KR101153845B1

KR101153845B1 - ＫｒａｓＧ１２ＶＲＮＡ를 특이적으로 인지할 수 있는 트랜스-스플라이싱 그룹 Ｉ 리보자임

Info

Publication number: KR101153845B1
Application number: KR1020080110937A
Authority: KR
Inventors: 이성욱; 김성진; 김주현; 정진숙
Original assignee: 단국대학교 산학협력단
Priority date: 2008-11-10
Filing date: 2008-11-10
Publication date: 2012-07-09
Anticipated expiration: 2028-11-10
Also published as: KR20100052070A

Abstract

본 발명은 단 염기 변이 전사체를 특이적으로 인지할 수 있는 트랜스-스플라이싱 그룹 I 리보자임에 관한 것으로서, 더욱 상세하게는 Kras 유전자의 12번째 코돈의 단 염기 변이 Ras RNA를 특이적으로 인식하여 치환할 수 있는 트랜스-스플라이싱 그룹 I 리보자임에 관한 것이다.

본 발명의 리보자임은 단 염기 변이된 유전자 전사체를 정상 유전자 전사체와 구별하여 특이적으로 인식하므로, 이러한 리보자임을 이용하여 돌연변이 전사체를 발현하는 암세포를 특이적으로 치료 및 진단이 가능하다.

단 염기 변이, 트랜스-스플라이싱 그룹 I 리보자임, Kras 유전자

Description

ＫｒａｓＧ１２ＶＲＮＡ를 특이적으로 인지할 수 있는 트랜스-스플라이싱 그룹 Ｉ 리보자임{trans―splicing group I ribozyme specific for Kras G12V RNA}

본 발명은 Kras 단 염기 변이 전사체를 특이적으로 인지할 수 있는 트랜스-스플라이싱 그룹 Ｉ 리보자임에 관한 것이다.

현재 항암치료로 주로 이용되고 있는 방사선 치료법과 항암화학요법 등은 이런 방법에 저항성을 보이는 환자에게는 비효과적이며 정상세포에도 부작용을 유발할 수 있는 한계점을 가지고 있었다. 따라서, 보다 특이적인 암 치료를 위해 암세포에서만 특이적인 세포사를 유도하는 방법이 필요하다.

라스 단백질(Ras protein)은 세포 성장, 증식과 분화를 조절하는 소 지티파제 단백질(small GTPase protein)으로 진화적으로 매우 보존되어 있고 각 조직마다 서로 다른 양을 발현한다.

이러한 라스의 아이소폼인 H-ras, N-ras, K-ras들은 서로 다른 위치에 의해 같은 활성자(activator)와 작동자(effector)를 가지고도 다른 기능을 한다(John F. Hancock. RAS proteins: different signals from diffrent locations. Nature Molecular Cell Biology 4; 373-384(2003)). RAS 아이소폼은 약 85%의 유사성을 가지고 있으며, H-ras와 N-ras의 하나 혹은 동시의 녹아웃 실험을 통해 정상 발달(normal development)에 필수적이지 않음이 알려졌다.

그러나, K-ras의 경우에는 배아 치사(embryonic lethal)가 되는 필수적인 단백질임이 보고되었다(Johnson, L. et al., K-ras is an essential gene in the mouse with partial functional overlap with N-ras. Genes Dev. 11; 2468-2481(1997)).

정상 세포에서의 RAS 단백질의 활성은 GTP와 GDP의 결합 비율에 의해 조절되어진다(Campbell, S. L. et al., Increasing complexity of Ras signaling. Oncogene 17;1395-1413(1998)). RAS 단백질의 활성 상태는 GTP나 GDP의 결합 여부에 의해 결정이 되는데, RAS-GTP는 활성 상태이고 RAS-GDP는 불활성 상태로 RAF, PI3K, RALGDS, PLCε등의 하위 단계 단백질들의 연속적인 인산화를 유도하여 외부로부터 신호전달을 하는 "분자 스위치(molecular switch)" 역할을 한다(Fabbro, D. et al., Protein tyrosine kinase inhibitors: new treatment modalities Curr. Opin. Pharmacol. 2; 374-381 (2002)).

비활성 상태의 RAS는 GEFs(guanine nucleotide exchange factors)에 의해 GDP를 방출하고 GTP 바인딩을 허용함으로써 활성화될 수 있다. Ras 신호화의 종결은 GAPs(GTPase activating proteins)에 의해 RAS의 내재성의 GTPase를 활성화시켜 GTP를 GDP로 전환함으로써 불활성화된다(John F. Hancock. RAS proteins: different signals from diffrent locations. Nature Molecular Cell Biology 4; 373-384(2003)).

RAS의 발현은 조직 특이적으로 발현되는데 각 조직의 암에서 RAS 돌연변이가 많이 발견되었다. Kras의 경우 대장, 흉선 그리고 폐에서 많이 발현이 되며 결장암, 췌장암, 폐암에서 Kras의 G12V 단 염기 돌연변이는 RAS 단백질의 구조적인 변화에 의해 GAP의 친화도와 활성도가 감소하게 된다. 따라서, RAS의 GTPase 활성이 감소되어 RAS는 계속적으로 단백질의 활성을 유지시킨다(Fabbro, D. et al., P A. Protein tyrosine kinase inhibitors: new treatment modalities, Curr. Opin. Pharmacol. 2; 374-381 (2002); Fahd Al-Mulla et al., Structural differences between valine-12 and aspartate-12 ras protein may modify carcinoma aggression. J. Pathol 18; 433-438(1999)).

Kras 유전자와 인간 암의 관계를 살펴보면 NSCLC(Non small cell carcinoma)의 33%, 결장암의 44% 그리고 췌장암의 90%에서 K-ras 유전자의 돌연변이가 발견된다. 다른 ras 종류의 경우 N-ras는 흑색종(13%), 간암(30%), 그리고 형질구성 백혈병(30%)에서 그 변이가 발견되며, H-ras는 방광(10%)그리고 신장암(10%)에서 발견된다(Mu D.Q. et al., Xualues of mutations of K-ras oncogene at codon 12 in detection of pancreatic cancer. World J Gastroenterol. 10(4); 471-5. (2004)).

이 중 특히 폐암의 경우 통증이 없어 발병의 진단이 늦고 또한 현재 폐암의 85% 이상이 사망하는 것으로 알려져 있다. 실제 대장암, 유방암 그리고 전립선암보다 더 많은 사람이 폐암으로 사망한다.

폐암은 암세포의 현미경적 형태에 따라 크게 비소세포성폐암(Non Small Cell Lung Cancer;NSCLC)와 소세포성폐암(Small Cell Lung Cancer;SCLC)으로 나뉘는데, 비소세포성폐암은 폐암의 65~75%, 소세포성폐암은 15~30%, 기타 폐암이 10%를 차지한다.

상기 비소세포성폐암은 편평상피암(squamous cell carcinoma)과 선암(adenocarcinoma) 그리고 대세포암종(large cell carcinoma)으로 분류한다(Minamoto T., Mai M. and Ronai Z. K-ras mutation: early detection in molecular diagnosis and risk assessment of colorectal, pancreas, and lung cancers a review. Cancer Detect Prev, 24(1); 1-12. (2004)).

상기 편평상피암은 폐암 중 가장 흔한 형태로 폐 중심부에 잘 생기며, 남자 폐암에 가장 맣은 형태로 흡연과 가장 관련이 많은 것으로 알려져 있다. 또한, 선암은 폐의 주변에서 주로 발견되며 간, 뇌, 뼈 등으로 전이가 잘 일어난다. 마지막으로 대세포암종은 전체 폐암의 4~10%를 차지하며, 폐 주변에 주로 발생하며, 암세포가 빠르게 증식하여 전이되는 속도가 빠른 경향이 있다.

상기 소세포성폐암은 주로 기도에서 처음 발생한다. 대개 기관의 표면이나 선을 따라서 생성되며 대부분은(80%) 폐 중앙부에 생기고 나머지(20%)는 주변부에서 발생한다. 전반적으로 악성도가 강해서 림프계통이나 혈액순환을 통해서 조기에 멀리 전이되는 경향이 있다. 소세포성폐암 암세포는 대개는 작고 동그란 형태로 자라는데, 그 모양을 본 따서 연맥세포암(oat cell)이라고도 한다.

나머지 기타 암으로, 세기관지폐포암(bronchioloalveoiar carcinoma)은 천천히 자라는 암세포이며, 나이가 많은 사람에게서 종종 발견되며 진행정도도 다양하 다. 또한, 전체 폐암의 1% 미만으로 발생하는 거대세포암(giant cell carcinoma)은 주로 주변부에서 크게 나타나며, 악성도가 심해서 빨리 진행된다.

이 중 편평상피암의 46%와 선암의 44%가 K-ras의 돌연변이로 인해 발병되는데, 그 중 40%가 K-ras의 단 염기 변이로 알려져 있다(Minamoto T., Mai M., and Ronai Z. K-ras mutation: early detection in molecular diagnosis and risk assessment of colorectal, pancreas, and lung cancers. Cancer Detect Prev. 24(1); 1-12. (2000)).

Kras는 세포 내에서 세포의 성장, 분열, 분화의 신호전달에 관여하는 상위단계의 단백질로, Kras의 돌연변이 중 12번, 13번, 61번 코돈이 핫 스팟(hot spot)으로 알려져 있다. 특히 Kras의 12, 13, 61번째 코돈의 돌연변이는 GTP의 가수분해에 관여하는 도메인을 암호화하는 부위에서 구조적 변화를 일으켜 세포의 계속적인 성장을 유도하여 암세포로 발전시키게 된다.

한편, Tetrahmena thermophila로부터의 그룹 I 인트론 리보자임이 실험관 내에서 자기 자신의 3'끝에 부착되어 있는 엑손 RNA를 트랜스하게 존재하는 5'엑손을 표적으로 한 후 부착시킴으로써 별도로 존재하는 전사체를 서로 연결시킬 수 있는 트랜스-스플라이싱 반응을 일으킬 수 있음이 보고되었다(Been, M. and Cech, T. One binding site determines sequence specificity of Tetrahymena pre-rRNA self-splicing, trans-splicing, and RNA enzyme activity. Cell 47; 207-216. (1986)). 이러한 리보자임은 포유류 세포 내에서도 일어날 수 있음이 밝혀졌 다(Jones, J.T., Lee, S.W., and Sullenger, B.A Tagging ribozyme reaction sites to follow trans-splicing in mammalian cells. Nat. Med. 2; 643-648. (1996); Lee, S.W. Effects of substrate RNA structure on the trans-splicing reaction by group I intron of Tetrahymena thermophila. Korean Journal of Microbiology 35; 211-217. (1999)).

이는 곧 그룹 I 리보자임의 트랜스-스플라이싱 반응을 통하여 여러 유전질환과 관련있는 유전자의 전사체도 보정될 수 있음을 시사하며 실제로 최근에 겸상적혈구(sickle cell) 질병을 갖은 환자의 적혈구 전구(erythrocyte precursor) 세포에서 β-글로빈 유전자 전사체가 트랜스-스플라이싱 리보자임에 의해 항-겸상적혈구화(anti-sickling), γ-글로빈 유전자 전사체로 전환될 수 있음이 밝혀졌다(Lan, N., Howrey, R.P., Lee, S.W., Howrey, R.P., Lee, S.W., Howrey, R.P., Smith, C.A., and Sullenger, B.A. Ribozyme-Mediated Repair of Sickle β-Globin mRNAs in Erythrocyte Precursors. Science 280; 1593-1596. (1998); Lan, N., Rooney, B.L., Lee, S.W., Howrey, R.P., Smith, C.A., and Sullenger, B.A. Enhancing RNA repair efficiency by combining trans-splicing ribozymes that recognize different accessible sites on a target RNA. Mol. Ther. 2; 245-255. (2000)).

또한, 여러 다른 부위를 인지하는 리보자임들을 함께 반응시켰을 경우 트랜스-스플라이싱 반응이 더욱 증가한다(Lan, N., Rooney, B.L., Lee, S.W., Howrey, R.P., Smith, C.A., and Sullenger, B.A. Enhancing RNA repair efficiency by combining trans-splicing ribozymes that recognize different accessible sites on a target RNA. Mol. Ther. 2; 245-255. (2000)).

그리고, 트랜스-스플라이싱 리보자임이 포유류 세포 내에 있는 근긴장성이양증 단백질 키나아제(myotonic dystrophy protein kinase) 유전자 전사체도 변형할 수 있다고 보고되었다(Phylactou, L.A., Darrah, C. and Wood, M.A.J.. Ribozyme-mediated trans-splicing of a trinucleotide repeat. Nat. Genet. 18; 378-381. (1998)). 선천성근긴장증(Myotonic congenita)을 유발하는 돌연변이 골격근 클로라이드 채널(mutant skeletal muscle chloride channel) RNA를 기능 면에서도 보정할 수 있었(Rogers, C.S., Vanoye, C.G., Sullenger, B.A., and George, Jr., A.L. Functional repair of a mutant chloride channel using a trans-splicing ribozyme. J. Clin. Invest. 110; 1783-1798. (2002)).

리보자임의 특이도 및 활성도를 증가하기 위해 표적 RNA 인지부위인 내부 가이드 염기서열(internal guide sequence;IGS)를 변형한 IGS를 갖는 리보자임을 식물 병원체인 오이 모자이크 바이러스 RNA를 표적으로 하여 사이토톡신을 트랜스-스플라이싱할 수 있도록 고안하였을 때, 효모 내에서 바이러스 RNA만 발현하는 세포만 특이적으로 그 성장을 방해할 수 있음이 발표되었다(Ayre, B.G., Kohler, U., Goodman, H.M., and Haseloff, J. Design of highly specific cytotoxins by using trans-splicing ribozymes. Proc. Natl. Acad. Sci. 96; 3507-3512. (1999)).

다른 예로 위와 같이 내부 가이드 염기서열이 변형되어 있고 돌연변이 p53 RNA를 표적으로 하여 트랜스-스플라이싱할 수 있는 리보자임을 인체의 결핍성 p53이 관련된 암세포에서 발현시킬 경우 돌연변이 p53 전사체를 효과적으로 보정함으 로써 정상적인 p53의 활성을 유도하였다(Watanabe, T., and Sullenger, B.A. Induction of wild-type p53 activity in human cancer cells by ribozymes that repair mutant p53 transcripts. Proc. Natl. Acad. Sci. 97; 8490-8494. (2000)). 이러한 연구는 트랜스-스플라이싱 리보자임이 돌연변이 유전자 RNA를 정상 RNA로 보정 가능함을 시사하고 있다.

또한, 최근에 C형 간염바이러스의 IRES RNA를 특이적으로 인지하는 리보자임을 개발하였으며 이러한 리보자임에 의해 HCV IRES RNA를 세포사 유발 RNA로 치환함으로써 표적 RNA를 발현하는 세포만 선택적으로 세포사가 유도할 수 있었다(Ryu, K.J., Kim, J.H., and Lee, S.W. Ribozyme-mediated selective induction of new gene activity in hepatitis C virus internal ribosome entry site-expressing cells by targeted trans-splicing. Mol. Ther. 7; 386-395. (2003)).

따라서, 이와 같은 결과는 트랜스-스플라이싱 리보자임에 의해 암 특이 유전자 전사체가 표적이 된 후 그 RNA가 다른 유전자 전사체로 치환되도록 재 프로그램될 수 있음을 시사한다.

지금까지 암 치료법에서 ras를 타겟으로 많은 연구들이 진행되어져 왔는데, RAS 단백질의 발현을 막고자 하는 안티센스 올리고뉴클레오타이드 등의 시도나 억제제를 이용하여 RAS의 멤브레인 로컬라이제이션(membrane localization)을 블록하려는 시도들이 보고되었으며, 또한 ras의 기능을 막기 위해 하위단계의 상호작용을 하는 단백질들의 억제제들을 이용한 시도 또한 연구되어왔다.

그러나, 이러한 시도들은 특정 RAS를 표적하거나 변이된 RAS를 특이적으로 제거 또는 억제하지 못하여 정상세포에까지 영향을 미치는 문제점들이 있었으며, 또한 발현되는 RAS의 일부분을 제거 억제하는 것만으로는 암이 치료되지는 않는다는 문제점이 있다. 그리고, 최근 많은 연구가 진행 중인 siRNA를 이용한 Kras G12V의 RNA 간섭은 돌연변이 Kras에 대해 특이적이지만 오프-타겟 효과와 변역반응을 건드리는 점에서 적당하지 않다.

이에, 본 발명자들은 상기와 같은 점을 감안하여 Tetrahymena thermophila의 그룹 I 인트론 리보자임을 이용한 유전자 치료 방법에 대해 예의 연구 노력한 결과, Kras의 12번째 코돈의 단 염기 변이를 선택적으로 인지하는 트랜스-스플라이싱 리보자임을 개발하고, 이러한 리보자임이 in vitro와 in vivo에서 돌연변이 Kras 전사체를 인지, 절단하고 타겟 부위의 하류(down-steam) 부위에 리보자임의 3' 엑손을 연결할 수 있으며, 이때 리보자임의 엑손을 치료 유전자 또는 리포터 유전자로 치환함으로써 암세포를 선택적으로 치료하거나 진단할 수 있음을 확인하고, 이를 토대로 본 발명을 완성하였다.

따라서, 본 발명의 목적은 단 염기 변이 Ras RNA를 특이적으로 인식하여 치환할 수 있는 트랜스-스플라이싱 그룹 I 리보자임 및 이의 용도를 제공하는 것이다.

상기 목적을 달성하기 위하여, 본 발명은

단 염기 변이 Ras RNA를 특이적으로 인식하여 치환할 수 있는 트랜스-스플라이싱 그룹 I 리보자임 및 이의 용도를 제공한다.

또한, 본 발명은 상술한 리보자임을 발현하는 벡터 및 이의 용도를 제공한다.

이때, 상기 리보자임은 Kras G12V RNA를 특이적으로 표적하며, 3' 엑손에는 치료 유전자 또는 리포터 유전자를 함유할 수 있다.

이하, 본 발명을 더욱 상세하게 설명하면 다음과 같다.

본 발명의 리보자임은 암 진행 여부를 진단할 수 있는 라스(Ras) 유전자의 단 염기 변이, 특히 Kras 유전자의 12번째 코돈의 단 염기 변이 Ras RNA를 특이적으로 인식하여 치환할 수 있는 트랜스-스플라이싱 그룹 I 리보자임이다.

도 1은 트랜스-스플라이싱 리보자임 매개 RNA 리프로그램을 나타낸 모식도이다.

도 1을 참조하면, 본 발명의 리보자임은 발암의 원인이 되는 온코진(oncogene)인 Kras의 돌연변이 중 12번째 코돈이 GGT에서 GTT로 변이된 KRas 전사체를 인지, 절단하고 타겟 부위의 하류(down-steam) 부위에 리보자임의 3'엑손을 연결한다. 이때 리보자임의 3' 엑손은 치료 유전자 또는 리포터 유전자를 함유함으로써 돌연변이 KRas 전사체를 보정하여 암세포만을 선택적으로 세포사를 유발하거나 진단하여, 치료 또는 진단할 수 있다.

그룹 I 인트론은 6개의 내부 가이드 염기서열(internal guide sequence; IGS)로 타겟을 인지하게 되는데, 이때 5' 끝의 G·U 와블 염기쌍(wobble base pair)은 필수이다. 따라서, 본 발명의 리보자임은 12번째 코돈이 GTT로 변이된 K-ras 12V를 타겟팅하는데 있어서 35번째 nt인 T와 G·U 와블 염기쌍을 이루도록 내부 가이드 염기서열로 'GCAGCU'을 갖거나, 36번째 nt인 T와 G·U 와블 염기쌍을 이루도록 내부 가이드 염기서열로 'GACAGC'를 갖게 하였다.

여기서, 치료 유전자(therapeutic gene)는 암 세포 내에서 발현되어 치료학적 효과를 나타내는 뉴클레오타이드 서열을 말한다. 예를 들면, 약제감수성 유전자, 세포사멸 유전자, 세포증식 억제 유전자, 세포독성 유전자, 종양 억제인자 유전자, 항원성 유전자, 항신생 혈관 생성 유전자, 사이토카인 유전자 등이 포함되 며, 이에 한정되는 것은 아니다.

상기 약제감수성 유전자(drug-sensitizing gene)는 독성이 없는 전구체(prodrug)를 독성물질로 전환시키는 효소에 대한 유전자로 유전자가 이입된 세포가 사멸하게 되므로 자살 유전자(suicide gene)로도 불린다. 즉, 정상세포에는 독성이 없는 전구체를 전신적으로 투여했을 때 암세포에만 전구체가 독성 대사체(toxic metabolite)로 전환되어 약제에 대한 감수성을 변화시킴으로써 암세포를 파괴시키는 방법이다. 대표적으로 HSV-tk(Herpes simplex virus-thymidine kinase) 유전자와 갠시클로비르(ganciclovir), 대장균의 사이토신 탈아미노효소 (cytosine deaminase, CD) 유전자와 5-플루오로시토신(5-fluorocytosine, 5-FC)가 가능하다.

상기 세포사멸 유전자(proapoptotic gene)는 발현되어 프로그램된 세포 소멸을 유도하는 뉴클레오타이드 서열을 말한다. 당업자에게 공지된 세포사멸 유전자로, p53, 아데노바이러스 E3-11.6K (Ad2 및 Ad5에서 유래) 또는 아데노바이러스 E3-10.5K (Ad에서 유래), 아데노바이러스 E4 유전자, p53 경로 유전자 및 카스파제를 코딩하는 유전자가 포함된다.

상기 세포증식 억제 유전자(cytostatic gene)는 세포 내에서 발현되어 세포 주기 도중에 세포 주기를 정지시키는 뉴클레오타이드 서열을 의미한다. 대표적으로 p21, 망막아세포종 유전자, E2F-Rb 융합 단백질 유전자, 사이클린-종속성 카이네이즈 억제인자를 코딩하는 유전자(예를 들면, p16, p15, p18 및 p19), 성장 중지 특이성 호메오박스(growth arrest specific homeobox, GAX) 유전자 (국제특허출원 공개 WO 97/16459호 및 WO 96/30385호) 등이 있다.

상기 세포독성 유전자(cytotoxic gene)는 세포 내에서 발현되어 독성 효과를 나타내는 뉴클레오타이드 서열을 말한다. 일예로, 슈도모나스 외독소(exotoxin), 리신 독소, 디프테리아 독소 등을 코딩하는 뉴클레오타이드 서열 등이 있다.

상기 종양 억제인자 유전자(tumor suppressor gene)는 표적 세포 내에서 발현되어 종양 표현형을 억제할 수 있거나 세포사멸을 유도할 수 있는 뉴클레오타이드 서열을 의미한다. 대표적으로 종양 괴사 인자(tumor necrosis factor-a, TNF-a), p53 유전자, APC 유전자, DPC-4/Smad4 유전자, BRCA-1 유전자, BRCA-2 유전자, WT-1 유전자, 망막아세포종 유전자(Lee et al., Nature, 329,642, 1987), MMAC-1 유전자, 선종양 폴립증 코일 단백질(adenomatous polyposis coil protein)(Albertsen et al. 미국특허공보 US 5,783,666호), 결손된 결장 종양 (DCC) 유전자, MMSC-2 유전자, NF-1 유전자, 염색체 3p21.3에 위치한 비인후 종양 억제인자 유전자(Cheng et al. Proc.Nat.Acad.Sci., 95,3042-3047, 1998), MTS1 유전자, CDK4 유전자, NF-1 유전자, NF-2 유전자 및 VHL 유전자가 포함된다.

상기 항원성 유전자(antigenic gene)는 표적 세포 내에서 발현되어 면역 시스템에서 인식할 수 있는 세포 표면 항원성 단백질을 생산하는 뉴클레오타이드 서열을 말한다. 당업자에게 공지된 항원성 유전자의 예에는 암태아성 항원 (carcinoembryonic antigen, CEA) 및 p53(Levine, A., 국제특허출원공개 WO 94/02167호)이 포함된다.

상기 사이토카인 유전자(cytokine gene)는 세포 내에서 발현되어 사이토카인 을 생성하는 뉴클레오타이드 서열을 의미한다. 대표적으로 GM-CSF, 인터류킨(IL-1, IL-2, IL-4, IL-12, IL-10, IL-19,IL-20), 인터페론 α, β, γ(인터페론 α-2β) 및 인터페론 α-2α-1과 같은 융합체 등이 포함된다.

상기 항신생혈관 생성 유전자(anti-angiogenic gene)는 발현되어 항-신생혈관 생성 인자를 세포밖으로 방출하는 뉴클레오타이드 서열을 말한다. 그 예로 안지오스타틴, 혈관 내피 성장 인자(VEGF)의 억제 인자, 엔도스타틴 등이 포함된다.

또한, 본 발명에서는 상술한 리보자임 활성을 통해 타겟 부위의 하류(down-steam) 부위에 리포터 유전자를 접합할 수 있다. 이렇게 접합된 리포터 유전자는 리포터 단백질로 발현되므로, 발현된 리포터 단백질의 활성 또는 양을 측정함으로써 암 세포를 진단할 수 있다.

이때 리포터 유전자는 본 발명이 속하는 분야에서 공지된 것을 사용할 수 있으며, 바람직하게는 LacZ, 클로람페니콜 아세틸 전이효소(CAT: chloramphenicol acetyl transferase), 레닐라 루시퍼라제(Renila luciferase), 반딧불이(firefly) 루시퍼라아제, 적색형광단백질(RFP), 녹색형광단백질(GFP), 분비성 태반 알칼라인 포스파타아제(secreted placental alkaline phosphatase, SEAP) 또는 HSV-tk(Herpes simplex virus-thymidine kinase)를 암호화하는 유전자를 사용한다. 이 중 활성 분석의 편이성 및 민감도 면에서 LacZ를 사용하는 것이 바람직하다.

이러한 리포터 단백질들의 활성은 하기와 같이 당업자에게 공지되어 있는 방법을 사용하여 측정할 수 있다:

반딧불이 루시퍼라아제(de Wet J. et al., Mol. Cell Biol., 7, 725-737, 1987 참조), 레닐라 루시퍼라아제([Lorenz W.W. et al., PNAS 88, 4438-42, 1991) 참조), 클로람페니콜 아세틸 전이효소(Gorman C. et al.,Mol. Cell Biol., 2, 1044-1051, 1982 참조), LacZ(Hall C.V. et al., J. Mol. Appl. Genet ., 2,101-109, 1983 참조), 인간 성장 호르몬(Selden R. et al.., Mol. Cell Biol., 6, 3173-3179, 1986 참조), 녹색 형광 단백질(Chalfie M. et al., Science, 263, 802-805, 1994 참조) 및 분비성 태반 알칼린 포스파타아제(Berger, J. et al., Gene, 66, 1-10, 1988 참조). 또한, 티미딘 키나제(thymidine kinase)를 리포터 단백질로 한 경우 PET(positron emission tomography) 이미징 법을 이용할 수 있다.

본 발명의 실시예 1에 따르면, 발암의 원인이 되는 온코진(oncogene)인 Kras의 돌연변이 중 12번째 코돈이 GGT에서 GTT로 변이된 Kras를 타겟으로 하여 KRib1, KRib2 트랜스-스플라이싱 리보자임을 제작하였다.

이때 12번째 코돈이 GTT로 변이된 K-ras 12V를 타겟팅하는데 있어서 35번째 nt인 T와 G·U 와블 염기쌍을 이루도록 디자인한 KRib1, 36번째 nt인 T와 G·U 와블 염기쌍을 이루도록 디자인한 KRib2를 제작하였다.

본 발명의 실시예 2에 따르면, 제작된 리보자임을 in vitro에서 트랜스-스플라이싱 반응 실험을 수행하였고, Kras G12V를 타겟팅하는 리보자임 KRib1, KRib2이 특이적으로 Kras G12V를 인지하여 절단하고 리보자임의 3' 엑손을 정확한 타겟 부위에 연결한다는 것을 확인할 수 있었다.

본 발명의 실시예 3에 따르면, IGS만 갖는 리보자임과 효율성을 높이기 위한 P1, P10, AS(안티센스) 구조를 추가한 변형(modification) 리보자임을 제작하고, 이들을 가지고 in vitro 트랜스-스플라이싱 반응을 수행하였다. 그 결과, P1, P10, AS가 있는 리보자임이 특이성과 효율성 면에서 우수하다는 것을 확인하였다.

본 발명의 실시예 4에 따르면, 세포 내에서도 트랜스-접합 산물 즉, 트랜스진의 발현이 유도되는지를 리포터 유전자를 사용하여 리포터 분석을 수행하였고, 그 결과 세포주에서 루시퍼라제의 발현을 확인하여, 세포 내에서도 본 발명의 리보자임이 특이적으로 트랜스-스플라이싱을 일으키며, 트랜스진의 발현이 유도된다는 것을 알 수 있었다.

또한, 트랜스-스플라이싱을 RNA 레벨에서 보기 위해 293 세포에 리보자임들과 Kras(G12V, 야생형) RNA를 코-트랜스펙션시킨 후 RT-PCR을 했다. 그 결과 1 세트에서 AS300KRib1-F.luc과 2 세트에서 AS100KRib2-F.luc이 in vitro 트랜스-스플라이싱 분석을 수행했던 것과 일치하는 결과인 트랜스-스플라이싱 밴드라고 예상 되어지는 크기의 밴드가 나왔다. 이 밴드가 비특이성 밴드가 아닌 트랜스-스플라이싱 산물임을 확인하기 위해 시퀀싱을 하여 Kras G12V의 타겟 부위에서 정확하게 트랜스-스플라이싱이 일어남을 확인하였다.

본 발명의 실시예 5에 따르면, 리보자임의 3'엑손에 TK 유전자(HSV-TK(herpes simplex virus thymidine kinase))를 도입하는 클로닝된 pAVQ CMV AS300KRib1-TK와 pAVQ CMV AS100KRib2-TK들 중 AS100KRib2-TK를 가지고 TK 분석을 통해 세포 생존력을 관찰하였다. TK 분석 결과, HT-29 세포에서는 GCV를 처리하거 나 미처리 시에 GFP의 발광에는 어떠한 변화도 없는 것으로 보아 세포의 생존력에 영향을 끼치지 않음을 볼 수가 있었다.

그러나, SW480 세포에서는 AS100KRib2-TK를 트랜스펙션한 세포에서 GCV를 처리하였을 때 GCV를 처리하지 않은 세포에 비해 GFP의 발광 수가 감소한 것을 볼 수가 있었다.

한편, 유전자 치료에서 중요시 되어야 할 점은 재조합 유전자를 세포에 전달시키는 방법이다.

일반적인 방법으로 크게 물리화학적인 방법으로 재조합 DNA를 직접 투여하는 방법과 생물학적인 방법으로 재조합 바이러스 벡터를 들 수가 있다. DNA를 직접 투여하는 방법은 간단하기 때문에 위험성이 낮고 경제적이라는 장점을 가지고 있지만, 재조합 DNA의 전달률이 낮은 단점을 가지고 있다.

이에 본 발명은 상기 리보자임의 세포 전달 방법으로서 위험성도 적고 DNA 전달율도 우수한 아데노 바이러스 전달 시스템을 포함한다.

아데노바이러스는 지름이 약 70~80 nm로 크며 내부는 20~26×10⁶ 달톤의 dsDNA를 가진 DNA 바이러스이다. 게놈의 양끝에는 ITR(inverted terminal repeat)이 있고, 이곳에서 바이러스 복제가 시작된다. 아데노바이러스의 게놈은 E1-E4의 초기부위, L1-L5의 후기부위로 구분된다.

아데노바이러스 벡터의 장점은 첫째, 인체 내 종양 발생에 무관한 바이러스로 알려져 있고, 오랜 바이러스 백신의 개발로 위험성이 낮은 바이러스이다. 둘째, 높은 역가의 바이러스를 제조할 수 있다. 셋째, 감수분열이 끝난 세포와 거의 모든 인체의 세포에 감염시킬 수 있다. 넷째, 아데노바이러스 벡터에 10kb에 가까운 DNA를 삽입할 수 있다.

단점은 첫째, 아데노바이러스가 숙주의 게놈에 통합하지 못함으로 영구적인 효과를 보지 못한다. 둘째, 바이러스에 대한 항원·항체반응으로 치료효과가 짧다. 셋째, 많은 적가 투여 시 부작용을 고려하지 않을 수 없다.

본 발명의 실시예 6에 따르면, BJ5183이라는 박테리아에서 리보자임 트랜스퍼 벡터와 pAdenovactor ΔE1/E3 백본 벡터의 코-트랜스포메이션을 하여 상동성 재조합을 시켜 재조합 리보자임을 만들었고, 이를 293 세포에 트랜스펙션함으로써 재조합 리보자임 바이러스를 얻게 되었다.

이렇게 얻은 재조합 리보자임 바이러스는 293 세포에 감염하여 증폭하는데 증폭하는 과정에서 재조합 리보자임 바이러스가 제대로 만들어지고 있는지 확인하기 위해서 바이러스 게놈 DNA를 추출하여 리보자임의 TK 유전자와 바이러스의 LITR을 PCR을 통해 확인하였다.

또한, SW480 세포에 감염하여 RNA 레벨에서도 TK 유전자를 RT-PCR을 통해 확인하였다. 이렇게 확인된 재조합 리보자임 바이러스들은 Vivapure^® AdenoPACK^TM 100 RT를 사용하여 정제하였다.

이렇게 정제된 Ad-AS300KRib1-TK와 Ad-AS100KRib2-TK 바이러스들은 종양 세포에 감염하여 종양 세포에서 Ad-리보자임의 특이성과 효울성을 보았다. SW480 세포에 감염한 결과 Ad-AS300KRib1-TK와 Ad-AS100KRib2-TK에서 트랜스-스플라이싱 밴 드를 보았고, 이것을 시퀀싱을 통해 타겟 부위에서 일어났는지를 확인하였다.

본 발명의 실시예 7에 따르면, Ad-리보자임들이 특이적으로 트랜스-스플라이싱을 일으켜 트랜스진을 유도함을 보기 위해서 MTS 분석을 진행하였다. 그 결과, Kras G12V 변이된 세포주인 SW480 세포와 CAPAN-1 세포에서는 대조군과 비교하였을 때 Ad-리보자임들만이 세포사가 유도됨을 볼 수 있었고, Kras 야생형인 HT-29 세포에서는 대조군과 비교하여 세포사가 일어나지 않음을 관찰하였다. 이는 Ad-리보자임이 Kras G12V를 특이적으로 인지하여 트랜스-스플라이싱하고 트랜스진을 유도하여 특이적으로 세포사를 일으킬 수 있음을 나타낸다.

본 발명의 실시예 8에 따르면, Ad-리보자임의 in vivo에서 특이성과 효율성을 알아보기 위해 이종이식 마우스 모델(xenograft mice model) 실험을 수행하였다.

그 결과, Kras 변이된 종양에서는 대조군과 비교하여 Ad-AS100KRib2-TK의 종양 무게가 감소하였다. 그러나, Kras 야생형 종양의 경우에는 음성 대조군인 Ad-CMV-LacZ와 양성 대조군인 Ad-CMV-TK와 비교했을 때, Ad-리보자임들이 Kras 야생형 종양에서는 작용을 하지 않는 것을 확인할 수 있었다.

이러한 결과는 본 발명에 따른 Ad-리보자임이 Kras G12V만을 발현하는 종양을 특이적으로 퇴행(regression)시킬 수 있다는 것을 보여준다.

이와 같이 본 발명의 리보자임은 암 특이적 유전자를 치료 유전자로 보정하여 유전자 발현을 유도한다. 그 결과 암 세포만을 선택적으로 세포사를 유발할 수 있다.

따라서, 본 발명에서는 상기 리보자임을 유효성분으로 포함하는 항암용 약학적 조성물을 제공한다.

본 발명의 항암용 약학적 조성물은 투여를 위해서 상기 기재한 유효 성분 이외에 추가로 약제학적으로 허용 가능한 담체를 1종 이상 포함하여 약제학적 조성물로 바람직하게 제제화할 수 있다.

본 발명의 조성물에 포함되는 약제학적으로 허용되는 담체는 제제시에 통상적으로 이용되는 것으로서, 액상 용액으로 제제화되는 조성물에 있어서 허용 가능한 약제학적 담체로는, 멸균 및 생체에 적합한 것으로서, 식염수, 멸균수, 링거액, 완충 식염수, 알부민 주사용액, 덱스트로즈 용액, 말토 덱스트린 용액, 글리세롤, 에탄올 및 이들 성분 중 1성분 이상을 혼합하여 사용할 수 있으며, 필요에 따라 항산화제, 완충액, 정균제 등 다른 통상의 첨가제를 첨가할 수 있다. 또한, 희석제, 분산제, 계면활성제, 결합제 및 윤활제를 부가적으로 첨가하여 수용액, 현탁액, 유탁액 등과 같은 주사용 제형, 환약, 캡슐, 과립 또는 정제로 제제화할 수 있으며, 표적 기관에 특이적으로 작용할 수 있도록 표적 기관 특이적 항체 또는 기타 리간드를 상기 담체와 결합시켜 사용할 수 있다.

본 발명의 약제학적 조성물은 비경구 투여가 바람직하고, 예컨대 정맥내 투여, 복강내 투여, 종양내 투여, 근육내 투여, 피하 투여, 또는 국부 투여를 이용하여 투여할 수 있으나, 이에 한정되는 것은 아니다.

본 발명 항암용 약학적 조성물의 투여량은 질환의 종류, 질환의 중증도, 조성물에 함유된 유효 성분 및 다른 성분의 종류 및 함량, 제형의 종류 및 환자의 연령, 체중, 일반 건강 상태, 성별 및 식이, 투여 시간, 투여 경로 및 조성물의 분비율, 치료기간, 동시 사용되는 약물을 비롯한 다양한 인자에 따라 조절될 수 있다.

