glu Glu Leu

110

Asp Glu Glu Lys Ser 125

Thr Leu Pro Leu Thr 140

Arg Lys Ile Arg Asn

160

Lys Val Tyr Val Gly

175

Gln Asn Met Glu Leu

190

Leu Ser Leu Leu Asp

205

Ile Ser Asn Lys Thr 220

Val Ser Phe Cys Leu

240

Arg Gly Ser Leu Pro

255

- 320 046157

Ala Glu His Gly Val Leu Ser Arg

260

Gln Leu Arg Ala Leu Pro Ser Glu

265 270

Asp Pro Tyr Gln Leu Glu Leu Pro Ala Leu Gln Ser Glu Val Pro Lys

275 280285

Asp Ser Thr His Gln Trp Leu Asp Gly Ser Asp Cys Val Leu Gln Ala

290 295300

Pro Gly Asn Thr Ser Cys Leu Leu His Tyr Met Pro Gln Ala Pro Ser

305 310 315320

Ala Glu Pro Pro Leu Glu Trp Pro Phe Pro Asp Leu Phe Ser Glu Pro

325 330335

Leu Cys Arg Gly Pro Ile Leu Pro Leu Gln Ala Asn Leu Thr Arg Lys

340 345350

Gly Gly Trp Leu Pro Thr Gly Ser Pro Ser Val Ile Leu Gln Asp Arg

355 360365

Tyr Ser Gly

370 <210> 212 <211> 1590 <212> DNA <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic polynucleotide <400> 212

atggagctgg aattggatgc tggtgaccaa gacctgctgg ccttcctgct aggaaagt 60 ggagattgg ggacggcacc cgatgaggcc gtgagggccc cactggactg ggcgctgccg 120 ctttctgagg tgccgagcga ctgggaagta gatgatttgc tgtgctccct gctgagtccc 180 ccagcgtcgt tgaacattct cagctcctcc aacccctgcc ttgtccacca tgaccacacc 240 tactccctcc cacgggaaac tgtctctatg gatctagaga gtgagagctg tagaaaagag 300 gggacccaga tgactccaca gcatatggag gagctggcag agcaggagat tgctaggcta 360 gtactgacag atgaggagaa gagtctattg gagaaggagg ggcttattct gcctgagaca 420 cttcctctca ctaagacaga ggaacaaatt ctgaaacgtg tgcggctcga accaggtgaa 480 aaaccttaca aatgtcctga atgtgggaaa tcattcagtc gcagcgacaa cctggtgaga 540 catcaacgca cccatacagg agaaaaacct tataaatgtc cagaatgtgg aaagtccttc 600

- 321 046157

tcacgagagg ataacttgca cactcatcaa cgaacacata ctggtgaaaa accataag 660 tgtcccgaat gtggtaaaag ttttagccgg agcgatgaac ttgtccgaca ccaacgaacc 720 catacaggcg agaagcctta caaatgtccc gagtgtggca agagcttctc acaatcaggg 780 aatctgactg agcatcaacg aactcatacc ggggaaaaac cttacaagtg tccagagtgt 840 gggaagagct tttccacaag tggacatctg gtacgccacc aggagacaca tacaggggag 900 aagccctaca aatgccccga atgcggtaaa agtttctctc agaatagtac cctgaccgaa 960 caccagcgaa cacacactgg gaaaaaaacg agtgtgtacg ttggggggttt agagagccgg 1020 gtcttgaaat acacagccca gaatatggag cttcagaaca aagtacagct tctggaggaa 1080 cagaatttgt cccttctaga tcaactgagg aaactccagg ccatggtgat tgagatctca 1140 aacaaaacca gcagcagcag cacctgcatc ttggtcctgc tagtctcctt ctgcctcctc 1200 cttgtacctg ctatgtactc ctctgacaca aggggggagcc tgccagctga gcatggagtg 1260 ttgtcccgcc agcttcgtgc cctccccagt gaggaccctt accagctgga gctgcctgcc 1320 ctgcagtcag aagtgccgaa agacagcaca caccagtggt tggacggctc agactgtgta 1380 ctccaggccc ctggcaacac ttcctgcctg ctgcattaca tgcctcaggc tccagtgca 1440 gagcctcccc tggagtggcc cttccctgac ctcttctcag agcctctctg ccgaggtccc 1500 atcctccccc tgcaggcaaa tctcacaagg aagggaggat ggcttcctac tggtagcccc 1560 tctgtcattt tgcaggacag atactcaggc 1590

<210> 213 <211> 530 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic polypeptide <400> 213