본 발명의 약학적 조성물은 단독으로, 또는 외과적 수술요법 등의 보조 치료 방법들과 병행하여 사용할 수 있다. 본 발명의 조성물과 함께 이용될 수 있는 화학 요법제(chemotherapeutic agent)는 시스플라틴(cisplatin), 카르보플라틴 carboplatin), 프로카르바진(procarbazine), 메클로레타민(mechlorethamine), 시클로포스파미드(cyclophosphamide), 이포스파미드 (ifosfamide), 멜팔란(melphalan), 클로라부실(chlorambucil), 비술판(bisulfan), 니트로소우레아 (nitrosourea), 디악티노마이신(dactinomycin), 다우노루비신(daunorubicin), 독소루비신(doxorubicin), 블레오마이신(bleomycin), 플리코마이신(plicomycin), 마이토마이신(mitomycin), 에토포시드(etoposide), 탁목시펜(tamoxifen), 택솔 (taxol), 트랜스플라티눔(transplatinum), 5-플루오로우라실(5-fluorouracil), 빈크리스틴(vincristin), 빈블라스틴(vinblastin) 및 메토트렉세이트(methotrexate) 등을 포함한다. 발명의 조성물과 함께 이용될 수 있는 방사 요법은 X-선 조사 및 -선 조사 등이다.

아울러, 본 발명의 리보자임은 암 특이적 유전자에 리포터 유전자를 접합하여 암세포만을 선택적으로 진단할 수 있다.

따라서, 본 발명에서는 상기 리보자임을 유효성분으로 포함하는 암 진단용 조성물을 제공한다.

이하 본 발명을 실시예를 통하여 보다 상세하게 설명한다. 그러나. 이들 실시예는 본 발명을 예시적으로 설명하기 위한 것으로 본 발명의 범위가 이들 실시예에 한정되는 것은 아니다.

실시예 1: Kras G12V RNA를 특이적으로 인지할 수 있는 트랜스-스플라이싱 그룹 Ｉ 리보자임 제작

1. K-ras 타겟팅 리보자임의 디자인

리보자임의 내부 가이드 염기서열(Internal guide sequence; IGS)이 타겟 주형 RNA를 인식하여 트랜스-스플라이싱 기작을 일으키는데 있어서 타겟 RNA의 2차 구조가 영향을 미치게 되므로 mfold program(RNA 2차 structure form program)을 이용하여 2차 구조를 예측하였으며, 타겟하려고 하는 12번 코돈 부위가 리보자임이 트랜스-스플라이싱을 일으키는데 용이한 구조인지를 확인하였다.

기존 맵핑 방법을 통하여 리보자임이 타겟팅 하기에 적합한 부분을 찾아 분 석한 결과와 비교하여 볼 때 Kras의 12번째 코돈이 Gly에서 Val으로 단 염기 변이가 일어난 경우의 이차구조가 타겟팅 가능한 구조라는 것을 확인하고 특이 리보자임을 제작하기로 결정하였다.

Kras의 12번째 코돈이 GGT에서 GTT로 치환된 변이를 타겟팅하는 리보자임의 내부 가이드 염기서열(internal guide sequence; IGS)을 디자인하였다. 이때, IGS가 변이된 단 염기와 G·U 염기쌍을 이루게 한 KRib1(GCAGCT)과 변이된 다음의 염기와 G·U 염기쌍을 이루게 한 KRib2(GACAGC) 두 가지의 리보자임을 디자인하였다(도 3 참조).

2. K-ras 12V(기질) 구조체 제작

2-1. In vitro 돌연변이 형성 및 클로닝

도 4는 in vitro 돌연변이 형성을 통해 Kras 야생형으로부터 제작된 Kras 12V의 구조를 나타낸 것이다.

타겟 구조체를 만들기 위하여 GeneEditor in vitro 부위 유도 돌연변이 형성 시스템(site-Directed mutagenesis System, Promega)를 사용하여 Kras 야생형으로부터 12번째 코돈이 발린으로 치환된 Kras 12V를 제작하였다.

서열번호 1의 프라이머(표 1 참조)를 사용하였으며, 이를 pUC19에 5' BamHI, 3' HindIII로 클로닝하였다.

[표 1]

서열번호	프라이머 서열
1	5'-GTTGGAGCTGGTGGCGTAGGC-3	Kras wild
2	5'-TAATACGACTCACTA-3'	Kras 5' BamHI
3	5'-GGGTAATACGACTCACTATAGGAGGTAAAAGTTATCAGGCATGC-3'	kras rib T7-5'
4	5'-GGGTAATACGACTCACTATAGACAGCAAAAGTTATCAGGCATGC-3'	RY-PCR
5	5'-TGTAAAACGACGGCCAGTG-3	RY-RT
6	5'-CGATGATCACGAAGACGCCAAAAACATA-3'	kras p1p10-rib1-5
7	5'-CGACATGATCACGAAGACGCCAAAAACATA-3'	kras p1p10-rib2-5
8	5'-AAGGAAAAAAGCGGCCGCTTATTACAATTTGGACTTT-3'	krib 3'
9	5'-GGGAAGCTTGCAAGAGTGCCTTGACGA-3	kras as 5'
10	5'-GGGAAGGCTTCCATCAATTACTACTTGC-3	kras as 3'
11	5'-GGGTAATACGACTCACTATAGGGCTACTAGGACCATAGGTA-3'	kras as300 5'
12	5'-CGCGGATCCACTGAATATAAACTTGTGGTA-3'	kras t/s BamHI5'
13	5'-CCCAAGCTTGCCCAACACCGGCATAAAG-3	cell t/s luci 3' nest
14	5'-GTGATCATCGCGATGATCACGCTTCGTACCCCTGCCAT-3'	TK 5' NruI 1set
15	5'-GTGATCATCGCGACATGATCACGCTTCGTACCCCTGCCAT-3'	TK 5' NruI 2setI
16	5'-AAGGAAAAAATTCGAATTAGCCTCCCCCAT-3'	3' TK BstBI
17	5'-TAATACGACTCACTATAGGG-3'	T7-R-Primer
18	5'-CCTTTCCCGCAATTTGACGGTCTT-3'	IGS rev seq
19	5'-CGGGAATTCGTGGTAATGACAAGC-3'	5' TK stable
20	5'-CGGGGATCCATCCAGGATAAAGACGTGCATGGG-3'	3' TK stable
21	5'-GGAATTCTGGAGTTTGTGACGTGGCG-3'	5' LITR EcoRI
22	5'-GCTCTAGATGGCCAAATCTTACTCGGTTACGC-3'	3' LITR XbaI
23	5'-GGAATTCATGGTCGTTTTACAACGTCGTG-3'	5' viral LacZ
24	5'-GGGAAGCTTCGGATTGACCGTAATGGGA-3'	cancer.lacZ-3

2-1-1. 벡터와 삽입물의 제조

Kras PCR 산물 DNA 1μg을 BamHI(0.3u)/ HindⅢ(0.3u)와 10X B 버퍼 (Roche)와 20 ㎕까지의 dH₂O 37 ℃에서 3시간 동안 반응시켜 2.5% TBE 겔에 전기영동을 하여 크기를 확인하였다. 그 다음 3μg의 pUC19 벡터도 동일하게 BamHI(0.3u)/ HindⅢ(0.3u) 처리하여 잘린 것을 확인한 후 페놀 & EtOH 침전 반응을 시킨 후 정량을 하였다.

2-1-2. DNA 용출(insert)

겔 추출(DNA)(Omega bio tec. E.Z.N.A Kit)를 이용하여 DNA 크기에 맞는 농도로 0.5 X TAE 아가로스 겔을 만들어 DNA를 로딩한 다음 롱 웨이브 UV로 밴드를 확인한 후 원하는 밴드 부분만을 정확히 잘라냈다.

각각의 겔 조각을 1.5㎕ 튜브에 50 ~ 300 ㎎이 되도록 넣고 용출 버퍼 0.5 ㎕씩을 넣고 60 ℃에서 10분간 완전히 녹이는데, 200㎎ 이상일 경우 10분 이상 녹였다. 겔을 완전히 녹인 후 검체 튜브에 겔 추출 컬럼을 끼운 후 녹은 용액을 컬럼에 옮겨 13000 rpm으로 60초간 돌린 후 가라앉은 용액을 버리고 워쉬Ⅰ버퍼 0.5 ㎕ 을 각기 컬럼에 넣고 60초간 돌린 후 다시 걸러진 버퍼를 버렸다.

컬럼을 새로운 1.5㎕ 튜브에 옮겨 끼운 후 dH₂O 30㎕를 넣고 1분간 보관 후 2분 정도 원심분리를 돌려 DNA를 얻어낸 후 정량 하여 -20 ℃에 보관하였다.

2-1-3. 연결

효소 처리된 pUC19과 삽입물(trans-splicing product)을 각각 벡터 : 삽입물(1:3) 비율로 T₄리가아제(Roche)로 4℃에서 16시간 동안 반응시켰다.

2-1-4. 형질전환

연결 산물을 대장균(JM109)에 형질전환하기 위해 수용성 세포(competent cell)로 제작된 JM109에 연결 산물 1㎕를 넣고 전기형질전환장치(electrophorator)를 25μF, 1.8Kv, 펄스 조절기(pulse controller) 400Ω으로 셋팅하여 형질전환한 후 1ml 후레쉬 배지에 넣어 잘 섞은 후 1시간 동안 배양하였다.

X-gal, IPTG를 넣어 만든 아가 플레이트에 골고루 깐 다음 37℃ 배양기에 16시간 동안 배양시킨 후 컬러 스크리닝을 통하여 백색 콜로니를 선택적으로 5ml 암피실린이 포함된 LB 배지에 접종하여 16시간 동안 37℃ 흔들면서 배양하였다.

2-2. 염기서열 확인

In vitro 돌연변이 형성을 통하여 제작된 Kras 12V의 염기서열을 확인하였 다. 종결자 준비 반응 혼합물(terminator ready reaction mixture, PE applied Biosystems) 3㎕, 정량된 DNA 100ng, 서열번호 2의 프라이머(표 1 참조) 3.2pmol을 총 10㎕ 반응으로 (96℃-10', 50℃-5', 60℃-4')× 25 사이클 반응하였다.

이후 정제를 위하여 dH₂O 40㎕ 첨가한 다음 3M NaoAC(1/10 volume), 100% EtOH(2 volume)를 넣고 보르텍스 후 13000 rpm, 4℃에서 30분간 원심분리하고 70% EtOH(400㎕)로 세척한 다음 EtOH를 제거하고 speed-vacuum으로 말린 후 주형 억제 시약(template suppression reagent) 15㎕에 녹였다. 이어서 보르텍스 & 스핀 다운 후 시퀀싱 튜브에 옮겨 자동 시퀀서(ABI 310 Genetic Analyzer)로 시퀀싱을 시행하였다.

3. 기질 RNA(K-ras)의 제조

3-1. K-ras 기질 제조

기질로 이용될 Kras 12V와 대조군으로 사용될 Kras 야생형의 DNA를 Hind III로 잘라서 페놀 추출, EtoH 침전하여 준비하였다.

3-2. 기질 전사

상기 2-1과 같은 방법으로 제조된 DNA를 전사를 통하여 RNA를 만들게 되는데 DNA 주형(1㎍), 1x 전사 버퍼, 10mM DTT (Sigma), 0.5mM ATP, GTP, CTP, UTP (Roche), 80U RNase 억제제(Kosco), T7 RNA 중합효소(Takara), DEPC-H₂O를 100㎕까지 넣어서 섞은 후 37℃에서 3시간동안 반응시켰다.

이를 위해 5'프라이머 제작 시에 T7 프로모터 부분을 붙이게 되며, 이후 남 아있는 DNA를 제거하기 위해 5U DNase I(promega)를 37℃에서 30분간 처리하여 DNA 주형을 완전히 제거한 후 페놀 추출(PH7.0), 에탄올 침전을 통하여 RNA만을 순수 분리 정제한 후 TE 버퍼(10mM Tris-Hcl PH7.5, 1mM EDTA)에 농축시켜 놓았다.

4. KRib1, KRib2 (Kras targeting ribozyme) 제조

4-1. 리보자임 기질 제조

Kras의 12번 코돈을 인지하여 트랜스-스플라이싱할 수 있는 내부 가이드 염기서열(Internal guide sequence;IGS)를 가진 리보자임을 제작하기 위해 서열번호 3, 4의 프라이머(표 1 참조)를 사용하여 pt7L21로부터 PCR하여 KRib1, KRib2를 제작하였다. 또한, d)pt7L21로부터 PCR을 통하여 촉매의 코어(catalytic core)가 제거된 d)KRib1, d)KRib2를 제작하여 대조군으로 사용하였다.

4-2. 리보자임 전사

기질과 같은 방법으로 전사를 하는데, 이때 dNTP 양을 1.5mM로 늘려서 자가-스플라이싱을 최대한 방지하며,1x 스플라이싱 버퍼(40Mm Tris (ph7.5), 5mM MgCl₂, 10mM DTT, 4mM spermidin)를 사용하였다.

5. RNA 용출

원하는 RNA만을 얻기 위해, RNA 용액의 1/2 볼륨의 변성 RNA 로딩 염료(10㎕ Formamide, 10mM EDTA, 0.01% Bromophenol blue, 0.01% Xylenecyanol)를 섞은 후 95℃에서 5분간 처리하여 변성시켰다.

변성된 RNA를 우레아가 들어있는 6% 폴리아크릴아마이드 겔에 로딩하여 0.5x TBE 버퍼에서 40분간 230V로 전기영동한 후 원하는 크기의 RNA 밴드 부분의 겔을 잘라서 용출 버퍼(0.6M Ammonium acetate, 1mM EDTA, 0.2% SDS) 400㎕에 넣어서 37 ℃에서 3시간 동안 반응시켰다. 그 다음 젤은 스핀 다운으로 가라앉은 후 그 상층액을 따서 페놀 추출과 EtOH 침전을 통하여 정제하고 TE 버퍼에 최종적으로 농축한 후 UV-분광광도계를 이용하여 RNA를 정량하였다. 이때 리보자임은 비-자가-스플라이싱 산물만을 용출하여 UV-분광광도계를 용출하여 정량하였다.

6. 페놀 추출 및 에탄올 침전

용액과 동량의 페놀 : 클로로포름 : 이소아밀알콜 (25:24:1)을 넣고 보르텍스를 이용하여 완전히 섞어준 뒤 4℃ 10000ⅹg에서 10분간 원심분리하였다. 상층 액만을 조심스럽게 1.5ml 튜브에 옮긴 후 2배 부피의 100% EtOH(-70℃)과 1/10 볼륨의 3M 소듐 아세테이트를 넣어 잘 섞어준 뒤 -70℃에서 40분간 넣어주었다.

4℃에서 30분간 원심분리 후 상층액을 조심스럽게 제거하고 70% EtOH (-70℃) 400㎕를 넣어 골고루 벽면을 씻은 다음 10분간 원심분리를 한 후 EtOH을 모두 제거한 후 10초 정도 말리고 TE 버퍼에 녹였다.

실시예 2: In vitro 에서 트랜스-스플라이싱 리보자임 KRib1, KRib2의 특이성 확인

1. 트랜스-스플라이싱 반응

상기 실시예 1에서 디자인한 리보자임 KRib1, KRib2의 특이성을 확인하기 위하여, 먼저 시험관 상에서 트랜스-스플라이싱 반응을 하였다. In vitro 돌연변이 형성을 통하여 제작된 Kras G12V RNA(10nM)와 KRib1과 KRib2 리보자임(50nM)의 RNA 를 시험관 내에서 트랜스-스플라이싱 반응을 하였다.

특이 IGS를 갖는 리보자임(KRib1, KRib2)과 Kras의 트랜스-스플라이싱 반응을 위하여 기질 RNA와 리보자임의 절대량을 10nΜ : 50nΜ로 정하였으며 기질 RNA(2x reaction buffer, substrate RNA, 0.2mM dGTP, DEPC-H₂O)는 37℃에서 미리 5분간 예열시켰다. 리보자임 RNA(2x reaction buffer, Ribozyme RNA, DEPC-H₂O)는 50℃에서 5분간 반응시킨 후 37℃에서 2분간 반응시킴으로써 올바른 3차 구조 형성을 유도한 후 두 반응물을 잘 섞어준 다음 37℃에서 3시간 동안 반응시켰다.

2. 역전사

트랜스-스플라이싱된 RNA 전사체의 3' 엑손(Ribozyme) 부분에 결합하는 서열번호 5의 3'프라이머(표 1 참조)와 5x RT 버퍼, 트랜스-스플라이싱 산물(10㎕)를 잘 섞은 다음 65℃에서 5분간 반응시킨 다음 상온에서 10분간 보관하였다.

상온 보관 후 0.5mM dNTP(BMS)와 MMLV 역전사효소 1U(Promega)와 DEPC dH₂0로 총 50㎕를 만들어 37℃에서 1시간 동안 반응시켰다. 이후 95℃에서 5분간 반응시켜 MMLV-RT의 활성을 완전히 제거 후 4℃에서 보관하였다.

3. PCR

RT 반응을 마친 반응물(20㎕)을 서열번호 2와 5의 프라이머(표 1 참조)를 사용하여 MgCl₂ 50mM, 5% DMSO를 넣고(63℃ 20 sec annealing/ 25 cycle) PCR을 수행하였다.

4. 클로닝(RT-PCR 산물) 및 염기서열 분석

pUC19에 RT-PCR 산물 클로닝(상기 실시예 1의 2-1과 동일)을 하였으며, 서열번호 2의 프라이머(표 1 참조)(3.2 pmol)를 사용하여 시퀀싱(상기 실시예 2의 2-2와 동일)을 수행하였다.

도 5는 in vitro에서 KRib1, KRib2의 Kras G12V 트랜스-스플라이싱 산물의 분석을 나타낸 것이다[A: RT-PCR 분석, B-C: 서열분석(B: KRib1, C: KRib2)].

도 5A를 참조하면, Kras에 결합하는 5' 프라이머와 리보자임 3' 엑손에 결합하는 3' 프라이머를 이용하여 RT-PCR을 수행한 결과, KRib1과 KRib2와 Kras G12V RNA를 반응시킨 샘플에서만 트랜스-스플라이싱이 일어났을 때 나올 수 있는 산물 크기의 밴드를 얻었다.

반면, 촉매성 코어(catalytic core) 부위를 제거한 불활성 리보자임인 (d)KRib1, d)KRib2)과 Kras G12V RNA를 반응시켰을 때 혹은 KRib1과 KRib2와 야생형 Kras RNA만을 각각 반응시켰을 때는 어떠한 산물도 볼 수 없었다.

또한, 도 5B 및 5C를 참조하면, PCR 산물을 클로닝 후 시퀀싱한 결과, 정확하게 타겟 부위에서 트랜스-스플라이싱이 일어나 5' 에는 Kras의 염기서열이 존재하고 3' 쪽으로는 리보자임의 3' 엑손 염기서열이 존재하는 것을 확인할 수 있었다.

이러한 결과를 통해 Kras G12V를 타겟팅하는 리보자임인 KRib1과 KRib2가 in vitro에서 특이적으로 Kras G12V를 인지하여 절단하고 리보자임의 3' 엑손을 정확한 타겟 부위에 연결한다는 것을 확인할 수 있었다.

실시예 3: 변형 정도에 따른 트랜스-스플라이싱 리보자임의 in vitro 에서 효율성 및 특이성 확인

타겟에 대한 특이적인 6개의 염기서열인 IGS 외에 P1/ P10 헬릭스와 안티센스 부위를 변형(modify)함으로써 트랜스-스플라이싱 리보자임의 특이성과 효율성을 높일 수 있는데, 이러한 강화된 리보자임을 디자인함에 있어서 각 부위를 점차 변형 정도에 따른 특이성과 효율성을 확인하고자 하였다.

도 6은 P1/P10 헬릭스와 안티센스 부위를 변형함으로써 특이성과 효율성이 증대된 트랜스-스플라이싱 리보자임을 나타낸 것이다.

도 6을 참조하면, IGS만을 가진 리보자임, P1/ P10 구조를 더해준 리보자임, 안티센스 100개 또는 300개를 가진 리보자임 이렇게 각 부위를 점차 더해준 변형된 리보자임 RNA 시리즈를 KRib1과 KRib2 두 종류의 리보자임에 대하여 각각 제조하였다.

1. 특이성 증가시킨 특이성 리보자임 제작

리보자임의 IGS에서 3' 엑손 부위를 서열번호 6의 프라이머와 서열번호 8의 프라이머/서열번호 7의 프라이머와 서열번호 8의 프라이머(표 1 참조)를 이용하여 PCR 한 후 Hind III와 Nru I으로 SEAP 프로모터 벡터에 클로닝하였다.

상기 벡터에 클로닝된 리보자임 뒤에 Nru I과 Xba I으로 루시퍼라제를 클로닝하였는데, 이때 3' 엑손인 루시퍼라제가 Kras에 연결되어 트랜스진의 단백질이 올바르게 발현되도록 루시퍼라제 유전자 앞에 여분의 뉴클레오타이드를 더하여 프레임을 맞춰 주었다.

그 후 Kras 서열상에서 IGS에 의해 인지되는 곳으로부터 3' 말단 쪽으로 상보적인 100nt, 300nt의 안티센스 스트랜드를 서열번호 9의 프라이머와 서열번호 10의 프라이머/ 서열번호 9의 프라이머와 서열번호 11의 프라이머(표 1 참조)를 이용하여 PCR을 수행하였으며, 이를 SEAP KRib 1, SEAP KRib 2 구조체의 IGS 앞에 Hind III 효소 부위로 클로닝하였다.

이렇게 완성된 SEAP-KRib 1, SEAP KRib 2, SEAP AS100 KRib1, SEAP AS100 KRib2, SEAP AS300 KRib1, SEAP AS300 KRib2를 pAV 3.0 벡터로 BamH I과 Not I 부위를 이용하여 각각 클로닝하는 작업을 통하여 CMV 프로모터를 가진 리보자임 구조체 셋트를 제작하였다.

2. 트랜스-스플라이싱 리보자임의 in vitro 에서 효율성 및 특이성 확인

상기에서 제작된 각각의 리보자임들(50nM)을 in vitro 상에서 Kras 야생형 RNA 또는 Kras G12V RNA(10nM)와 트랜스-스플라이싱 반응하였으며 Kras에 결합하는 5' 프라이머와 리보자임 3' 엑손에 결합하는 3' 프라이머를 이용하여 상기 실시예 2와 같이 RT-PCR을 수행하였다.

도 7은 in vitro에서 변형된 리보자임의 트랜스-스플라이싱 효율성 및 특이성 분석 결과를 나타낸 것이다.

도 7A 및 7B를 참조하면, KRib1 또는 KRib2와 Kras G12V RNA를 반응시킨 샘플에서 트랜스-스플라이싱이 일어났을 때 나올 수 있는 산물 크기의 밴드를 얻었다.

in vitro 상에서의 변형 정도에 따른 KRib1의 2 셋트의 트랜스-스플라이싱 반응 실험의 결과 KRib1 셋트의 경우 안티센스 100개를 갖는 리보자임 보다 300개를 갖는 리보자임의 트랜스-접합(t/s) 산물 양이 많음을 PCR로 확인할 수 있었으며, KRib2의 경우 안티센스 100개를 갖는 리보자임과 300개를 갖는 리보자임의 트랜스-접합 산물 양이 같음을 PCR로 확인할 수 있었다.

또한, 변형된 정도에 따라 Kras 야생형 RNA와 트랜스-스플라이싱한 샘플에서도 트랜스-스플라이싱 산물과 같은 크기의 PCR 밴드가 흐리게 나왔으나, 이를 클로닝 후 시퀀싱한 결과 트랜스-스플라이싱되지 않은 리보자임이 PCR되었거나 기질이 PCR된 비특이적 PCR 산물임을 확인할 수 있었다.

그리고, 촉매성 코어 부위를 제거한 불활성 리보자임인 (d)KRib1, d)KRib2)과 Kras RNA를 반응시켰을 때 혹은 리보자임과 야행형 Kras RNA만을 각각 반응시켰을 때는 어떠한 산물도 볼 수 없었다.

도 7C 및 도 7D를 참조하면, PCR 산물을 클로닝 후 시퀀싱한 결과, 정확하게 타겟 부위에서 트랜스-스플라이싱이 일어나 5' 에는 Kras의 염기서열이 존재하고 3' 쪽으로는 리보자임의 3' 엑손에 루시퍼라제(Luciferase) 염기서열이 존재하는 것을 확인할 수 있었다.

실시예 4: 세포주에서의 트랜스-스플라이싱 특이성, 효율성 확인

특이성 리보자임이 세포내에서 단 염기 변이된 Kras G12V RNA를 표적으로 리보자임에 부착된 특정유전자를 트랜스-스플라이싱하는지 알아보기 위해서, Kras 야생형을 발현하는 293 세포에서 3' 엑손에 반딧불의 루시퍼라제를 태깅한 리보자임 을 디자인하고 리포터 에세이를 수행하였다.

루시퍼라제를 태깅한 리보자임은 12번째 코돈이 Gly(ggt)에서 Val(gtt)로 단 염기 변이된 Kras G12V RNA를 타겟팅하는데, 이때 IGS를 변이된 단 염기와 G·U 염기쌍을 이루게 한 KRib1(GCAGCT) 1 셋트와 변이된 다음의 염기와 G·U 염기쌍을 이루게 한 KRib2(GACAGC) 2 셋트 두 가지의 리보자임을 디자인하였다.

이렇게 디자인된 1 셋트와 2 셋트 리보자임들은 pAV3.0 벡터에 AS100KRib/ AS300KRib의 1 셋트, 2 셋트의 구조체, pAV AS100 KRib1, pAV as300 KRib1의 구조체를 제작하였다. 클로닝된 리보자임들을 이용하여 Kras 야생형을 발현하는 293 세포에서 트랜스-접합 산물 분석을 수행하였다. 기질인 Kras G12V는 RNA 상태로, 리보자임은 DNA 플라스미드 상태로 코-트랜스펙션시켰다.

1. 세포 배양

293 세포는 10% FBS(fetal bovine serum), DMEM(Dulbecco's modified eagle medium; HyClone), 0.5% antibiotic/antimycotic solution이 첨가된 배지로 37℃, 5% CO₂배양기에서 배양하였으며, 3~4일마다 계대배양하였다.

이때, 1X PBS(8g NaCl, 0.2g KCl, 1.14g Na₂HPO₄, 0.2g KH₄PO₄/L)로 세포를 씻어준 후, 1x 트립신/EDTA(8.2g NaCl, 0.2g KCl, 1.14g Na₂HPO₄, 0.2g KH₂PO₄, 0.029g Na₂EDTA·dH₂0, 1g trypsin, pH 7.35/L) 1㎖를 처리하여 CO₂배양기에 1~3분간 두었다. 배지 9ml로 트립신을 불활성화시킨 뒤 1500×g로 2분 30초간 원심분리하여 상청액을 제거하고 배지로 재현탁하였다. 새 100mm 디쉬에 이중 1/3을 씨딩 하여 배양하였다.

2. DMRIE-C에 의한 DNA, RNA 코트랜스펙션

클로닝으로 얻은 도 8A 및 도 9A 구조체들의 DNA와 Kras RNA를(상기 실시예 1의 3-2와 동일한 방법으로 제조) 293 세포에 코트랜스펙션시켰다. 환원된 혈청 배지(Reduced serum media) Opti-MEMⅠ 300㎕에 구조체 DNA 1㎍을 넣고 Opti-MEMⅠ300㎕에 DMRIE-C (Invitrogen) 3㎕를 넣은 튜브와 섞어 30분간 상온에서 반응시켰다.

Opti-MEMⅠ 600㎕와 DMRIE-C 3㎕에 Kras 야생형, Kras 12V RNA 4㎍을 넣고 섞어준 뒤 세척한 세포에 오버레이하였다. 4시간 동안 5% CO₂ 배양기에서 배양한 후 콤플리트 성장 배지(complete growth media)를 교체해 주었다.

3. 듀얼 루시퍼라제 분석

35mm 디쉬에 트랜스펙션한 각각 세포의 배지를 제거하고 1x PBS로 잘 닦아준다. 여기서 1x 수동 용해 버퍼(passive lysis buffer)를 200㎕ 넣고 상온에서 15분간 용해시킨 후 세포를 수확한 뒤 13000rpm으로 30초간 원심분리하여 상층액 만을 새 튜브에 옮겼다.

루미노미터 튜브에 LARII(Luciferase assay reagent II)을 100 ㎕를 넣고 여기에 세포 용해물 20㎕를 넣어 섞은 후 루미노미터로 읽었다. 다시 여기에 Stop & Glo reagent mix(Stop & Glo 20㎕ + Stop & Glo buffer 1㎖)를 100㎕넣고 섞은 후 루미노미터(TD+20/20)로 읽었다. 지연 시간은 3초, 통합 시간(integrate time)은 12초로 하고 감도 단계(sensitivity level)는 각각의 세포에 알맞게 60%로 설정하 였다.

4. RT-PCR을 통한 세포내에서의 트랜스-스플라이싱 산물의 발현 확인

4-1. RNA 분리

세포에서 총 RNA를 분리하기 위해, 20mM EDTA가 첨가된 Trizol(Invitrogen)을 사용하였다. 매뉴얼을 기본으로 Trizol 1㎖을 처리한 후 5분간 배양하고 세포를 떼어낸 후 클로로포름을 200㎕ 첨가한 후 보르텍스를 하여 4℃ 원심분리기에서 10분간 원심분리하였다. 상층액을 새로운 튜브에 옮긴 후 EtOH 침전하였다. 그 후 DNase(Promega)를 처리하였다.

4-2. 역전사

상기 2에서 분리한 총 RNA 중 5㎕를 L-아르기닌아마이드가 첨가된 OligodT(Invitrigen)를 사용하여 총 RNA를 잘 섞은 다음 65℃에서 5분간 배양시킨 다음 4℃에서 5분간 보관하였다. 그 후 5x RT buffer, 0.5mM dNTP(BMS)와 MMLV 역전사효소 1U (Promega)와 DEPC dH₂0로 총 25㎕를 만들어 37℃에서 1시간 동안 반응시켰다. 이후, 95℃에서 5분간 반응 시켜 MMLV-RT의 활성을 완전히 제거 후 4℃에서 보관했다.

4-3. PCR

RT 반응을 마친 cDNA는 서열번호 12,13의 프라이머(표 1 참조)를 사용하여 PCR 버퍼, dNTP, PCR 태그 중합효소, 5% DMSO를 넣고 (58℃ 30sec annealing/ 30 cycle) PCR하였다. 로딩 대조군으로 사용된 GAPDH는 프라이머를 사용하여 PCR 버퍼, dNTP, PCR 태그 중합효소를 넣고 PCR하였다.

4-4. 클로닝(RT-PCR 산물) 및 염기서열 분석

pGEM-T 이지 벡터 시스템(Promega)을 이용하여 TA-벡터에 RT-PCR 산물 클로닝을 하였으며, 서열번호 17의 프라이머(표 1 참조) 3.2 pmol을 사용하여 시퀀싱을 수행하였다.

4-4-1. RT-PCR 산물 용출

DNA 겔 추출(Bio basic inc.)을 이용하여 상기 실시예 1의 2-1-2와 같은 방법으로 DNA를 용출을 했다.

4-4-2. 연결

TA-벡터와 RT-PCR 산물 용출한 DNA(trans-splicing product)를 상기 실시예 1의 2-1-3과 같은 방법으로 연결했다.

4-4-3. 형질전환

연결 산물을 대장균(JM109)에 상기 실시예 1의 2-1-4와 같은 방법으로 형질전환했다.

4-4-4. 시퀀싱을 통한 염기 서열 분석

콜로니를 접종하여 미니-준비로 얻어진 DNA를 EcoRI으로 잘라서 삽입물이 들어간 클론을 얻었다. 상기 실시예 1의 2-2와 같은 방법으로 시퀀싱을 통해 서열을 확인하였다.

5. 트랜스-스플라이싱 특이성, 효율성 확인

도 8B는 트랜스-스플라이싱 리보자임에 의한 트랜스진의 리포터 에세이 결과를 나타낸 것이다.

도 8B를 참조하면, pAV AS100 KRib1을 Kras 야생형 또는 Kras G12V와 코-트랜스펙션한 경우 Kras 야생형을 넣어준 샘플에서는 루시퍼라제의 발현이 없었고 Kras G12V를 넣어준 샘플에서는 루시퍼라제가 적은 양이지만 발현된다는 것을 확인할 수 있었다.

그런데, pAV AS300 KRib1은 Kras 야생형을 넣어준 샘플 보다 Kras G12V를 넣어준 샘플에서 더 높은 루시퍼라제 값을 관찰할 수 있었으나, Kras G12V에 특이적으로 트랜스진이 발현되지 않음을 관찰하였다. 대조군으로 사용한 Mock이나 기질(Kras G12V)만 넣어준 샘플, 벡터 대조군의 샘플에서는 루시퍼라제가 발현되지 않았다.

이러한 결과로부터, AS100KRib1이 실험관 내에서 확인된 결과와 동일하게 세포에서도 Kras G12V에 특이적으로 트랜스진을 발현하는 것을 관찰할 수 있었다.

도 9B는 트랜스-스플라이싱 리보자임에 의한 트랜스진의 선별적 발현을 확인하기 위한 RT-PCR 분석 결과를 것이다

도 9B를 참조하면, Krib 1 셋트에서는 KRAS G12V RNA와 AS300KRib1-F,luc(레인 8)에서만 특이적으로 T/S 산물의 밴드를 볼 수가 있었다. 또한, 2 셋트에서는 KRAS G12V RNA와 AS100KRib2-F.luc(레인 6)에서만 T/S 산물 밴드를 특이적으로 볼 수 있었다.

이러한 특이적인 밴드가 트랜스-접합 산물인지 그리고 정확한 부위에서 스플라이싱이 일어났는지 알아보기 위해 트랜스-스플라이싱 밴드를 시퀀싱하여 확인하였다. 그 결과, 정확한 부위에서 트랜스-스플라이싱이 일어났음을 알 수 있었다. 이로부터 본 발명의 리보자임은 특이적으로 Kras 야생형과 Kras G12V인 단 염기 변이를 구별하여 트랜스-스플라이싱을 일으킨다는 것을 확인할 수 있었다.

실시예 5: In vivo 에서 트랜스-스플라이싱 트랜스진 발현을 통한 세포 사멸 유도 확인

그룹 I 리보자임은 특이적인 부위를 인지하여 절단하는 것 이외에도 자신의 3' 엑손을 타겟 부위의 5' 엑손에 연결하는 역할을 함으로써 새로운 트래스진의 발현을 유도할 수가 있다. 따라서, Kras 리보자임의 3'엑손에 HSV-TK(thymidine kinase) 유전자를 삽입함으로써 전구 약물인 GCV을 처리하면 특이적으로 트랜스-스플라이싱이 된 새로운 트랜스진을 발현하는 세포에서 세포사를 유도하게 된다.

도 10A는 SW480 세포에서 TK 분석을 위한 구조체를 나타낸 것이다.

도 10A를 참조하면, TK 유전자를 도입한 리보자임이 세포사를 유도할 수 있는지 알아보기 위해 먼저 pAVQ 트랜스퍼 벡터에 TK 유전자가 삽입된 구조체를 제작하였다.

1. Kras-리보자임-TK 구조체 클로닝

1-1. 벡터와 삽입물 준비

AS300KRib1-F.luc와 AS100KRib2-F.luc를 가지고 3' 엑손에 F.루시퍼라제 대신 HSV-TK의 TK 유전자를 삽입하기 위해 클로닝을 하였다.

pAVQ 벡터였던 리보자임 구조체들 3μg을 BglII(3U, Rocho)와 BstBI(3U, NEB) 제한효소를 이용하여 37℃ 3시간 자른 후 1% TAE 겔에 걸어 잘린 것을 확인 한 후, 플라스미드를 용출하였다.

TK 유전자를 얻기 위해 pCDNA3.1(+) CMV-TK의 구조체를 이용하여 TK 유전자를 서열번호 14, 15의 프라이머와 서열번호 16의 프라이머(표 1 참조)를 가지고 PCR[96℃-5'→(96℃-30", 58℃-1'30", 72℃-1'30") × 30 cycles → 72℃-7']로 증폭하였다.

이렇게 만들어진 TK DNA를 페놀 추출 및 에탄올 침전 후 3μg에 NruI(3U;Rocho)과 BstBI(3U;NEB) 제한효소를 이용하여 dH₂O를 최종 50㎕까지 넣고 섞어 37℃에 3시간 동안 반응시킨 후 페놀 추출 및 에탄올 침전을 통하여 DNA를 정제하고 dH₂O에 녹였다.

1-2. 연결

효소 처리된 pAVQ KRib1/KRib2 벡터와 삽입물(TK gene)을 각각 벡터:삽입물(1:3) 비율로 T₄리가아제(Roche)로 4℃에서 16시간 동안 반응시켰다.

1-3. 형질전환

연결한 벡터와 삽입물을 DH5α 수용성 세포에 42℃에서 1분 30초간 열 쇼크를 주어 형질전환을 한 후 5분간 얼음에 보관하고 1㎖의 새로운 LB를 첨가하여 37℃에서 30분간 배양시켰다. 그 후 카나마이신 아가 플레이트에 골고루 깐 다음 37℃ 배양기에 16시간 동안 배양시킨 후 콜로니를 암피실린이 포함된 5㎖ LB 배지에 접종하여 16시간 동안 37℃에서 흔들면서 배양하였다.

1-4. 시퀀싱을 통한 클로닝 확인

콜로니를 접종하여 미니-준비로 얻어진 DNA를 상기 실시예 1의 2-2와 같은 방법으로 서열번호 18의 프라이머(표 1 참조) 시퀀싱을 하여 확인하였다.

2. 자살유전자 활성 분석

실험을 하기 하루 전에 SW480(Kras G12V) 세포과 HT-29(wild type Kras) 세포를 35mm 디쉬에 2×10⁵ 세포를 배양하였다. 클로닝된 도 10A의 구조체와 함께 pEGFP-NI(250ng)을 코-트랜스펙션하였다.

2-1. DNA 트랜스펙션

구조체와 함께 pEGFP-NI(250ng)을 welfec M Gold 트랜스펙션 시약(Welgen)을 사용하여 코-트랜스펙션하였다. PBS로 세포를 세척한 후 환원된 혈청 배지 OPTI-MEM 2㎖을 첨가하여 37℃ 5% CO₂ 배양기에서 배양하였다. 환원된 혈청 배지 OPTI-MEM 100㎕에 GFP 250ng와 리보자임 1μg을 섞은 후 골드 인핸서 4㎕를 OPTI-MEM 100ul에 희석시킨 후 DNA 혼합물에 섞어 15분간 배양했다.

이 DNA 혼합물과 OPTI-MEM 100㎕에 희석시킨 M gold 6㎕를 반응시킨 후 15분 배양했다. 배양 후 세포 위에 골고루 첨가해주었다. 그 후 37℃ 5% CO₂ 배양기에서 5시간 배양 후 콤플리트 메디아로 교체해 주었다.

2-2. 세포 생존율 관찰

24시간 후 200 μM GCV(Cymevene^®, Roche)을 각 디쉬에 첨가해 주고 3일 동안 배양했다. AxioCam HR4 version 5.05.10. Zeiss 현미경을 이용하여 현광파장을 9 필터(488~509nm)로 하여 트랜스펙션된 세포에서 GFP의 발광 수를 측정하였다. 세 포의 생존율(%)은 아래와 같은 공식을 이용하였다.

[수학식 1]

3. 세포 사멸 유도 확인

Kras 야생형을 내생적으로 발현하는 세포주 HT-29와 Kras G12V를 내생적으로 발현하는 세포주 SW480에 AS100KRib2-TK를 GFP 유전자와 함께 플라스미드 상태로 트랜스펙션하였다.

도 10B를 참조하면, HT-29 세포에서는 GCV를 처리하거나 미처리 시에 GFP의 발광에는 어떠한 변화도 않은 것으로 보아 세포의 생존력에 영향을 끼치지 않음을 볼 수가 있었다.

그러나, SW480 세포에서는 AS100KRib2-TK를 트랜스펙션한 세포에서 GCV를 처리하였을 때 GCV를 처리하지 않은 세포에 비해 GFP의 발광 수가 감소한 것을 볼 수가 있었다. 이는 AS100KRib2-TK가 특이적인 트랜스-스플라이싱에 의한 TK 유전자의 발현으로 전구약물인 GCV를 톡신으로 바꾸어 세포 생존율을 감소시켰음을 알 수가 있다.

이러한 결과로 부터, 본 발명의 리보자임은 야생형 Kras와 변이된 Kras를 특이적으로 구별하여 트랜스-스플라이싱하는 것을 확인할 수 있었다.

실시예 6: 아데노바이러스 전달 시스템을 위한 바이러스 구조체 제작

pAVQ 트랜스퍼 벡터에 TK 유전자가 삽입된 구조체인 pAVQ 트랜스퍼 벡터 리 보자임을 pAdenovactor ΔE1/E3 백본 벡터와 함께 BJ5183 균주의 박테리아에 코-트랜스펙션시켜 트랜스퍼 벡터 리보자임과 pAdenovactor ΔE1/E3 백본 벡터를 상동 재조합시켰다. 이러한 방법으로 도 11B의 아데노바이러스 재조합 트랜스-스플라이싱 리보자임을 제작하였다.

1. 전달 시스템의 개발을 위한 재조합 아데노바이러스 벡터

클로닝된 도 10A의 pAdenovactor-CMV5-IRES-GFP 트랜스퍼 벡터(Qbiogene)들을 박테리아(BJ5183)에서 pAdenovactor ΔE1/E3 백본 벡터(Qbiogene)와 함께 코-트랜스포메이션시켜서 상동 재조합시켰다(도 11A 참조).

먼저, 트랜스퍼 벡터를 제한효소 PmeI으로 선형화시킨 후 페놀 추출 및 에탄올 침전을 통해 정제한 후 정량하여 1μg과 100ng의 pAdenovactor ΔE1/E3 백본 벡터를 BJ5183에 코-트랜스포메이션했다.

이렇게 얻은 도 11B의 BJ5183에서 상동 재조합된 재조합 벡터들은 PacI으로 선형화하여 페놀 추출 및 에탄올 침전을 통해 정제한 후 정량하여 293 세포에 트랜스펙션하였다. 293 세포에 트랜스펙션된 선형화된 재조합 벡터들은 3번의 플라크 정제를 통해 재조합 아데노바이러스를 만들었다.

293 세포를 이용하여 증폭시킨 재조합 바이러스들은 Vivapure^® AdenoPACK^TM 100 RT(Sartorius AG.)를 사용하여 정제하고 농축시켰다. 재조합 아데노바이러스들의 적가를 결정하기 위해 TCID50(tissue culture infectious dose for 50% of the cells) 방법을 이용하였다. 바이랄 스톡(Viral stock)들은 10%의 수크로스와 함께 -80℃에 보관하였다.

1-1. 정제 전에 Ad-AS300KRib1-TK의 분석

상동 재조합시킨 바이러스 재조합 리보자임들은 293 세포에 트랜스펙션함으로써 감염 능력을 가진 Ad-리보자임-TK 바이러스를 얻게 되었다. 이러한 바이러스들은 적가가 낮기 때문에 293 세포에 여러 번의 감염을 통해 고 적가를 얻어야한다. 고 적가의 바이러스를 얻게 되면 정제 과정을 통해 순수한 바이러스들을 정제하게 된다.

따라서, 바이러스들을 정제하기 전에 바이러스가 잘 생성되고 있는지, 올바른 Ad-리보자임들이 만들어지고 있는지 확인을 해야 한다. 또한, 여러 클론들 중 효율이 좋은 클론을 선별해야 한다. 이러한 확인을 위해 여러 번의 감염 중 저 적가에서 바이러스의 게놈을 얻어 PCR을 통해 리보자임의 3'엑손의 TK(thymidine kinase) 유전자와 pAdenovactor ΔE1/E3 백본 벡터의 LITR(left inverted terminal repeat)을 PCR을 통해 확인하였다(도 12A).

또한, 저 적가의 Ad-AS300KRib1-TK를 Kras G12V 변이된 세포주인 SW480 세포에 감염하여 Ad-AS300KRib1-TK의 발현을 확인하였다(도 12B). 바이러스는 클론들의 밴드를 비교하여 클론 18번을 가지고 바이러스 정제를 진행하였다.

1-2. 정제 전에 Ad-AS100KRib2-TK의 분석

상기 1-1과 같은 방식으로 Ad-AS100KRib2-TK을 분석하였다(도 13). 바이러스의 게놈을 뽑아 PCR을 해본 결과 리보자임의 3'엑손의 TK(thymidine kinase) 유전자와 pAdenovactor ΔE1/E3 백본 벡터의 LITR(left inverted terminal repeat)을 확인하였다(도 13A). 그리고 SW480 세포에 감염하여 Ad-AS100KRib2-TK의 발현을 확인하였다(도 13B). 바이러스는 클론들의 밴드를 비교한 결과 클론 1번을 가지고 바이러스 정제를 진행하였다.

1-3. 정제 전에 Ad-AS100KRib2-LacZ의 분석

상기 1-1과 같은 방식으로 Ad-AS100KRib2-LacZ를 분석하였다(도 14). 바이러스의 게놈을 뽑아 PCR을 해본 결과 리보자임의 3'엑손의 LacZ 유전자와 pAdenovactor ΔE1/E3 백본 벡터의 LITR(left inverted terminal repeat)을 확인하였다(도 14A). 그리고 SW480 세포에 감염하여 Ad-AS100KRib2-LacZ의 발현을 확인하였다(도 14B). 바이러스는 클론들의 밴드를 비교한 결과 클론 30번을 가지고 바이러스 정제를 진행하였다.

2. 재조합 바이러스의 확인

3번의 플라크 정제를 통해 재조합 아데노바이러스를 만드는 중간에 바이러스를 감염한 293 세포에서 바이러스가 제대로 만들어지고 있는지 확인하기 위해 바이러스 게놈 DNA를 추출하여 서열번호 19 내지 24의 프라이머(표 1 참조)를 가지고 PCR[96℃-5'→(96℃-30", 58℃-1'30", 72℃-1') × 30 cycles → 72℃-7']을 통해 증폭하였고, 2% TBE 아가로스 겔에 로딩하여 크기를 확인하였다. 또한, SW480 세포에 주입하여 상기 실시예 4의 5, 6과 같은 방법으로 RT-PCR을 통해 재조합 바이러스들을 확인하였다.

3. 바이러스 재조합들의 트랜스-스플라이싱 확인

상기 1에서 정제된 Ad-리보자임들을 Kras G12V 변이된 세포주인 SW480에 50 M.O.I의 바이러스를 감염한 후, 48시간 후에 11-3,4와 같은 방법으로 서열번호 1과 14의 프라이머(표 1 참조)를 가지고 RT-PCR과 서열번호 17의 프라이머(표 1 참조)를 가지고 시퀀싱을 통해 트랜스-스플라이싱을 확인하였다.

4. 종양 세포에서 Ad-리보자임을 이용한 T/S 산물 확인

이렇게 제작된 Ad-리보자임은 종양세포에서 특이적으로 트랜스-스플라이싱 일어나 T/S 산물을 생성하는지 확인하기 위해 Kras G12V 변이된 세포주인 SW480 세포에 Ad-리보자임을 감염하였다. 대조군으로 Ad-CMV-TK와 혼합하여 사용하였다. Ad-CMV-TK는 리보자임의 3'엑손의 TK 유전자와 같은 유전자로 리보자임이 없는 구조체이다. 혼합(Mix)은 RNA 추출 시 in vitro 상에서 트랜스-스플라이싱이 일어나지 않는다는 것을 보여주는 것으로 SW480 세포의 내생적인 돌연변이 Kras와 내생적인 Kras 야생형인 HT-29 세포에 Ad-AS100KRib2-TK를 감염한 것을 튜브 상에서 반응시킨 것이다.

도 15는 Kras G12V 변이된 SW480 세포에서 아데노바이러스 전달 시스템을 이용한 트랜스-접합 RNA 산물의 분석 결과를 나타낸 것이다.

도 15를 참조하면, 대조군들은 밴드가 없는 반면에 Ad-AS300KRib1-TK와 Ad0AS100KRib2-TK를 감염한 레인에서만 특이적으로 밴드가 보이는 것을 알 수가 있다.

이러한 특이적 밴드가 리보자임에 의한 특이적인 T/S 산물인지를 확인하기 위해서 시퀀싱을 통해 확인하였다. 그 결과, Ad-리보자임들이 특이적이고 올바르게 작용을 하고 있음을 알 수 있었다.

실시예 7: 아데노바이러스 Kras G12V-특이성 리보자임에 의한 Kras G12V 돌열변이된 암 세포에서 특이적인 세포 사멸

Kras G12V 변이된 암 세포에서 Kras G12V를 타겟팅하는 특이성 리보자임이 세포사를 유도하는지를 실험하기 위해 MTS 분석을 진행하였다. Kras G12V 변이된 세포주인 SW480 세포, CAPAN-1 세포와 Kras 야생형인 HT-29 세포에 Ad-리보자임들을 감염시키고 다음날 GCV를 처리함으로써 암 세포의 세포사를 관찰하였다.

1. 세포 배양

HT-29(colorectal adenocarcinoma)는 10% FBS(fetal bovine serum), DMEM(Dulbecco's modified eagle medium; HyClone), 0.5% antibiotic/antimycotic solution이 첨가된 배지로 SW480(colorectal adenocarcinoma)는 10% FBS(fetal bovine serum), RPMI 1640(Roswell Park Memorial Institute 1640; HyClone), 0.5% 항생/항진균 용액이 첨가된 배지로 CAPAN-1(pancreatic adenocarcinoma)은 10% FBS(fetal bovine serum), RPMI 1640(Roswell Park Memorial Institute 1640; HyClone), 0.5% antibiotic/antimycotic solution이 첨가된 배지로 37℃, 5% CO₂배양기에서 배양하였으며, 3~4일마다 계대배양하였다.

이때, 1X PBS(8g NaCl, 0.2g KCl, 1.14g Na₂HPO₄, 0.2g KH₄PO₄/L)로 세포를 씻어준 후, 1x 트립신/EDTA(8.2g NaCl, 0.2g KCl, 1.14g Na₂HPO₄, 0.2g KH₂PO₄, 0.029g Na₂EDTA·dH₂0, 1g trypsin, pH 7.35/L) 1㎖를 처리하여 CO₂배양기에 1~3분간 두었다. 배지 9ml로 트립신을 불활성화시킨 뒤 1500×g로 2분 30초간 원심분리 하여 상청액을 제거하고 배지로 재현탁하였다. 새 100mm 디쉬에 이중 1/3을 씨딩하여 배양하였다.

2. 세포 증식 분석

세포 증식(MTS) 분석법은 CellTiter 96Aqueous one solution reagent(MTS, Promega)의 매뉴얼을 기초로 하여 수행하였다.

96-웰 플레이트에 세포를 3×10³ cells/well(0.1 ㎖)로 씨딩하였다. 37℃ 5% CO₂배양기에서 밤새도록 배양한 후, 각 웰에 다양한 M.O.I의 바이러스를 감염시켰다. 16시간 후에 각 플레이트 100 uM GCV(Cymevene^®, Roche)을 처리하여 5일 동안 배양시켰다. Opti-MEM 100ul에 CellTiter 96Aqueous one solution reagent(MTS, Promega)를 20% 추가하여 각 웰에 첨가한 후 1~4시간 동안 37℃ 5% CO₂ 배양기에서 배양한 후 세포 생존율을 490nm에서 흡수도를 측정하였다.

3. Kras G12V 돌열변이된 암 세포에서 특이적인 세포 사멸 확인

도 16A는 SW480 세포에서 MTS 분석 결과를 나타낸 것이다.

도 16A의 왼쪽 그래프를 참조하면, 동일하게 GCV를 100uM로 처리하였을 때 Ad-리보자임들은 M.O.I 의존적으로 감염하였다. 도 16A의 오른쪽 그래프를 참조하면, 동일하게 Ad-리보자임들을 150 M.O.I로 감염하고 GCV 복용별로 처리를 하였다.

Ad-CMV-TK과 이미 보고된 논문에서 특이성과 효율이 인정된 Ad-CMV-리보자임-TK가 양성 대조군으로 사용되었으며, Ad-CMV-LacZ는 음성 대조군으로 사용되었다.

그 결과, 양성 대조군과 음성 대조군과 비교하였을 때 Ad-리보자임들은 특이 적인 트랜스-스플라이싱과 트랜스진이 유도되어 세포사가 일어남을 볼 수 있었다.

도 16B 및 16C는 각각 CAPAN-1 세포, HT-29 세포에서 MTS 분석 결과를 나타낸 것이다.

도 16B의 경우도 동일하게 GCV를 100uM로 처리하고, Ad-리보자임들은 M.O.I 의존적으로 감염한 결과, Ad-리보자임들에서 특이적인 세포사가 일어남을 볼 수 있었다. 그러나, 도 16C 야생형 Kras인 HT-29 세포에서는 양성 바이러스들을 제외하고는 세포사가 일어나지 않음을 볼 수 있었다.

이러한 결과로부터 Ad-리보자임들이 특이적으로 변이된 Kras G12V를 인지하여 트랜스-스플라이싱을 해서 트랜스진을 유도했음을 알 수가 있다.

실시예 8: Ad-리보자임에 의한 이종이식 모델에서의 종양 퇴행 확인

1. 실험동물

특이적으로 병원체 부재 조건인 4주에서 5주의 수컷 BALB/cAnNCrI 누드 마우스(Orient Bio Inc.)를 실험을 위해 사용하였다. 이종이식 마우스 모델을 만들기 위해, 먼저 마우스 8마리에 CAPAN-1(1×10⁷cells)을 복강 내 주사한 후 3주 후에 종양 생성시킨 다음 아데노바이러스 리보자임들을 이틀 간격으로 3회(1×10⁹ PFU) 감염시켰다. GCV(50mg/kg)는 바이러스를 처음 감염시킨 날 다음날부터 하루에 2번씩 10일 동안 복강을 통해 처리하였다. 그리고 야생형의 Kras 피하 종양 모델을 만들기 위해서 HT-29을 이용해서 종양 모델을 제작하였다. 수컷 BALB/c 누드 마우스 7마리에 1×10⁷cells을 피하 주사하여 2주 후에 종양을 생성시켰다. Ad-리보자임은 정맥 내로 이틀 간격으로 3번, 1×10⁹cell을 처리하였다. GCV(50mg/kg)는 10일 동안 2번씩 복강으로 처리한 후 관찰하였다.

2. 이종이식 모델에서의 종양 퇴행 확인

In vivo에서 Ad-리보자임의 특이성과 효율성을 확인하기 위해서 MTS 분석을 통해 확인된 Ad-리보자임들을 인간 종양이 이식된 이종이식 마우스 모델에 감염했다.

Kras G12V 변이된 CAPAN-1을 i,p로 주사하여 종양을 생성하였고, Ad-리보자임을 처리했다(도 17A 참조). 그 후 GCV를 처리하여 Ad-리보자임에 의해 특이적으로 종양이 퇴행(regression)되는지를 확인했다. 음성 바이러스로는 Ad-CMV-LacZ를 감염했고 양성 바이러스로는 Ad-CMV-TK를 감염했다.

Ad-CMV-LacZ에 비해 Ad-CMV-TK와 Ad-AS100KRib2-TK는 유사하게 Kras G12V 변이된 종양에서 종양 퇴행이 일어남을 알 수가 있었다(P=0.467). 또한, 야생형 Kras를 발현하는 HT-29로 피하의 종양 마우스 모델을 만들었다.

도 17B는 H&E 염색하여 괴사를 제외한 종양을 관찰하여 종양의 중량을 그래프로 나타낸 것이다.

도 17B를 참조하면, 종양의 중량은 음성 대조군과 비슷하게 Ad-리보자임들도 감소하지 않은 것을 알 수 있다(P=0.5137). 그러나, 양성 바이러스인 Ad-CMV-TK인 경우에는 종양의 중량이 감소한 것을 볼 수 있었다. 또한, Kras G12V 변이된 종양 과 야생형 Kras 종양의 결과를 비교해 보면 Ad-리보자임은 Kras G12V를 특이적으로 인지하여 작용한다는 것을 알 수가 있다.

도 2는 Kras RNA의 이차 구조를 나타낸 것이다[화살표: Kras의 12번째 코돈].

도 3은 Kras RNA의 모식도와 특이성 트랜스-스플라이싱 리보자임을 나타낸 것이다.

도 6은 P1/P10 헬릭스와 안티센스 부위를 변형함으로써 특이성과 효율성이 증대된 트랜스-스플라이싱 리보자임을 나타낸 것이다[A. 변형된 리보자임의 모식도, 화살표: trans-splicing junction; B. 변형된 KRib1 , KRib2의 구조체].

도 7은 in vitro에서 변형된 리보자임의 트랜스-스플라이싱 효율성 및 특이성 분석 결과를 나타낸 것이다[A-B: 트랜스-접합 산물의 RT-PCR 분석, C-D: 트랜스-접합 RT-PCR 산물의 서열분석(C: AS300KRib1, D: AS100KRib2)].

도 8은 트랜스-스플라이싱 리보자임에 의한 트랜스진의 선별적 유도를 확인하기 위한 리포터 에세이 결과를 나타낸 것이다[A: 293 세포에서 리포터 분석을 위한 구조체, B: Kras (야생형 또는 12V) RNA 및 레닐라 루시퍼라제가 함유된 리보 자임(KRib1 세트 / KRib2 세트)의 트랜스펙션].

도 9는 트랜스-스플라이싱 리보자임에 의한 트랜스진의 선별적 발현을 확인하기 위한 RT-PCR 분석 결과를 것이다[A: 293 세포에서 리포터 분석을 위한 구조체, B: Kras (wild or 12V) RNA가 함유된 리보자임(KRib1 세트 / KRib2 세트)의 RT-PCR; W: 야생형 Kras, 12V: Kras G12V, IGS: 내부 가이드 염기서열, D)KRib: 불활성 리보자임].

도 10은 Kras G12V를 내생적으로 발현하는 세포주 SW480에서 TK 유전자 유도에 의한 세포 퇴행을 나타낸 것이다[A: SW480 세포에서 TK 분석을 위한 구조체, B: GFP 유전자 함유 AS100KRib2-TK의 트랜스펙션]

도 11은 박테리아에서 상동 재조합에 의한 재조합 아데노바이러스의 세대를 나타낸 것이다[A: 재조합 아데노바이러스 전달 시스템의 모식도, B: 아데노바이러스 리보자임 재조합의 구조체].

도 12는 재조합 Ad-AS300KRib1-TK의 분석을 나타낸 것이다[A. 바이러스 정제 전 PCR에 의한 바이러스 게놈 확인, B: Kras G12V 변이된 SW480에서 바이러스 정제 전 RT-PCR에 의한 Ad-AS300KRib1-TK 발현 확인; TK : Thymidine Kinase, LITR : Left Inverted Terminal Repeat].

도 13은 재조합 Ad-AS100KRib2-TK의 분석을 나타낸 것이다[A. 바이러스 정제 전 PCR에 의한 바이러스 게놈 확인, B: Kras G12V 변이된 SW480에서 바이러스 정제 전 RT-PCR에 의한 Ad-AS100KRib2-TK 발현 확인; TK : Thymidine Kinase, LITR : Left Inverted Terminal Repeat].

도 14는 재조합 Ad-AS100KRib2-LacZ의 분석을 나타낸 것이다[A. 바이러스 정제 전 PCR에 의한 바이러스 게놈 확인, B: Kras G12V 변이된 SW480에서 바이러스 정제 전 RT-PCR에 의한 Ad-AS100KRib2-LacZ 발현 확인; LITR : Left Inverted Terminal Repeat].

도 15는 Kras G12V 변이된 암 세포 SW480에서 아데노바이러스 전달 시스템을 이용한 트랜스-접합 RNA 산물의 분석을 나타낸 것이다.

도 16은 종양세포주에서 아데노바이러스 전달 시스템에 의한 특이적 트랜스-스플라이싱 리보자임의 세포 생존율을 분석한 것이다[A: SW480(KrasG12V)에서 세포 생존율 MTS 분석([left panel: M.O.I dependent / 100uM GCV , right panel GCV dose dependent / virus 150M.O.I), B: CAPAN-1(KrasG12V)에서 세포 생존율 MTS 분석(M.O.I dependent / 100uM GCV), C: HT-29(wild Kras)에서 세포 생존율 MTS 분석(left panel: M.O.I dependent / 100uM GCV , right panel GCV dose dependent / virus 150M.O.I)].

도 17은 이종이식 마우스 모델에서 아데노바이러스 인코딩 Kras G12V-특이적 리보자임의 분석을 나타낸 것이다[A: 복강 내 CAPAN-1(Kras G12V) 종양 모델(n=6 또는 7)에서 Ad-리보자임에 의한 효율적이고 특이적인 종양 퇴행 보임, B: 피하 내 HT-29(wild type Kras) 종양 모델(n=7)에서 Ad-리보자임에 의한 종양 퇴행 없음].