Met Glu Leu Glu Leu Asp Ala Gly Asp Gln Asp Leu Leu Ala Phe Leu

5 1015

Leu Glu Glu Ser Gly Asp Leu Gly Thr Ala Pro Asp Glu Ala ValArg

2530

Ala Pro Leu Asp Trp Ala Leu Pro Leu Ser Glu Val Pro Ser Asp Trp 35 4045

Glu Val Asp Asp Leu Leu Cys Ser Leu Leu Ser Pro Pro Ala Ser Leu 50 5560

Asn Ile Leu Ser Ser Ser Asn Pro Cys Leu Val His His Asp His Thr 65 70 7580

- 322 046157

Tyr

Cys

Ala

Leu

Lys 145

Lys

Asn

Cys

His

Gly 225

His

Ser

Lys

His

Cys 305

His

Ser Leu Pro Arg Glu

Thr Val Ser Met Asp

Leu Glu Ser Glu Ser

Arg Lys Glu Gly Thr 100

Glu Gln Glu Ile Ala

115

Leu Glu Lys Glu Gly 130

Thr Glu Glu Gln Ile

150

Pro Tyr Lys Cys Pro

165

Leu Val Arg His Gln 180

Pro Glu Cys Gly Lys 195

Gln Arg Thr His Thr 210

Lys Ser Phe Ser Arg

230

Thr Gly Glu Lys Pro

245

Gln Ser Gly Asn Leu

260

Pro Tyr Lys Cys Pro

275

Leu Val Arg His Gln 290

Pro Glu Cys Gly Lys

310

Gln Arg Thr His Thr

Gln Met Thr Pro Gln

105

His Met Glu Glu Leu

110

Arg Leu Val Leu Thr 120

Asp Glu Glu Lys Ser 125

Leu Ile Leu Pro Glu 135

Thr Leu Pro Leu Thr 140

Leu Lys Arg Val Arg

155

Glu Cys Gly Lys Ser

170

Arg Thr His Thr Gly 185

Ser Phe Ser Arg Glu

200

Gly Glu Lys Pro Tyr 215

Ser Asp Glu Leu Val

235

Tyr Lys Cys Pro Glu

250

Thr Glu His Gln Arg

265

Glu Cys Gly Lys Ser 280

Arg Thr His Thr Gly 295

Ser Phe Ser Gln Asn

315

Gly Lys Lys Thr Ser

Leu Glu Pro Gly Glu

160

Phe Ser Arg Ser Asp

175

Glu Lys Pro Tyr Lys

190

Asp Asn Leu His Thr 205

Lys Cys Pro Glu Cys 220

Arg His Gln Arg Thr

240

Cys Gly Lys Ser Phe

255

Thr His Thr Gly Glu

270

Phe Ser Thr Ser Gly 285

Glu Lys Pro Tyr Lys 300

Ser Thr Leu Thr Glu

320

Val Tyr Val Gly Gly

- 323 046157

325 330

335

Leu Glu Ser Arg Val Leu Lys Tyr Thr Ala Gln Asn

340 345

Met Glu Leu Gln

350

Asn Lys Val Gln Leu Leu Glu Glu Gln Asn Leu Ser

355 360

Leu Leu Asp Gln 365

Leu Arg Lys Leu Gln Ala Met Val Ile Glu Ile Ser

370 375 380

Asn Lys Thr Ser

Ser Ser Ser Thr Cys Ile Leu Val Leu Leu Val Ser

385 390 395

Phe Cys Leu Leu

400

Leu Val Pro Ala Met Tyr Ser Ser Asp Thr Arg Gly

405 410

Ser Leu Pro Ala

415

Glu His Gly Val Leu Ser Arg Gln Leu Arg Ala Leu

420 425

Pro Ser Glu Asp

430

Pro Tyr Gln Leu Glu Leu Pro Ala Leu Gln Ser Glu

435 440

Val Pro Lys Asp 445

Ser Thr His Gln Trp Leu Asp Gly Ser Asp Cys Val

450 455 460

Leu Gln Ala Pro

Gly Asn Thr Ser Cys Leu Leu His Tyr Met Pro Gln

465 470 475

Ala Pro Ser Ala

480

Glu Pro Pro Leu Glu Trp Pro Phe Pro Asp Leu Phe

485 490

Ser Glu Pro Leu

495

Cys Arg Gly Pro Ile Leu Pro Leu Gln Ala Asn Leu

500 505

Thr Arg Lys Gly

510

Gly Trp Leu Pro Thr Gly Ser Pro Ser Val Ile Leu

515 520

Gln Asp Arg Tyr 525

Ser Gly

530 <210> 214 <211> 1590 <212> DNA <213> Artificial sequence <220>

<223> Description of the artificial polynucleotide sequence

Synthetic

- 324 046157 <400> 214 atggagctgg aattggatgc tggtgaccaa gacctgctgg ccttcctgct agaggaaagt 60 ggagatttgg ggacggcacc cgatgaggcc gtgaggccc cactggactg ggcgctgccg120 ctttctgagg tgccgagcga ctgggaagta gat gatttgc tgtgctccct gctgagtccc180 ccagcgtcgt tgaacattct cagctcctcc aacccctgcc ttgtccacca tgaccacacc240 tactccctcc cacgggaaac tgtctctatg gatctagaga gtgagagctg tagaaaagag300 gggacccaga tgactccaca gcatatggag gagctggcag agca ggagat tgctaggcta360 gtactgacag atgaggagaa gagtctattg gagaaggagg ggcttattct gcctgagaca420 cttcctctca ctaagacaga ggaacaaatt ctgaaacgtg tgcggcttga gcccggagag480 aagccgtaca agtgccctga gtgcggcaag tcttttagca gaagagacga acttaatgtc540 caccagcgaa cgcatactgg tgaaaagccc tataaatgtc ctgaatgtgg gaaatcattc60 0 tccagccgca gaacctgtag ggctcaccag cgaacacaca ccggcgaaaa accatacaaa660 tgtccagaat gcgggaaatc cttttctcag tcatccaact tggtgagaca tcaacgcacg720 cacactggag aaaagcctta caaatgcccg gaatgtggaa agtctcttttc ccaattggcc7 80 catttgcgag cccatcagag gactcacacg ggcgagaaac cttacaaatg cccggaatgc840 gggaaatctt tttcaacgag tggcaacctc gtaagacacc aaagaacgca tacaggcgaa900 aagccatata agtgtcctga gtgtggtaaa tcattctcac acaggaccac cctgacaaat960 caccagcgca cgcacaccgg caagaagaca agcgtgtacg ttgggggttt agagagccgg1020 gtcttgaaat acacagccca gaatatggag cttcagaaca aagtacagct tctggaggaa1080 cagaatttgt cccttctaga tcaactgagg aaactccagg ccatggtgat tgagatctca1140 aacaaaacca gcagcagcag cacctgcatc ttggtcctgc tagtctcctt ctgcctcctc1200 cttgtacctg ctatgtactc ctctgacaca agggggagcc tgccagctga gcatggagtg1260 ttgtcccgcc ag cttcgtgc cctccccagt gaggaccctt accagctgga gctgcctgcc1320 ctgcagtcag aagtgccgaa agacagcaca caccagtggt tggacggctc agactgtgta1380 ctccaggccc ctggcaacac ttcctgcctg ctgcattaca tgcctcaggc tcccagtgca1440 gagcctcccc tggagtggcc cttccctgac ctcttctcag agcctctctg ccgaggtccc1500 atcctccccc tgcaggcaaa tctcacaagg aagggaggat ggcttcctac tggtagcccc1560 tctgtcattt tgcaggacag atactcaggc159 0 <210> 215 <211> 530 <212> Protein <213> Artificial sequence <220>