<110> INDUSTRY-ACADEMIC COOPERATION FOUNDATION, DANKOOK UNIVERSITY <120> TRANS-SPLICING GROUP I RIBOZYME SPECIFIC FOR KRAS G12V RNA <160> 44 <170> KopatentIn 1.71 <210> 1 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Kras wild primer <400> 1 gttggagctg gtggcgtagg c 21 <210> 2 <211> 15 <212> DNA <213> Artificial Sequence <220> <223> Kras 5' BamHI primer <400> 2 taatacgact cacta 15 <210> 3 <211> 44 <212> DNA <213> Artificial Sequence <220> <223> kras rib T7-5' primer <400> 3 gggtaatacg actcactata ggaggtaaaa gttatcaggc atgc 44 <210> 4 <211> 44 <212> DNA <213> Artificial Sequence <220> <223> RY-PCR primer <400> 4 gggtaatacg actcactata gacagcaaaa gttatcaggc atgc 44 <210> 5 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> RY-RT primer <400> 5 tgtaaaacga cggccagtg 19 <210> 6 <211> 28 <212> DNA <213> Artificial Sequence <220> <223> kras p1p10-rib1-5 primer <400> 6 cgatgatcac gaagacgcca aaaacata 28 <210> 7 <211> 30 <212> DNA <213> Artificial Sequence <220> <223> kras p1p10-rib2-5 primer <400> 7 cgacatgatc acgaagacgc caaaaacata 30 <210> 8 <211> 37 <212> DNA <213> Artificial Sequence <220> <223> krib 3' primer <400> 8 aaggaaaaaa gcggccgctt attacaattt ggacttt 37 <210> 9 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> kras as 5' primer <400> 9 gggaagcttg caagagtgcc ttgacga 27 <210> 10 <211> 28 <212> DNA <213> Artificial Sequence <220> <223> kras as 3' primer <400> 10 gggaaggctt ccatcaatta ctacttgc 28 <210> 11 <211> 41 <212> DNA <213> Artificial Sequence <220> <223> kras as300 5' priemr <400> 11 gggtaatacg actcactata gggctactag gaccataggt a 41 <210> 12 <211> 30 <212> DNA <213> Artificial Sequence <220> <223> kras t/s BamHI5' primer <400> 12 cgcggatcca ctgaatataa acttgtggta 30 <210> 13 <211> 28 <212> DNA <213> Artificial Sequence <220> <223> cell t/s luci 3' nest primer <400> 13 cccaagcttg cccaacaccg gcataaag 28 <210> 14 <211> 38 <212> DNA <213> Artificial Sequence <220> <223> TK 5' NruI 1set primer <400> 14 gtgatcatcg cgatgatcac gcttcgtacc cctgccat 38 <210> 15 <211> 40 <212> DNA <213> Artificial Sequence <220> <223> TK 5' NruI 2setI primer <400> 15 gtgatcatcg cgacatgatc acgcttcgta cccctgccat 40 <210> 16 <211> 30 <212> DNA <213> Artificial Sequence <220> <223> 3' TK BstBI primer <400> 16 aaggaaaaaa ttcgaattag cctcccccat 30 <210> 17 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> T7-R-Primer <400> 17 taatacgact cactataggg 20 <210> 18 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> IGS rev seq <400> 18 cctttcccgc aatttgacgg tctt 24 <210> 19 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> 5' TK stable primer <400> 19 cgggaattcg tggtaatgac aagc 24 <210> 20 <211> 33 <212> DNA <213> Artificial Sequence <220> <223> 3' TK stable primer <400> 20 cggggatcca tccaggataa agacgtgcat ggg 33 <210> 21 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> 5' LITR EcoRI primer <400> 21 ggaattctgg agtttgtgac gtggcg 26 <210> 22 <211> 32 <212> DNA <213> Artificial Sequence <220> <223> 3' LITR XbaI primer <400> 22 gctctagatg gccaaatctt actcggttac gc 32 <210> 23 <211> 29 <212> DNA <213> Artificial Sequence <220> <223> 5' viral LacZ primer <400> 23 ggaattcatg gtcgttttac aacgtcgtg 29 <210> 24 <211> 28 <212> DNA <213> Artificial Sequence <220> <223> cancer.lacZ-3 primer <400> 24 gggaagcttc ggattgaccg taatggga 28 <210> 25 <211> 513 <212> RNA <213> Artificial Sequence <220> <223> AS100KRib1 <400> 25 aagcuuccau caauuacuac uugcuuccug uaggaauccu cuauuguugg aucauauucg 60 uccacaaaau gauucugaau uagcuguauc gucaaggcac ucuugcaagc uucauuauca 120 gcagcuaaaa guuaucaggc augcaccugg uagcuagucu uuaaaccaau agauugcauc 180 gguuuaaaag gcaagaccgu caaauugcgg gaaagggguc aacagccguu caguaccaag 240 ucucagggga aacuuugaga uggccuugca aaggguaugg uaauaagcug acggacaugg 300 uccuaaccac gcagccaagu ccuaagucaa cagaucuucu guugauaugg augcaguuca 360 cagacuaaau gucggucggg gaagauguau ucuucucaua agauauaguc ggaccucucc 420 uuaaugggag cuagcggaug aagugaugca acacuggagc cgcugggaac uaauuuguau 480 gcgaaaguau auugauuagu uuuggaguac ucg 513 <210> 26 <211> 713 <212> RNA <213> Artificial Sequence <220> <223> AS300KRib1 <400> 26 aagcuucuac uaggaccaua gguacaucuu cagaguccuu aacucuuuua auuuguucuc 60 uauaauggug aauaucuuca aaugauuuag uauuauuuau ggcaaauaca caaagaaagc 120 ccuccccagu ccucauguac uggucccuca uugcacugua cuccucuuga ccugcugugu 180 cgagaauauc caagagacag guuucuccau caauuacuac uugcuuccug uaggaauccu 240 cuauuguugg aucauauucg uccacaaaau gauucugaau uagcuguauc gucaaggcac 300 ucuugcaagc uucauuauca gcagcuaaaa guuaucaggc augcaccugg uagcuagucu 360 uuaaaccaau agauugcauc gguuuaaaag gcaagaccgu caaauugcgg gaaagggguc 420 aacagccguu caguaccaag ucucagggga aacuuugaga uggccuugca aaggguaugg 480 uaauaagcug acggacaugg uccuaaccac gcagccaagu ccuaagucaa cagaucuucu 540 guugauaugg augcaguuca cagacuaaau gucggucggg gaagauguau ucuucucaua 600 agauauaguc ggaccucucc uuaaugggag cuagcggaug aagugaugca acacuggagc 660 cgcugggaac uaauuuguau gcgaaaguau auugauuagu uuuggaguac ucg 713 <210> 27 <211> 515 <212> RNA <213> Artificial Sequence <220> <223> AS100KRib2 <400> 27 aagcuuccau caauuacuac uugcuuccug uaggaauccu cuauuguugg aucauauucg 60 uccacaaaau gauucugaau uagcuguauc gucaaggcac ucuugcaagc uucaucaugg 120 ccgacagcaa aaguuaucag gcaugcaccu gguagcuagu cuuuaaacca auagauugca 180 ucgguuuaaa aggcaagacc gucaaauugc gggaaagggg ucaacagccg uucaguacca 240 agucucaggg gaaacuuuga gauggccuug caaaggguau gguaauaagc ugacggacau 300 gguccuaacc acgcagccaa guccuaaguc aacagaucuu cuguugauau ggaugcaguu 360 cacagacuaa augucggucg gggaagaugu auucuucuca uaagauauag ucggaccucu 420 ccuuaauggg agcuagcgga ugaagugaug caacacugga gccgcuggga acuaauuugu 480 augcgaaagu auauugauua guuuuggagu acucg 515 <210> 28 <211> 715 <212> RNA <213> Artificial Sequence <220> <223> AS300KRib2 <400> 28 aagcuucuac uaggaccaua gguacaucuu cagaguccuu aacucuuuua auuuguucuc 60 uauaauggug aauaucuuca aaugauuuag uauuauuuau ggcaaauaca caaagaaagc 120 ccuccccagu ccucauguac uggucccuca uugcacugua cuccucuuga ccugcugugu 180 cgagaauauc caagagacag guuucuccau caauuacuac uugcuuccug uaggaauccu 240 cuauuguugg aucauauucg uccacaaaau gauucugaau uagcuguauc gucaaggcac 300 ucuugcaagc uucaucaugg ccgacagcaa aaguuaucag gcaugcaccu gguagcuagu 360 cuuuaaacca auagauugca ucgguuuaaa aggcaagacc gucaaauugc gggaaagggg 420 ucaacagccg uucaguacca agucucaggg gaaacuuuga gauggccuug caaaggguau 480 gguaauaagc ugacggacau gguccuaacc acgcagccaa guccuaaguc aacagaucuu 540 cuguugauau ggaugcaguu cacagacuaa augucggucg gggaagaugu auucuucuca 600 uaagauauag ucggaccucu ccuuaauggg agcuagcgga ugaagugaug caacacugga 660 gccgcuggga acuaauuugu augcgaaagu auauugauua guuuuggagu acucg 715 <210> 29 <211> 2184 <212> DNA <213> Artificial Sequence <220> <223> AS100KRib1-F.luci <400> 29 aagcttccat caattactac ttgcttcctg taggaatcct ctattgttgg atcatattcg 60 tccacaaaat gattctgaat tagctgtatc gtcaaggcac tcttgcaagc ttcattatca 120 gcagctaaaa gttatcaggc atgcacctgg tagctagtct ttaaaccaat agattgcatc 180 ggtttaaaag gcaagaccgt caaattgcgg gaaaggggtc aacagccgtt cagtaccaag 240 tctcagggga aactttgaga tggccttgca aagggtatgg taataagctg acggacatgg 300 tcctaaccac gcagccaagt cctaagtcaa cagatcttct gttgatatgg atgcagttca 360 cagactaaat gtcggtcggg gaagatgtat tcttctcata agatatagtc ggacctctcc 420 ttaatgggag ctagcggatg aagtgatgca acactggagc cgctgggaac taatttgtat 480 gcgaaagtat attgattagt tttggagtac tcgcgatgat cacgaagacg ccaaaaacat 540 aaagaaaggc ccggcgccat tctatcctct agaggatgga accgctggag agcaactgca 600 taaggctatg aagagatacg ccctggttcc tggaacaatt gcttttacag atgcacatat 660 cgaggtgaac atcacgtacg cggaatactt cgaaatgtcc gttcggttgg cagaagctat 720 gaaacgatat gggctgaata caaatcacag aatcgtcgta tgcagtgaaa actctcttca 780 attctttatg ccggtgttgg gcgcgttatt tatcggagtt gcagttgcgc ccgcgaacga 840 catttataat gaacgtgaat tgctcaacag tatgaacatt tcgcagccta ccgtagtgtt 900 tgtttccaaa aaggggttgc aaaaaatttt gaacgtgcaa aaaaaattac caataatcca 960 gaaaattatt atcatggatt ctaaaacgga ttaccaggga tttcagtcga tgtacacgtt 1020 cgtcacatct catctacctc ccggttttaa tgaatacgat tttgtaccag agtcctttga 1080 tcgtgacaaa acaattgcac tgataatgaa ttcctctgga tctactgggt tacctaaggg 1140 tgtggccctt ccgcatagaa ctgcctgcgt cagattctcg catgccagag atcctatttt 1200 tggcaatcaa atcattccgg atactgcgat tttaagtgtt gttccattcc atcacggttt 1260 tggaatgttt actacactcg gatatttgat atgtggattt cgagtcgtct taatgtatag 1320 atttgaagaa gagctgtttt tacgatccct tcaggattac aaaattcaaa gtgcgttgct 1380 agtaccaacc ctattttcat tcttcgccaa aagcactctg attgacaaat acgatttatc 1440 taatttacac gaaattgctt ctgggggcgc acctctttcg aaagaagtcg gggaagcggt 1500 tgcaaaacgc ttccatcttc cagggatacg acaaggatat gggctcactg agactacatc 1560 agctattctg attacacccg agggggatga taaaccgggc gcggtcggta aagttgttcc 1620 attttttgaa gcgaaggttg tggatctgga taccgggaaa acgctgggcg ttaatcagag 1680 aggcgaatta tgtgtcagag gacctatgat tatgtccggt tatgtaaaca atccggaagc 1740 gaccaacgcc ttgattgaca aggatggatg gctacattct ggagacatag cttactggga 1800 cgaagacgaa cacttcttca tagttgaccg cttgaagtct ttaattaaat acaaaggata 1860 tcaggtggcc cccgctgaat tggaatcgat attgttacaa caccccaaca tcttcgacgc 1920 gggcgtggca ggtcttcccg acgatgacgc cggtgaactt cccgccgccg ttgttgtttt 1980 ggagcacgga aagacgatga cggaaaaaga gatcgtggat tacgtggcca gtcaagtaac 2040 aaccgcgaaa aagttgcgcg gaggagttgt gtttgtggac gaagtaccga aaggtcttac 2100 cggaaaactc gacgcaagaa aaatcagaga gatcctcata aaggccaaga agggcggaaa 2160 gtccaaattg taataagcgg ccgc 2184 <210> 30 <211> 2384 <212> DNA <213> Artificial Sequence <220> <223> AS300KRib1-F.luci <400> 30 aagcttctac taggaccata ggtacatctt cagagtcctt aactctttta atttgttctc 60 tataatggtg aatatcttca aatgatttag tattatttat ggcaaataca caaagaaagc 120 cctccccagt cctcatgtac tggtccctca ttgcactgta ctcctcttga cctgctgtgt 180 cgagaatatc caagagacag gtttctccat caattactac ttgcttcctg taggaatcct 240 ctattgttgg atcatattcg tccacaaaat gattctgaat tagctgtatc gtcaaggcac 300 tcttgcaagc ttcattatca gcagctaaaa gttatcaggc atgcacctgg tagctagtct 360 ttaaaccaat agattgcatc ggtttaaaag gcaagaccgt caaattgcgg gaaaggggtc 420 aacagccgtt cagtaccaag tctcagggga aactttgaga tggccttgca aagggtatgg 480 taataagctg acggacatgg tcctaaccac gcagccaagt cctaagtcaa cagatcttct 540 gttgatatgg atgcagttca cagactaaat gtcggtcggg gaagatgtat tcttctcata 600 agatatagtc ggacctctcc ttaatgggag ctagcggatg aagtgatgca acactggagc 660 cgctgggaac taatttgtat gcgaaagtat attgattagt tttggagtac tcgcgatgat 720 cacgaagacg ccaaaaacat aaagaaaggc ccggcgccat tctatcctct agaggatgga 780 accgctggag agcaactgca taaggctatg aagagatacg ccctggttcc tggaacaatt 840 gcttttacag atgcacatat cgaggtgaac atcacgtacg cggaatactt cgaaatgtcc 900 gttcggttgg cagaagctat gaaacgatat gggctgaata caaatcacag aatcgtcgta 960 tgcagtgaaa actctcttca attctttatg ccggtgttgg gcgcgttatt tatcggagtt 1020 gcagttgcgc ccgcgaacga catttataat gaacgtgaat tgctcaacag tatgaacatt 1080 tcgcagccta ccgtagtgtt tgtttccaaa aaggggttgc aaaaaatttt gaacgtgcaa 1140 aaaaaattac caataatcca gaaaattatt atcatggatt ctaaaacgga ttaccaggga 1200 tttcagtcga tgtacacgtt cgtcacatct catctacctc ccggttttaa tgaatacgat 1260 tttgtaccag agtcctttga tcgtgacaaa acaattgcac tgataatgaa ttcctctgga 1320 tctactgggt tacctaaggg tgtggccctt ccgcatagaa ctgcctgcgt cagattctcg 1380 catgccagag atcctatttt tggcaatcaa atcattccgg atactgcgat tttaagtgtt 1440 gttccattcc atcacggttt tggaatgttt actacactcg gatatttgat atgtggattt 1500 cgagtcgtct taatgtatag atttgaagaa gagctgtttt tacgatccct tcaggattac 1560 aaaattcaaa gtgcgttgct agtaccaacc ctattttcat tcttcgccaa aagcactctg 1620 attgacaaat acgatttatc taatttacac gaaattgctt ctgggggcgc acctctttcg 1680 aaagaagtcg gggaagcggt tgcaaaacgc ttccatcttc cagggatacg acaaggatat 1740 gggctcactg agactacatc agctattctg attacacccg agggggatga taaaccgggc 1800 gcggtcggta aagttgttcc attttttgaa gcgaaggttg tggatctgga taccgggaaa 1860 acgctgggcg ttaatcagag aggcgaatta tgtgtcagag gacctatgat tatgtccggt 1920 tatgtaaaca atccggaagc gaccaacgcc ttgattgaca aggatggatg gctacattct 1980 ggagacatag cttactggga cgaagacgaa cacttcttca tagttgaccg cttgaagtct 2040 ttaattaaat acaaaggata tcaggtggcc cccgctgaat tggaatcgat attgttacaa 2100 caccccaaca tcttcgacgc gggcgtggca ggtcttcccg acgatgacgc cggtgaactt 2160 cccgccgccg ttgttgtttt ggagcacgga aagacgatga cggaaaaaga gatcgtggat 2220 tacgtggcca gtcaagtaac aaccgcgaaa aagttgcgcg gaggagttgt gtttgtggac 2280 gaagtaccga aaggtcttac cggaaaactc gacgcaagaa aaatcagaga gatcctcata 2340 aaggccaaga agggcggaaa gtccaaattg taataagcgg ccgc 2384 <210> 31 <211> 2188 <212> DNA <213> Artificial Sequence <220> <223> AS100KRib2-F.luci <400> 31 aagcttccat caattactac ttgcttcctg taggaatcct ctattgttgg atcatattcg 60 tccacaaaat gattctgaat tagctgtatc gtcaaggcac tcttgcaagc ttcatcatgg 120 ccgacagcaa aagttatcag gcatgcacct ggtagctagt ctttaaacca atagattgca 180 tcggtttaaa aggcaagacc gtcaaattgc gggaaagggg tcaacagccg ttcagtacca 240 agtctcaggg gaaactttga gatggccttg caaagggtat ggtaataagc tgacggacat 300 ggtcctaacc acgcagccaa gtcctaagtc aacagatctt ctgttgatat ggatgcagtt 360 cacagactaa atgtcggtcg gggaagatgt attcttctca taagatatag tcggacctct 420 ccttaatggg agctagcgga tgaagtgatg caacactgga gccgctggga actaatttgt 480 atgcgaaagt atattgatta gttttggagt actcgcgaca tgatcacgaa gacgccaaaa 540 acataaagaa aggcccggcg ccattctatc ctctagagga tggaaccgct ggagagcaac 600 tgcataaggc tatgaagaga tacgccctgg ttcctggaac aattgctttt acagatgcac 660 atatcgaggt gaacatcacg tacgcggaat acttcgaaat gtccgttcgg ttggcagaag 720 ctatgaaacg atatgggctg aatacaaatc acagaatcgt cgtatgcagt gaaaactctc 780 ttcaattctt tatgccggtg ttgggcgcgt tatttatcgg agttgcagtt gcgcccgcga 840 acgacattta taatgaacgt gaattgctca acagtatgaa catttcgcag cctaccgtag 900 tgtttgtttc caaaaagggg ttgcaaaaaa ttttgaacgt gcaaaaaaaa ttaccaataa 960 tccagaaaat tattatcatg gattctaaaa cggattacca gggatttcag tcgatgtaca 1020 cgttcgtcac atctcatcta cctcccggtt ttaatgaata cgattttgta ccagagtcct 1080 ttgatcgtga caaaacaatt gcactgataa tgaattcctc tggatctact gggttaccta 1140 agggtgtggc ccttccgcat agaactgcct gcgtcagatt ctcgcatgcc agagatccta 1200 tttttggcaa tcaaatcatt ccggatactg cgattttaag tgttgttcca ttccatcacg 1260 gttttggaat gtttactaca ctcggatatt tgatatgtgg atttcgagtc gtcttaatgt 1320 atagatttga agaagagctg tttttacgat cccttcagga ttacaaaatt caaagtgcgt 1380 tgctagtacc aaccctattt tcattcttcg ccaaaagcac tctgattgac aaatacgatt 1440 tatctaattt acacgaaatt gcttctgggg gcgcacctct ttcgaaagaa gtcggggaag 1500 cggttgcaaa acgcttccat cttccaggga tacgacaagg atatgggctc actgagacta 1560 catcagctat tctgattaca cccgaggggg atgataaacc gggcgcggtc ggtaaagttg 1620 ttccattttt tgaagcgaag gttgtggatc tggataccgg gaaaacgctg ggcgttaatc 1680 agagaggcga attatgtgtc agaggaccta tgattatgtc cggttatgta aacaatccgg 1740 aagcgaccaa cgccttgatt gacaaggatg gatggctaca ttctggagac atagcttact 1800 gggacgaaga cgaacacttc ttcatagttg accgcttgaa gtctttaatt aaatacaaag 1860 gatatcaggt ggcccccgct gaattggaat cgatattgtt acaacacccc aacatcttcg 1920 acgcgggcgt ggcaggtctt cccgacgatg acgccggtga acttcccgcc gccgttgttg 1980 ttttggagca cggaaagacg atgacggaaa aagagatcgt ggattacgtg gccagtcaag 2040 taacaaccgc gaaaaagttg cgcggaggag ttgtgtttgt ggacgaagta ccgaaaggtc 2100 ttaccggaaa actcgacgca agaaaaatca gagagatcct cataaaggcc aagaagggcg 2160 gaaagtccaa attgtaataa gcggccgc 2188 <210> 32 <211> 2377 <212> DNA <213> Artificial Sequence <220> <223> AS300KRib2-F.luci <400> 32 aagcttctac taggaccata ggtacatctt cagagtcctt aactctttta atttgttctc 60 tataatggtg aatatcttca aatgatttag tattatttat ggcaaataca caaagaaagc 120 cctccccagt cctcatgtac tggtccctca ttgcactgta ctcctcttga cctgctgtgt 180 cgagaatatc caagagacag gtttctccat caattactac ttgcttcctg taggaatcct 240 ctattgttgg atcatattcg tccacaaaat gattctgaat tagctgtatc gtcaaggcac 300 tcttgcaagc ttcatcatgg ccgacagcaa aagttatcag gcatgcacct ggtagctagt 360 ctttaaacca atagattgca tcggtttaaa aggcaagacc gtcaaattgc gggaaagggg 420 tcaacagccg ttcagtacca agtctcaggg gaaactttga gatggccttg caaagggtat 480 ggtaataagc tgacggacat ggtcctaacc acgcagccaa gtcctaagtc aacagatctt 540 ctgttgatat ggatgcagtt cacagactaa atgtcggtcg gggaagatgt attcttctca 600 taagatatag tcggacctct ccttaatggg agctagcgga tgaagtgatg caacactgga 660 gccgctggga actaatttgt atgcgaaagt atattgatta gttttggagt actcgcgaca 720 tgatcacgaa gacgccaaaa acataaagaa aggcccggcg ccattctatc ctctagagga 780 tggaaccgct ggagagcaac tgcataaggc tatgaagaga tacgccctgg ttcctggaac 840 aattgctttt acagatgcac atatcgaggt gaacatcacg tacgcggaat acttcgaaat 900 gtccgttcgg ttggcagaag ctatgaaacg atatgggctg aatacaaatc acagaatcgt 960 cgtatgcagt gaaaactctc ttcaattctt tatgccggtg ttgggcgcgt tatttatcgg 1020 agttgcagtt gcgcccgcga acgacattta taatgaacgt gaattgctca acagtatgaa 1080 catttcgcag cctaccgtag tgtttgtttc caaaaagggg ttgcaaaaaa ttttgaacgt 1140 gcaaaaaaaa ttaccaataa tccagaaaat tattatcatg gattctaaaa cggattacca 1200 gggatttcag tcgatgtaca cgttcgtcac atctcatcta cctcccggtt ttaatgaata 1260 cgattttgta ccagagtcct ttgatcgtga caaaacaatt gcactgataa tgaattcctc 1320 tggatctact gggttaccta agggtgtggc ccttccgcat agaactgcct gcgtcagatt 1380 ctcgcatgcc agagatccta tttttggcaa tcaaatcatt ccggatactg cgattttaag 1440 tgttgttcca ttccatcacg gttttggaat gtttactaca ctcggatatt tgatatgtgg 1500 atttcgagtc gtcttaatgt atagatttga agaagagctg tttttacgat cccttcagga 1560 ttacaaaatt caaagtgcgt tgctagtacc aaccctattt tcattcttcg ccaaaagcac 1620 tctgattgac aaatacgatt tatctaattt acacgaaatt gcttctgggg gcgcacctct 1680 ttcgaaagaa gtcggggaag cggttgcaaa acgcttccat cttccaggga tacgacaagg 1740 atatgggctc actgagacta catcagctat tctgattaca cccgaggggg atgataaacc 1800 gggcgcggtc ggtaaagttg ttccattttt tgaagcgaag gttgtggatc tggataccgg 1860 gaaaacgctg ggcgttaatc agagaggcga attatgtgtc agaggaccta tgattatgtc 1920 cggttatgta aacaatccgg aagcgaccaa cgccttgatt gacaaggatg gatggctaca 1980 ttctggagac atagcttact gggacgaaga cgaacacttc ttcatagttg accgcttgaa 2040 gtctttaatt aaatacaaag gatatcaggt ggcccccgct gaattggaat cgatattgtt 2100 acaacacccc aacatcttcg acgcgggcgt ggcaggtctt cccgacgatg acgccggtga 2160 acttcccgcc gccgttgttg ttttggagca cggaaagacg atgacggaaa aagagatcgt 2220 ggattacgtg gccagtcaag taacaaccgc gaaaaagttg cgcggaggag ttgtgtttgt 2280 ggacgaagta ccgaaaggtc ttaccggaaa actcgacgca agaaaaatca gagagatcct 2340 cataaaggcc aagaagggcg gaaagtccaa attgtaa 2377 <210> 33 <211> 1650 <212> DNA <213> Artificial Sequence <220> <223> AS100KRib1-TK <400> 33 aagcttccat caattactac ttgcttcctg taggaatcct ctattgttgg atcatattcg 60 tccacaaaat gattctgaat tagctgtatc gtcaaggcac tcttgcaagc ttcattatca 120 gcagctaaaa gttatcaggc atgcacctgg tagctagtct ttaaaccaat agattgcatc 180 ggtttaaaag gcaagaccgt caaattgcgg gaaaggggtc aacagccgtt cagtaccaag 240 tctcagggga aactttgaga tggccttgca aagggtatgg taataagctg acggacatgg 300 tcctaaccac gcagccaagt cctaagtcaa cagatcttct gttgatatgg atgcagttca 360 cagactaaat gtcggtcggg gaagatgtat tcttctcata agatatagtc ggacctctcc 420 ttaatgggag ctagcggatg aagtgatgca acactggagc cgctgggaac taatttgtat 480 gcgaaagtat attgattagt tttggagtac tcgcgatgat cacgcttcgt acccctgcca 540 tcaacacgcg tctgcgttcg accaggctgc gcgttctcgc ggccatagca accgacgtac 600 ggcgttgcgc cctcgccggc agcaagaagc cacggaagtc cgcctggagc agaaaatgcc 660 cacgctactg cgggtttata tagacggtcc tcacgggatg gggaaaacca ccaccacgca 720 actgctggtg gccctgggtt cgcgcgacga tatcgtctac gtacccgagc cgatgactta 780 ctggcaggtg ctgggggctt ccgagacaat cgcgaacatc tacaccacac aacaccgcct 840 cgaccagggt gagatatcgg ccggggacgc ggcggtggta atgacaagcg cccagataac 900 aatgggcatg ccttatgccg tgaccgacgc cgttctggct cctcatgtcg ggggggaggc 960 tgggagttca catgccccgc ccccggccct caccctcatc ttcgaccgcc atcccatcgc 1020 cgccctcctg tgctacccgg ccgcgcgata ccttatgggc agcatgaccc cccaggccgt 1080 gctggcgttc gtggccctca tcccgccgac cttgcccggc acaaacatcg tgttgggggc 1140 ccttccggag gacagacaca tcgaccgcct ggccaaacgc cagcgccccg gcgagcggct 1200 tgacctggct atgctggccg cgattcgccg cgtttacggg ctgcttgcca atacggtgcg 1260 gtatctgcag ggcggcgggt cgtggtggga ggattgggga cagctttcgg ggacggccgt 1320 gccgccccag ggtgccgagc cccagagcaa cgcgggccca cgaccccata tcggggacac 1380 gttatttacc ctgtttcggg cccccgagtt gctggccccc aacggcgacc tgtataacgt 1440 gtttgcctgg gccttggacg tcttggccaa acgcctccgt cccatgcacg tctttatcct 1500 ggattacgac caatcgcccg ccggctgccg ggacgccctg ctgcaactta cctccgggat 1560 ggtccagacc cacgtcacca ccccaggctc cataccgacg atctgcgacc tggcgcgcac 1620 gtttgcccgg gagatggggg aggctaataa 1650 <210> 34 <211> 1850 <212> DNA <213> Artificial Sequence <220> <223> AS300KRib1-TK <400> 34 aagcttctac taggaccata ggtacatctt cagagtcctt aactctttta atttgttctc 60 tataatggtg aatatcttca aatgatttag tattatttat ggcaaataca caaagaaagc 120 cctccccagt cctcatgtac tggtccctca ttgcactgta ctcctcttga cctgctgtgt 180 cgagaatatc caagagacag gtttctccat caattactac ttgcttcctg taggaatcct 240 ctattgttgg atcatattcg tccacaaaat gattctgaat tagctgtatc gtcaaggcac 300 tcttgcaagc ttcattatca gcagctaaaa gttatcaggc atgcacctgg tagctagtct 360 ttaaaccaat agattgcatc ggtttaaaag gcaagaccgt caaattgcgg gaaaggggtc 420 aacagccgtt cagtaccaag tctcagggga aactttgaga tggccttgca aagggtatgg 480 taataagctg acggacatgg tcctaaccac gcagccaagt cctaagtcaa cagatcttct 540 gttgatatgg atgcagttca cagactaaat gtcggtcggg gaagatgtat tcttctcata 600 agatatagtc ggacctctcc ttaatgggag ctagcggatg aagtgatgca acactggagc 660 cgctgggaac taatttgtat gcgaaagtat attgattagt tttggagtac tcgcgatgat 720 cacgcttcgt acccctgcca tcaacacgcg tctgcgttcg accaggctgc gcgttctcgc 780 ggccatagca accgacgtac ggcgttgcgc cctcgccggc agcaagaagc cacggaagtc 840 cgcctggagc agaaaatgcc cacgctactg cgggtttata tagacggtcc tcacgggatg 900 gggaaaacca ccaccacgca actgctggtg gccctgggtt cgcgcgacga tatcgtctac 960 gtacccgagc cgatgactta ctggcaggtg ctgggggctt ccgagacaat cgcgaacatc 1020 tacaccacac aacaccgcct cgaccagggt gagatatcgg ccggggacgc ggcggtggta 1080 atgacaagcg cccagataac aatgggcatg ccttatgccg tgaccgacgc cgttctggct 1140 cctcatgtcg ggggggaggc tgggagttca catgccccgc ccccggccct caccctcatc 1200 ttcgaccgcc atcccatcgc cgccctcctg tgctacccgg ccgcgcgata ccttatgggc 1260 agcatgaccc cccaggccgt gctggcgttc gtggccctca tcccgccgac cttgcccggc 1320 acaaacatcg tgttgggggc ccttccggag gacagacaca tcgaccgcct ggccaaacgc 1380 cagcgccccg gcgagcggct tgacctggct atgctggccg cgattcgccg cgtttacggg 1440 ctgcttgcca atacggtgcg gtatctgcag ggcggcgggt cgtggtggga ggattgggga 1500 cagctttcgg ggacggccgt gccgccccag ggtgccgagc cccagagcaa cgcgggccca 1560 cgaccccata tcggggacac gttatttacc ctgtttcggg cccccgagtt gctggccccc 1620 aacggcgacc tgtataacgt gtttgcctgg gccttggacg tcttggccaa acgcctccgt 1680 cccatgcacg tctttatcct ggattacgac caatcgcccg ccggctgccg ggacgccctg 1740 ctgcaactta cctccgggat ggtccagacc cacgtcacca ccccaggctc cataccgacg 1800 atctgcgacc tggcgcgcac gtttgcccgg gagatggggg aggctaataa 1850 <210> 35 <211> 1654 <212> DNA <213> Artificial Sequence <220> <223> AS100KRib2-TK <400> 35 aagcttccat caattactac ttgcttcctg taggaatcct ctattgttgg atcatattcg 60 tccacaaaat gattctgaat tagctgtatc gtcaaggcac tcttgcaagc ttcatcatgg 120 ccgacagcaa aagttatcag gcatgcacct ggtagctagt ctttaaacca atagattgca 180 tcggtttaaa aggcaagacc gtcaaattgc gggaaagggg tcaacagccg ttcagtacca 240 agtctcaggg gaaactttga gatggccttg caaagggtat ggtaataagc tgacggacat 300 ggtcctaacc acgcagccaa gtcctaagtc aacagatctt ctgttgatat ggatgcagtt 360 cacagactaa atgtcggtcg gggaagatgt attcttctca taagatatag tcggacctct 420 ccttaatggg agctagcgga tgaagtgatg caacactgga gccgctggga actaatttgt 480 atgcgaaagt atattgatta gttttggagt actcgcgaca tgatcacgct tcgtacccct 540 gccatcaaca cgcgtctgcg ttcgaccagg ctgcgcgttc tcgcggccat agcaaccgac 600 gtacggcgtt gcgccctcgc cggcagcaag aagccacgga agtccgcctg gagcagaaaa 660 tgcccacgct actgcgggtt tatatagacg gtcctcacgg gatggggaaa accaccacca 720 cgcaactgct ggtggccctg ggttcgcgcg acgatatcgt ctacgtaccc gagccgatga 780 cttactggca ggtgctgggg gcttccgaga caatcgcgaa catctacacc acacaacacc 840 gcctcgacca gggtgagata tcggccgggg acgcggcggt ggtaatgaca agcgcccaga 900 taacaatggg catgccttat gccgtgaccg acgccgttct ggctcctcat gtcggggggg 960 aggctgggag ttcacatgcc ccgcccccgg ccctcaccct catcttcgac cgccatccca 1020 tcgccgccct cctgtgctac ccggccgcgc gataccttat gggcagcatg accccccagg 1080 ccgtgctggc gttcgtggcc ctcatcccgc cgaccttgcc cggcacaaac atcgtgttgg 1140 gggcccttcc ggaggacaga cacatcgacc gcctggccaa acgccagcgc cccggcgagc 1200 ggcttgacct ggctatgctg gccgcgattc gccgcgttta cgggctgctt gccaatacgg 1260 tgcggtatct gcagggcggc gggtcgtggt gggaggattg gggacagctt tcggggacgg 1320 ccgtgccgcc ccagggtgcc gagccccaga gcaacgcggg cccacgaccc catatcgggg 1380 acacgttatt taccctgttt cgggcccccg agttgctggc ccccaacggc gacctgtata 1440 acgtgtttgc ctgggccttg gacgtcttgg ccaaacgcct ccgtcccatg cacgtcttta 1500 tcctggatta cgaccaatcg cccgccggct gccgggacgc cctgctgcaa cttacctccg 1560 ggatggtcca gacccacgtc accaccccag gctccatacc gacgatctgc gacctggcgc 1620 gcacgtttgc ccgggagatg ggggaggcta ataa 1654 <210> 36 <211> 1854 <212> DNA <213> Artificial Sequence <220> <223> AS300KRib2-TK <400> 36 aagcttctac taggaccata ggtacatctt cagagtcctt aactctttta atttgttctc 60 tataatggtg aatatcttca aatgatttag tattatttat ggcaaataca caaagaaagc 120 cctccccagt cctcatgtac tggtccctca ttgcactgta ctcctcttga cctgctgtgt 180 cgagaatatc caagagacag gtttctccat caattactac ttgcttcctg taggaatcct 240 ctattgttgg atcatattcg tccacaaaat gattctgaat tagctgtatc gtcaaggcac 300 tcttgcaagc ttcatcatgg ccgacagcaa aagttatcag gcatgcacct ggtagctagt 360 ctttaaacca atagattgca tcggtttaaa aggcaagacc gtcaaattgc gggaaagggg 420 tcaacagccg ttcagtacca agtctcaggg gaaactttga gatggccttg caaagggtat 480 ggtaataagc tgacggacat ggtcctaacc acgcagccaa gtcctaagtc aacagatctt 540 ctgttgatat ggatgcagtt cacagactaa atgtcggtcg gggaagatgt attcttctca 600 taagatatag tcggacctct ccttaatggg agctagcgga tgaagtgatg caacactgga 660 gccgctggga actaatttgt atgcgaaagt atattgatta gttttggagt actcgcgaca 720 tgatcacgct tcgtacccct gccatcaaca cgcgtctgcg ttcgaccagg ctgcgcgttc 780 tcgcggccat agcaaccgac gtacggcgtt gcgccctcgc cggcagcaag aagccacgga 840 agtccgcctg gagcagaaaa tgcccacgct actgcgggtt tatatagacg gtcctcacgg 900 gatggggaaa accaccacca cgcaactgct ggtggccctg ggttcgcgcg acgatatcgt 960 ctacgtaccc gagccgatga cttactggca ggtgctgggg gcttccgaga caatcgcgaa 1020 catctacacc acacaacacc gcctcgacca gggtgagata tcggccgggg acgcggcggt 1080 ggtaatgaca agcgcccaga taacaatggg catgccttat gccgtgaccg acgccgttct 1140 ggctcctcat gtcggggggg aggctgggag ttcacatgcc ccgcccccgg ccctcaccct 1200 catcttcgac cgccatccca tcgccgccct cctgtgctac ccggccgcgc gataccttat 1260 gggcagcatg accccccagg ccgtgctggc gttcgtggcc ctcatcccgc cgaccttgcc 1320 cggcacaaac atcgtgttgg gggcccttcc ggaggacaga cacatcgacc gcctggccaa 1380 acgccagcgc cccggcgagc ggcttgacct ggctatgctg gccgcgattc gccgcgttta 1440 cgggctgctt gccaatacgg tgcggtatct gcagggcggc gggtcgtggt gggaggattg 1500 gggacagctt tcggggacgg ccgtgccgcc ccagggtgcc gagccccaga gcaacgcggg 1560 cccacgaccc catatcgggg acacgttatt taccctgttt cgggcccccg agttgctggc 1620 ccccaacggc gacctgtata acgtgtttgc ctgggccttg gacgtcttgg ccaaacgcct 1680 ccgtcccatg cacgtcttta tcctggatta cgaccaatcg cccgccggct gccgggacgc 1740 cctgctgcaa cttacctccg ggatggtcca gacccacgtc accaccccag gctccatacc 1800 gacgatctgc gacctggcgc gcacgtttgc ccgggagatg ggggaggcta ataa 1854 <210> 37 <211> 8793 <212> DNA <213> Artificial Sequence <220> <223> pAVQ-AS300KRib1 <400> 37 taacatcatc aataatatac cttattttgg attgaagcca atatgataat gagggggtgg 60 agtttgtgac gtggcgcggg gcgtgggaac ggggcgggtg acgtagtagt gtggcggaag 120 tgtgatgttg caagtgtggc ggaacacatg taagcgacgg atgtggcaaa agtgacgttt 180 ttggtgtgcg ccggtgtaca caggaagtga caattttcgc gcggttttag gcggatgttg 240 tagtaaattt gggcgtaacc gagtaagatt tggccatttt cgcgggaaaa ctgaataaga 300 ggaagtgaaa tctgaataat tttgtgttac tcatagcgcg taatactgcg atctatacat 360 tgaatcaata ttggcaatta gccatattag tcattggtta tatagcataa atcaatattg 420 gctattggcc attgcatacg ttgtatctat atcataatat gtacatttat attggctcat 480 gtccaatatg accgccatgt tgacattgat tattgactag ttattaatag taatcaatta 540 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg 600 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc 660 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa 720 ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca 780 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta 840 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt 900 acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg 960 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata 1020 accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca 1080 gagctcgttt agtgaaccgt cagatcctca ctctcttccg catcgctgtc tgcgagggcc 1140 agctgttggg ctcgcggttg aggacaaact cttcgcggtc tttccagtac tcttggatcg 1200 gaaacccgtc ggcctccgaa cggtactccg ccaccgaggg acctgagcca gtccgcatcg 1260 accggatcgg aaaacctctc gagaaaggcg tctaaccagt cacagtcgca aggtaggctg 1320 agcaccgtgg cgggcggcag cgggtggcgg tcggggttgt ttctggcgga ggtgctgctg 1380 atgatgtaat taaagtaggc ggtcttgagc cggcggatgg tcgaggtgag gtgtggcagg 1440 cttgagatcc agctgttggg gtgagtactc cctctcaaaa gcgggcatga cttctgcgct 1500 aagattgtca gtttccaaaa acgaggagga tttgatattc acctggcccg atctggccat 1560 acacttgagt gacaatgaca tccactttgc ctttctctcc acaggtgtcc actcccaggt 1620 ccaagtttgg aagatcccta ctaggaccat aggtacatct tcagagtcct taactctttt 1680 aatttgttct ctataatggt gaatatcttc aaatgattta gtattattta tggcaaatac 1740 acaaagaaag ccctccccag tcctcatgta ctggtccctc attgcactgt actcctcttg 1800 acctgctgtg tcgagaatat ccaagagaca ggtttctcca tcaattacta cttgcttcct 1860 gtaggaatcc tctattgttg gatcatattc gtccacaaaa tgattctgaa ttagctgtat 1920 cgtcaaggca ctcttgcaag cttcattatc agcagctaaa agttatcagg catgcacctg 1980 gtagctagtc tttaaaccaa tagattgcat cggtttaaaa ggcaagaccg tcaaattgcg 2040 ggaaaggggt caacagccgt tcagtaccaa gtctcagggg aaactttgag atggccttgc 2100 aaagggtatg gtaataagct gacggacatg gtcctaacca cgcagccaag tcctaagtca 2160 acagatcttc tgttgatatg gatgcagttc acagactaaa tgtcggtcgg ggaagatgta 2220 ttcttctcat aagatatagt cggacctctc cttaatggga gctagcggat gaagtgatgc 2280 aacactggag ccgctgggaa ctaatttgta tgcgaaagta tattgattag ttttggagta 2340 ctccgaaaga tcccaacgaa aagagagacc acatggtcct tcttgagttt gtaacagctg 2400 ctgggattac acatggcatg gatgaactgt acaactgagg atcccccgac ctcgacctct 2460 ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca 2520 ctcggaagga catatgggag ggcaaatcat ttggtcgaga tccctcggag atcggatctg 2580 ggcgtggtta