- 325 046157 <223> Description of the artificial sequence polypeptide <400> 215

Met Glu Leu Glu Leu Asp Ala Gly Asp Gln Asp Leu 1 5 10

Synthetic

Leu Ala Phe Leu

Leu Glu Glu Ser Gly Asp Leu Gly Thr Ala Pro Asp 20 25

Glu Ala Val Arg 30

Ala Pro Leu Asp Trp Ala Leu Pro Leu Ser Glu Val

40

Pro Ser Asp Trp 45

Glu Val Asp Asp Leu Leu Cys Ser Leu Leu Ser Pro

55 60

Pro Ala Ser Leu

Asn Ile Leu Ser Ser Ser Asn Pro Cys Leu Val His 65 70 75

His Asp His Thr

Tyr Ser Leu Pro Arg Glu Thr Val Ser Met Asp Leu

90

Glu Ser Glu Ser

Cys Arg Lys Glu Gly Thr Gln Met Thr Pro Gln His

100 105

Met Glu Glu Leu

110

Ala Glu Gln Glu Ile Ala Arg Leu Val Leu Thr Asp

115 120

Glu Glu Lys Ser 125

Leu Leu Glu Lys Glu Gly Leu Ile Leu Pro Glu Thr

130 135 140

Leu Pro Leu Thr

Lys Thr Glu Glu Gln Ile Leu Lys Arg Val Arg Leu

145 150 155

Glu Pro Gly Glu

160

Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe

165 170

Ser Arg Arg Asp

175

Glu Leu Asn Val His Gln Arg Thr His Thr Gly Glu

180 185

Lys Pro Tyr Lys

190

Cys Pro Glu Cys Gly Lys Ser Phe Ser Ser Arg Arg

195 200

Thr Cys Arg Ala 205

His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys

210 215 220

Cys Pro Glu Cys

Gly Lys Ser Phe Ser Gln Ser Ser Asn Leu Val Arg

225 230 235

His Gln Arg Thr

240

- 326 046157

His

Ser

Lys

Asn

Cys 305

His

Leu

Asn

Leu

Ser 385

Leu

Glu

Pro

Ser

Gly 465

Glu

Thr Gly Glu Lys Pro

245

Gln Leu Ala His Leu

260

Pro Tyr Lys Cys Pro

275

Leu Val Arg His Gln 290

Pro Glu Cys Gly Lys

310

Gln Arg Thr His Thr

325

Glu Ser Arg Val Leu

340

Lys Val Gln Leu Leu

355

Arg Lys Leu Gln Ala 370

Ser Ser Thr Cys Ile

390

Val Pro Ala Met Tyr

405

His Gly Val Leu Ser

420

Tyr Gln Leu Glu Leu

435

Thr His Gln Trp Leu 450

Asn Thr Ser Cys Leu

470

Pro Pro Leu Glu Trp

485

Tyr Lys Cys Pro Glu

250

Cys Gly Lys Ser Phe

255

Arg Ala His Gln Arg 265

Glu Cys Gly Lys Ser

280

Arg Thr His Thr Gly 295

Ser Phe Ser His Arg

315

Gly Lys Lys Thr Ser

330

Lys Tyr Thr Ala Gln

345

Glu Glu Gln Asn Leu

360

Met Val Ile Glu Ile 375

Leu Val Leu Leu Val

395

Ser Ser Asp Thr Arg

410

Arg Gln Leu Arg Ala 425

Pro Ala Leu Gln Ser

440

Asp Gly Ser Asp Cys 455

Leu His Tyr Met Pro

475

Pro Phe Pro Asp Leu

490

Thr His Thr Gly Glu

270

Phe Ser Thr Ser Gly 285

Glu Lys Pro Tyr Lys 300

Thr Thr Leu Thr Asn

320

Val Tyr Val Gly Gly

335

Asn Met Glu Leu Gln

350

Ser Leu Leu Asp Gln

365

Ser Asn Lys Thr Ser 380

Ser Phe Cys Leu Leu

400

Gly Ser Leu Pro Ala

415

Leu Pro Ser Glu Asp 430

Glu Val Pro Lys Asp 445

Val Leu Gln Ala Pro 460

Gln Ala Pro Ser Ala

480

Phe Ser Glu Pro Leu

495

- 327 046157

Cys Arg Gly Pro Ile Leu Pro Leu Gln Ala Asn Leu Thr Arg Lys Gly 500 505 510

Gly Trp Leu Pro Thr Gly Ser Pro Ser Val Ile Leu Gln Asp Arg Tyr 515 520 525

Ser Gly 530 <210> 216 <211> 1590 <212> DNA <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic polynucleotide

<400> 216 atggagctgg aattggatgc tggtgaccaa gacctgctgg ccttcctgct aggaaagt 60 ggagattgg ggacggcacc cgatgaggcc gtgagggccc cactggactg ggcgctgccg 120 ctttctgagg tgccgagcga ctgggaagta gatgatttgc tgtgctccct gctgagtccc 180 ccagcgtcgt tgaacattct cagctcctcc aacccctgcc ttgtccacca tgaccacacc 240 tactccctcc cacgggaaac tgtctctatg gatctagaga gtgagagctg tagaaaagag 300 gggacccaga tgactccaca gcatatggag gagctggcag agcaggagat tgctaggcta 360 gtactgacag atgaggagaa gagtctattg gagaaggagg ggcttattct gcctgagaca 420 cttcctctca ctaagacaga ggaacaaatt ctgaaacgtg tgcggcgccc ttacgcttgc 480 ccagtggagt cctgtgatcg ccgcttctcc cgcagcgaca acctggtgag acacatccgc 540 atccacacag gccagaagcc cttccagtgc cgcatctgca tgagaaactt cagccgagag 600 gataacttgc acactcacat ccgcaccac acaggcgaaa agcccttcgc ctgcgacatc 660 tgtggaagaa agtttgcccg gagcgatgaa cttgtccgac ataccaagat ccacttgcgg 720 cagaaggacc gcccttacgc ttgcccagtg gagtcctgtg atcgccgctt ctcccaatca 780 gggaatctga ctgagcacat ccgcatccac acaggccaga agcccttcca gtgccgcatc 840 tgcatgagaa acttcagcac aagtggacat ctggtacgcc acatccgcac ccacacaggc 900 gaaaagccct tcgcctgcga catctgtgga agaaagtttg cccagaatag taccctgacc 960 gaacatacca agatccactt gcggcagaag gacgtgtacg ttggggggttt agagagccgg 1020 gtcttgaaat acacagccca gaatatggag cttcagaaca aagtacagct tctggaggaa 1080 cagaatttgt cccttctaga tcaactgagg aaactccagg ccatggtgat tgagatctca 1140 aacaaaacca gcagcagcag cacctgcatc ttggtcctgc tagtctcctt ctgcctcctc 1200

- 328 046157

cttgtacctg ctatgtactc ctctgacaca aggggggagcc tgccagctga gcatggagtg 1260 ttgtcccgcc agcttcgtgc cctccccagt gaggaccctt accagctgga gctgcctgcc 1320 ctgcagtcag aagtgccgaa agacagcaca caccagtggt tggacggctc agactgtgta 1380 ctccaggccc ctggcaacac ttcctgcctg ctgcattaca tgcctcaggc tccagtgca 1440 gagcctcccc tggagtggcc cttccctgac ctcttctcag agcctctctg ccgaggtccc 1500 atcctccccc tgcaggcaaa tctcacaagg aagggaggat ggcttcctac tggtagcccc 1560 tctgtcattt tgcaggacag atactcaggc 1590

<210> 217 <211> 530 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic polypeptide <400> 217

Met Glu Leu Glu Leu Asp Ala Gly Asp Gln Asp Leu Leu Ala Phe Leu 1 5 1015

Leu Glu Glu Ser Gly Asp Leu Gly Thr Ala Pro Asp Glu Ala Val Arg 20 2530

Ala Pro Leu Asp Trp Ala Leu Pro Leu Ser Glu Val Pro Ser Asp Trp 35 4045

Glu Val Asp Asp Leu Leu Cys Ser Leu Leu Ser Pro Pro Ala Ser Leu 50 5560

Asn Ile Leu Ser Ser Ser Asn Pro Cys Leu Val His His Asp His Thr 65 70 7580

Tyr Ser Leu Pro Arg Glu Thr Val Ser Met Asp Leu Glu Ser Glu Ser 85 9095

Cys Arg Lys Glu Gly Thr Gln Met Thr Pro Gln His Met Glu Glu Leu

100 105110

Ala Glu Gln Glu Ile Ala Arg Leu Val Leu Thr Asp Glu Glu Lys Ser

115 120125

Leu Leu Glu Lys Glu Gly Leu Ile Leu Pro Glu Thr Leu Pro Leu Thr

130 135140

Lys Thr Glu Glu Gln Ile Leu Lys Arg Val Arg Arg Pro Tyr Ala Cys

- 329 046157

145

Pro

Arg

Cys

Thr

Phe 225

Gln

Phe

Gln

Gly

Ala 305

Glu

Leu

Asn

Leu

Ser 385

150

155

160

Val Glu Ser Cys Asp

165

Arg Arg Phe Ser Arg

170

Ser Asp Asn Leu Val

175

His Ile Arg Ile His

180

Met Arg Asn Phe Ser 195

His Thr Gly Glu Lys 210

Ala Arg Ser Asp Glu

230

Lys Asp Arg Pro Tyr

245

Ser Gln Ser Gly Asn 260

Lys Pro Phe Gln Cys

275

His Leu Val Arg His 290

Cys Asp Ile Cys Gly

310

His Thr Lys Ile His

325

Glu Ser Arg Val Leu

340

Lys Val Gln Leu Leu

355

Arg Lys Leu Gln Ala 370

Ser Ser Thr Cys Ile

390

Thr Gly Gln Lys Pro 185

Phe Gln Cys Arg Ile

190

Arg Glu Asp Asn Leu

200

Pro Phe Ala Cys Asp 215

Leu Val Arg His Thr

235

Ala Cys Pro Val Glu

250

Leu Thr Glu His Ile

265

Arg Ile Cys Met Arg 280

Ile Arg Thr His Thr 295

Arg Lys Phe Ala Gln

315

Leu Arg Gln Lys Asp

330

Lys Tyr Thr Ala Gln

345

Glu Glu Gln Asn Leu

360

Met Val Ile Glu Ile 375

Leu Val Leu Leu Val

395

His Thr His Ile Arg

205

Ile Cys Gly Arg Lys 220

Lys Ile His Leu Arg

240

Ser Cys Asp Arg Arg

255

Arg Ile His Thr Gly 270

Asn Phe Ser Thr Ser

285

Gly Glu Lys Pro Phe 300

Asn Ser Thr Leu Thr

320

Val Tyr Val Gly Gly

335

Asn Met Glu Leu Gln

350

Ser Leu Leu Asp Gln

365

Ser Asn Lys Thr Ser 380

Ser Phe Cys Leu Leu

400

- 330 046157

Leu Val

Pro Ala

Glu His

Gly Val

420

Pro Tyr

Gln Leu 435

Ser Thr

450

His Gln

Met Tyr 405

Leu Ser

Glu Leu

Trp Leu

Gly Asn 465

Thr Ser

Cys Leu

470

Glu Pro

ProLeu

Glu Trp 485

CysArg

Gly Pro

500

Ile Leu

Gly Trp

Leu Pro 515

Thr Gly

Ser Ser

Arg Gln

Pro Ala 440

Asp Gly 455

Leu His

Pro Phe

ProLeu

Ser Pro

520

Asp Thr

410

Leu Arg 425

Leu Gln

Ser Asp

Tyr Met

Pro Asp

490

Gln Ala 505

Ser Val

Arg Gly

Ala Leu

Ser Glu

Cys Val

460

Pro Gln 475

Leu Phe

Asn Leu

Ile Leu

Ser Leu

Pro Ser

430

Val Pro 445

Leu Gln

Ala Pro

Ser Glu

Thr Arg

510

Gln Asp 525

Pro Ala 415

Glu Asp

Lys Asp

Ala Pro

Ser Ala

480

Pro Leu 495

Lys Gly

Arg Tyr

Ser Gly

530 <210> 218 <211> 1590 <212> DNA <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic polynucleotide