agggtgggaa agaatatata aggtgggggt cttatgtagt tttgtatctg 2640 ttttgcagca gccgccgccg ccatgagcac caactcgttt gatggaagca ttgtgagctc 2700 atatttgaca acgcgcatgc ccccatgggc cggggtgcgt cagaatgtga tgggctccag 2760 cattgatggt cgccccgtcc tgcccgcaaa ctctactacc ttgacctacg agaccgtgtc 2820 tggaacgccg ttggagactg cagcctccgc cgccgcttca gccgctgcag ccaccgcccg 2880 cgggattgtg actgactttg ctttcctgag cccgcttgca agcagtgcag cttcccgttc 2940 atccgcccgc gatgacaagt tgacggctct tttggcacaa ttggattctt tgacccggga 3000 acttaatgtc gtttctcagc agctgttgga tctgcgccag caggtttctg ccctgaaggc 3060 ttcctcccct cccaatgcgg tttaaaacat aaataaaaaa ccagactctg tttggatttg 3120 gatcaagcaa gtgtcttgct gtctttattt aggggttttg cgcgcgcggt aggcccggga 3180 ccagcggtct cggtcgttga gggtcctgtg tattttttcc aggacgtggt aaaggtgact 3240 ctggatgttc agatacatgg gcataagccc gtctctgggg tggaggtagc accactgcag 3300 agcttcatgc tgcggggtgg tgttgtagat gatccagtcg tagcaggagc gctgggcgtg 3360 gtgcctaaaa atgtctttca gtagcaagct gattgccagg ggcaggccct tggtgtaagt 3420 gtttacaaag cggttaagct gggatgggtg catacgtggg gatatgagat gcatcttgga 3480 ctgtattttt aggttggcta tgttcccagc catatccctc cggggattca tgttgtgcag 3540 aaccaccagc acagtgtatc cggtgcactt gggaaatttg tcatgtagct tagaaggaaa 3600 tgcgtggaag aacttggaga cgcccttgtg acctccaaga ttttccatgc attcgtccat 3660 aatgatggca atgggcccac gggcggcggc ctgggcgaag atatttctgg gatcactaac 3720 gtcatagttg tgttccagga tgagatcgtc ataggccatt tttacaaagc gcgggcggag 3780 ggtgccagac tgcggtataa tggttccatc cggcccaggg gcgtagttac cctcacagat 3840 ttgcatttcc cacgctttga gttcagatgg ggggatcatg tctacctgcg gggcgatgaa 3900 gaaaacggtt tccggggtag gggagatcag ctgggaagaa agcaggttcc tgagcagctg 3960 cgacttaccg cagccggtgg gcccgtaaat cacacctatt accgggtgca actggtagtt 4020 aagagagctg cagctgccgt catccctgag caggggggcc acttcgttaa gcatgtccct 4080 gactcgcatg ttttccctga ccaaatccgc cagaaggcgc tcgccgccca gcgatagcag 4140 ttcttgcaag gaagcaaagt ttttcaacgg tttgagaccg tccgccgtag gcatgctttt 4200 gagcgtttga ccaagcagtt ccaggcggtc ccacagctcg gtcacctgct ctacggcatc 4260 tcgatccagc atatctcctc gtttcgcggg ttggggcggc tttcgctgta cggcagtagt 4320 cggtgctcgt ccagacgggc cagggtcatg tctttccacg ggcgcagggt cctcgtcagc 4380 gtagtctggg tcacggtgaa ggggtgcgct ccgggctgcg cgctggccag ggtgcgcttg 4440 aggctggtcc tgctggtgct gaagcgctgc cggtcttcgc cctgcgcgtc ggccaggtag 4500 catttgacca tggtgtcata gtccagcccc tccgcggcgt ggcccttggc gcgcagcttg 4560 cccttggagg aggcgccgca cgaggggcag tgcagacttt tgagggcgta gagcttgggc 4620 gcgagaaata ccgattccgg ggagtaggca tccgcgccgc aggccccgca gacggtctcg 4680 cattccacga gccaggtgag ctctggccgt tcggggtcaa aaaccaggtt tcccccatgc 4740 tttttgatgc gtttcttacc tctggtttcc atgagccggt gtccacgctc ggtgacgaaa 4800 aggctgtccg tgtccccgta tacagacttg agagggagtt taaacgaatt caatagcttg 4860 ttgcatgggc ggcgatataa aatgcaaggt gctgctcaaa aaatcaggca aagcctcgcg 4920 caaaaaagaa agcacatcgt agtcatgctc atgcagataa aggcaggtaa gctccggaac 4980 caccacagaa aaagacacca tttttctctc aaacatgtct gcgggtttct gcataaacac 5040 aaaataaaat aacaaaaaaa catttaaaca ttagaagcct gtcttacaac aggaaaaaca 5100 acccttataa gcataagacg gactacggcc atgccggcgt gaccgtaaaa aaactggtca 5160 ccgtgattaa aaagcaccac cgacagctcc tcggtcatgt ccggagtcat aatgtaagac 5220 tcggtaaaca catcaggttg attcatcggt cagtgctaaa aagcgaccga aatagcccgg 5280 gggaatacat acccgcaggc gtagagacaa cattacagcc cccataggag gtataacaaa 5340 attaatagga gagaaaaaca cataaacacc tgaaaaaccc tcctgcctag gcaaaatagc 5400 accctcccgc tccagaacaa catacagcgc ttcacagcgg cagcctaaca gtcagcctta 5460 ccagtaaaaa agaaaaccta ttaaaaaaac accactcgac acggcaccag ctcaatcagt 5520 cacagtgtaa aaaagggcca agtgcagagc gagtatatat aggactaaaa aatgacgtaa 5580 cggttaaagt ccacaaaaaa cacccagaaa accgcacgcg aacctacgcc cagaaacgaa 5640 agccaaaaaa cccacaactt cctcaaatcg tcacttccgt tttcccacgt tacgtaactt 5700 cccattttaa gaaaactaca attcccaaca catacaagtt actccgccct aaaacctacg 5760 tcacccgccc cgttcccacg ccccgcgcca cgtcacaaac tccaccccct cattatcata 5820 ttggcttcaa tccaaaataa ggtatattat tgatgatgtt aattaacatg catggatcca 5880 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgctcttc 5940 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc 6000 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat 6060 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt 6120 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg 6180 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc 6240 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt 6300 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa 6360 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta 6420 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa 6480 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa 6540 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt 6600 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt 6660 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat 6720 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat 6780 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc 6840 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc 6900 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta 6960 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga 7020 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg 7080 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc 7140 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctgcagccat 7200 gagattatca aaaaggatct tcacctagat ccttttcacg tagaaagcca gtccgcagaa 7260 acggtgctga ccccggatga atgtcagcta ctgggctatc tggacaaggg aaaacgcaag 7320 cgcaaagaga aagcaggtag cttgcagtgg gcttacatgg cgatagctag actgggcggt 7380 tttatggaca gcaagcgaac cggaattgcc agctggggcg ccctctggta aggttgggaa 7440 gccctgcaaa gtaaactgga tggctttctc gccgccaagg atctgatggc gcaggggatc 7500 aagctctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca 7560 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac 7620 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt 7680 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc 7740 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg 7800 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc 7860 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc 7920 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat 7980 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc 8040 cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca 8100 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga 8160 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat 8220 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc 8280 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gaattttgtt 8340 aaaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aacatccctt 8400 ataaatcaaa agaatagacc gcgatagggt tgagtgttgt tccagtttgg aacaagagtc 8460 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg 8520 gcccactacg tgaaccatca cccaaatcaa gttttttgcg gtcgaggtgc cgtaaagctc 8580 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg 8640 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag 8700 cggtcacgct gcgcgtaacc accacacccg cgcgcttaat gcgccgctac agggcgcgtc 8760 cattcgccat tcaggatcga attaattctt aat 8793 <210> 38 <211> 9939 <212> DNA <213> Artificial Sequence <220> <223> pAVQ-AS300KRib1-TK <400> 38 taacatcatc aataatatac cttattttgg attgaagcca atatgataat gagggggtgg 60 agtttgtgac gtggcgcggg gcgtgggaac ggggcgggtg acgtagtagt gtggcggaag 120 tgtgatgttg caagtgtggc ggaacacatg taagcgacgg atgtggcaaa agtgacgttt 180 ttggtgtgcg ccggtgtaca caggaagtga caattttcgc gcggttttag gcggatgttg 240 tagtaaattt gggcgtaacc gagtaagatt tggccatttt cgcgggaaaa ctgaataaga 300 ggaagtgaaa tctgaataat tttgtgttac tcatagcgcg taatactgcg atctatacat 360 tgaatcaata ttggcaatta gccatattag tcattggtta tatagcataa atcaatattg 420 gctattggcc attgcatacg ttgtatctat atcataatat gtacatttat attggctcat 480 gtccaatatg accgccatgt tgacattgat tattgactag ttattaatag taatcaatta 540 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg 600 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc 660 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa 720 ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca 780 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta 840 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt 900 acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg 960 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata 1020 accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca 1080 gagctcgttt agtgaaccgt cagatcctca ctctcttccg catcgctgtc tgcgagggcc 1140 agctgttggg ctcgcggttg aggacaaact cttcgcggtc tttccagtac tcttggatcg 1200 gaaacccgtc ggcctccgaa cggtactccg ccaccgaggg acctgagcca gtccgcatcg 1260 accggatcgg aaaacctctc gagaaaggcg tctaaccagt cacagtcgca aggtaggctg 1320 agcaccgtgg cgggcggcag cgggtggcgg tcggggttgt ttctggcgga ggtgctgctg 1380 atgatgtaat taaagtaggc ggtcttgagc cggcggatgg tcgaggtgag gtgtggcagg 1440 cttgagatcc agctgttggg gtgagtactc cctctcaaaa gcgggcatga cttctgcgct 1500 aagattgtca gtttccaaaa acgaggagga tttgatattc acctggcccg atctggccat 1560 acacttgagt gacaatgaca tccactttgc ctttctctcc acaggtgtcc actcccaggt 1620 ccaagtttgg aagatcccta ctaggaccat aggtacatct tcagagtcct taactctttt 1680 aatttgttct ctataatggt gaatatcttc aaatgattta gtattattta tggcaaatac 1740 acaaagaaag ccctccccag tcctcatgta ctggtccctc attgcactgt actcctcttg 1800 acctgctgtg tcgagaatat ccaagagaca ggtttctcca tcaattacta cttgcttcct 1860 gtaggaatcc tctattgttg gatcatattc gtccacaaaa tgattctgaa ttagctgtat 1920 cgtcaaggca ctcttgcaag cttcattatc agcagctaaa agttatcagg catgcacctg 1980 gtagctagtc tttaaaccaa tagattgcat cggtttaaaa ggcaagaccg tcaaattgcg 2040 ggaaaggggt caacagccgt tcagtaccaa gtctcagggg aaactttgag atggccttgc 2100 aaagggtatg gtaataagct gacggacatg gtcctaacca cgcagccaag tcctaagtca 2160 acagatcttc tgttgatatg gatgcagttc acagactaaa tgtcggtcgg ggaagatgta 2220 ttcttctcat aagatatagt cggacctctc cttaatggga gctagcggat gaagtgatgc 2280 aacactggag ccgctgggaa ctaatttgta tgcgaaagta tattgattag ttttggagta 2340 ctcgcgatga tcacgcttcg tacccctgcc atcaacacgc gtctgcgttc gaccaggctg 2400 cgcgttctcg cggccatagc aaccgacgta cggcgttgcg ccctcgccgg cagcaagaag 2460 ccacggaagt ccgcctggag cagaaaatgc ccacgctact gcgggtttat atagacggtc 2520 ctcacgggat ggggaaaacc accaccacgc aactgctggt ggccctgggt tcgcgcgacg 2580 atatcgtcta cgtacccgag ccgatgactt actggcaggt gctgggggct tccgagacaa 2640 tcgcgaacat ctacaccaca caacaccgcc tcgaccaggg tgagatatcg gccggggacg 2700 cggcggtggt aatgacaagc gcccagataa caatgggcat gccttatgcc gtgaccgacg 2760 ccgttctggc tcctcatgtc gggggggagg ctgggagttc acatgccccg cccccggccc 2820 tcaccctcat cttcgaccgc catcccatcg ccgccctcct gtgctacccg gccgcgcgat 2880 accttatggg cagcatgacc ccccaggccg tgctggcgtt cgtggccctc atcccgccga 2940 ccttgcccgg cacaaacatc gtgttggggg cccttccgga ggacagacac atcgaccgcc 3000 tggccaaacg ccagcgcccc ggcgagcggc ttgacctggc tatgctggcc gcgattcgcc 3060 gcgtttacgg gctgcttgcc aatacggtgc ggtatctgca gggcggcggg tcgtggtggg 3120 aggattgggg acagctttcg gggacggccg tgccgcccca gggtgccgag ccccagagca 3180 acgcgggccc acgaccccat atcggggaca cgttatttac cctgtttcgg gcccccgagt 3240 tgctggcccc caacggcgac ctgtataacg tgtttgcctg ggccttggac gtcttggcca 3300 aacgcctccg tcccatgcac gtctttatcc tggattacga ccaatcgccc gccggctgcc 3360 gggacgccct gctgcaactt acctccggga tggtccagac ccacgtcacc accccaggct 3420 ccataccgac gatctgcgac ctggcgcgca cgtttgcccg ggagatgggg gaggctaact 3480 gaagcttttc gaaagatccc aacgaaaaga gagaccacat ggtccttctt gagtttgtaa 3540 cagctgctgg gattacacat ggcatggatg aactgtacaa ctgaggatcc cccgacctcg 3600 acctctggct aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt 3660 ctctcactcg gaaggacata tgggagggca aatcatttgg tcgagatccc tcggagatcg 3720 gatctgggcg tggttaaggg tgggaaagaa tatataaggt gggggtctta tgtagttttg 3780 tatctgtttt gcagcagccg ccgccgccat gagcaccaac tcgtttgatg gaagcattgt 3840 gagctcatat ttgacaacgc gcatgccccc atgggccggg gtgcgtcaga atgtgatggg 3900 ctccagcatt gatggtcgcc ccgtcctgcc cgcaaactct actaccttga cctacgagac 3960 cgtgtctgga acgccgttgg agactgcagc ctccgccgcc gcttcagccg ctgcagccac 4020 cgcccgcggg attgtgactg actttgcttt cctgagcccg cttgcaagca gtgcagcttc 4080 ccgttcatcc gcccgcgatg acaagttgac ggctcttttg gcacaattgg attctttgac 4140 ccgggaactt aatgtcgttt ctcagcagct gttggatctg cgccagcagg tttctgccct 4200 gaaggcttcc tcccctccca atgcggttta aaacataaat aaaaaaccag actctgtttg 4260 gatttggatc aagcaagtgt cttgctgtct ttatttaggg gttttgcgcg cgcggtaggc 4320 ccgggaccag cggtctcggt cgttgagggt cctgtgtatt ttttccagga cgtggtaaag 4380 gtgactctgg atgttcagat acatgggcat aagcccgtct ctggggtgga ggtagcacca 4440 ctgcagagct tcatgctgcg gggtggtgtt gtagatgatc cagtcgtagc aggagcgctg 4500 ggcgtggtgc ctaaaaatgt ctttcagtag caagctgatt gccaggggca ggcccttggt 4560 gtaagtgttt acaaagcggt taagctggga tgggtgcata cgtggggata tgagatgcat 4620 cttggactgt atttttaggt tggctatgtt cccagccata tccctccggg gattcatgtt 4680 gtgcagaacc accagcacag tgtatccggt gcacttggga aatttgtcat gtagcttaga 4740 aggaaatgcg tggaagaact tggagacgcc cttgtgacct ccaagatttt ccatgcattc 4800 gtccataatg atggcaatgg gcccacgggc ggcggcctgg gcgaagatat ttctgggatc 4860 actaacgtca tagttgtgtt ccaggatgag atcgtcatag gccattttta caaagcgcgg 4920 gcggagggtg ccagactgcg gtataatggt tccatccggc ccaggggcgt agttaccctc 4980 acagatttgc atttcccacg ctttgagttc agatgggggg atcatgtcta cctgcggggc 5040 gatgaagaaa acggtttccg gggtagggga gatcagctgg gaagaaagca ggttcctgag 5100 cagctgcgac ttaccgcagc cggtgggccc gtaaatcaca cctattaccg ggtgcaactg 5160 gtagttaaga gagctgcagc tgccgtcatc cctgagcagg ggggccactt cgttaagcat 5220 gtccctgact cgcatgtttt ccctgaccaa atccgccaga aggcgctcgc cgcccagcga 5280 tagcagttct tgcaaggaag caaagttttt caacggtttg agaccgtccg ccgtaggcat 5340 gcttttgagc gtttgaccaa gcagttccag gcggtcccac agctcggtca cctgctctac 5400 ggcatctcga tccagcatat ctcctcgttt cgcgggttgg ggcggctttc gctgtacggc 5460 agtagtcggt gctcgtccag acgggccagg gtcatgtctt tccacgggcg cagggtcctc 5520 gtcagcgtag tctgggtcac ggtgaagggg tgcgctccgg gctgcgcgct ggccagggtg 5580 cgcttgaggc tggtcctgct ggtgctgaag cgctgccggt cttcgccctg cgcgtcggcc 5640 aggtagcatt tgaccatggt gtcatagtcc agcccctccg cggcgtggcc cttggcgcgc 5700 agcttgccct tggaggaggc gccgcacgag gggcagtgca gacttttgag ggcgtagagc 5760 ttgggcgcga gaaataccga ttccggggag taggcatccg cgccgcaggc cccgcagacg 5820 gtctcgcatt ccacgagcca ggtgagctct ggccgttcgg ggtcaaaaac caggtttccc 5880 ccatgctttt tgatgcgttt cttacctctg gtttccatga gccggtgtcc acgctcggtg 5940 acgaaaaggc tgtccgtgtc cccgtataca gacttgagag ggagtttaaa cgaattcaat 6000 agcttgttgc atgggcggcg atataaaatg caaggtgctg ctcaaaaaat caggcaaagc 6060 ctcgcgcaaa aaagaaagca catcgtagtc atgctcatgc agataaaggc aggtaagctc 6120 cggaaccacc acagaaaaag acaccatttt tctctcaaac atgtctgcgg gtttctgcat 6180 aaacacaaaa taaaataaca aaaaaacatt taaacattag aagcctgtct tacaacagga 6240 aaaacaaccc ttataagcat aagacggact acggccatgc cggcgtgacc gtaaaaaaac 6300 tggtcaccgt gattaaaaag caccaccgac agctcctcgg tcatgtccgg agtcataatg 6360 taagactcgg taaacacatc aggttgattc atcggtcagt gctaaaaagc gaccgaaata 6420 gcccggggga atacataccc gcaggcgtag agacaacatt acagccccca taggaggtat 6480 aacaaaatta ataggagaga aaaacacata aacacctgaa aaaccctcct gcctaggcaa 6540 aatagcaccc tcccgctcca gaacaacata cagcgcttca cagcggcagc ctaacagtca 6600 gccttaccag taaaaaagaa aacctattaa aaaaacacca ctcgacacgg caccagctca 6660 atcagtcaca gtgtaaaaaa gggccaagtg cagagcgagt atatatagga ctaaaaaatg 6720 acgtaacggt taaagtccac aaaaaacacc cagaaaaccg cacgcgaacc tacgcccaga 6780 aacgaaagcc aaaaaaccca caacttcctc aaatcgtcac ttccgttttc ccacgttacg 6840 taacttccca ttttaagaaa actacaattc ccaacacata caagttactc cgccctaaaa 6900 cctacgtcac ccgccccgtt cccacgcccc gcgccacgtc acaaactcca ccccctcatt 6960 atcatattgg cttcaatcca aaataaggta tattattgat gatgttaatt aacatgcatg 7020 gatccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg 7080 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 7140 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 7200 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 7260 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 7320 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 7380 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 7440 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 7500 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 7560 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 7620 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 7680 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt 7740 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 7800 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 7860 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 7920 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 7980 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 8040 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 8100 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 8160 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 8220 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 8280 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctgc 8340 agccatgaga ttatcaaaaa ggatcttcac ctagatcctt ttcacgtaga aagccagtcc 8400 gcagaaacgg tgctgacccc ggatgaatgt cagctactgg gctatctgga caagggaaaa 8460 cgcaagcgca aagagaaagc aggtagcttg cagtgggctt acatggcgat agctagactg 8520 ggcggtttta tggacagcaa gcgaaccgga attgccagct ggggcgccct ctggtaaggt 8580 tgggaagccc tgcaaagtaa actggatggc tttctcgccg ccaaggatct gatggcgcag 8640 gggatcaagc tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg 8700 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca 8760 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt 8820 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg 8880 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga 8940 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca 9000 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct 9060 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac 9120 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc 9180 gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt 9240 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt 9300 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg 9360 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat 9420 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgaat 9480 tttgttaaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaca 9540 tcccttataa atcaaaagaa tagaccgcga tagggttgag tgttgttcca gtttggaaca 9600 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg 9660 gcgatggccc actacgtgaa ccatcaccca aatcaagttt tttgcggtcg aggtgccgta 9720 aagctctaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg 9780 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa 9840 gtgtagcggt cacgctgcgc gtaaccacca cacccgcgcg cttaatgcgc cgctacaggg 9900 cgcgtccatt cgccattcag gatcgaatta attcttaat 9939 <210> 39 <211> 8607 <212> DNA <213> Artificial Sequence <220> <223> pAVQ-AS100KRib2 <400> 39 taacatcatc aataatatac cttattttgg attgaagcca atatgataat gagggggtgg 60 agtttgtgac gtggcgcggg gcgtgggaac ggggcgggtg acgtagtagt gtggcggaag 120 tgtgatgttg caagtgtggc ggaacacatg taagcgacgg atgtggcaaa agtgacgttt 180 ttggtgtgcg ccggtgtaca caggaagtga caattttcgc gcggttttag gcggatgttg 240 tagtaaattt gggcgtaacc gagtaagatt tggccatttt cgcgggaaaa ctgaataaga 300 ggaagtgaaa tctgaataat tttgtgttac tcatagcgcg taatactgcg atctatacat 360 tgaatcaata ttggcaatta gccatattag tcattggtta tatagcataa atcaatattg 420 gctattggcc attgcatacg ttgtatctat atcataatat gtacatttat attggctcat 480 gtccaatatg accgccatgt tgacattgat tattgactag ttattaatag taatcaatta 540 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg 600 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc 660 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa 720 ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca 780 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta 840 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt 900 acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg 960 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata 1020 accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca 1080 gagctcgttt agtgaaccgt cagatcctca ctctcttccg catcgctgtc tgcgagggcc 1140 agctgttggg ctcgcggttg aggacaaact cttcgcggtc tttccagtac tcttggatcg 1200 gaaacccgtc ggcctccgaa cggtactccg ccaccgaggg acctgagcca gtccgcatcg 1260 accggatcgg aaaacctctc gagaaaggcg tctaaccagt cacagtcgca aggtaggctg 1320 agcaccgtgg cgggcggcag cgggtggcgg tcggggttgt ttctggcgga ggtgctgctg 1380 atgatgtaat taaagtaggc ggtcttgagc cggcggatgg tcgaggtgag gtgtggcagg 1440 cttgagatcc agctgttggg gtgagtactc cctctcaaaa gcgggcatga cttctgcgct 1500 aagattgtca gtttccaaaa acgaggagga tttgatattc acctggcccg atctggccat 1560 acacttgagt gacaatgaca tccactttgc ctttctctcc acaggtgtcc actcccaggt 1620 ccaagtttgg aagatcccca tcaattacta cttgcttcct gtaggaatcc tctattgttg 1680 gatcatattc gtccacaaaa tgattctgaa ttagctgtat cgtcaaggca ctcttgcaag 1740 cttcatcatg gccgacagca aaagttatca ggcatgcacc tggtagctag tctttaaacc 1800 aatagattgc atcggtttaa aaggcaagac cgtcaaattg cgggaaaggg gtcaacagcc 1860 gttcagtacc aagtctcagg ggaaactttg agatggcctt gcaaagggta tggtaataag 1920 ctgacggaca tggtcctaac cacgcagcca agtcctaagt caacagatct tctgttgata 1980 tggatgcagt tcacagacta aatgtcggtc ggggaagatg tattcttctc ataagatata 2040 gtcggacctc tccttaatgg gagctagcgg atgaagtgat gcaacactgg agccgctggg 2100 aactaatttg tatgcgaaag tatattgatt agttttggag tactcgcgac atgatcacga 2160 aagatcccaa cgaaaagaga gaccacatgg tccttcttga gtttgtaaca gctgctggga 2220 ttacacatgg catggatgaa ctgtacaact gaggatcccc cgacctcgac ctctggctaa 2280 taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga 2340 aggacatatg ggagggcaaa tcatttggtc gagatccctc ggagatcgga tctgggcgtg 2400 gttaagggtg ggaaagaata tataaggtgg gggtcttatg tagttttgta tctgttttgc 2460 agcagccgcc gccgccatga gcaccaactc gtttgatgga agcattgtga gctcatattt 2520 gacaacgcgc atgcccccat gggccggggt gcgtcagaat gtgatgggct ccagcattga 2580 tggtcgcccc gtcctgcccg caaactctac taccttgacc tacgagaccg tgtctggaac 2640 gccgttggag actgcagcct ccgccgccgc ttcagccgct gcagccaccg cccgcgggat 2700 tgtgactgac tttgctttcc tgagcccgct tgcaagcagt gcagcttccc gttcatccgc 2760 ccgcgatgac aagttgacgg ctcttttggc acaattggat tctttgaccc gggaacttaa 2820 tgtcgtttct cagcagctgt tggatctgcg ccagcaggtt tctgccctga aggcttcctc 2880 ccctcccaat gcggtttaaa acataaataa aaaaccagac tctgtttgga tttggatcaa 2940 gcaagtgtct tgctgtcttt atttaggggt tttgcgcgcg cggtaggccc gggaccagcg 3000 gtctcggtcg ttgagggtcc tgtgtatttt ttccaggacg tggtaaaggt gactctggat 3060 gttcagatac atgggcataa gcccgtctct ggggtggagg tagcaccact gcagagcttc 3120 atgctgcggg gtggtgttgt agatgatcca gtcgtagcag gagcgctggg cgtggtgcct 3180 aaaaatgtct ttcagtagca agctgattgc caggggcagg cccttggtgt aagtgtttac 3240 aaagcggtta agctgggatg ggtgcatacg tggggatatg agatgcatct tggactgtat 3300 ttttaggttg gctatgttcc cagccatatc cctccgggga ttcatgttgt gcagaaccac 3360 cagcacagtg tatccggtgc acttgggaaa tttgtcatgt agcttagaag gaaatgcgtg 3420 gaagaacttg gagacgccct tgtgacctcc aagattttcc atgcattcgt ccataatgat 3480 ggcaatgggc ccacgggcgg cggcctgggc gaagatattt ctgggatcac taacgtcata 3540 gttgtgttcc aggatgagat cgtcataggc catttttaca aagcgcgggc ggagggtgcc 3600 agactgcggt ataatggttc catccggccc aggggcgtag ttaccctcac agatttgcat 3660 ttcccacgct ttgagttcag atggggggat catgtctacc tgcggggcga tgaagaaaac 3720 ggtttccggg gtaggggaga tcagctggga agaaagcagg ttcctgagca gctgcgactt 3780 accgcagccg gtgggcccgt aaatcacacc tattaccggg tgcaactggt agttaagaga 3840 gctgcagctg ccgtcatccc tgagcagggg ggccacttcg ttaagcatgt ccctgactcg 3900 catgttttcc ctgaccaaat ccgccagaag gcgctcgccg cccagcgata gcagttcttg 3960 caaggaagca aagtttttca acggtttgag accgtccgcc gtaggcatgc ttttgagcgt 4020 ttgaccaagc agttccaggc ggtcccacag ctcggtcacc tgctctacgg catctcgatc 4080 cagcatatct cctcgtttcg cgggttgggg cggctttcgc tgtacggcag tagtcggtgc 4140 tcgtccagac gggccagggt catgtctttc cacgggcgca gggtcctcgt cagcgtagtc 4200 tgggtcacgg tgaaggggtg cgctccgggc tgcgcgctgg ccagggtgcg cttgaggctg 4260 gtcctgctgg tgctgaagcg ctgccggtct tcgccctgcg cgtcggccag gtagcatttg 4320 accatggtgt catagtccag cccctccgcg gcgtggccct tggcgcgcag cttgcccttg 4380 gaggaggcgc cgcacgaggg gcagtgcaga cttttgaggg cgtagagctt gggcgcgaga 4440 aataccgatt ccggggagta ggcatccgcg ccgcaggccc cgcagacggt ctcgcattcc 4500 acgagccagg tgagctctgg ccgttcgggg tcaaaaacca ggtttccccc atgctttttg 4560 atgcgtttct tacctctggt ttccatgagc cggtgtccac gctcggtgac gaaaaggctg 4620 tccgtgtccc cgtatacaga cttgagaggg agtttaaacg aattcaatag cttgttgcat 4680 gggcggcgat ataaaatgca aggtgctgct caaaaaatca ggcaaagcct cgcgcaaaaa 4740 agaaagcaca tcgtagtcat gctcatgcag ataaaggcag gtaagctccg gaaccaccac 4800 agaaaaagac accatttttc tctcaaacat gtctgcgggt ttctgcataa acacaaaata 4860 aaataacaaa aaaacattta aacattagaa gcctgtctta caacaggaaa aacaaccctt 4920 ataagcataa gacggactac ggccatgccg gcgtgaccgt aaaaaaactg gtcaccgtga 4980 ttaaaaagca ccaccgacag ctcctcggtc atgtccggag tcataatgta agactcggta 5040 aacacatcag gttgattcat cggtcagtgc taaaaagcga ccgaaatagc ccgggggaat 5100 acatacccgc aggcgtagag acaacattac agcccccata ggaggtataa caaaattaat 5160 aggagagaaa aacacataaa cacctgaaaa accctcctgc ctaggcaaaa tagcaccctc 5220 ccgctccaga acaacataca gcgcttcaca gcggcagcct aacagtcagc cttaccagta 5280 aaaaagaaaa cctattaaaa aaacaccact cgacacggca ccagctcaat cagtcacagt 5340 gtaaaaaagg gccaagtgca gagcgagtat atataggact aaaaaatgac gtaacggtta 5400 aagtccacaa aaaacaccca gaaaaccgca cgcgaaccta cgcccagaaa cgaaagccaa 5460 aaaacccaca acttcctcaa atcgtcactt ccgttttccc acgttacgta acttcccatt 5520 ttaagaaaac tacaattccc aacacataca agttactccg ccctaaaacc tacgtcaccc 5580 gccccgttcc cacgccccgc gccacgtcac aaactccacc ccctcattat catattggct 5640 tcaatccaaa ataaggtata ttattgatga tgttaattaa catgcatgga tccatatgcg 5700 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct cttccgcttc 5760 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc 5820 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc 5880 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag 5940 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc 6000 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt 6060 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct 6120 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg 6180 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct 6240 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat 6300 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg 6360 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa 6420 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt 6480 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc 6540 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt 6600 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta 6660 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat 6720 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac 6780 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg 6840 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag 6900 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt 6960 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctgcag ccatgagatt 7020 atcaaaaagg atcttcacct agatcctttt cacgtagaaa gccagtccgc agaaacggtg 7080 ctgaccccgg atgaatgtca gctactgggc tatctggaca agggaaaacg caagcgcaaa 7140 gagaaagcag gtagcttgca gtgggcttac atggcgatag ctagactggg cggttttatg 7200 gacagcaagc gaaccggaat tgccagctgg ggcgccctct ggtaaggttg ggaagccctg 7260 caaagtaaac tggatggctt tctcgccgcc aaggatctga tggcgcaggg gatcaagctc 7320 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg 7380 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg 7440 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa 7500 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct 7560 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga 7620 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc 7680 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac 7740 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc 7800 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact 7860 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga 7920 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg 7980 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga 8040 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga 8100 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgaattt tgttaaaatt 8160 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaacatc ccttataaat 8220 caaaagaata gaccgcgata gggttgagtg ttgttccagt ttggaacaag agtccactat 8280 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac 8340 tacgtgaacc atcacccaaa tcaagttttt tgcggtcgag gtgccgtaaa gctctaaatc 8400 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga 8460 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca 8520 cgctgcgcgt aaccaccaca cccgcgcgct taatgcgccg ctacagggcg cgtccattcg 8580 ccattcagga tcgaattaat tcttaat 8607 <210> 40 <211> 9737 <212> DNA <213> Artificial Sequence <220> <223> pAVQ-AS100KRib2-TK <400> 40 taacatcatc aataatatac cttattttgg attgaagcca atatgataat gagggggtgg 60 agtttgtgac gtggcgcggg gcgtgggaac ggggcgggtg acgtagtagt gtggcggaag 120 tgtgatgttg caagtgtggc ggaacacatg taagcgacgg atgtggcaaa agtgacgttt 180 ttggtgtgcg ccggtgtaca caggaagtga caattttcgc gcggttttag gcggatgttg 240 tagtaaattt gggcgtaacc gagtaagatt tggccatttt cgcgggaaaa ctgaataaga 300 ggaagtgaaa tctgaataat tttgtgttac tcatagcgcg taatactgcg atctatacat 360 tgaatcaata ttggcaatta gccatattag tcattggtta tatagcataa atcaatattg 420 gctattggcc attgcatacg ttgtatctat atcataatat gtacatttat attggctcat 480 gtccaatatg accgccatgt tgacattgat tattgactag ttattaatag taatcaatta 540 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg 600 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc 660 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa 720 ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca 780 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta 840 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt 900 acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg 960 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata 1020 accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca 1080 gagctcgttt agtgaaccgt cagatcctca ctctcttccg catcgctgtc tgcgagggcc 1140 agctgttggg ctcgcggttg aggacaaact cttcgcggtc tttccagtac tcttggatcg 1200 gaaacccgtc ggcctccgaa cggtactccg ccaccgaggg acctgagcca gtccgcatcg 1260 accggatcgg aaaacctctc gagaaaggcg tctaaccagt cacagtcgca aggtaggctg 1320 agcaccgtgg cgggcggcag cgggtggcgg tcggggttgt ttctggcgga ggtgctgctg 1380 atgatgtaat taaagtaggc ggtcttgagc cggcggatgg tcgaggtgag gtgtggcagg 1440 cttgagatcc agctgttggg gtgagtactc cctctcaaaa gcgggcatga cttctgcgct 1500 aagattgtca gtttccaaaa acgaggagga tttgatattc acctggcccg atctggccat 1560 acacttgagt gacaatgaca tccactttgc ctttctctcc acaggtgtcc actcccaggt 1620 ccaagtttgg aagatcccca tcaattacta cttgcttcct gtaggaatcc tctattgttg 1680 gatcatattc gtccacaaaa tgattctgaa ttagctgtat cgtcaaggca ctcttgcaag 1740 cttcatcatg gccgacagca aaagttatca ggcatgcacc tggtagctag tctttaaacc 1800 aatagattgc atcggtttaa aaggcaagac cgtcaaattg cgggaaaggg gtcaacagcc 1860 gttcagtacc aagtctcagg ggaaactttg agatggcctt gcaaagggta tggtaataag 1920 ctgacggaca tggtcctaac cacgcagcca agtcctaagt caacagatct tctgttgata 1980 tggatgcagt tcacagacta aatgtcggtc ggggaagatg tattcttctc ataagatata 2040 gtcggacctc tccttaatgg gagctagcgg atgaagtgat gcaacactgg agccgctggg 2100 aactaatttg tatgcgaaag tatattgatt agttttggag tactcgcgac atgatcacgc 2160 ttcgtacccc tgccatcaac acgcgtctgc gttcgaccag gctgcgcgtt ctcgcggcca 2220 tagcaaccga cgtacggcgt tgcgccctcg ccggcagcaa gaagccacgg aagtccgcct 2280 ggagcagaaa atgcccacgc tactgcgggt ttatatagac ggtcctcacg ggatggggaa 2340 aaccaccacc acgcaactgc tggtggccct gggttcgcgc gacgatatcg tctacgtacc 2400 cgagccgatg acttactggc aggtgctggg ggcttccgag acaatcgcga acatctacac 2460 cacacaacac cgcctcgacc agggtgagat atcggccggg gacgcggcgg tggtaatgac 2520 aagcgcccag ataacaatgg gcatgcctta tgccgtgacc gacgccgttc tggctcctca 2580 tgtcgggggg gaggctggga gttcacatgc cccgcccccg gccctcaccc tcatcttcga 2640 ccgccatccc atcgccgccc tcctgtgcta cccggccgcg cgatacctta tgggcagcat 2700 gaccccccag gccgtgctgg cgttcgtggc cctcatcccg ccgaccttgc ccggcacaaa 2760 catcgtgttg ggggcccttc cggaggacag acacatcgac cgcctggcca aacgccagcg 2820 ccccggcgag cggcttgacc tggctatgct ggccgcgatt cgccgcgttt acgggctgct 2880 tgccaatacg gtgcggtatc tgcagggcgg cgggtcgtgg tgggaggatt ggggacagct 2940 ttcggggacg gccgtgccgc cccagggtgc cgagccccag agcaacgcgg gcccacgacc 3000 ccatatcggg gacacgttat ttaccctgtt tcgggccccc gagttgctgg cccccaacgg 3060 cgacctgtat aacgtgtttg cctgggcctt ggacgtcttg gccaaacgcc tccgtcccat 3120 gcacgtcttt atcctggatt acgaccaatc gcccgccggc tgccgggacg ccctgctgca 3180 acttacctcc gggatggtcc agacccacgt caccacccca ggctccatac cgacgatctg 3240 cgacctggcg cgcacgtttg cccgggagat gggggaggct aataattcga aagatcccaa 3300 cgaaaagaga gaccacatgg tccttcttga gtttgtaaca gctgctggga ttacacatgg 3360 catggatgaa ctgtacaact gaggatcccc cgacctcgac ctctggctaa taaaggaaat 3420 ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg 3480 ggagggcaaa tcatttggtc gagatccctc ggagatcgga tctgggcgtg gttaagggtg 3540 ggaaagaata tataaggtgg gggtcttatg tagttttgta tctgttttgc agcagccgcc 3600 gccgccatga gcaccaactc gtttgatgga agcattgtga gctcatattt gacaacgcgc 3660 atgcccccat gggccggggt gcgtcagaat gtgatgggct ccagcattga tggtcgcccc 3720 gtcctgcccg caaactctac taccttgacc tacgagaccg tgtctggaac gccgttggag 3780 actgcagcct ccgccgccgc ttcagccgct gcagccaccg cccgcgggat tgtgactgac 3840 tttgctttcc tgagcccgct tgcaagcagt gcagcttccc gttcatccgc ccgcgatgac 3900 aagttgacgg ctcttttggc acaattggat tctttgaccc gggaacttaa tgtcgtttct 3960 cagcagctgt tggatctgcg ccagcaggtt tctgccctga aggcttcctc ccctcccaat 4020 gcggtttaaa acataaataa aaaaccagac tctgtttgga tttggatcaa gcaagtgtct 4080 tgctgtcttt atttaggggt tttgcgcgcg cggtaggccc gggaccagcg gtctcggtcg 4140 ttgagggtcc tgtgtatttt ttccaggacg tggtaaaggt gactctggat gttcagatac 4200 atgggcataa gcccgtctct ggggtggagg tagcaccact gcagagcttc atgctgcggg 4260 gtggtgttgt agatgatcca gtcgtagcag gagcgctggg cgtggtgcct aaaaatgtct 4320 ttcagtagca agctgattgc caggggcagg cccttggtgt aagtgtttac aaagcggtta 4380 agctgggatg ggtgcatacg tggggatatg agatgcatct tggactgtat ttttaggttg 4440 gctatgttcc cagccatatc cctccgggga ttcatgttgt gcagaaccac cagcacagtg 4500 tatccggtgc acttgggaaa tttgtcatgt agcttagaag gaaatgcgtg gaagaacttg 4560 gagacgccct tgtgacctcc aagattttcc atgcattcgt ccataatgat ggcaatgggc 4620 ccacgggcgg cggcctgggc gaagatattt ctgggatcac taacgtcata gttgtgttcc 4680 aggatgagat cgtcataggc catttttaca aagcgcgggc ggagggtgcc agactgcggt 4740 ataatggttc catccggccc aggggcgtag ttaccctcac agatttgcat ttcccacgct 4800 ttgagttcag atggggggat catgtctacc tgcggggcga tgaagaaaac ggtttccggg 4860 gtaggggaga tcagctggga agaaagcagg ttcctgagca gctgcgactt accgcagccg 4920 gtgggcccgt aaatcacacc tattaccggg tgcaactggt agttaagaga gctgcagctg 4980 ccgtcatccc tgagcagggg ggccacttcg ttaagcatgt ccctgactcg catgttttcc 5040 ctgaccaaat ccgccagaag gcgctcgccg cccagcgata gcagttcttg caaggaagca 5100 aagtttttca acggtttgag accgtccgcc gtaggcatgc ttttgagcgt ttgaccaagc 5160 agttccaggc ggtcccacag ctcggtcacc tgctctacgg catctcgatc cagcatatct 5220 cctcgtttcg cgggttgggg cggctttcgc tgtacggcag tagtcggtgc tcgtccagac 5280 gggccagggt catgtctttc cacgggcgca gggtcctcgt cagcgtagtc tgggtcacgg 5340 tgaaggggtg cgctccgggc tgcgcgctgg ccagggtgcg cttgaggctg gtcctgctgg 5400 tgctgaagcg ctgccggtct tcgccctgcg cgtcggccag gtagcatttg accatggtgt 5460 catagtccag cccctccgcg gcgtggccct tggcgcgcag cttgcccttg gaggaggcgc 5520 cgcacgaggg gcagtgcaga cttttgaggg cgtagagctt gggcgcgaga aataccgatt 5580 ccggggagta ggcatccgcg ccgcaggccc cgcagacggt ctcgcattcc acgagccagg 5640 tgagctctgg ccgttcgggg tcaaaaacca ggtttccccc atgctttttg atgcgtttct 5700 tacctctggt ttccatgagc cggtgtccac gctcggtgac gaaaaggctg tccgtgtccc 5760 cgtatacaga cttgagaggg agtttaaacg aattcaatag cttgttgcat gggcggcgat 5820 ataaaatgca aggtgctgct caaaaaatca ggcaaagcct cgcgcaaaaa agaaagcaca 5880 tcgtagtcat gctcatgcag ataaaggcag gtaagctccg gaaccaccac agaaaaagac 5940 accatttttc tctcaaacat gtctgcgggt ttctgcataa acacaaaata aaataacaaa 6000 aaaacattta aacattagaa gcctgtctta caacaggaaa aacaaccctt ataagcataa 6060 gacggactac ggccatgccg gcgtgaccgt aaaaaaactg gtcaccgtga ttaaaaagca 6120 ccaccgacag ctcctcggtc atgtccggag tcataatgta agactcggta aacacatcag 6180 gttgattcat cggtcagtgc taaaaagcga ccgaaatagc ccgggggaat acatacccgc 6240 aggcgtagag acaacattac agcccccata ggaggtataa caaaattaat aggagagaaa 6300 aacacataaa cacctgaaaa accctcctgc ctaggcaaaa tagcaccctc ccgctccaga 6360 acaacataca gcgcttcaca gcggcagcct aacagtcagc cttaccagta aaaaagaaaa 6420 cctattaaaa aaacaccact cgacacggca ccagctcaat cagtcacagt gtaaaaaagg 6480 gccaagtgca gagcgagtat atataggact aaaaaatgac gtaacggtta aagtccacaa 6540 aaaacaccca gaaaaccgca cgcgaaccta cgcccagaaa cgaaagccaa aaaacccaca 6600 acttcctcaa atcgtcactt ccgttttccc acgttacgta acttcccatt ttaagaaaac 6660 tacaattccc aacacataca agttactccg ccctaaaacc tacgtcaccc gccccgttcc 6720 cacgccccgc gccacgtcac aaactccacc ccctcattat catattggct tcaatccaaa 6780 ataaggtata ttattgatga tgttaattaa catgcatgga tccatatgcg gtgtgaaata 6840 ccgcacagat gcgtaaggag aaaataccgc atcaggcgct cttccgcttc ctcgctcact 6900 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta 6960 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag 7020 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc 7080 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta 7140 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg 7200 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc 7260 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac 7320 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac 7380 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg 7440 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga 7500 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt 7560 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag 7620 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct 7680 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg 7740 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat 7800 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc 7860 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg 7920 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct 7980 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca 8040 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg 8100 ccagttaata gtttgcgcaa cgttgttgcc attgctgcag ccatgagatt atcaaaaagg 8160 atcttcacct agatcctttt cacgtagaaa gccagtccgc agaaacggtg ctgaccccgg 8220 atgaatgtca gctactgggc tatctggaca agggaaaacg caagcgcaaa gagaaagcag 8280 gtagcttgca gtgggcttac atggcgatag ctagactggg cggttttatg gacagcaagc 8340 gaaccggaat tgccagctgg ggcgccctct ggtaaggttg ggaagccctg caaagtaaac 8400 tggatggctt tctcgccgcc aaggatctga tggcgcaggg gatcaagctc tgatcaagag 8460 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc 8520 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat 8580 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg 8640 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg 8700 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta 8760 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta 8820 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc 8880 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc 8940 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg 9000 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg 9060 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt 9120 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc 9180 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc 9240 atcgccttct atcgccttct tgacgagttc ttctgaattt tgttaaaatt tttgttaaat 9300 cagctcattt tttaaccaat aggccgaaat cggcaacatc ccttataaat caaaagaata 9360 gaccgcgata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt 9420 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc 9480 atcacccaaa tcaagttttt tgcggtcgag gtgccgtaaa gctctaaatc ggaaccctaa 9540 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg 9600 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt 9660 aaccaccaca cccgcgcgct taatgcgccg ctacagggcg cgtccattcg ccattcagga 9720 tcgaattaat tcttaat 9737 <210> 41 <211> 11671 <212> DNA <213> Artificial Sequence <220> <223> pAVQ-AS100KRib2-LacZ <400> 41 taacatcatc aataatatac cttattttgg attgaagcca atatgataat gagggggtgg 60 agtttgtgac gtggcgcggg gcgtgggaac ggggcgggtg acgtagtagt gtggcggaag 120 tgtgatgttg caagtgtggc ggaacacatg taagcgacgg atgtggcaaa agtgacgttt 180 ttggtgtgcg ccggtgtaca caggaagtga caattttcgc gcggttttag gcggatgttg 240 tagtaaattt gggcgtaacc gagtaagatt tggccatttt cgcgggaaaa ctgaataaga 300 ggaagtgaaa tctgaataat tttgtgttac tcatagcgcg taatactgcg atctatacat 360 tgaatcaata ttggcaatta gccatattag tcattggtta tatagcataa atcaatattg 420 gctattggcc attgcatacg ttgtatctat atcataatat gtacatttat attggctcat 480 gtccaatatg accgccatgt tgacattgat tattgactag ttattaatag taatcaatta 540 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg 600 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc 660 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa 720 ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca 780 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta 840 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt 900 acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg 960 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata 1020 accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca 1080 gagctcgttt agtgaaccgt cagatcctca ctctcttccg catcgctgtc tgcgagggcc 1140 agctgttggg ctcgcggttg aggacaaact cttcgcggtc tttccagtac tcttggatcg 1200 gaaacccgtc ggcctccgaa cggtactccg ccaccgaggg acctgagcca gtccgcatcg 1260 accggatcgg aaaacctctc gagaaaggcg tctaaccagt cacagtcgca aggtaggctg 1320 agcaccgtgg cgggcggcag cgggtggcgg tcggggttgt ttctggcgga ggtgctgctg 1380 atgatgtaat taaagtaggc ggtcttgagc cggcggatgg tcgaggtgag gtgtggcagg 1440 cttgagatcc agctgttggg gtgagtactc cctctcaaaa gcgggcatga cttctgcgct 1500 aagattgtca gtttccaaaa acgaggagga tttgatattc acctggcccg atctggccat 1560 acacttgagt gacaatgaca tccactttgc ctttctctcc acaggtgtcc actcccaggt 1620 ccaagtttgg aagatcccca tcaattacta cttgcttcct gtaggaatcc tctattgttg 1680 gatcatattc gtccacaaaa tgattctgaa ttagctgtat cgtcaaggca ctcttgcaag 1740 cttcatcatg gccgacagca aaagttatca ggcatgcacc tggtagctag tctttaaacc 1800 aatagattgc atcggtttaa aaggcaagac cgtcaaattg cgggaaaggg gtcaacagcc 1860 gttcagtacc aagtctcagg ggaaactttg agatggcctt gcaaagggta tggtaataag 1920 ctgacggaca tggtcctaac cacgcagcca agtcctaagt caacagatct tctgttgata 1980 tggatgcagt tcacagacta aatgtcggtc ggggaagatg tattcttctc ataagatata 2040 gtcggacctc tccttaatgg gagctagcgg atgaagtgat gcaacactgg agccgctggg 2100 aactaatttg tatgcgaaag tatattgatt agttttggag tactcgcgac atcatcacgt 2160 cgttttacaa cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta 2220 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg 2280 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgctttgcc tggtttccgg 2340 caccagaagc ggtgccggaa agctggctgg agtgcgatct tcctgaggcc gatactgtcg 2400 tcgtcccctc aaactggcag atgcacggtt acgatgcgcc catctacacc aacgtgacct 2460 atcccattac ggtcaatccg ccgtttgttc ccacggagaa tccgacgggt tgttactcgc 2520 tcacatttaa tgttgatgaa agctggctac aggaaggcca gacgcgaatt atttttgatg 2580 gcgttaactc ggcgtttcat ctgtggtgca acgggcgctg ggtcggttac ggccaggaca 2640 gtcgtttgcc gtctgaattt gacctgagcg catttttacg cgccggagaa aaccgcctcg 2700 cggtgatggt gctgcgttgg agtgacggca gttatctgga agatcaggat atgtggcgga 2760 tgagcggcat tttccgtgac gtctcgttgc tgcataaacc gactacacaa atcagcgatt 2820 tccatgttgc cactcgcttt aatgatgatt tcagccgcgc tgtactggag gctgaagttc 2880 agatgtgcgg cgagttgcgt gactacctac gggtaacagt ttctttatgg cagggtgaaa 2940 cgcaggtcgc cagcggcacc gcgcctttcg gcggtgaaat tatcgatgag cgtggtggtt 3000 atgccgatcg cgtcacacta cgtctgaacg tcgaaaaccc gaaactgtgg agcgccgaaa 3060 tcccgaatct ctatcgtgcg gtggttgaac tgcacaccgc cgacggcacg ctgattgaag 3120 cagaagcctg cgatgtcggt ttccgcgagg tgcggattga aaatggtctg ctgctgctga 3180 acggcaagcc gttgctgatt cgaggcgtta accgtcacga gcatcatcct ctgcatggtc 3240 aggtcatgga tgagcagacg atggtgcagg atatcctgct gatgaagcag aacaacttta 3300 acgccgtgcg ctgttcgcat tatccgaacc atccgctgtg gtacacgctg tgcgaccgct 3360 acggcctgta tgtggtggat gaagccaata ttgaaaccca cggcatggtg ccaatgaatc 3420 gtctgaccga tgatccgcgc tggctaccgg cgatgagcga acgcgtaacg cgaatggtgc 3480 agcgcgatcg taatcacccg agtgtgatca tctggtcgct ggggaatgaa tcaggccacg 3540 gcgctaatca cgacgcgctg tatcgctgga tcaaatctgt cgatccttcc cgcccggtgc 3600 agtatgaagg cggcggagcc gacaccacgg ccaccgatat tatttgcccg atgtacgcgc 3660 gcgtggatga agaccagccc ttcccggctg tgccgaaatg gtccatcaaa aaatggcttt 3720 cgctacctgg agagacgcgc ccgctgatcc tttgcgaata cgcccacgcg atgggtaaca 3780 gtcttggcgg tttcgctaaa tactggcagg cgtttcgtca gtatccccgt ttacagggcg 3840 gcttcgtctg ggactgggtg gatcagtcgc tgattaaata tgatgaaaac ggcaacccgt 3900 ggtcggctta cggcggtgat tttggcgata cgccgaacga tcgccagttc tgtatgaacg 3960 gtctggtctt tgccgaccgc acgccgcatc cagcgctgac ggaagcaaaa caccagcagc 4020 agtttttcca gttccgttta tccgggcaaa ccatcgaagt gaccagcgaa tacctgttcc 4080 gtcatagcga taacgagctc ctgcactgga tggtggcgct ggatggtaag ccgctggcaa 4140 gcggtgaagt gcctctggat gtcgctccac aaggtaaaca gttgattgaa ctgcctgaac 4200 taccgcagcc ggagagcgcc gggcaactct ggctcacagt acgcgtagtg caaccgaacg 4260 cgaccgcatg gtcagaagcc gggcacatca gcgcctggca gcagtggcgt ctggcggaaa 4320 acctcagtgt gacgctcccc gccgcgtccc acgccatccc gcatctgacc accagcgaaa 4380 tggatttttg catcgagctg ggtaataagc gttggcaatt taaccgccag tcaggctttc 4440 tttcacagat gtggattggc gataaaaaac aactgctgac gccgctgcgc gatcagttca 4500 cccgtgcacc gctggataac gacattggcg taagtgaagc gacccgcatt gaccctaacg 4560 cctgggtcga acgctggaag gcggcgggcc attaccaggc cgaagcagcg ttgttgcagt 4620 gcacggcaga tacacttgct gatgcggtgc tgattacgac cgctcacgcg tggcagcatc 4680 aggggaaaac cttatttatc agccggaaaa cctaccggat tgatggtagt ggtcaaatgg 4740 cgattaccgt tgatgttgaa gtggcgagcg atacaccgca tccggcgcgg attggcctga 4800 actgccagct ggcgcaggta gcagagcggg taaactggct cggattaggg ccgcaagaaa 4860 actatcccga ccgccttact gccgcctgtt ttgaccgctg ggatctgcca ttgtcagaca 4920 tgtatacccc gtacgtcttc ccgagcgaaa acggtctgcg ctgcgggacg cgcgaattga 4980 attatggccc acaccagtgg cgcggcgact tccagttcaa catcagccgc tacagtcaac 5040 agcaactgat ggaaaccagc catcgccatc tgctgcacgc ggaagaaggc acatggctga 5100 atatcgacgg tttccatatg gggattggtg gcgacgactc ctggagcccg tcagtatcgg 5160 cggaattcca gctgagcgcc ggtcgctacc attaccagtt ggtctggtgt caaaaataat 5220 tcgaaagatc ccaacgaaaa gagagaccac atggtccttc ttgagtttgt aacagctgct 5280 gggattacac atggcatgga tgaactgtac aactgaggat cccccgacct cgacctctgg 5340 ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact 5400 cggaaggaca tatgggaggg caaatcattt ggtcgagatc cctcggagat cggatctggg 5460 cgtggttaag ggtgggaaag aatatataag gtgggggtct tatgtagttt tgtatctgtt 5520 ttgcagcagc cgccgccgcc atgagcacca actcgtttga tggaagcatt gtgagctcat 5580 atttgacaac gcgcatgccc ccatgggccg gggtgcgtca gaatgtgatg ggctccagca 5640 ttgatggtcg ccccgtcctg cccgcaaact ctactacctt gacctacgag accgtgtctg 5700 gaacgccgtt ggagactgca gcctccgccg ccgcttcagc cgctgcagcc accgcccgcg 5760 ggattgtgac tgactttgct ttcctgagcc cgcttgcaag cagtgcagct tcccgttcat 5820 ccgcccgcga tgacaagttg acggctcttt tggcacaatt ggattctttg acccgggaac 5880 ttaatgtcgt ttctcagcag ctgttggatc tgcgccagca ggtttctgcc ctgaaggctt 5940 cctcccctcc caatgcggtt taaaacataa ataaaaaacc agactctgtt tggatttgga 6000 tcaagcaagt gtcttgctgt ctttatttag gggttttgcg cgcgcggtag gcccgggacc 6060 agcggtctcg gtcgttgagg gtcctgtgta ttttttccag gacgtggtaa aggtgactct 6120 ggatgttcag atacatgggc ataagcccgt ctctggggtg gaggtagcac cactgcagag 6180 cttcatgctg cggggtggtg ttgtagatga tccagtcgta gcaggagcgc tgggcgtggt 6240 gcctaaaaat gtctttcagt agcaagctga ttgccagggg caggcccttg gtgtaagtgt 6300 ttacaaagcg gttaagctgg gatgggtgca tacgtgggga tatgagatgc atcttggact 6360 gtatttttag gttggctatg ttcccagcca tatccctccg gggattcatg ttgtgcagaa 6420 ccaccagcac agtgtatccg gtgcacttgg gaaatttgtc atgtagctta gaaggaaatg 6480 cgtggaagaa cttggagacg cccttgtgac ctccaagatt ttccatgcat tcgtccataa 6540 tgatggcaat gggcccacgg gcggcggcct gggcgaagat atttctggga tcactaacgt 6600 catagttgtg ttccaggatg agatcgtcat aggccatttt tacaaagcgc gggcggaggg 6660 tgccagactg cggtataatg gttccatccg gcccaggggc gtagttaccc tcacagattt 6720 gcatttccca cgctttgagt tcagatgggg ggatcatgtc tacctgcggg gcgatgaaga 6780 aaacggtttc cggggtaggg gagatcagct gggaagaaag caggttcctg agcagctgcg 6840 acttaccgca gccggtgggc ccgtaaatca cacctattac cgggtgcaac tggtagttaa 6900 gagagctgca gctgccgtca tccctgagca ggggggccac ttcgttaagc atgtccctga 6960 ctcgcatgtt ttccctgacc aaatccgcca gaaggcgctc gccgcccagc gatagcagtt 7020 cttgcaagga agcaaagttt ttcaacggtt tgagaccgtc cgccgtaggc atgcttttga 7080 gcgtttgacc aagcagttcc aggcggtccc acagctcggt cacctgctct acggcatctc 7140 gatccagcat atctcctcgt ttcgcgggtt ggggcggctt tcgctgtacg gcagtagtcg 7200 gtgctcgtcc agacgggcca gggtcatgtc tttccacggg cgcagggtcc tcgtcagcgt 7260 agtctgggtc acggtgaagg ggtgcgctcc gggctgcgcg ctggccaggg tgcgcttgag 7320 gctggtcctg ctggtgctga agcgctgccg gtcttcgccc tgcgcgtcgg ccaggtagca 7380 tttgaccatg gtgtcatagt ccagcccctc cgcggcgtgg cccttggcgc gcagcttgcc 7440 cttggaggag gcgccgcacg aggggcagtg cagacttttg agggcgtaga gcttgggcgc 7500 gagaaatacc gattccgggg agtaggcatc cgcgccgcag gccccgcaga cggtctcgca 7560 ttccacgagc caggtgagct ctggccgttc ggggtcaaaa accaggtttc ccccatgctt 7620 tttgatgcgt ttcttacctc tggtttccat gagccggtgt ccacgctcgg tgacgaaaag 7680 gctgtccgtg tccccgtata cagacttgag agggagttta aacgaattca atagcttgtt 7740 gcatgggcgg cgatataaaa tgcaaggtgc tgctcaaaaa atcaggcaaa gcctcgcgca 7800 aaaaagaaag cacatcgtag tcatgctcat gcagataaag gcaggtaagc tccggaacca 7860 ccacagaaaa agacaccatt tttctctcaa acatgtctgc gggtttctgc ataaacacaa 7920 aataaaataa caaaaaaaca tttaaacatt agaagcctgt cttacaacag gaaaaacaac 7980 ccttataagc ataagacgga ctacggccat gccggcgtga ccgtaaaaaa actggtcacc 8040 gtgattaaaa agcaccaccg acagctcctc ggtcatgtcc ggagtcataa tgtaagactc 8100 ggtaaacaca tcaggttgat tcatcggtca gtgctaaaaa gcgaccgaaa tagcccgggg 8160 gaatacatac ccgcaggcgt agagacaaca ttacagcccc cataggaggt ataacaaaat 8220 taataggaga gaaaaacaca taaacacctg aaaaaccctc ctgcctaggc aaaatagcac 8280 cctcccgctc cagaacaaca tacagcgctt cacagcggca gcctaacagt cagccttacc 8340 agtaaaaaag aaaacctatt aaaaaaacac cactcgacac ggcaccagct caatcagtca 8400 cagtgtaaaa aagggccaag tgcagagcga gtatatatag gactaaaaaa tgacgtaacg 8460 gttaaagtcc acaaaaaaca cccagaaaac cgcacgcgaa cctacgccca gaaacgaaag 8520 ccaaaaaacc cacaacttcc tcaaatcgtc acttccgttt tcccacgtta cgtaacttcc 8580 cattttaaga aaactacaat tcccaacaca tacaagttac tccgccctaa aacctacgtc 8640 acccgccccg ttcccacgcc ccgcgccacg tcacaaactc caccccctca ttatcatatt 8700 ggcttcaatc caaaataagg tatattattg atgatgttaa ttaacatgca tggatccata 8760 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg 8820 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc 8880 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt 8940 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc 9000 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa 9060 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc 9120 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg 9180 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc 9240 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc 9300 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca 9360 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact 9420 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg 9480 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt 9540 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct 9600 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga 9660 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa 9720 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac 9780 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga 9840 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc 9900 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca 9960 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta 10020 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct gcagccatga 10080 gattatcaaa aaggatcttc acctagatcc ttttcacgta gaaagccagt ccgcagaaac 10140 ggtgctgacc ccggatgaat gtcagctact gggctatctg gacaagggaa aacgcaagcg 10200 caaagagaaa gcaggtagct tgcagtgggc ttacatggcg atagctagac tgggcggttt 10260 tatggacagc aagcgaaccg gaattgccag ctggggcgcc ctctggtaag gttgggaagc 10320 cctgcaaagt aaactggatg gctttctcgc cgccaaggat ctgatggcgc aggggatcaa 10380 gctctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg 10440 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa 10500 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg 10560 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt 10620 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa 10680 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc 10740 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg 10800 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg 10860 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg 10920 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg 10980 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact 11040 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg 11100 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc 11160 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga attttgttaa 11220 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa catcccttat 11280 aaatcaaaag aatagaccgc gatagggttg agtgttgttc cagtttggaa caagagtcca 11340 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc 11400 ccactacgtg aaccatcacc caaatcaagt tttttgcggt cgaggtgccg taaagctcta 11460 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg 11520 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg 11580 gtcacgctgc gcgtaaccac cacacccgcg cgcttaatgc gccgctacag ggcgcgtcca 11640 ttcgccattc aggatcgaat taattcttaa t 11671 <210> 42 <211> 36428 <212> DNA <213> Artificial Sequence <220> <223> Ad-AS300KRib1-TK <400> 42 ttaagggtgg gaaagaatat ataaggtggg ggtcttatgt agttttgtat ctgttttgca 60 gcagccgccg ccgccatgag caccaactcg tttgatggaa gcattgtgag ctcatatttg 120 acaacgcgca tgcccccatg ggccggggtg cgtcagaatg tgatgggctc cagcattgat 180 ggtcgccccg tcctgcccgc aaactctact accttgacct acgagaccgt gtctggaacg 240 ccgttggaga ctgcagcctc cgccgccgct tcagccgctg cagccaccgc ccgcgggatt 300 gtgactgact ttgctttcct gagcccgctt gcaagcagtg cagcttcccg ttcatccgcc 360 cgcgatgaca agttgacggc tcttttggca caattggatt ctttgacccg ggaacttaat 420 gtcgtttctc agcagctgtt ggatctgcgc cagcaggttt ctgccctgaa ggcttcctcc 480 cctcccaatg cggtttaaaa cataaataaa aaaccagact ctgtttggat ttggatcaag 540 caagtgtctt gctgtcttta tttaggggtt ttgcgcgcgc ggtaggcccg ggaccagcgg 600 tctcggtcgt tgagggtcct gtgtattttt tccaggacgt ggtaaaggtg actctggatg 660 ttcagataca tgggcataag cccgtctctg gggtggaggt agcaccactg cagagcttca 720 tgctgcgggg tggtgttgta gatgatccag tcgtagcagg agcgctgggc gtggtgccta 780 aaaatgtctt tcagtagcaa gctgattgcc aggggcaggc ccttggtgta agtgtttaca 840 aagcggttaa gctgggatgg gtgcatacgt ggggatatga gatgcatctt ggactgtatt 900 tttaggttgg ctatgttccc agccatatcc ctccggggat tcatgttgtg cagaaccacc 960 agcacagtgt atccggtgca cttgggaaat ttgtcatgta gcttagaagg aaatgcgtgg 1020 aagaacttgg agacgccctt gtgacctcca agattttcca tgcattcgtc cataatgatg 1080 gcaatgggcc cacgggcggc ggcctgggcg aagatatttc tgggatcact aacgtcatag 1140 ttgtgttcca ggatgagatc gtcataggcc atttttacaa agcgcgggcg gagggtgcca 1200 gactgcggta taatggttcc atccggccca ggggcgtagt taccctcaca gatttgcatt 1260 tcccacgctt tgagttcaga tggggggatc atgtctacct gcggggcgat gaagaaaacg 1320 gtttccgggg taggggagat cagctgggaa gaaagcaggt tcctgagcag ctgcgactta 1380 ccgcagccgg tgggcccgta aatcacacct attaccgggt gcaactggta gttaagagag 1440 ctgcagctgc cgtcatccct gagcaggggg gccacttcgt taagcatgtc cctgactcgc 1500 atgttttccc tgaccaaatc cgccagaagg cgctcgccgc ccagcgatag cagttcttgc 1560 aaggaagcaa agtttttcaa cggtttgaga ccgtccgccg taggcatgct tttgagcgtt 1620 tgaccaagca gttccaggcg gtcccacagc tcggtcacct gctctacggc atctcgatcc 1680 agcatatctc ctcgtttcgc gggttggggc ggctttcgct gtacggcagt agtcggtgct 1740 cgtccagacg ggccagggtc atgtctttcc acgggcgcag ggtcctcgtc agcgtagtct 1800 gggtcacggt gaaggggtgc gctccgggct gcgcgctggc cagggtgcgc ttgaggctgg 1860 tcctgctggt gctgaagcgc tgccggtctt cgccctgcgc gtcggccagg tagcatttga 1920 ccatggtgtc atagtccagc ccctccgcgg cgtggccctt ggcgcgcagc ttgcccttgg 1980 aggaggcgcc gcacgagggg cagtgcagac ttttgagggc gtagagcttg ggcgcgagaa 2040 ataccgattc cggggagtag gcatccgcgc cgcaggcccc gcagacggtc tcgcattcca 2100 cgagccaggt gagctctggc cgttcggggt caaaaaccag gtttccccca tgctttttga 2160 tgcgtttctt acctctggtt tccatgagcc ggtgtccacg ctcggtgacg aaaaggctgt 2220 ccgtgtcccc gtatacagac ttgagaggcc tgtcctcgag cggtgttccg cggtcctcct 2280 cgtatagaaa ctcggaccac tctgagacaa aggctcgcgt ccaggccagc acgaaggagg 2340 ctaagtggga ggggtagcgg tcgttgtcca ctagggggtc cactcgctcc agggtgtgaa 2400 gacacatgtc gccctcttcg gcatcaagga aggtgattgg tttgtaggtg taggccacgt 2460 gaccgggtgt tcctgaaggg gggctataaa agggggtggg ggcgcgttcg tcctcactct 2520 cttccgcatc gctgtctgcg agggccagct gttggggtga gtactccctc tgaaaagcgg 2580 gcatgacttc tgcgctaaga ttgtcagttt ccaaaaacga ggaggatttg atattcacct 2640 ggcccgcggt gatgcctttg agggtggccg catccatctg gtcagaaaag acaatctttt 2700 tgttgtcaag cttggtggca aacgacccgt agagggcgtt ggacagcaac ttggcgatgg 2760 agcgcagggt ttggtttttg tcgcgatcgg cgcgctcctt ggccgcgatg tttagctgca 2820 cgtattcgcg cgcaacgcac cgccattcgg gaaagacggt ggtgcgctcg tcgggcacca 2880 ggtgcacgcg ccaaccgcgg ttgtgcaggg tgacaaggtc aacgctggtg gctacctctc 2940 cgcgtaggcg ctcgttggtc cagcagaggc ggccgccctt gcgcgagcag aatggcggta 3000 gggggtctag ctgcgtctcg tccggggggt ctgcgtccac ggtaaagacc ccgggcagca 3060 ggcgcgcgtc gaagtagtct atcttgcatc cttgcaagtc tagcgcctgc tgccatgcgc 3120 gggcggcaag cgcgcgctcg tatgggttga gtgggggacc ccatggcatg gggtgggtga 3180 gcgcggaggc gtacatgccg caaatgtcgt aaacgtagag gggctctctg agtattccaa 3240 gatatgtagg gtagcatctt ccaccgcgga tgctggcgcg cacgtaatcg tatagttcgt 3300 gcgagggagc gaggaggtcg ggaccgaggt tgctacgggc gggctgctct gctcggaaga 3360 ctatctgcct gaagatggca tgtgagttgg atgatatggt tggacgctgg aagacgttga 3420 agctggcgtc tgtgagacct accgcgtcac gcacgaagga ggcgtaggag tcgcgcagct 3480 tgttgaccag ctcggcggtg acctgcacgt ctagggcgca gtagtccagg gtttccttga 3540 tgatgtcata cttatcctgt cccttttttt tccacagctc gcggttgagg acaaactctt 3600 cgcggtcttt ccagtactct tggatcggaa acccgtcggc ctccgaacgg taagagccta 3660 gcatgtagaa ctggttgacg gcctggtagg cgcagcatcc cttttctacg ggtagcgcgt 3720 atgcctgcgc ggccttccgg agcgaggtgt gggtgagcgc aaaggtgtcc ctgaccatga 3780 ctttgaggta ctggtatttg aagtcagtgt cgtcgcatcc gccctgctcc cagagcaaaa 3840 agtccgtgcg ctttttggaa cgcggatttg gcagggcgaa ggtgacatcg ttgaagagta 3900 tctttcccgc gcgaggcata aagttgcgtg tgatgcggaa gggtcccggc acctcggaac 3960 ggttgttaat tacctgggcg gcgagcacga tctcgtcaaa gccgttgatg ttgtggccca 4020 caatgtaaag ttccaagaag cgcgggatgc ccttgatgga aggcaatttt ttaagttcct 4080 cgtaggtgag ctcttcaggg gagctgagcc cgtgctctga aagggcccag tctgcaagat 4140 gagggttgga agcgacgaat gagctccaca ggtcacgggc cattagcatt tgcaggtggt 4200 cgcgaaaggt cctaaactgg cgacctatgg ccattttttc tggggtgatg cagtagaagg 4260 taagcgggtc ttgttcccag cggtcccatc caaggttcgc ggctaggtct cgcgcggcag 4320 tcactagagg ctcatctccg ccgaacttca tgaccagcat gaagggcacg agctgcttcc 4380 caaaggcccc catccaagta taggtctcta catcgtaggt gacaaagaga cgctcggtgc 4440 gaggatgcga gccgatcggg aagaactgga tctcccgcca ccaattggag gagtggctat 4500 tgatgtggtg aaagtagaag tccctgcgac gggccgaaca ctcgtgctgg cttttgtaaa 4560 aacgtgcgca gtactggcag cggtgcacgg gctgtacatc ctgcacgagg ttgacctgac 4620 gaccgcgcac aaggaagcag agtgggaatt tgagcccctc gcctggcggg tttggctggt 4680 ggtcttctac ttcggctgct tgtccttgac cgtctggctg ctcgagggga gttacggtgg 4740 atcggaccac cacgccgcgc gagcccaaag tccagatgtc cgcgcgcggc ggtcggagct 4800 tgatgacaac atcgcgcaga tgggagctgt ccatggtctg gagctcccgc ggcgtcaggt 4860 