<400> 218 atggagctgg aattggatgc tggtgaccaa gacctgctgg ccttcctgct aggaaagt ggagattgg ggacggcacc cgatgaggcc gtgagggccc cactggactg ggcgctgccg ctttctgagg tgccgagcga ctgggaagta gatgatttgc tgtgctccct gctgagtccc ccagcgtcgt tgaacattct cagctcctcc aacccctgcc ttgtccacca tgaccacacc tactccctcc cacgggaaac tgtctctatg gatctagaga gtgagagctg tagaaaagag gggacccaga tgactccaca gcatatggag gagctggcag agcaggagat tgctaggcta gtactgacag atgaggagaa gagtctattg gagaaggagg ggcttattct gcctgagaca cttcctctca ctaagacaga ggaacaaatt ctgaaacgtg tgcggcgccc ttacgcttgc ccagtggagt cctgtgatcg ccgcttctcc cgctcagaca acctcgttcg acacatccgc

120

180

240

300

360

420

480

540

- 331 046157

atccacacag gccagaagcc cttccagtgc cgcatctgca tgagaaactt cagccaccgg 600 actacactca cgaaccacat ccgcaccac acaggcgaaa agcccttcgc ctgcgacatc 660 tgtggaagaa agtttgccag agaagacaat ctccatactc ataccaagat ccacttgcgg 720 cagaaggacc gcccttacgc ttgcccagtg gagtcctgtg atcgccgctt ctccaccagc 780 cattctctca ctgaacacat ccgcatccac acaggccaga agcccttcca gtgccgcatc 840 tgcatgagaa acttcagcca gtctagctca ctggtgaggc acatccgcac ccacacaggc 900 gaaaagccct tcgcctgcga catctgtgga agaaagtttg ccaggagaga taacctgcat 960 acgcatacca agatccactt gcggcagaag gacgtgtacg ttggggggttt agagagccgg 1020 gtcttgaaat acacagccca gaatatggag cttcagaaca aagtacagct tctggaggaa 1080 cagaatttgt cccttctaga tcaactgagg aaactccagg ccatggtgat tgagatctca 1140 aacaaaacca gcagcagcag cacctgcatc ttggtcctgc tagtctcctt ctgcctcctc 1200 cttgtacctg ctatgtactc ctctgacaca aggggggagcc tgccagctga gcatggagtg 1260 ttgtcccgcc agcttcgtgc cctccccagt gaggaccctt accagctgga gctgcctgcc 1320 ctgcagtcag aagtgccgaa agacagcaca caccagtggt tggacggctc agactgtgta 1380 ctccaggccc ctggcaacac ttcctgcctg ctgcattaca tgcctcaggc tccagtgca 1440 gagcctcccc tggagtggcc cttccctgac ctcttctcag agcctctctg ccgaggtccc 1500 atcctccccc tgcaggcaaa tctcacaagg aagggaggat ggcttcctac tggtagcccc 1560 tctgtcattt tgcaggacag atactcaggc 1590

<210> 219 <211> 530 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic polypeptide <400> 219