caggcgggag ctcctgcagg tttacctcgc atagacgggt cagggcgcgg gctagatcca 4920 ggtgatacct aatttccagg ggctggttgg tggcggcgtc gatggcttgc aagaggccgc 4980 atccccgcgg cgcgactacg gtaccgcgcg gcgggcggtg ggccgcgggg gtgtccttgg 5040 atgatgcatc taaaagcggt gacgcgggcg agcccccgga ggtagggggg gctccggacc 5100 cgccgggaga gggggcaggg gcacgtcggc gccgcgcgcg ggcaggagct ggtgctgcgc 5160 gcgtaggttg ctggcgaacg cgacgacgcg gcggttgatc tcctgaatct ggcgcctctg 5220 cgtgaagacg acgggcccgg tgagcttgag cctgaaagag agttcgacag aatcaatttc 5280 ggtgtcgttg acggcggcct ggcgcaaaat ctcctgcacg tctcctgagt tgtcttgata 5340 ggcgatctcg gccatgaact gctcgatctc ttcctcctgg agatctccgc gtccggctcg 5400 ctccacggtg gcggcgaggt cgttggaaat gcgggccatg agctgcgaga aggcgttgag 5460 gcctccctcg ttccagacgc ggctgtagac cacgccccct tcggcatcgc gggcgcgcat 5520 gaccacctgc gcgagattga gctccacgtg ccgggcgaag acggcgtagt ttcgcaggcg 5580 ctgaaagagg tagttgaggg tggtggcggt gtgttctgcc acgaagaagt acataaccca 5640 gcgtcgcaac gtggattcgt tgatatcccc caaggcctca aggcgctcca tggcctcgta 5700 gaagtccacg gcgaagttga aaaactggga gttgcgcgcc gacacggtta actcctcctc 5760 cagaagacgg atgagctcgg cgacagtgtc gcgcacctcg cgctcaaagg ctacaggggc 5820 ctcttcttct tcttcaatct cctcttccat aagggcctcc ccttcttctt cttctggcgg 5880 cggtggggga ggggggacac ggcggcgacg acggcgcacc gggaggcggt cgacaaagcg 5940 ctcgatcatc tccccgcggc gacggcgcat ggtctcggtg acggcgcggc cgttctcgcg 6000 ggggcgcagt tggaagacgc cgcccgtcat gtcccggtta tgggttggcg gggggctgcc 6060 atgcggcagg gatacggcgc taacgatgca tctcaacaat tgttgtgtag gtactccgcc 6120 gccgagggac ctgagcgagt ccgcatcgac cggatcggaa aacctctcga gaaaggcgtc 6180 taaccagtca cagtcgcaag gtaggctgag caccgtggcg ggcggcagcg ggcggcggtc 6240 ggggttgttt ctggcggagg tgctgctgat gatgtaatta aagtaggcgg tcttgagacg 6300 gcggatggtc gacagaagca ccatgtcctt gggtccggcc tgctgaatgc gcaggcggtc 6360 ggccatgccc caggcttcgt tttgacatcg gcgcaggtct ttgtagtagt cttgcatgag 6420 cctttctacc ggcacttctt cttctccttc ctcttgtcct gcatctcttg catctatcgc 6480 tgcggcggcg gcggagtttg gccgtaggtg gcgccctctt cctcccatgc gtgtgacccc 6540 gaagcccctc atcggctgaa gcagggctag gtcggcgaca acgcgctcgg ctaatatggc 6600 ctgctgcacc tgcgtgaggg tagactggaa gtcatccatg tccacaaagc ggtggtatgc 6660 gcccgtgttg atggtgtaag tgcagttggc cataacggac cagttaacgg tctggtgacc 6720 cggctgcgag agctcggtgt acctgagacg cgagtaagcc ctcgagtcaa atacgtagtc 6780 gttgcaagtc cgcaccaggt actggtatcc caccaaaaag tgcggcggcg gctggcggta 6840 gaggggccag cgtagggtgg ccggggctcc gggggcgaga tcttccaaca taaggcgatg 6900 atatccgtag atgtacctgg acatccaggt gatgccggcg gcggtggtgg aggcgcgcgg 6960 aaagtcgcgg acgcggttcc agatgttgcg cagcggcaaa aagtgctcca tggtcgggac 7020 gctctggccg gtcaggcgcg cgcaatcgtt gacgctctac cgtgcaaaag gagagcctgt 7080 aagcgggcac tcttccgtgg tctggtggat aaattcgcaa gggtatcatg gcggacgacc 7140 ggggttcgag ccccgtatcc ggccgtccgc cgtgatccat gcggttaccg cccgcgtgtc 7200 gaacccaggt gtgcgacgtc agacaacggg ggagtgctcc ttttggcttc cttccaggcg 7260 cggcggctgc tgcgctagct tttttggcca ctggccgcgc gcagcgtaag cggttaggct 7320 ggaaagcgaa agcattaagt ggctcgctcc ctgtagccgg agggttattt tccaagggtt 7380 gagtcgcggg acccccggtt cgagtctcgg accggccgga ctgcggcgaa cgggggtttg 7440 cctccccgtc atgcaagacc ccgcttgcaa attcctccgg aaacagggac gagccccttt 7500 tttgcttttc ccagatgcat ccggtgctgc ggcagatgcg cccccctcct cagcagcggc 7560 aagagcaaga gcagcggcag acatgcaggg caccctcccc tcctcctacc gcgtcaggag 7620 gggcgacatc cgcggttgac gcggcagcag atggtgatta cgaacccccg cggcgccggg 7680 cccggcacta cctggacttg gaggagggcg agggcctggc gcggctagga gcgccctctc 7740 ctgagcggta cccaagggtg cagctgaagc gtgatacgcg tgaggcgtac gtgccgcggc 7800 agaacctgtt tcgcgaccgc gagggagagg agcccgagga gatgcgggat cgaaagttcc 7860 acgcagggcg cgagctgcgg catggcctga atcgcgagcg gttgctgcgc gaggaggact 7920 ttgagcccga cgcgcgaacc gggattagtc ccgcgcgcgc acacgtggcg gccgccgacc 7980 tggtaaccgc atacgagcag acggtgaacc aggagattaa ctttcaaaaa agctttaaca 8040 accacgtgcg tacgcttgtg gcgcgcgagg aggtggctat aggactgatg catctgtggg 8100 actttgtaag cgcgctggag caaaacccaa atagcaagcc gctcatggcg cagctgttcc 8160 ttatagtgca gcacagcagg gacaacgagg cattcaggga tgcgctgcta aacatagtag 8220 agcccgaggg ccgctggctg ctcgatttga taaacatcct gcagagcata gtggtgcagg 8280 agcgcagctt gagcctggct gacaaggtgg ccgccatcaa ctattccatg cttagcctgg 8340 gcaagtttta cgcccgcaag atataccata ccccttacgt tcccatagac aaggaggtaa 8400 agatcgaggg gttctacatg cgcatggcgc tgaaggtgct taccttgagc gacgacctgg 8460 gcgtttatcg caacgagcgc atccacaagg ccgtgagcgt gagccggcgg cgcgagctca 8520 gcgaccgcga gctgatgcac agcctgcaaa gggccctggc tggcacgggc agcggcgata 8580 gagaggccga gtcctacttt gacgcgggcg ctgacctgcg ctgggcccca agccgacgcg 8640 ccctggaggc agctggggcc ggacctgggc tggcggtggc acccgcgcgc gctggcaacg 8700 tcggcggcgt ggaggaatat gacgaggacg atgagtacga gccagaggac ggcgagtact 8760 aagcggtgat gtttctgatc agatgatgca agacgcaacg gacccggcgg tgcgggcggc 8820 gctgcagagc cagccgtccg gccttaactc cacggacgac tggcgccagg tcatggaccg 8880 catcatgtcg ctgactgcgc gcaatcctga cgcgttccgg cagcagccgc aggccaaccg 8940 gctctccgca attctggaag cggtggtccc ggcgcgcgca aaccccacgc acgagaaggt 9000 gctggcgatc gtaaacgcgc tggccgaaaa cagggccatc cggcccgacg aggccggcct 9060 ggtctacgac gcgctgcttc agcgcgtggc tcgttacaac agcggcaacg tgcagaccaa 9120 cctggaccgg ctggtggggg atgtgcgcga ggccgtggcg cagcgtgagc gcgcgcagca 9180 gcagggcaac ctgggctcca tggttgcact aaacgccttc ctgagtacac agcccgccaa 9240 cgtgccgcgg ggacaggagg actacaccaa ctttgtgagc gcactgcggc taatggtgac 9300 tgagacaccg caaagtgagg tgtaccagtc tgggccagac tattttttcc agaccagtag 9360 acaaggcctg cagaccgtaa acctgagcca ggctttcaaa aacttgcagg ggctgtgggg 9420 ggtgcgggct cccacaggcg accgcgcgac cgtgtctagc ttgctgacgc ccaactcgcg 9480 cctgttgctg ctgctaatag cgcccttcac ggacagtggc agcgtgtccc gggacacata 9540 cctaggtcac ttgctgacac tgtaccgcga ggccataggt caggcgcatg tggacgagca 9600 tactttccag gagattacaa gtgtcagccg cgcgctgggg caggaggaca cgggcagcct 9660 ggaggcaacc ctaaactacc tgctgaccaa ccggcggcag aagatcccct cgttgcacag 9720 tttaaacagc gaggaggagc gcattttgcg ctacgtgcag cagagcgtga gccttaacct 9780 gatgcgcgac ggggtaacgc ccagcgtggc gctggacatg accgcgcgca acatggaacc 9840 gggcatgtat gcctcaaacc ggccgtttat caaccgccta atggactact tgcatcgcgc 9900 ggccgccgtg aaccccgagt atttcaccaa tgccatcttg aacccgcact ggctaccgcc 9960 ccctggtttc tacaccgggg gattcgaggt gcccgagggt aacgatggat tcctctggga 10020 cgacatagac gacagcgtgt tttccccgca accgcagacc ctgctagagt tgcaacagcg 10080 cgagcaggca gaggcggcgc tgcgaaagga aagcttccgc aggccaagca gcttgtccga 10140 tctaggcgct gcggccccgc ggtcagatgc tagtagccca tttccaagct tgatagggtc 10200 tcttaccagc actcgcacca cccgcccgcg cctgctgggc gaggaggagt acctaaacaa 10260 ctcgctgctg cagccgcagc gcgaaaaaaa cctgcctccg gcatttccca acaacgggat 10320 agagagccta gtggacaaga tgagtagatg gaagacgtac gcgcaggagc acagggacgt 10380 gccaggcccg cgcccgccca cccgtcgtca aaggcacgac cgtcagcggg gtctggtgtg 10440 ggaggacgat gactcggcag acgacagcag cgtcctggat ttgggaggga gtggcaaccc 10500 gtttgcgcac cttcgcccca ggctggggag aatgttttaa aaaaaaaaaa gcatgatgca 10560 aaataaaaaa ctcaccaagg ccatggcacc gagcgttggt tttcttgtat tccccttagt 10620 atgcggcgcg cggcgatgta tgaggaaggt cctcctccct cctacgagag tgtggtgagc 10680 gcggcgccag tggcggcggc gctgggttct cccttcgatg ctcccctgga cccgccgttt 10740 gtgcctccgc ggtacctgcg gcctaccggg gggagaaaca gcatccgtta ctctgagttg 10800 gcacccctat tcgacaccac ccgtgtgtac ctggtggaca acaagtcaac ggatgtggca 10860 tccctgaact accagaacga ccacagcaac tttctgacca cggtcattca aaacaatgac 10920 tacagcccgg gggaggcaag cacacagacc atcaatcttg acgaccggtc gcactggggc 10980 ggcgacctga aaaccatcct gcataccaac atgccaaatg tgaacgagtt catgtttacc 11040 aataagttta aggcgcgggt gatggtgtcg cgcttgccta ctaaggacaa tcaggtggag 11100 ctgaaatacg agtgggtgga gttcacgctg cccgagggca actactccga gaccatgacc 11160 atagacctta tgaacaacgc gatcgtggag cactacttga aagtgggcag acagaacggg 11220 gttctggaaa gcgacatcgg ggtaaagttt gacacccgca acttcagact ggggtttgac 11280 cccgtcactg gtcttgtcat gcctggggta tatacaaacg aagccttcca tccagacatc 11340 attttgctgc caggatgcgg ggtggacttc acccacagcc gcctgagcaa cttgttgggc 11400 atccgcaagc ggcaaccctt ccaggagggc tttaggatca cctacgatga tctggagggt 11460 ggtaacattc ccgcactgtt ggatgtggac gcctaccagg cgagcttgaa agatgacacc 11520 gaacagggcg ggggtggcgc aggcggcagc aacagcagtg gcagcggcgc ggaagagaac 11580 tccaacgcgg cagccgcggc aatgcagccg gtggaggaca tgaacgatca tgccattcgc 11640 ggcgacacct ttgccacacg ggctgaggag aagcgcgctg aggccgaagc agcggccgaa 11700 gctgccgccc ccgctgcgca acccgaggtc gagaagcctc agaagaaacc ggtgatcaaa 11760 cccctgacag aggacagcaa gaaacgcagt tacaacctaa taagcaatga cagcaccttc 11820 acccagtacc gcagctggta ccttgcatac aactacggcg accctcagac cggaatccgc 11880 tcatggaccc tgctttgcac tcctgacgta acctgcggct cggagcaggt ctactggtcg 11940 ttgccagaca tgatgcaaga ccccgtgacc ttccgctcca cgcgccagat cagcaacttt 12000 ccggtggtgg gcgccgagct gttgcccgtg cactccaaga gcttctacaa cgaccaggcc 12060 gtctactccc aactcatccg ccagtttacc tctctgaccc acgtgttcaa tcgctttccc 12120 gagaaccaga ttttggcgcg cccgccagcc cccaccatca ccaccgtcag tgaaaacgtt 12180 cctgctctca cagatcacgg gacgctaccg ctgcgcaaca gcatcggagg agtccagcga 12240 gtgaccatta ctgacgccag acgccgcacc tgcccctacg tttacaaggc cctgggcata 12300 gtctcgccgc gcgtcctatc gagccgcact ttttgagcaa gcatgtccat ccttatatcg 12360 cccagcaata acacaggctg gggcctgcgc ttcccaagca agatgtttgg cggggccaag 12420 aagcgctccg accaacaccc agtgcgcgtg cgcgggcact accgcgcgcc ctggggcgcg 12480 cacaaacgcg gccgcactgg gcgcaccacc gtcgatgacg ccatcgacgc ggtggtggag 12540 gaggcgcgca actacacgcc cacgccgcca ccagtgtcca cagtggacgc ggccattcag 12600 accgtggtgc gcggagcccg gcgctatgct aaaatgaaga gacggcggag gcgcgtagca 12660 cgtcgccacc gccgccgacc cggcactgcc gcccaacgcg cggcggcggc cctgcttaac 12720 cgcgcacgtc gcaccggccg acgggcggcc atgcgggccg ctcgaaggct ggccgcgggt 12780 attgtcactg tgccccccag gtccaggcga cgagcggccg ccgcagcagc cgcggccatt 12840 agtgctatga ctcagggtcg caggggcaac gtgtattggg tgcgcgactc ggttagcggc 12900 ctgcgcgtgc ccgtgcgcac ccgccccccg cgcaactaga ttgcaagaaa aaactactta 12960 gactcgtact gttgtatgta tccagcggcg gcggcgcgca acgaagctat gtccaagcgc 13020 aaaatcaaag aagagatgct ccaggtcatc gcgccggaga tctatggccc cccgaagaag 13080 gaagagcagg attacaagcc ccgaaagcta aagcgggtca aaaagaaaaa gaaagatgat 13140 gatgatgaac ttgacgacga ggtggaactg ctgcacgcta ccgcgcccag gcgacgggta 13200 cagtggaaag gtcgacgcgt aaaacgtgtt ttgcgacccg gcaccaccgt agtctttacg 13260 cccggtgagc gctccacccg cacctacaag cgcgtgtatg atgaggtgta cggcgacgag 13320 gacctgcttg agcaggccaa cgagcgcctc ggggagtttg cctacggaaa gcggcataag 13380 gacatgctgg cgttgccgct ggacgagggc aacccaacac ctagcctaaa gcccgtaaca 13440 ctgcagcagg tgctgcccgc gcttgcaccg tccgaagaaa agcgcggcct aaagcgcgag 13500 tctggtgact tggcacccac cgtgcagctg atggtaccca agcgccagcg actggaagat 13560 gtcttggaaa aaatgaccgt ggaacctggg ctggagcccg aggtccgcgt gcggccaatc 13620 aagcaggtgg cgccgggact gggcgtgcag accgtggacg ttcagatacc cactaccagt 13680 agcaccagta ttgccaccgc cacagagggc atggagacac aaacgtcccc ggttgcctca 13740 gcggtggcgg atgccgcggt gcaggcggtc gctgcggccg cgtccaagac ctctacggag 13800 gtgcaaacgg acccgtggat gtttcgcgtt tcagcccccc ggcgcccgcg cggttcgagg 13860 aagtacggcg ccgccagcgc gctactgccc gaatatgccc tacatccttc cattgcgcct 13920 acccccggct atcgtggcta cacctaccgc cccagaagac gagcaactac ccgacgccga 13980 accaccactg gaacccgccg ccgccgtcgc cgtcgccagc ccgtgctggc cccgatttcc 14040 gtgcgcaggg tggctcgcga aggaggcagg accctggtgc tgccaacagc gcgctaccac 14100 cccagcatcg tttaaaagcc ggtctttgtg gttcttgcag atatggccct cacctgccgc 14160 ctccgtttcc cggtgccggg attccgagga agaatgcacc gtaggagggg catggccggc 14220 cacggcctga cgggcggcat gcgtcgtgcg caccaccggc ggcggcgcgc gtcgcaccgt 14280 cgcatgcgcg gcggtatcct gcccctcctt attccactga tcgccgcggc gattggcgcc 14340 gtgcccggaa ttgcatccgt ggccttgcag gcgcagagac actgattaaa aacaagttgc 14400 atgtggaaaa atcaaaataa aaagtctgga ctctcacgct cgcttggtcc tgtaactatt 14460 ttgtagaatg gaagacatca actttgcgtc tctggccccg cgacacggct cgcgcccgtt 14520 catgggaaac tggcaagata tcggcaccag caatatgagc ggtggcgcct tcagctgggg 14580 ctcgctgtgg agcggcatta aaaatttcgg ttccaccgtt aagaactatg gcagcaaggc 14640 ctggaacagc agcacaggcc agatgctgag ggataagttg aaagagcaaa atttccaaca 14700 aaaggtggta gatggcctgg cctctggcat tagcggggtg gtggacctgg ccaaccaggc 14760 agtgcaaaat aagattaaca gtaagcttga tccccgccct cccgtagagg agcctccacc 14820 ggccgtggag acagtgtctc cagaggggcg tggcgaaaag cgtccgcgcc ccgacaggga 14880 agaaactctg gtgacgcaaa tagacgagcc tccctcgtac gaggaggcac taaagcaagg 14940 cctgcccacc acccgtccca tcgcgcccat ggctaccgga gtgctgggcc agcacacacc 15000 cgtaacgctg gacctgcctc cccccgccga cacccagcag aaacctgtgc tgccaggccc 15060 gaccgccgtt gttgtaaccc gtcctagccg cgcgtccctg cgccgcgccg ccagcggtcc 15120 gcgatcgttg cggcccgtag ccagtggcaa ctggcaaagc acactgaaca gcatcgtggg 15180 tctgggggtg caatccctga agcgccgacg atgcttctga atagctaacg tgtcgtatgt 15240 gtgtcatgta tgcgtccatg tcgccgccag aggagctgct gagccgccgc gcgcccgctt 15300 tccaagatgg ctaccccttc gatgatgccg cagtggtctt acatgcacat ctcgggccag 15360 gacgcctcgg agtacctgag ccccgggctg gtgcagtttg cccgcgccac cgagacgtac 15420 ttcagcctga ataacaagtt tagaaacccc acggtggcgc ctacgcacga cgtgaccaca 15480 gaccggtccc agcgtttgac gctgcggttc atccctgtgg accgtgagga tactgcgtac 15540 tcgtacaagg cgcggttcac cctagctgtg ggtgataacc gtgtgctgga catggcttcc 15600 acgtactttg acatccgcgg cgtgctggac aggggcccta cttttaagcc ctactctggc 15660 actgcctaca acgccctggc tcccaagggt gccccaaatc cttgcgaatg ggatgaagct 15720 gctactgctc ttgaaataaa cctagaagaa gaggacgatg acaacgaaga cgaagtagac 15780 gagcaagctg agcagcaaaa aactcacgta tttgggcagg cgccttattc tggtataaat 15840 attacaaagg agggtattca aataggtgtc gaaggtcaaa cacctaaata tgccgataaa 15900 acatttcaac ctgaacctca aataggagaa tctcagtggt acgaaactga aattaatcat 15960 gcagctggga gagtccttaa aaagactacc ccaatgaaac catgttacgg ttcatatgca 16020 aaacccacaa atgaaaatgg agggcaaggc attcttgtaa agcaacaaaa tggaaagcta 16080 gaaagtcaag tggaaatgca atttttctca actactgagg cgaccgcagg caatggtgat 16140 aacttgactc ctaaagtggt attgtacagt gaagatgtag atatagaaac cccagacact 16200 catatttctt acatgcccac tattaaggaa ggtaactcac gagaactaat gggccaacaa 16260 tctatgccca acaggcctaa ttacattgct tttagggaca attttattgg tctaatgtat 16320 tacaacagca cgggtaatat gggtgttctg gcgggccaag catcgcagtt gaatgctgtt 16380 gtagatttgc aagacagaaa cacagagctt tcataccagc ttttgcttga ttccattggt 16440 gatagaacca ggtacttttc tatgtggaat caggctgttg acagctatga tccagatgtt 16500 agaattattg aaaatcatgg aactgaagat gaacttccaa attactgctt tccactggga 16560 ggtgtgatta atacagagac tcttaccaag gtaaaaccta aaacaggtca ggaaaatgga 16620 tgggaaaaag atgctacaga attttcagat aaaaatgaaa taagagttgg aaataatttt 16680 gccatggaaa tcaatctaaa tgccaacctg tggagaaatt tcctgtactc caacatagcg 16740 ctgtatttgc ccgacaagct aaagtacagt ccttccaacg taaaaatttc tgataaccca 16800 aacacctacg actacatgaa caagcgagtg gtggctcccg ggttagtgga ctgctacatt 16860 aaccttggag cacgctggtc ccttgactat atggacaacg tcaacccatt taaccaccac 16920 cgcaatgctg gcctgcgcta ccgctcaatg ttgctgggca atggtcgcta tgtgcccttc 16980 cacatccagg tgcctcagaa gttctttgcc attaaaaacc tccttctcct gccgggctca 17040 tacacctacg agtggaactt caggaaggat gttaacatgg ttctgcagag ctccctagga 17100 aatgacctaa gggttgacgg agccagcatt aagtttgata gcatttgcct ttacgccacc 17160 ttcttcccca tggcccacaa caccgcctcc acgcttgagg ccatgcttag aaacgacacc 17220 aacgaccagt cctttaacga ctatctctcc gccgccaaca tgctctaccc tatacccgcc 17280 aacgctacca acgtgcccat atccatcccc tcccgcaact gggcggcttt ccgcggctgg 17340 gccttcacgc gccttaagac taaggaaacc ccatcactgg gctcgggcta cgacccttat 17400 tacacctact ctggctctat accctaccta gatggaacct tttacctcaa ccacaccttt 17460 aagaaggtgg ccattacctt tgactcttct gtcagctggc ctggcaatga ccgcctgctt 17520 acccccaacg agtttgaaat taagcgctca gttgacgggg agggttacaa cgttgcccag 17580 tgtaacatga ccaaagactg gttcctggta caaatgctag ctaactacaa cattggctac 17640 cagggcttct atatcccaga gagctacaag gaccgcatgt actccttctt tagaaacttc 17700 cagcccatga gccgtcaggt ggtggatgat actaaataca aggactacca acaggtgggc 17760 atcctacacc aacacaacaa ctctggattt gttggctacc ttgcccccac catgcgcgaa 17820 ggacaggcct accctgctaa cttcccctat ccgcttatag gcaagaccgc agttgacagc 17880 attacccaga aaaagtttct ttgcgatcgc accctttggc gcatcccatt ctccagtaac 17940 tttatgtcca tgggcgcact cacagacctg ggccaaaacc ttctctacgc caactccgcc 18000 cacgcgctag acatgacttt tgaggtggat cccatggacg agcccaccct tctttatgtt 18060 ttgtttgaag tctttgacgt ggtccgtgtg caccggccgc accgcggcgt catcgaaacc 18120 gtgtacctgc gcacgccctt ctcggccggc aacgccacaa cataaagaag caagcaacat 18180 caacaacagc tgccgccatg ggctccagtg agcaggaact gaaagccatt gtcaaagatc 18240 ttggttgtgg gccatatttt ttgggcacct atgacaagcg ctttccaggc tttgtttctc 18300 cacacaagct cgcctgcgcc atagtcaata cggccggtcg cgagactggg ggcgtacact 18360 ggatggcctt tgcctggaac ccgcactcaa aaacatgcta cctctttgag ccctttggct 18420 tttctgacca gcgactcaag caggtttacc agtttgagta cgagtcactc ctgcgccgta 18480 gcgccattgc ttcttccccc gaccgctgta taacgctgga aaagtccacc caaagcgtac 18540 aggggcccaa ctcggccgcc tgtggactat tctgctgcat gtttctccac gcctttgcca 18600 actggcccca aactcccatg gatcacaacc ccaccatgaa ccttattacc ggggtaccca 18660 actccatgct caacagtccc caggtacagc ccaccctgcg tcgcaaccag gaacagctct 18720 acagcttcct ggagcgccac tcgccctact tccgcagcca cagtgcgcag attaggagcg 18780 ccacttcttt ttgtcacttg aaaaacatgt aaaaataatg tactagagac actttcaata 18840 aaggcaaatg cttttatttg tacactctcg ggtgattatt tacccccacc cttgccgtct 18900 gcgccgttta aaaatcaaag gggttctgcc gcgcatcgct atgcgccact ggcagggaca 18960 cgttgcgata ctggtgttta gtgctccact taaactcagg cacaaccatc cgcggcagct 19020 cggtgaagtt ttcactccac aggctgcgca ccatcaccaa cgcgtttagc aggtcgggcg 19080 ccgatatctt gaagtcgcag ttggggcctc cgccctgcgc gcgcgagttg cgatacacag 19140 ggttgcagca ctggaacact atcagcgccg ggtggtgcac gctggccagc acgctcttgt 19200 cggagatcag atccgcgtcc aggtcctccg cgttgctcag ggcgaacgga gtcaactttg 19260 gtagctgcct tcccaaaaag ggcgcgtgcc caggctttga gttgcactcg caccgtagtg 19320 gcatcaaaag gtgaccgtgc ccggtctggg cgttaggata cagcgcctgc ataaaagcct 19380 tgatctgctt aaaagccacc tgagcctttg cgccttcaga gaagaacatg ccgcaagact 19440 tgccggaaaa ctgattggcc ggacaggccg cgtcgtgcac gcagcacctt gcgtcggtgt 19500 tggagatctg caccacattt cggccccacc ggttcttcac gatcttggcc ttgctagact 19560 gctccttcag cgcgcgctgc ccgttttcgc tcgtcacatc catttcaatc acgtgctcct 19620 tatttatcat aatgcttccg tgtagacact taagctcgcc ttcgatctca gcgcagcggt 19680 gcagccacaa cgcgcagccc gtgggctcgt gatgcttgta ggtcacctct gcaaacgact 19740 gcaggtacgc ctgcaggaat cgccccatca tcgtcacaaa ggtcttgttg ctggtgaagg 19800 tcagctgcaa cccgcggtgc tcctcgttca gccaggtctt gcatacggcc gccagagctt 19860 ccacttggtc aggcagtagt ttgaagttcg cctttagatc gttatccacg tggtacttgt 19920 ccatcagcgc gcgcgcagcc tccatgccct tctcccacgc agacacgatc ggcacactca 19980 gcgggttcat caccgtaatt tcactttccg cttcgctggg ctcttcctct tcctcttgcg 20040 tccgcatacc acgcgccact gggtcgtctt cattcagccg ccgcactgtg cgcttacctc 20100 ctttgccatg cttgattagc accggtgggt tgctgaaacc caccatttgt agcgccacat 20160 cttctctttc ttcctcgctg tccacgatta cctctggtga tggcgggcgc tcgggcttgg 20220 gagaagggcg cttctttttc ttcttgggcg caatggccaa atccgccgcc gaggtcgatg 20280 gccgcgggct gggtgtgcgc ggcaccagcg cgtcttgtga tgagtcttcc tcgtcctcgg 20340 actcgatacg ccgcctcatc cgcttttttg ggggcgcccg gggaggcggc ggcgacgggg 20400 acggggacga cacgtcctcc atggttgggg gacgtcgcgc cgcaccgcgt ccgcgctcgg 20460 gggtggtttc gcgctgctcc tcttcccgac tggccatttc cttctcctat aggcagaaaa 20520 agatcatgga gtcagtcgag aagaaggaca gcctaaccgc cccctctgag ttcgccacca 20580 ccgcctccac cgatgccgcc aacgcgccta ccaccttccc cgtcgaggca cccccgcttg 20640 aggaggagga agtgattatc gagcaggacc caggttttgt aagcgaagac gacgaggacc 20700 gctcagtacc aacagaggat aaaaagcaag accaggacaa cgcagaggca aacgaggaac 20760 aagtcgggcg gggggacgaa aggcatggcg actacctaga tgtgggagac gacgtgctgt 20820 tgaagcatct gcagcgccag tgcgccatta tctgcgacgc gttgcaagag cgcagcgatg 20880 tgcccctcgc catagcggat gtcagccttg cctacgaacg ccacctattc tcaccgcgcg 20940 taccccccaa acgccaagaa aacggcacat gcgagcccaa cccgcgcctc aacttctacc 21000 ccgtatttgc cgtgccagag gtgcttgcca cctatcacat ctttttccaa aactgcaaga 21060 tacccctatc ctgccgtgcc aaccgcagcc gagcggacaa gcagctggcc ttgcggcagg 21120 gcgctgtcat acctgatatc gcctcgctca acgaagtgcc aaaaatcttt gagggtcttg 21180 gacgcgacga gaagcgcgcg gcaaacgctc tgcaacagga aaacagcgaa aatgaaagtc 21240 actctggagt gttggtggaa ctcgagggtg acaacgcgcg cctagccgta ctaaaacgca 21300 gcatcgaggt cacccacttt gcctacccgg cacttaacct accccccaag gtcatgagca 21360 cagtcatgag tgagctgatc gtgcgccgtg cgcagcccct ggagagggat gcaaatttgc 21420 aagaacaaac agaggagggc ctacccgcag ttggcgacga gcagctagcg cgctggcttc 21480 aaacgcgcga gcctgccgac ttggaggagc gacgcaaact aatgatggcc gcagtgctcg 21540 ttaccgtgga gcttgagtgc atgcagcggt tctttgctga cccggagatg cagcgcaagc 21600 tagaggaaac attgcactac acctttcgac agggctacgt acgccaggcc tgcaagatct 21660 ccaacgtgga gctctgcaac ctggtctcct accttggaat tttgcacgaa aaccgccttg 21720 ggcaaaacgt gcttcattcc acgctcaagg gcgaggcgcg ccgcgactac gtccgcgact 21780 gcgtttactt atttctatgc tacacctggc agacggccat gggcgtttgg cagcagtgct 21840 tggaggagtg caacctcaag gagctgcaga aactgctaaa gcaaaacttg aaggacctat 21900 ggacggcctt caacgagcgc tccgtggccg cgcacctggc ggacatcatt ttccccgaac 21960 gcctgcttaa aaccctgcaa cagggtctgc cagacttcac cagtcaaagc atgttgcaga 22020 actttaggaa ctttatccta gagcgctcag gaatcttgcc cgccacctgc tgtgcacttc 22080 ctagcgactt tgtgcccatt aagtaccgcg aatgccctcc gccgctttgg ggccactgct 22140 accttctgca gctagccaac taccttgcct accactctga cataatggaa gacgtgagcg 22200 gtgacggtct actggagtgt cactgtcgct gcaacctatg caccccgcac cgctccctgg 22260 tttgcaattc gcagctgctt aacgaaagtc aaattatcgg tacctttgag ctgcagggtc 22320 cctcgcctga cgaaaagtcc gcggctccgg ggttgaaact cactccgggg ctgtggacgt 22380 cggcttacct tcgcaaattt gtacctgagg actaccacgc ccacgagatt aggttctacg 22440 aagaccaatc ccgcccgcca aatgcggagc ttaccgcctg cgtcattacc cagggccaca 22500 ttcttggcca attgcaagcc atcaacaaag cccgccaaga gtttctgcta cgaaagggac 22560 ggggggttta cttggacccc cagtccggcg aggagctcaa cccaatcccc ccgccgccgc 22620 agccctatca gcagcagccg cgggcccttg cttcccagga tggcacccaa aaagaagctg 22680 cagctgccgc cgccacccac ggacgaggag gaatactggg acagtcaggc agaggaggtt 22740 ttggacgagg aggaggagga catgatggaa gactgggaga gcctagacga ggaagcttcc 22800 gaggtcgaag aggtgtcaga cgaaacaccg tcaccctcgg tcgcattccc ctcgccggcg 22860 ccccagaaat cggcaaccgg ttccagcatg gctacaacct ccgctcctca ggcgccgccg 22920 gcactgcccg ttcgccgacc caaccgtaga tgggacacca ctggaaccag ggccggtaag 22980 tccaagcagc cgccgccgtt agcccaagag caacaacagc gccaaggcta ccgctcatgg 23040 cgcgggcaca agaacgccat agttgcttgc ttgcaagact gtgggggcaa catctccttc 23100 gcccgccgct ttcttctcta ccatcacggc gtggccttcc cccgtaacat cctgcattac 23160 taccgtcatc tctacagccc atactgcacc ggcggcagcg gcagcggcag caacagcagc 23220 ggccacacag aagcaaaggc gaccggatag caagactctg acaaagccca agaaatccac 23280 agcggcggca gcagcaggag gaggagcgct gcgtctggcg cccaacgaac ccgtatcgac 23340 ccgcgagctt agaaacagga tttttcccac tctgtatgct atatttcaac agagcagggg 23400 ccaagaacaa gagctgaaaa taaaaaacag gtctctgcga tccctcaccc gcagctgcct 23460 gtatcacaaa agcgaagatc agcttcggcg cacgctggaa gacgcggagg ctctcttcag 23520 taaatactgc gcgctgactc ttaaggacta gtttcgcgcc ctttctcaaa tttaagcgcg 23580 aaaactacgt catctccagc ggccacaccc ggcgccagca cctgtcgtca gcgccattat 23640 gagcaaggaa attcccacgc cctacatgtg gagttaccag ccacaaatgg gacttgcggc 23700 tggagctgcc caagactact caacccgaat aaactacatg agcgcgggac cccacatgat 23760 atcccgggtc aacggaatcc gcgcccaccg aaaccgaatt ctcttggaac aggcggctat 23820 taccaccaca cctcgtaata accttaatcc ccgtagttgg cccgctgccc tggtgtacca 23880 ggaaagtccc gctcccacca ctgtggtact tcccagagac gcccaggccg aagttcagat 23940 gactaactca ggggcgcagc ttgcgggcgg ctttcgtcac agggtgcggt cgcccgggca 24000 gggtataact cacctgacaa tcagagggcg aggtattcag ctcaacgacg agtcggtgag 24060 ctcctcgctt ggtctccgtc cggacgggac atttcagatc ggcggcgccg gccgtccttc 24120 attcacgcct cgtcaggcaa tcctaactct gcagacctcg tcctctgagc cgcgctctgg 24180 aggcattgga actctgcaat ttattgagga gtttgtgcca tcggtctact ttaacccctt 24240 ctcgggacct cccggccact atccggatca atttattcct aactttgacg cggtaaagga 24300 ctcggcggac ggctacgact gaatgttaag tggagaggca gagcaactgc gcctgaaaca 24360 cctggtccac tgtcgccgcc acaagtgctt tgcccgcgac tccggtgagt tttgctactt 24420 tgaattgccc gaggatcata tcgagggccc ggcgcacggc gtccggctta ccgcccaggg 24480 agagcttgcc cgtagcctga ttcgggagtt tacccagcgc cccctgctag ttgagcggga 24540 caggggaccc tgtgttctca ctgtgatttg caactgtcct aaccttggat tacatcaaga 24600 tcctctagtt ataactagag tacccgggga tcttattccc tttaactaat aaaaaaaaat 24660 aataaagcat cacttactta aaatcagtta gcaaatttct gtccagttta ttcagcagca 24720 cctccttgcc ctcctcccag ctctggtatt gcagcttcct cctggctgca aactttctcc 24780 acaatctaaa tggaatgtca gtttcctcct gttcctgtcc atccgcaccc actatcttca 24840 tgttgttgca gatgaagcgc gcaagaccgt ctgaagatac cttcaacccc gtgtatccat 24900 atgacacgga aaccggtcct ccaactgtgc cttttcttac tcctcccttt gtatccccca 24960 atgggtttca agagagtccc cctggggtac tctctttgcg cctatccgaa cctctagtta 25020 cctccaatgg catgcttgcg ctcaaaatgg gcaacggcct ctctctggac gaggccggca 25080 accttacctc ccaaaatgta accactgtga gcccacctct caaaaaaacc aagtcaaaca 25140 taaacctgga aatatctgca cccctcacag ttacctcaga agccctaact gtggctgccg 25200 ccgcacctct aatggtcgcg ggcaacacac tcaccatgca atcacaggcc ccgctaaccg 25260 tgcacgactc caaacttagc attgccaccc aaggacccct cacagtgtca gaaggaaagc 25320 tagccctgca aacatcaggc cccctcacca ccaccgatag cagtaccctt actatcactg 25380 cctcaccccc tctaactact gccactggta gcttgggcat tgacttgaaa gagcccattt 25440 atacacaaaa tggaaaacta ggactaaagt acggggctcc tttgcatgta acagacgacc 25500 taaacacttt gaccgtagca actggtccag gtgtgactat taataatact tccttgcaaa 25560 ctaaagttac tggagccttg ggttttgatt cacaaggcaa tatgcaactt aatgtagcag 25620 gaggactaag gattgattct caaaacagac gccttatact tgatgttagt tatccgtttg 25680 atgctcaaaa ccaactaaat ctaagactag gacagggccc tctttttata aactcagccc 25740 acaacttgga tattaactac aacaaaggcc tttacttgtt tacagcttca aacaattcca 25800 aaaagcttga ggttaaccta agcactgcca aggggttgat gtttgacgct acagccatag 25860 ccattaatgc aggagatggg cttgaatttg gttcacctaa tgcaccaaac acaaatcccc 25920 tcaaaacaaa aattggccat ggcctagaat ttgattcaaa caaggctatg gttcctaaac 25980 taggaactgg ccttagtttt gacagcacag gtgccattac agtaggaaac aaaaataatg 26040 ataagctaac tttgtggacc acaccagctc catctcctaa ctgtagacta aatgcagaga 26100 aagatgctaa actcactttg gtcttaacaa aatgtggcag tcaaatactt gctacagttt 26160 cagttttggc tgttaaaggc agtttggctc caatatctgg aacagttcaa agtgctcatc 26220 ttattataag atttgacgaa aatggagtgc tactaaacaa ttccttcctg gacccagaat 26280 attggaactt tagaaatgga gatcttactg aaggcacagc ctatacaaac gctgttggat 26340 ttatgcctaa cctatcagct tatccaaaat ctcacggtaa aactgccaaa agtaacattg 26400 tcagtcaagt ttacttaaac ggagacaaaa ctaaacctgt aacactaacc attacactaa 26460 acggtacaca ggaaacagga gacacaactc caagtgcata ctctatgtca ttttcatggg 26520 actggtctgg ccacaactac attaatgaaa tatttgccac atcctcttac actttttcat 26580 acattgccca agaataaaga atcgtttgtg ttatgtttca acgtgtttat ttttcaattg 26640 cagaaaattt caagtcattt ttcattcagt agtatagccc caccaccaca tagcttatac 26700 agatcaccgt accttaatca aactcacaga accctagtat tcaacctgcc acctccctcc 26760 caacacacag agtacacagt cctttctccc cggctggcct taaaaagcat catatcatgg 26820 gtaacagaca tattcttagg tgttatattc cacacggttt cctgtcgagc caaacgctca 26880 tcagtgatat taataaactc cccgggcagc tcacttaagt tcatgtcgct gtccagctgc 26940 tgagccacag gctgctgtcc aacttgcggt tgcttaacgg gcggcgaagg agaagtccac 27000 gcctacatgg gggtagagtc ataatcgtgc atcaggatag ggcggtggtg ctgcagcagc 27060 gcgcgaataa actgctgccg ccgccgctcc gtcctgcagg aatacaacat ggcagtggtc 27120 tcctcagcga tgattcgcac cgcccgcagc ataaggcgcc ttgtcctccg ggcacagcag 27180 cgcaccctga tctcacttaa atcagcacag taactgcagc acagcaccac aatattgttc 27240 aaaatcccac agtgcaaggc gctgtatcca aagctcatgg cggggaccac agaacccacg 27300 tggccatcat accacaagcg caggtagatt aagtggcgac ccctcataaa cacgctggac 27360 ataaacatta cctcttttgg catgttgtaa ttcaccacct cccggtacca tataaacctc 27420 tgattaaaca tggcgccatc caccaccatc ctaaaccagc tggccaaaac ctgcccgccg 27480 gctatacact gcagggaacc gggactggaa caatgacagt ggagagccca ggactcgtaa 27540 ccatggatca tcatgctcgt catgatatca atgttggcac aacacaggca cacgtgcata 27600 cacttcctca ggattacaag ctcctcccgc gttagaacca tatcccaggg aacaacccat 27660 tcctgaatca gcgtaaatcc cacactgcag ggaagacctc gcacgtaact cacgttgtgc 27720 attgtcaaag tgttacattc gggcagcagc ggatgatcct ccagtatggt agcgcgggtt 27780 tctgtctcaa aaggaggtag acgatcccta ctgtacggag tgcgccgaga caaccgagat 27840 cgtgttggtc gtagtgtcat gccaaatgga acgccggacg tagtcatatt tcctgaagca 27900 aaaccaggtg cgggcgtgac aaacagatct gcgtctccgg tctcgccgct tagatcgctc 27960 tgtgtagtag ttgtagtata tccactctct caaagcatcc aggcgccccc tggcttcggg 28020 ttctatgtaa actccttcat gcgccgctgc cctgataaca tccaccaccg cagaataagc 28080 cacacccagc caacctacac attcgttctg cgagtcacac acgggaggag cgggaagagc 28140 tggaagaacc atgttttttt ttttattcca aaagattatc caaaacctca aaatgaagat 28200 ctattaagtg aacgcgctcc cctccggtgg cgtggtcaaa ctctacagcc aaagaacaga 28260 taatggcatt tgtaagatgt tgcacaatgg cttccaaaag gcaaacggcc ctcacgtcca 28320 agtggacgta aaggctaaac ccttcagggt gaatctcctc tataaacatt ccagcacctt 28380 caaccatgcc caaataattc tcatctcgcc accttctcaa tatatctcta agcaaatccc 28440 gaatattaag tccggccatt gtaaaaatct gctccagagc gccctccacc ttcagcctca 28500 agcagcgaat catgattgca aaaattcagg ttcctcacag acctgtataa gattcaaaag 28560 cggaacatta acaaaaatac cgcgatcccg taggtccctt cgcagggcca gctgaacata 28620 atcgtgcagg tctgcacgga ccagcgcggc cacttccccg ccaggaacct tgacaaaaga 28680 acccacactg attatgacac gcatactcgg agctatgcta accagcgtag ccccgatgta 28740 agctttcaat agcttgttgc atgggcggcg atataaaatg caaggtgctg ctcaaaaaat 28800 caggcaaagc ctcgcgcaaa aaagaaagca catcgtagtc atgctcatgc agataaaggc 28860 aggtaagctc cggaaccacc acagaaaaag acaccatttt tctctcaaac atgtctgcgg 28920 gtttctgcat aaacacaaaa taaaataaca aaaaaacatt taaacattag aagcctgtct 28980 tacaacagga aaaacaaccc ttataagcat aagacggact acggccatgc cggcgtgacc 29040 gtaaaaaaac tggtcaccgt gattaaaaag caccaccgac agctcctcgg tcatgtccgg 29100 agtcataatg taagactcgg taaacacatc aggttgattc atcggtcagt gctaaaaagc 29160 gaccgaaata gcccggggga atacataccc gcaggcgtag agacaacatt acagccccca 29220 taggaggtat aacaaaatta ataggagaga aaaacacata aacacctgaa aaaccctcct 29280 gcctaggcaa aatagcaccc tcccgctcca