Met Glu Leu Glu Leu Asp Ala Gly Asp Gln Asp Leu Leu Ala Phe Leu 1 5 1015

Leu Glu Glu Ser Gly Asp Leu Gly Thr Ala Pro Asp Glu Ala Val Arg 20 2530

Ala Pro Leu Asp Trp Ala Leu Pro Leu Ser Glu Val Pro Ser Asp Trp 35 4045

Glu Val Asp Asp Leu Leu Cys Ser Leu Leu Ser Pro Pro Ala Ser Leu 50 5560

- 332 046157

Asn Ile Leu Ser Ser Ser Asn Pro Cys Leu Val 65 7075

His His Asp His Thr

Tyr Ser Leu Pro Arg Glu Thr Val Ser Met Asp Leu Glu Ser Glu Ser 85 9095

Cys Arg Lys Glu Gly Thr Gln Met Thr Pro Gln His Met Glu Glu Leu 100 105110

Ala Glu Gln Glu Ile Ala Arg Leu Val Leu Thr Asp Glu Glu Lys Ser 115 120125

Leu Leu Glu Lys Glu Gly Leu Ile Leu Pro Glu Thr Leu Pro Leu Thr 130 135140

Lys Thr Glu Glu Gln Ile Leu Lys Arg Val Arg Arg Pro Tyr Ala Cys

145 150 155160

Pro Val Glu Ser Cys Asp Arg Arg Phe Ser Arg Ser Asp Asn Leu Val

165 170175

Arg His Ile Arg Ile His Thr Gly Gln Lys Pro Phe Gln Cys Arg Ile

180 185190

Cys Met Arg Asn Phe Ser His Arg Thr Thr Leu Thr Asn His Ile Arg

195 200205

Thr His Thr Gly Glu Lys Pro Phe Ala Cys Asp Ile Cys Gly Arg Lys

210 215220

Phe Ala Arg Glu Asp Asn Leu His Thr His Thr Lys Ile His Leu Arg

225 230 235240

Gln Lys Asp Arg Pro Tyr Ala Cys Pro Val Glu Ser Cys Asp Arg Arg

245 250255

Phe Ser Thr Ser His Ser Leu Thr Glu His Ile Arg Ile His Thr Gly

260 265270

Gln Lys Pro Phe Gln Cys Arg Ile Cys Met Arg Asn Phe Ser Gln Ser

275 280285

Ser Ser Leu Val Arg His Ile Arg Thr His Thr Gly Glu Lys Pro Phe

290 295300

Ala Cys Asp Ile Cys Gly Arg Lys Phe Ala Arg Glu Asp Asn Leu His

305 310 315320

- 333 046157

Thr His Thr Lys Ile His Leu Arg

325

Gln Lys Asp Val Tyr Val Gly Gly

330 335

Leu Glu

Ser Arg Val

340

Leu Lys Tyr Thr Ala Gln Asn Met Glu Leu Gln

345 350

Asn Lys Val Gln

355

Leu Leu Glu Glu Gln Asn Leu Ser Leu Leu Asp Gln

360 365

Leu Arg Lys Leu Gln Ala Met Val

370 375

Ile Glu Ile Ser Asn Lys Thr Ser

380

Ser Ser Ser Thr Cys

385

Ile Leu Val Leu Leu Val Ser Phe Cys Leu Leu

390 395400

Leu Val Pro Ala Met Tyr Ser Ser Asp Thr Arg Gly Ser Leu Pro Ala

405 410415

Glu His Gly Val Leu Ser Arg Gln Leu Arg Ala Leu Pro Ser Glu Asp

420 425430

Pro Tyr Gln Leu Glu Leu Pro Ala Leu Gln Ser Glu Val Pro Lys Asp

435 440445

Ser Thr His Gln Trp Leu Asp Gly Ser Asp Cys Val Leu Gln Ala Pro

450 455460

Gly Asn Thr Ser Cys Leu Leu His Tyr Met Pro Gln Ala Pro Ser Ala

465 470 475480

Glu Pro Pro Leu Glu Trp Pro Phe Pro Asp Leu Phe Ser Glu Pro Leu

485 490495

Cys Arg Gly Pro Ile Leu Pro Leu Gln Ala Asn Leu Thr Arg Lys Gly

500 505510

Gly Trp Leu Pro Thr Gly Ser Pro Ser Val Ile Leu Gln Asp Arg Tyr

515 520525

Ser Gly

530 <210> 220 <211> 1140 <212> DNA <213> Artificial sequence

- 334 046157 <220>

<223> Description of the artificial sequence: Synthetic polynucleotide <400> 220 atggagctgg aattggatgc tggtgaccaa gacctgctgg ccttcctgct agaggaaagt60 ggagatttgg ggacggcacc cgatgaggcc gtgaggccc cactggactg ggcgctgccg120 ctttctgagg tgccgagcga ctgggaag ta gatgatttgc tgtgctccct gctgagtccc180 ccagcgtcgt tgaacattct cagctcctcc aacccctgcc ttgtccacca tgaccacacc240 tactccctcc cacgggaaac tgtctctatg gatctagaga gtgagagctg tagaaaagag300 gggacccaga tgactccaca gcatatggag gagctggcag agcaggagat tgctaggcta360 gtactgacag atgaggagaa gagtctattg gagaaggagg ggcttattct gcctgagaca420 cttcctctca ctaagacaga ggaacaaatt ctgaaacgtg tgcggctcga accaggtgaa480 aaaccttaca aatgtcctga atgtgggaaa tcattcagtc gcagcgacaa cctggtgaga540 catcaacgca cccata cagg agaaaaacct tataaatgtc cagaatgtgg aaagtccttc600 tcacgagagg ataacttgca cactcatcaa cgaacacata ctggtgaaaa accatacaag660 tgtcccgaat gtggtaaaag ttttagccgg agcgatgaac ttgtccgaca ccaacgaacc720 catacaggcg agaagcctta caaatgtccc gagtgtggca agagcttctc acaatcaggg780 aatctgactg agcatcaacg aactcatacc ggggaaaaac cttacaagtg tccagagtgt840 gggaagagct tttccacaag tggacatctg gtacgccacc agaggacaca tacaggggag900 aagccctaca aatgccccga atgcggtaaa agtttctctc agaatagtac cctgaccgaa960 caccagcgaa cacacactgg gaaaaaaacg agtgtgtacg ttggggggttt agagagccgg1020 gtcttgaaat acacagccca ga atatggag cttcagaaca aagtacagct tctggaggaa1080 cagaatttgt cccttctaga tcaactgagg aaactccagg ccatggtgat tgagatatca1140 <210> 221 <211> 380 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic polypeptide <400> 221

Met Glu Leu Glu Leu Asp Ala Gly Asp Gln Asp Leu Leu Ala Phe Leu

5 10 15

Leu Glu Glu Ser Gly Asp Leu Gly Thr Ala Pro Asp Glu Ala Val Arg 20 25 30

Ala Pro Leu Asp Trp Ala Leu Pro Leu Ser Glu Val Pro Ser Asp Trp

- 335 046157

40 45

Glu

Asn 65

Tyr

Cys

Ala

Leu

Lys 145

Lys

Asn

Cys

His

Gly 225

His

Ser

Lys

Val Asp Asp Leu Leu Cys Ser Leu Leu Ser Pro Pro Ala Ser 50 55 60

Ile Leu Ser Ser Ser Asn Pro Cys Leu Val His His Asp His 70 75

Ser Leu Pro Arg Glu Thr Val Ser Met Asp Leu Glu Ser Glu 85 90 95

Arg Lys Glu Gly Thr Gln Met Thr Pro Gln His Met Glu Glu

100 105 110

Glu Gln Glu Ile Ala Arg Leu Val Leu Thr Asp Glu Glu Lys

115 120 125

Leu Glu Lys Glu Gly Leu Ile Leu Pro Glu Thr Leu Pro Leu

130 135 140

Thr Glu Glu Gln Ile Leu Lys Arg Val Arg Leu Glu Pro Gly

150 155

Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Ser

165 170 175

Leu Val Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr

180 185 190

Pro Glu Cys Gly Lys Ser Phe Ser Arg Glu Asp Asn Leu His

195 200 205

Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu

210 215 220

Lys Ser Phe Ser Arg Ser Asp Glu Leu Val Arg His Gln Arg

230 235

Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser

245 250 255

Gln Ser Gly Asn Leu Thr Glu His Gln Arg Thr His Thr Gly

260 265 270

Pro Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Ser

275 280 285

Leu

Thr 80

Ser

Leu

Ser

Thr

Glu 160

Asp

Lys

Thr

Cys

Thr 240

Phe

Glu

Gly

- 336 046157

His Leu Val Arg His Gln Arg Thr

290 295

His Thr Gly Glu Lys Pro Tyr Lys

300

Cys Pro Glu Cys Gly Lys Ser Phe

305 310

Ser Gln Asn Ser Thr Leu Thr Glu

315 320

His Gln Arg Thr His Thr Gly Lys

325

Lys Thr Ser Val Tyr Val Gly Gly

330 335

Leu Glu Ser Arg Val Leu Lys Tyr

340

Thr Ala Gln Asn Met Glu Leu Gln

345 350

Asn Lys Val Gln Leu Leu Glu Glu

355 360

Gln Asn Leu Ser Leu Leu Asp Gln

365

Leu Arg Lys Leu Gln Ala Met Val

370 375

Ile Glu Ile Ser

380 <210> 222 <211> 1140 <212> DNA <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic polynucleotide

<400> 222 atggagctgg aattggatgc tggtgaccaa gacctgctgg ccttcctgct aggaaagt 60 ggagattgg ggacggcacc cgatgaggcc gtgagggccc cactggactg ggcgctgccg 120 ctttctgagg tgccgagcga ctgggaagta gatgatttgc tgtgctccct gctgagtccc 180 ccagcgtcgt tgaacattct cagctcctcc aacccctgcc ttgtccacca tgaccacacc 240 tactccctcc cacgggaaac tgtctctatg gatctagaga gtgagagctg tagaaaagag 300 gggacccaga tgactccaca gcatatggag gagctggcag agcaggagat tgctaggcta 360 gtactgacag atgaggagaa gagtctattg gagaaggagg ggcttattct gcctgagaca 420 cttcctctca ctaagacaga ggaacaaatt ctgaaacgtg tgcggcgccc ttacgcttgc 480 ccagtggagt cctgtgatcg ccgcttctcc cgcagcgaca acctggtgag acacatccgc 540 atccacacag gccagaagcc cttccagtgc cgcatctgca tgagaaactt cagccgagag 600 gataacttgc acactcacat ccgcaccac acaggcgaaa agcccttcgc ctgcgacatc 660 tgtggaagaa agtttgcccg gagcgatgaa cttgtccgac ataccaagat ccacttgcgg 720 cagaaggacc gcccttacgc ttgcccagtg gagtcctgtg atcgccgctt ctcccaatca 780 gggaatctga ctgagcacat ccgcatccac acaggccaga agcccttcca gtgccgcatc 840 tgcatgagaa acttcagcac aagtggacat ctggtacgcc acatccgcac ccacacaggc 900

- 337 046157 gaaaagccct gaacatacca gtcttgaaat cagaatttgt tcgcctgcga agatccactt acacagccca cccttctaga catctgtgga gcggcagaag gaatatggag tcaactgagg agaaagtttg gacgtgtacg cttcagaaca aaactccagg cccagaatag ttggggggt tt aagtacagct ccatggtgat taccctgacc agagagccgg tctggaggaa tgagatctca

960

1020

1080

1140 <210> 223 <211> 380 <212> Protein <213> Artificial sequence <220>

<223> Description of artificial sequence: Synthetic polypeptide <400> 223

Met Glu Leu Glu Leu Asp Ala Gly Asp Gln Asp Leu Leu Ala Phe Leu

5 10 15

Leu Glu Glu

Ser Gly Asp Leu Gly Thr Ala Pro Asp Glu Ala Val Arg 20 25 30

Ala Pro Leu Asp Trp Ala Leu Pro Leu

40

Ser Glu Val Pro Ser Asp Trp 45

Glu Val Asp Asp Leu Leu Cys

55

Ser Leu Leu Ser Pro Pro Ala Ser Leu

Asn Ile Leu Ser Ser Ser Asn Pro Cys Leu Val His His Asp His Thr 65 70 75 80

Tyr Ser

Leu Pro

CysArg

Lys Glu

100

Ala Glu

Gln Glu 115

Leu Leu

130

Glu Lys

Lys Thr 145

Glu Glu

Pro Val

Glu Ser

Arg Glu 85

Gly Thr

Ile Ala

Glu Gly

Gln Ile

150

Cys Asp 165

Thr Val

SerMet

Asp Leu

Glu Ser

Gln Met

Thr Pro 105

Gln His

Met Glu

110

Arg Leu

120

Val Leu

Thr Asp

Glu Glu 125

Leu Ile 135

Leu Pro

Glu Thr

140

Leu Pro

Leu Lys

Arg Val

Arg Arg 155

Pro Tyr

Arg Arg

Phe Ser

170

Arg Ser

Asp Asn

Glu Ser 95

Glu Leu

Lys Ser

Leu Thr

Ala Cys 160

Leu Val 175

- 338 046157

Arg His Ile Arg Ile His Thr Gly Gln Lys Pro Phe

180 185

Gln Cys Arg Ile

190

Cys Met Arg Asn Phe Ser Arg Glu Asp Asn Leu His

195 200

Thr His Ile Arg 205

Thr His Thr Gly Glu Lys Pro Phe Ala Cys Asp Ile

210 215 220

Cys Gly Arg Lys

Phe Ala Arg Ser Asp Glu Leu Val Arg His 225 230

Thr Lys 235

Ile His Leu Arg

240

Gln Lys Asp Arg Pro Tyr Ala Cys Pro Val Glu Ser

245 250

Cys Asp Arg Arg

255

Phe Ser Gln Ser Gly Asn Leu Thr Glu His Ile Arg

260 265

Ile His Thr Gly 270

Gln Lys Pro Phe Gln Cys Arg Ile Cys Met Arg Asn

275 280

Phe Ser Thr Ser 285

Gly His Leu Val Arg His Ile Arg Thr His Thr Gly

290 295 300

Glu Lys Pro Phe

Ala Cys Asp Ile Cys Gly Arg Lys Phe Ala Gln Asn

305 310 315

Ser Thr Leu Thr

320

Glu His Thr Lys Ile His Leu Arg Gln Lys Asp Val

325 330

Tyr Val Gly Gly

335

Leu Glu Ser Arg Val Leu Lys Tyr Thr Ala Gln Asn

340 345

Met Glu Leu Gln

350

Asn Lys Val Gln Leu Leu Glu Glu Gln Asn Leu Ser

355 360

Leu Leu Asp Gln 365

Leu Arg Lys Leu Gln Ala Met Val Ile Glu Ile Ser

370 375 380 <210> 224 <211> 155 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial sequence polypeptide <400> 224

Synthetic

- 339 046157

Met Glu 1

Leu Glu

Leu Asp 5

Ala Gly

Asp Gln

Asp Leu

Leu Ala Phe Leu

Leu Glu

Glu Ser

Gly Asp

Leu Gly

Thr Ala 25

Pro Asp

Glu Ala Val Arg 30

Ala Pro

Leu Asp 35

Trp Ala

Leu Pro

Leu Ser

Glu Val

Pro Ser Asp Trp 45

Glu Val

Asp Asp

Leu Leu

Cys Ser 55

Leu Leu

Ser Pro

Pro Ala Ser Leu

Asn Ile 65

Leu Ser

Ser Ser

Asn Pro

Cys Leu

Val His 75

His Asp His Thr

Tyr Ser

Leu Pro

Arg Glu 85

Thr Val

SerMet

Asp Leu

Glu Ser Glu Ser

CysArg

Lys Glu

100

Gly Thr

Gln Met

Thr Pro 105

Gln His

Met Glu Glu Leu

110

Ala Glu

Gln Glu 115

Ile Ala

Arg Leu

120

Val Leu

Thr Asp

Glu Glu Lys Ser 125

Leu Leu

130

Glu Lys

Glu Gly

Leu Ile 135

Leu Pro

Glu Thr

140

Leu Pro Leu Thr

Lys Thr 145

Glu Glu

Gln Ile

150

Leu Lys

Arg Val

Arg 155 <210> 225 <211> 199 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial sequence polypeptide <400> 225

Synthetic

Val Tyr Val Gly Gly Leu 1 5

Glu Ser Arg Val Leu Lys 10

Tyr Thr Ala Gln 15

Asn Met Glu Leu Gln Asn

Lys Val Gln Leu Leu Glu 25

Glu Gln Asn Leu

Ser Leu Leu Asp Gln Leu 35

Arg Lys Leu Gln Ala Met 40

Val Ile Glu Ile 45

Ser Asn Lys Thr Ser Ser

Ser Ser Thr Cys Ile Leu

Val Leu Leu Val

- 340 046157

55 60

Ser Phe 65

Gly Ser

Leu Pro

Glu Val

Cys Leu

Leu Pro

Ser Glu

100

Pro Lys 115

Leu Leu

Val Pro

Ala Met

Ala Glu 85

Asp Pro

Asp Ser

His Gly

Val Leu

Tyr Gln

Leu Glu 105

Thr His

120

Gln Trp

Val Leu

130

Gln Ala

Pro Gly

Asn Thr 135

Ser Cys

Gln Ala 145

Pro Ser

Ala Glu

150

Pro Pro

Leu Glu

Phe Ser

Glu Pro

Leu Cys 165

Arg Gly

Pro Ile

170

Leu Thr

Arg Lys

180

Gly Gly

Trp Leu

Pro Thr 185

Tyr Ser 75

Ser Arg

Leu Pro

Leu Asp

Leu Leu

140

Trp Pro 155

Leu Pro

Gly Ser

Ser Asp Thr Arg 80

Gln Leu Arg Ala 95

Ala Leu Gln Ser

110

Gly Ser Asp Cys 125

His Tyr Met Pro

Phe Pro Asp Leu

160

Leu Gln Ala Asn

175

Pro Ser Val Ile

190

Leu Gln

Asp Arg 195

Tyr Ser

Gly <210> 226 <211> 49 <212> Protein <213> Artificial sequence <220>

<223> Description of the artificial sequence polypeptide <400> 226

Synthetic

Val Tyr Val Gly Gly Leu 1 5

Asn Met Glu Leu Gln Asn

Ser Leu Leu Asp Gln Leu 35

Glu Ser Arg Val Leu Lys 10

Lys Val Gln Leu Leu Glu 25

Arg Lys Leu Gln Ala Met 40

Tyr Thr Ala Gln 15

Glu Gln Asn Leu

Val Ile Glu Ile 45

Ser

Claims (24)

1. A polynucleotide comprising a nucleic acid sequence encoding a non-naturally occurring transcription factor, wherein the non-naturally occurring transcription factor comprises an amino acid sequence having at least 97% identity to the sequence of SEQ ID NO: 127. 2. The polynucleotide according to claim 1, wherein the non-naturally occurring transcription factor contains the amino acid sequence of SEQ ID NO: 127. 3. The polynucleotide of claim 1 or 2, wherein the polynucleotide further comprises a regulatory element that controls expression of the transcription factor at a higher level in parvalbumin (PV)-containing neurons than in other cell types, and wherein the regulatory element comprises SEQ ID NO: 1 , 2, 3 or 4. 4. The polynucleotide according to claim 3, wherein the regulatory element contains SEQ ID NO: 2. 5. Polynucleotide according to any one of claims 1-4, additionally containing a microRNA binding site. 6. The polynucleotide according to claim 5, in which the microRNA binding site contains SEQ ID NO: 9, 11 or 13. 7. The polynucleotide according to claim 5, wherein the microRNA binding site contains SEQ ID NO: 7, 14 or 15. 8. The polynucleotide of claim 5, wherein the polynucleotide comprises a nucleic acid sequence having at least 95% identity to SEQ ID NO: 71. 9. The polynucleotide according to claim 5, where the polynucleotide contains SEQ ID NO: 71. 10. The polynucleotide according to any one of claims 1 to 9, wherein the non-naturally occurring transcription factor increases expression of the SCNIA gene in the cell. 11. An expression vector containing a polynucleotide according to any one of claims 1 to 10. 12. The expression vector of claim 11, wherein the expression vector is a viral vector. 13. The expression vector of claim 11, wherein the viral vector is an adeno-associated virus (AAV) vector. 14. The expression vector according to claim 13, where the AAV vector has a serotype of AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, AAV12, rh10, avian AAV, cattle AAV, canine AAV , equine AAV, primate AAV, non-primate AAV, ovine AAV, scAAV, scAAV1, scAAV2, scAAV5, scAAV8 or scAAV9, or hybrids thereof. 15. The expression vector of claim 14, wherein the polynucleotide further comprises a 5' AAV inverted terminal repeat (ITR) sequence and a 3' AAV ITR sequence. 16. The expression vector of claim 15, wherein each of the 5' AAV ITR and 3' AAV ITR sequences is independently AAV1, AAV2, AAV5, AAV8 or AAV9. 17. A pharmaceutical composition containing a polynucleotide according to any of claims 1-10 or an expression vector according to any of claims 11-16. 18. A cell containing a polynucleotide according to any of claims 1-10 or an expression vector according to any of claims 11-16. 19. Use of a polynucleotide according to any one of claims 1 to 10 in the manufacture of a medicament for the treatment of epilepsy in a subject in need thereof. 20. Use of a polynucleotide according to any one of claims 1 to 10 in the manufacture of a medicament for the treatment of Dravet syndrome in a subject in need thereof. 21. Use of a polynucleotide according to any one of claims 1 to 10 in the production of a medicinal product for reducing the frequency or duration of seizures. 22. Use of the expression vector according to any one of claims 11 to 16 in the production of a medicament for the treatment of epilepsy in a subject in need thereof. 23. Use of the expression vector according to any one of claims 11 to 16 in the production of a medicament for the treatment of Dravet syndrome in a subject in need thereof. 24. Use of the expression vector according to any one of claims 11 to 16 in the production of a medicinal product for reducing the frequency or duration of seizures.