gaacaacata cagcgcttca cagcggcagc 29340 ctaacagtca gccttaccag taaaaaagaa aacctattaa aaaaacacca ctcgacacgg 29400 caccagctca atcagtcaca gtgtaaaaaa gggccaagtg cagagcgagt atatatagga 29460 ctaaaaaatg acgtaacggt taaagtccac aaaaaacacc cagaaaaccg cacgcgaacc 29520 tacgcccaga aacgaaagcc aaaaaaccca caacttcctc aaatcgtcac ttccgttttc 29580 ccacgttacg taacttccca ttttaagaaa actacaattc ccaacacata caagttactc 29640 cgccctaaaa cctacgtcac ccgccccgtt cccacgcccc gcgccacgtc acaaactcca 29700 ccccctcatt atcatattgg cttcaatcca aaataaggta tattattgat gatgttaatt 29760 aacatgcatg gatccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc 29820 gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 29880 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 29940 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 30000 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 30060 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 30120 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 30180 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 30240 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 30300 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 30360 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 30420 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc 30480 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 30540 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 30600 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 30660 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 30720 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 30780 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 30840 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 30900 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 30960 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 31020 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 31080 ccattgctgc agccatgaga ttatcaaaaa ggatcttcac ctagatcctt ttcacgtaga 31140 aagccagtcc gcagaaacgg tgctgacccc ggatgaatgt cagctactgg gctatctgga 31200 caagggaaaa cgcaagcgca aagagaaagc aggtagcttg cagtgggctt acatggcgat 31260 agctagactg ggcggtttta tggacagcaa gcgaaccgga attgccagct ggggcgccct 31320 ctggtaaggt tgggaagccc tgcaaagtaa actggatggc tttctcgccg ccaaggatct 31380 gatggcgcag gggatcaagc tctgatcaag agacaggatg aggatcgttt cgcatgattg 31440 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg 31500 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg 31560 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg 31620 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg 31680 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc 31740 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc 31800 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc 31860 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc 31920 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg 31980 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct 32040 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt 32100 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc 32160 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt 32220 tcttctgaat tttgttaaaa tttttgttaa atcagctcat tttttaacca ataggccgaa 32280 atcggcaaca tcccttataa atcaaaagaa tagaccgcga tagggttgag tgttgttcca 32340 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc 32400 gtctatcagg gcgatggccc actacgtgaa ccatcaccca aatcaagttt tttgcggtcg 32460 aggtgccgta aagctctaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg 32520 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg 32580 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgcgcg cttaatgcgc 32640 cgctacaggg cgcgtccatt cgccattcag gatcgaatta attcttaatt aacatcatca 32700 ataatatacc ttattttgga ttgaagccaa tatgataatg agggggtgga gtttgtgacg 32760 tggcgcgggg cgtgggaacg gggcgggtga cgtagtagtg tggcggaagt gtgatgttgc 32820 aagtgtggcg gaacacatgt aagcgacgga tgtggcaaaa gtgacgtttt tggtgtgcgc 32880 cggtgtacac aggaagtgac aattttcgcg cggttttagg cggatgttgt agtaaatttg 32940 ggcgtaaccg agtaagattt ggccattttc gcgggaaaac tgaataagag gaagtgaaat 33000 ctgaataatt ttgtgttact catagcgcgt aatactgcga tctatacatt gaatcaatat 33060 tggcaattag ccatattagt cattggttat atagcataaa tcaatattgg ctattggcca 33120 ttgcatacgt tgtatctata tcataatatg tacatttata ttggctcatg tccaatatga 33180 ccgccatgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta 33240 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc 33300 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg 33360 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg 33420 gcagtacatc aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa 33480 tggcccgcct ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac 33540 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg 33600 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg 33660 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg 33720 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta 33780 gtgaaccgtc agatcctcac tctcttccgc atcgctgtct gcgagggcca gctgttgggc 33840 tcgcggttga ggacaaactc ttcgcggtct ttccagtact cttggatcgg aaacccgtcg 33900 gcctccgaac ggtactccgc caccgaggga cctgagccag tccgcatcga ccggatcgga 33960 aaacctctcg agaaaggcgt ctaaccagtc acagtcgcaa ggtaggctga gcaccgtggc 34020 gggcggcagc gggtggcggt cggggttgtt tctggcggag gtgctgctga tgatgtaatt 34080 aaagtaggcg gtcttgagcc ggcggatggt cgaggtgagg tgtggcaggc ttgagatcca 34140 gctgttgggg tgagtactcc ctctcaaaag cgggcatgac ttctgcgcta agattgtcag 34200 tttccaaaaa cgaggaggat ttgatattca cctggcccga tctggccata cacttgagtg 34260 acaatgacat ccactttgcc tttctctcca caggtgtcca ctcccaggtc caagtttgga 34320 agatccctac taggaccata ggtacatctt cagagtcctt aactctttta atttgttctc 34380 tataatggtg aatatcttca aatgatttag tattatttat ggcaaataca caaagaaagc 34440 cctccccagt cctcatgtac tggtccctca ttgcactgta ctcctcttga cctgctgtgt 34500 cgagaatatc caagagacag gtttctccat caattactac ttgcttcctg taggaatcct 34560 ctattgttgg atcatattcg tccacaaaat gattctgaat tagctgtatc gtcaaggcac 34620 tcttgcaagc ttcattatca gcagctaaaa gttatcaggc atgcacctgg tagctagtct 34680 ttaaaccaat agattgcatc ggtttaaaag gcaagaccgt caaattgcgg gaaaggggtc 34740 aacagccgtt cagtaccaag tctcagggga aactttgaga tggccttgca aagggtatgg 34800 taataagctg acggacatgg tcctaaccac gcagccaagt cctaagtcaa cagatcttct 34860 gttgatatgg atgcagttca cagactaaat gtcggtcggg gaagatgtat tcttctcata 34920 agatatagtc ggacctctcc ttaatgggag ctagcggatg aagtgatgca acactggagc 34980 cgctgggaac taatttgtat gcgaaagtat attgattagt tttggagtac tcgcgatgat 35040 cacgcttcgt acccctgcca tcaacacgcg tctgcgttcg accaggctgc gcgttctcgc 35100 ggccatagca accgacgtac ggcgttgcgc cctcgccggc agcaagaagc cacggaagtc 35160 cgcctggagc agaaaatgcc cacgctactg cgggtttata tagacggtcc tcacgggatg 35220 gggaaaacca ccaccacgca actgctggtg gccctgggtt cgcgcgacga tatcgtctac 35280 gtacccgagc cgatgactta ctggcaggtg ctgggggctt ccgagacaat cgcgaacatc 35340 tacaccacac aacaccgcct cgaccagggt gagatatcgg ccggggacgc ggcggtggta 35400 atgacaagcg cccagataac aatgggcatg ccttatgccg tgaccgacgc cgttctggct 35460 cctcatgtcg ggggggaggc tgggagttca catgccccgc ccccggccct caccctcatc 35520 ttcgaccgcc atcccatcgc cgccctcctg tgctacccgg ccgcgcgata ccttatgggc 35580 agcatgaccc cccaggccgt gctggcgttc gtggccctca tcccgccgac cttgcccggc 35640 acaaacatcg tgttgggggc ccttccggag gacagacaca tcgaccgcct ggccaaacgc 35700 cagcgccccg gcgagcggct tgacctggct atgctggccg cgattcgccg cgtttacggg 35760 ctgcttgcca atacggtgcg gtatctgcag ggcggcgggt cgtggtggga ggattgggga 35820 cagctttcgg ggacggccgt gccgccccag ggtgccgagc cccagagcaa cgcgggccca 35880 cgaccccata tcggggacac gttatttacc ctgtttcggg cccccgagtt gctggccccc 35940 aacggcgacc tgtataacgt gtttgcctgg gccttggacg tcttggccaa acgcctccgt 36000 cccatgcacg tctttatcct ggattacgac caatcgcccg ccggctgccg ggacgccctg 36060 ctgcaactta cctccgggat ggtccagacc cacgtcacca ccccaggctc cataccgacg 36120 atctgcgacc tggcgcgcac gtttgcccgg gagatggggg aggctaactg aagcttttcg 36180 aaggcgaaag atcccaacga aaagagagac cacatggtcc ttcttgagtt tgtaacagct 36240 gctgggatta cacatggcat ggatgaactg tacaactgag gatcccccga cctcgacctc 36300 tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc 36360 actcggaagg acatatggga gggcaaatca tttggtcgag atccctcgga gatcggatct 36420 gggcgtgg 36428 <210> 43 <211> 36226 <212> DNA <213> Artificial Sequence <220> <223> Ad-AS100KRib2-TK <400> 43 ttaagggtgg gaaagaatat ataaggtggg ggtcttatgt agttttgtat ctgttttgca 60 gcagccgccg ccgccatgag caccaactcg tttgatggaa gcattgtgag ctcatatttg 120 acaacgcgca tgcccccatg ggccggggtg cgtcagaatg tgatgggctc cagcattgat 180 ggtcgccccg tcctgcccgc aaactctact accttgacct acgagaccgt gtctggaacg 240 ccgttggaga ctgcagcctc cgccgccgct tcagccgctg cagccaccgc ccgcgggatt 300 gtgactgact ttgctttcct gagcccgctt gcaagcagtg cagcttcccg ttcatccgcc 360 cgcgatgaca agttgacggc tcttttggca caattggatt ctttgacccg ggaacttaat 420 gtcgtttctc agcagctgtt ggatctgcgc cagcaggttt ctgccctgaa ggcttcctcc 480 cctcccaatg cggtttaaaa cataaataaa aaaccagact ctgtttggat ttggatcaag 540 caagtgtctt gctgtcttta tttaggggtt ttgcgcgcgc ggtaggcccg ggaccagcgg 600 tctcggtcgt tgagggtcct gtgtattttt tccaggacgt ggtaaaggtg actctggatg 660 ttcagataca tgggcataag cccgtctctg gggtggaggt agcaccactg cagagcttca 720 tgctgcgggg tggtgttgta gatgatccag tcgtagcagg agcgctgggc gtggtgccta 780 aaaatgtctt tcagtagcaa gctgattgcc aggggcaggc ccttggtgta agtgtttaca 840 aagcggttaa gctgggatgg gtgcatacgt ggggatatga gatgcatctt ggactgtatt 900 tttaggttgg ctatgttccc agccatatcc ctccggggat tcatgttgtg cagaaccacc 960 agcacagtgt atccggtgca cttgggaaat ttgtcatgta gcttagaagg aaatgcgtgg 1020 aagaacttgg agacgccctt gtgacctcca agattttcca tgcattcgtc cataatgatg 1080 gcaatgggcc cacgggcggc ggcctgggcg aagatatttc tgggatcact aacgtcatag 1140 ttgtgttcca ggatgagatc gtcataggcc atttttacaa agcgcgggcg gagggtgcca 1200 gactgcggta taatggttcc atccggccca ggggcgtagt taccctcaca gatttgcatt 1260 tcccacgctt tgagttcaga tggggggatc atgtctacct gcggggcgat gaagaaaacg 1320 gtttccgggg taggggagat cagctgggaa gaaagcaggt tcctgagcag ctgcgactta 1380 ccgcagccgg tgggcccgta aatcacacct attaccgggt gcaactggta gttaagagag 1440 ctgcagctgc cgtcatccct gagcaggggg gccacttcgt taagcatgtc cctgactcgc 1500 atgttttccc tgaccaaatc cgccagaagg cgctcgccgc ccagcgatag cagttcttgc 1560 aaggaagcaa agtttttcaa cggtttgaga ccgtccgccg taggcatgct tttgagcgtt 1620 tgaccaagca gttccaggcg gtcccacagc tcggtcacct gctctacggc atctcgatcc 1680 agcatatctc ctcgtttcgc gggttggggc ggctttcgct gtacggcagt agtcggtgct 1740 cgtccagacg ggccagggtc atgtctttcc acgggcgcag ggtcctcgtc agcgtagtct 1800 gggtcacggt gaaggggtgc gctccgggct gcgcgctggc cagggtgcgc ttgaggctgg 1860 tcctgctggt gctgaagcgc tgccggtctt cgccctgcgc gtcggccagg tagcatttga 1920 ccatggtgtc atagtccagc ccctccgcgg cgtggccctt ggcgcgcagc ttgcccttgg 1980 aggaggcgcc gcacgagggg cagtgcagac ttttgagggc gtagagcttg ggcgcgagaa 2040 ataccgattc cggggagtag gcatccgcgc cgcaggcccc gcagacggtc tcgcattcca 2100 cgagccaggt gagctctggc cgttcggggt caaaaaccag gtttccccca tgctttttga 2160 tgcgtttctt acctctggtt tccatgagcc ggtgtccacg ctcggtgacg aaaaggctgt 2220 ccgtgtcccc gtatacagac ttgagaggcc tgtcctcgag cggtgttccg cggtcctcct 2280 cgtatagaaa ctcggaccac tctgagacaa aggctcgcgt ccaggccagc acgaaggagg 2340 ctaagtggga ggggtagcgg tcgttgtcca ctagggggtc cactcgctcc agggtgtgaa 2400 gacacatgtc gccctcttcg gcatcaagga aggtgattgg tttgtaggtg taggccacgt 2460 gaccgggtgt tcctgaaggg gggctataaa agggggtggg ggcgcgttcg tcctcactct 2520 cttccgcatc gctgtctgcg agggccagct gttggggtga gtactccctc tgaaaagcgg 2580 gcatgacttc tgcgctaaga ttgtcagttt ccaaaaacga ggaggatttg atattcacct 2640 ggcccgcggt gatgcctttg agggtggccg catccatctg gtcagaaaag acaatctttt 2700 tgttgtcaag cttggtggca aacgacccgt agagggcgtt ggacagcaac ttggcgatgg 2760 agcgcagggt ttggtttttg tcgcgatcgg cgcgctcctt ggccgcgatg tttagctgca 2820 cgtattcgcg cgcaacgcac cgccattcgg gaaagacggt ggtgcgctcg tcgggcacca 2880 ggtgcacgcg ccaaccgcgg ttgtgcaggg tgacaaggtc aacgctggtg gctacctctc 2940 cgcgtaggcg ctcgttggtc cagcagaggc ggccgccctt gcgcgagcag aatggcggta 3000 gggggtctag ctgcgtctcg tccggggggt ctgcgtccac ggtaaagacc ccgggcagca 3060 ggcgcgcgtc gaagtagtct atcttgcatc cttgcaagtc tagcgcctgc tgccatgcgc 3120 gggcggcaag cgcgcgctcg tatgggttga gtgggggacc ccatggcatg gggtgggtga 3180 gcgcggaggc gtacatgccg caaatgtcgt aaacgtagag gggctctctg agtattccaa 3240 gatatgtagg gtagcatctt ccaccgcgga tgctggcgcg cacgtaatcg tatagttcgt 3300 gcgagggagc gaggaggtcg ggaccgaggt tgctacgggc gggctgctct gctcggaaga 3360 ctatctgcct gaagatggca tgtgagttgg atgatatggt tggacgctgg aagacgttga 3420 agctggcgtc tgtgagacct accgcgtcac gcacgaagga ggcgtaggag tcgcgcagct 3480 tgttgaccag ctcggcggtg acctgcacgt ctagggcgca gtagtccagg gtttccttga 3540 tgatgtcata cttatcctgt cccttttttt tccacagctc gcggttgagg acaaactctt 3600 cgcggtcttt ccagtactct tggatcggaa acccgtcggc ctccgaacgg taagagccta 3660 gcatgtagaa ctggttgacg gcctggtagg cgcagcatcc cttttctacg ggtagcgcgt 3720 atgcctgcgc ggccttccgg agcgaggtgt gggtgagcgc aaaggtgtcc ctgaccatga 3780 ctttgaggta ctggtatttg aagtcagtgt cgtcgcatcc gccctgctcc cagagcaaaa 3840 agtccgtgcg ctttttggaa cgcggatttg gcagggcgaa ggtgacatcg ttgaagagta 3900 tctttcccgc gcgaggcata aagttgcgtg tgatgcggaa gggtcccggc acctcggaac 3960 ggttgttaat tacctgggcg gcgagcacga tctcgtcaaa gccgttgatg ttgtggccca 4020 caatgtaaag ttccaagaag cgcgggatgc ccttgatgga aggcaatttt ttaagttcct 4080 cgtaggtgag ctcttcaggg gagctgagcc cgtgctctga aagggcccag tctgcaagat 4140 gagggttgga agcgacgaat gagctccaca ggtcacgggc cattagcatt tgcaggtggt 4200 cgcgaaaggt cctaaactgg cgacctatgg ccattttttc tggggtgatg cagtagaagg 4260 taagcgggtc ttgttcccag cggtcccatc caaggttcgc ggctaggtct cgcgcggcag 4320 tcactagagg ctcatctccg ccgaacttca tgaccagcat gaagggcacg agctgcttcc 4380 caaaggcccc catccaagta taggtctcta catcgtaggt gacaaagaga cgctcggtgc 4440 gaggatgcga gccgatcggg aagaactgga tctcccgcca ccaattggag gagtggctat 4500 tgatgtggtg aaagtagaag tccctgcgac gggccgaaca ctcgtgctgg cttttgtaaa 4560 aacgtgcgca gtactggcag cggtgcacgg gctgtacatc ctgcacgagg ttgacctgac 4620 gaccgcgcac aaggaagcag agtgggaatt tgagcccctc gcctggcggg tttggctggt 4680 ggtcttctac ttcggctgct tgtccttgac cgtctggctg ctcgagggga gttacggtgg 4740 atcggaccac cacgccgcgc gagcccaaag tccagatgtc cgcgcgcggc ggtcggagct 4800 tgatgacaac atcgcgcaga tgggagctgt ccatggtctg gagctcccgc ggcgtcaggt 4860 caggcgggag ctcctgcagg tttacctcgc atagacgggt cagggcgcgg gctagatcca 4920 ggtgatacct aatttccagg ggctggttgg tggcggcgtc gatggcttgc aagaggccgc 4980 atccccgcgg cgcgactacg gtaccgcgcg gcgggcggtg ggccgcgggg gtgtccttgg 5040 atgatgcatc taaaagcggt gacgcgggcg agcccccgga ggtagggggg gctccggacc 5100 cgccgggaga gggggcaggg gcacgtcggc gccgcgcgcg ggcaggagct ggtgctgcgc 5160 gcgtaggttg ctggcgaacg cgacgacgcg gcggttgatc tcctgaatct ggcgcctctg 5220 cgtgaagacg acgggcccgg tgagcttgag cctgaaagag agttcgacag aatcaatttc 5280 ggtgtcgttg acggcggcct ggcgcaaaat ctcctgcacg tctcctgagt tgtcttgata 5340 ggcgatctcg gccatgaact gctcgatctc ttcctcctgg agatctccgc gtccggctcg 5400 ctccacggtg gcggcgaggt cgttggaaat gcgggccatg agctgcgaga aggcgttgag 5460 gcctccctcg ttccagacgc ggctgtagac cacgccccct tcggcatcgc gggcgcgcat 5520 gaccacctgc gcgagattga gctccacgtg ccgggcgaag acggcgtagt ttcgcaggcg 5580 ctgaaagagg tagttgaggg tggtggcggt gtgttctgcc acgaagaagt acataaccca 5640 gcgtcgcaac gtggattcgt tgatatcccc caaggcctca aggcgctcca tggcctcgta 5700 gaagtccacg gcgaagttga aaaactggga gttgcgcgcc gacacggtta actcctcctc 5760 cagaagacgg atgagctcgg cgacagtgtc gcgcacctcg cgctcaaagg ctacaggggc 5820 ctcttcttct tcttcaatct cctcttccat aagggcctcc ccttcttctt cttctggcgg 5880 cggtggggga ggggggacac ggcggcgacg acggcgcacc gggaggcggt cgacaaagcg 5940 ctcgatcatc tccccgcggc gacggcgcat ggtctcggtg acggcgcggc cgttctcgcg 6000 ggggcgcagt tggaagacgc cgcccgtcat gtcccggtta tgggttggcg gggggctgcc 6060 atgcggcagg gatacggcgc taacgatgca tctcaacaat tgttgtgtag gtactccgcc 6120 gccgagggac ctgagcgagt ccgcatcgac cggatcggaa aacctctcga gaaaggcgtc 6180 taaccagtca cagtcgcaag gtaggctgag caccgtggcg ggcggcagcg ggcggcggtc 6240 ggggttgttt ctggcggagg tgctgctgat gatgtaatta aagtaggcgg tcttgagacg 6300 gcggatggtc gacagaagca ccatgtcctt gggtccggcc tgctgaatgc gcaggcggtc 6360 ggccatgccc caggcttcgt tttgacatcg gcgcaggtct ttgtagtagt cttgcatgag 6420 cctttctacc ggcacttctt cttctccttc ctcttgtcct gcatctcttg catctatcgc 6480 tgcggcggcg gcggagtttg gccgtaggtg gcgccctctt cctcccatgc gtgtgacccc 6540 gaagcccctc atcggctgaa gcagggctag gtcggcgaca acgcgctcgg ctaatatggc 6600 ctgctgcacc tgcgtgaggg tagactggaa gtcatccatg tccacaaagc ggtggtatgc 6660 gcccgtgttg atggtgtaag tgcagttggc cataacggac cagttaacgg tctggtgacc 6720 cggctgcgag agctcggtgt acctgagacg cgagtaagcc ctcgagtcaa atacgtagtc 6780 gttgcaagtc cgcaccaggt actggtatcc caccaaaaag tgcggcggcg gctggcggta 6840 gaggggccag cgtagggtgg ccggggctcc gggggcgaga tcttccaaca taaggcgatg 6900 atatccgtag atgtacctgg acatccaggt gatgccggcg gcggtggtgg aggcgcgcgg 6960 aaagtcgcgg acgcggttcc agatgttgcg cagcggcaaa aagtgctcca tggtcgggac 7020 gctctggccg gtcaggcgcg cgcaatcgtt gacgctctac cgtgcaaaag gagagcctgt 7080 aagcgggcac tcttccgtgg tctggtggat aaattcgcaa gggtatcatg gcggacgacc 7140 ggggttcgag ccccgtatcc ggccgtccgc cgtgatccat gcggttaccg cccgcgtgtc 7200 gaacccaggt gtgcgacgtc agacaacggg ggagtgctcc ttttggcttc cttccaggcg 7260 cggcggctgc tgcgctagct tttttggcca ctggccgcgc gcagcgtaag cggttaggct 7320 ggaaagcgaa agcattaagt ggctcgctcc ctgtagccgg agggttattt tccaagggtt 7380 gagtcgcggg acccccggtt cgagtctcgg accggccgga ctgcggcgaa cgggggtttg 7440 cctccccgtc atgcaagacc ccgcttgcaa attcctccgg aaacagggac gagccccttt 7500 tttgcttttc ccagatgcat ccggtgctgc ggcagatgcg cccccctcct cagcagcggc 7560 aagagcaaga gcagcggcag acatgcaggg caccctcccc tcctcctacc gcgtcaggag 7620 gggcgacatc cgcggttgac gcggcagcag atggtgatta cgaacccccg cggcgccggg 7680 cccggcacta cctggacttg gaggagggcg agggcctggc gcggctagga gcgccctctc 7740 ctgagcggta cccaagggtg cagctgaagc gtgatacgcg tgaggcgtac gtgccgcggc 7800 agaacctgtt tcgcgaccgc gagggagagg agcccgagga gatgcgggat cgaaagttcc 7860 acgcagggcg cgagctgcgg catggcctga atcgcgagcg gttgctgcgc gaggaggact 7920 ttgagcccga cgcgcgaacc gggattagtc ccgcgcgcgc acacgtggcg gccgccgacc 7980 tggtaaccgc atacgagcag acggtgaacc aggagattaa ctttcaaaaa agctttaaca 8040 accacgtgcg tacgcttgtg gcgcgcgagg aggtggctat aggactgatg catctgtggg 8100 actttgtaag cgcgctggag caaaacccaa atagcaagcc gctcatggcg cagctgttcc 8160 ttatagtgca gcacagcagg gacaacgagg cattcaggga tgcgctgcta aacatagtag 8220 agcccgaggg ccgctggctg ctcgatttga taaacatcct gcagagcata gtggtgcagg 8280 agcgcagctt gagcctggct gacaaggtgg ccgccatcaa ctattccatg cttagcctgg 8340 gcaagtttta cgcccgcaag atataccata ccccttacgt tcccatagac aaggaggtaa 8400 agatcgaggg gttctacatg cgcatggcgc tgaaggtgct taccttgagc gacgacctgg 8460 gcgtttatcg caacgagcgc atccacaagg ccgtgagcgt gagccggcgg cgcgagctca 8520 gcgaccgcga gctgatgcac agcctgcaaa gggccctggc tggcacgggc agcggcgata 8580 gagaggccga gtcctacttt gacgcgggcg ctgacctgcg ctgggcccca agccgacgcg 8640 ccctggaggc agctggggcc ggacctgggc tggcggtggc acccgcgcgc gctggcaacg 8700 tcggcggcgt ggaggaatat gacgaggacg atgagtacga gccagaggac ggcgagtact 8760 aagcggtgat gtttctgatc agatgatgca agacgcaacg gacccggcgg tgcgggcggc 8820 gctgcagagc cagccgtccg gccttaactc cacggacgac tggcgccagg tcatggaccg 8880 catcatgtcg ctgactgcgc gcaatcctga cgcgttccgg cagcagccgc aggccaaccg 8940 gctctccgca attctggaag cggtggtccc ggcgcgcgca aaccccacgc acgagaaggt 9000 gctggcgatc gtaaacgcgc tggccgaaaa cagggccatc cggcccgacg aggccggcct 9060 ggtctacgac gcgctgcttc agcgcgtggc tcgttacaac agcggcaacg tgcagaccaa 9120 cctggaccgg ctggtggggg atgtgcgcga ggccgtggcg cagcgtgagc gcgcgcagca 9180 gcagggcaac ctgggctcca tggttgcact aaacgccttc ctgagtacac agcccgccaa 9240 cgtgccgcgg ggacaggagg actacaccaa ctttgtgagc gcactgcggc taatggtgac 9300 tgagacaccg caaagtgagg tgtaccagtc tgggccagac tattttttcc agaccagtag 9360 acaaggcctg cagaccgtaa acctgagcca ggctttcaaa aacttgcagg ggctgtgggg 9420 ggtgcgggct cccacaggcg accgcgcgac cgtgtctagc ttgctgacgc ccaactcgcg 9480 cctgttgctg ctgctaatag cgcccttcac ggacagtggc agcgtgtccc gggacacata 9540 cctaggtcac ttgctgacac tgtaccgcga ggccataggt caggcgcatg tggacgagca 9600 tactttccag gagattacaa gtgtcagccg cgcgctgggg caggaggaca cgggcagcct 9660 ggaggcaacc ctaaactacc tgctgaccaa ccggcggcag aagatcccct cgttgcacag 9720 tttaaacagc gaggaggagc gcattttgcg ctacgtgcag cagagcgtga gccttaacct 9780 gatgcgcgac ggggtaacgc ccagcgtggc gctggacatg accgcgcgca acatggaacc 9840 gggcatgtat gcctcaaacc ggccgtttat caaccgccta atggactact tgcatcgcgc 9900 ggccgccgtg aaccccgagt atttcaccaa tgccatcttg aacccgcact ggctaccgcc 9960 ccctggtttc tacaccgggg gattcgaggt gcccgagggt aacgatggat tcctctggga 10020 cgacatagac gacagcgtgt tttccccgca accgcagacc ctgctagagt tgcaacagcg 10080 cgagcaggca gaggcggcgc tgcgaaagga aagcttccgc aggccaagca gcttgtccga 10140 tctaggcgct gcggccccgc ggtcagatgc tagtagccca tttccaagct tgatagggtc 10200 tcttaccagc actcgcacca cccgcccgcg cctgctgggc gaggaggagt acctaaacaa 10260 ctcgctgctg cagccgcagc gcgaaaaaaa cctgcctccg gcatttccca acaacgggat 10320 agagagccta gtggacaaga tgagtagatg gaagacgtac gcgcaggagc acagggacgt 10380 gccaggcccg cgcccgccca cccgtcgtca aaggcacgac cgtcagcggg gtctggtgtg 10440 ggaggacgat gactcggcag acgacagcag cgtcctggat ttgggaggga gtggcaaccc 10500 gtttgcgcac cttcgcccca ggctggggag aatgttttaa aaaaaaaaaa gcatgatgca 10560 aaataaaaaa ctcaccaagg ccatggcacc gagcgttggt tttcttgtat tccccttagt 10620 atgcggcgcg cggcgatgta tgaggaaggt cctcctccct cctacgagag tgtggtgagc 10680 gcggcgccag tggcggcggc gctgggttct cccttcgatg ctcccctgga cccgccgttt 10740 gtgcctccgc ggtacctgcg gcctaccggg gggagaaaca gcatccgtta ctctgagttg 10800 gcacccctat tcgacaccac ccgtgtgtac ctggtggaca acaagtcaac ggatgtggca 10860 tccctgaact accagaacga ccacagcaac tttctgacca cggtcattca aaacaatgac 10920 tacagcccgg gggaggcaag cacacagacc atcaatcttg acgaccggtc gcactggggc 10980 ggcgacctga aaaccatcct gcataccaac atgccaaatg tgaacgagtt catgtttacc 11040 aataagttta aggcgcgggt gatggtgtcg cgcttgccta ctaaggacaa tcaggtggag 11100 ctgaaatacg agtgggtgga gttcacgctg cccgagggca actactccga gaccatgacc 11160 atagacctta tgaacaacgc gatcgtggag cactacttga aagtgggcag acagaacggg 11220 gttctggaaa gcgacatcgg ggtaaagttt gacacccgca acttcagact ggggtttgac 11280 cccgtcactg gtcttgtcat gcctggggta tatacaaacg aagccttcca tccagacatc 11340 attttgctgc caggatgcgg ggtggacttc acccacagcc gcctgagcaa cttgttgggc 11400 atccgcaagc ggcaaccctt ccaggagggc tttaggatca cctacgatga tctggagggt 11460 ggtaacattc ccgcactgtt ggatgtggac gcctaccagg cgagcttgaa agatgacacc 11520 gaacagggcg ggggtggcgc aggcggcagc aacagcagtg gcagcggcgc ggaagagaac 11580 tccaacgcgg cagccgcggc aatgcagccg gtggaggaca tgaacgatca tgccattcgc 11640 ggcgacacct ttgccacacg ggctgaggag aagcgcgctg aggccgaagc agcggccgaa 11700 gctgccgccc ccgctgcgca acccgaggtc gagaagcctc agaagaaacc ggtgatcaaa 11760 cccctgacag aggacagcaa gaaacgcagt tacaacctaa taagcaatga cagcaccttc 11820 acccagtacc gcagctggta ccttgcatac aactacggcg accctcagac cggaatccgc 11880 tcatggaccc tgctttgcac tcctgacgta acctgcggct cggagcaggt ctactggtcg 11940 ttgccagaca tgatgcaaga ccccgtgacc ttccgctcca cgcgccagat cagcaacttt 12000 ccggtggtgg gcgccgagct gttgcccgtg cactccaaga gcttctacaa cgaccaggcc 12060 gtctactccc aactcatccg ccagtttacc tctctgaccc acgtgttcaa tcgctttccc 12120 gagaaccaga ttttggcgcg cccgccagcc cccaccatca ccaccgtcag tgaaaacgtt 12180 cctgctctca cagatcacgg gacgctaccg ctgcgcaaca gcatcggagg agtccagcga 12240 gtgaccatta ctgacgccag acgccgcacc tgcccctacg tttacaaggc cctgggcata 12300 gtctcgccgc gcgtcctatc gagccgcact ttttgagcaa gcatgtccat ccttatatcg 12360 cccagcaata acacaggctg gggcctgcgc ttcccaagca agatgtttgg cggggccaag 12420 aagcgctccg accaacaccc agtgcgcgtg cgcgggcact accgcgcgcc ctggggcgcg 12480 cacaaacgcg gccgcactgg gcgcaccacc gtcgatgacg ccatcgacgc ggtggtggag 12540 gaggcgcgca actacacgcc cacgccgcca ccagtgtcca cagtggacgc ggccattcag 12600 accgtggtgc gcggagcccg gcgctatgct aaaatgaaga gacggcggag gcgcgtagca 12660 cgtcgccacc gccgccgacc cggcactgcc gcccaacgcg cggcggcggc cctgcttaac 12720 cgcgcacgtc gcaccggccg acgggcggcc atgcgggccg ctcgaaggct ggccgcgggt 12780 attgtcactg tgccccccag gtccaggcga cgagcggccg ccgcagcagc cgcggccatt 12840 agtgctatga ctcagggtcg caggggcaac gtgtattggg tgcgcgactc ggttagcggc 12900 ctgcgcgtgc ccgtgcgcac ccgccccccg cgcaactaga ttgcaagaaa aaactactta 12960 gactcgtact gttgtatgta tccagcggcg gcggcgcgca acgaagctat gtccaagcgc 13020 aaaatcaaag aagagatgct ccaggtcatc gcgccggaga tctatggccc cccgaagaag 13080 gaagagcagg attacaagcc ccgaaagcta aagcgggtca aaaagaaaaa gaaagatgat 13140 gatgatgaac ttgacgacga ggtggaactg ctgcacgcta ccgcgcccag gcgacgggta 13200 cagtggaaag gtcgacgcgt aaaacgtgtt ttgcgacccg gcaccaccgt agtctttacg 13260 cccggtgagc gctccacccg cacctacaag cgcgtgtatg atgaggtgta cggcgacgag 13320 gacctgcttg agcaggccaa cgagcgcctc ggggagtttg cctacggaaa gcggcataag 13380 gacatgctgg cgttgccgct ggacgagggc aacccaacac ctagcctaaa gcccgtaaca 13440 ctgcagcagg tgctgcccgc gcttgcaccg tccgaagaaa agcgcggcct aaagcgcgag 13500 tctggtgact tggcacccac cgtgcagctg atggtaccca agcgccagcg actggaagat 13560 gtcttggaaa aaatgaccgt ggaacctggg ctggagcccg aggtccgcgt gcggccaatc 13620 aagcaggtgg cgccgggact gggcgtgcag accgtggacg ttcagatacc cactaccagt 13680 agcaccagta ttgccaccgc cacagagggc atggagacac aaacgtcccc ggttgcctca 13740 gcggtggcgg atgccgcggt gcaggcggtc gctgcggccg cgtccaagac ctctacggag 13800 gtgcaaacgg acccgtggat gtttcgcgtt tcagcccccc ggcgcccgcg cggttcgagg 13860 aagtacggcg ccgccagcgc gctactgccc gaatatgccc tacatccttc cattgcgcct 13920 acccccggct atcgtggcta cacctaccgc cccagaagac gagcaactac ccgacgccga 13980 accaccactg gaacccgccg ccgccgtcgc cgtcgccagc ccgtgctggc cccgatttcc 14040 gtgcgcaggg tggctcgcga aggaggcagg accctggtgc tgccaacagc gcgctaccac 14100 cccagcatcg tttaaaagcc ggtctttgtg gttcttgcag atatggccct cacctgccgc 14160 ctccgtttcc cggtgccggg attccgagga agaatgcacc gtaggagggg catggccggc 14220 cacggcctga cgggcggcat gcgtcgtgcg caccaccggc ggcggcgcgc gtcgcaccgt 14280 cgcatgcgcg gcggtatcct gcccctcctt attccactga tcgccgcggc gattggcgcc 14340 gtgcccggaa ttgcatccgt ggccttgcag gcgcagagac actgattaaa aacaagttgc 14400 atgtggaaaa atcaaaataa aaagtctgga ctctcacgct cgcttggtcc tgtaactatt 14460 ttgtagaatg gaagacatca actttgcgtc tctggccccg cgacacggct cgcgcccgtt 14520 catgggaaac tggcaagata tcggcaccag caatatgagc ggtggcgcct tcagctgggg 14580 ctcgctgtgg agcggcatta aaaatttcgg ttccaccgtt aagaactatg gcagcaaggc 14640 ctggaacagc agcacaggcc agatgctgag ggataagttg aaagagcaaa atttccaaca 14700 aaaggtggta gatggcctgg cctctggcat tagcggggtg gtggacctgg ccaaccaggc 14760 agtgcaaaat aagattaaca gtaagcttga tccccgccct cccgtagagg agcctccacc 14820 ggccgtggag acagtgtctc cagaggggcg tggcgaaaag cgtccgcgcc ccgacaggga 14880 agaaactctg gtgacgcaaa tagacgagcc tccctcgtac gaggaggcac taaagcaagg 14940 cctgcccacc acccgtccca tcgcgcccat ggctaccgga gtgctgggcc agcacacacc 15000 cgtaacgctg gacctgcctc cccccgccga cacccagcag aaacctgtgc tgccaggccc 15060 gaccgccgtt gttgtaaccc gtcctagccg cgcgtccctg cgccgcgccg ccagcggtcc 15120 gcgatcgttg cggcccgtag ccagtggcaa ctggcaaagc acactgaaca gcatcgtggg 15180 tctgggggtg caatccctga agcgccgacg atgcttctga atagctaacg tgtcgtatgt 15240 gtgtcatgta tgcgtccatg tcgccgccag aggagctgct gagccgccgc gcgcccgctt 15300 tccaagatgg ctaccccttc gatgatgccg cagtggtctt acatgcacat ctcgggccag 15360 gacgcctcgg agtacctgag ccccgggctg gtgcagtttg cccgcgccac cgagacgtac 15420 ttcagcctga ataacaagtt tagaaacccc acggtggcgc ctacgcacga cgtgaccaca 15480 gaccggtccc agcgtttgac gctgcggttc atccctgtgg accgtgagga tactgcgtac 15540 tcgtacaagg cgcggttcac cctagctgtg ggtgataacc gtgtgctgga catggcttcc 15600 acgtactttg acatccgcgg cgtgctggac aggggcccta cttttaagcc ctactctggc 15660 actgcctaca acgccctggc tcccaagggt gccccaaatc cttgcgaatg ggatgaagct 15720 gctactgctc ttgaaataaa cctagaagaa gaggacgatg acaacgaaga cgaagtagac 15780 gagcaagctg agcagcaaaa aactcacgta tttgggcagg cgccttattc tggtataaat 15840 attacaaagg agggtattca aataggtgtc gaaggtcaaa cacctaaata tgccgataaa 15900 acatttcaac ctgaacctca aataggagaa tctcagtggt acgaaactga aattaatcat 15960 gcagctggga gagtccttaa aaagactacc ccaatgaaac catgttacgg ttcatatgca 16020 aaacccacaa atgaaaatgg agggcaaggc attcttgtaa agcaacaaaa tggaaagcta 16080 gaaagtcaag tggaaatgca atttttctca actactgagg cgaccgcagg caatggtgat 16140 aacttgactc ctaaagtggt attgtacagt gaagatgtag atatagaaac cccagacact 16200 catatttctt acatgcccac tattaaggaa ggtaactcac gagaactaat gggccaacaa 16260 tctatgccca acaggcctaa ttacattgct tttagggaca attttattgg tctaatgtat 16320 tacaacagca cgggtaatat gggtgttctg gcgggccaag catcgcagtt gaatgctgtt 16380 gtagatttgc aagacagaaa cacagagctt tcataccagc ttttgcttga ttccattggt 16440 gatagaacca ggtacttttc tatgtggaat caggctgttg acagctatga tccagatgtt 16500 agaattattg aaaatcatgg aactgaagat gaacttccaa attactgctt tccactggga 16560 ggtgtgatta atacagagac tcttaccaag gtaaaaccta aaacaggtca ggaaaatgga 16620 tgggaaaaag atgctacaga attttcagat aaaaatgaaa taagagttgg aaataatttt 16680 gccatggaaa tcaatctaaa tgccaacctg tggagaaatt tcctgtactc caacatagcg 16740 ctgtatttgc ccgacaagct aaagtacagt ccttccaacg taaaaatttc tgataaccca 16800 aacacctacg actacatgaa caagcgagtg gtggctcccg ggttagtgga ctgctacatt 16860 aaccttggag cacgctggtc ccttgactat atggacaacg tcaacccatt taaccaccac 16920 cgcaatgctg gcctgcgcta ccgctcaatg ttgctgggca atggtcgcta tgtgcccttc 16980 cacatccagg tgcctcagaa gttctttgcc attaaaaacc tccttctcct gccgggctca 17040 tacacctacg agtggaactt caggaaggat gttaacatgg ttctgcagag ctccctagga 17100 aatgacctaa gggttgacgg agccagcatt aagtttgata gcatttgcct ttacgccacc 17160 ttcttcccca tggcccacaa caccgcctcc acgcttgagg ccatgcttag aaacgacacc 17220 aacgaccagt cctttaacga ctatctctcc gccgccaaca tgctctaccc tatacccgcc 17280 aacgctacca acgtgcccat atccatcccc tcccgcaact gggcggcttt ccgcggctgg 17340 gccttcacgc gccttaagac taaggaaacc ccatcactgg gctcgggcta cgacccttat 17400 tacacctact ctggctctat accctaccta gatggaacct tttacctcaa ccacaccttt 17460 aagaaggtgg ccattacctt tgactcttct gtcagctggc ctggcaatga ccgcctgctt 17520 acccccaacg agtttgaaat taagcgctca gttgacgggg agggttacaa cgttgcccag 17580 tgtaacatga ccaaagactg gttcctggta caaatgctag ctaactacaa cattggctac 17640 cagggcttct atatcccaga gagctacaag gaccgcatgt actccttctt tagaaacttc 17700 cagcccatga gccgtcaggt ggtggatgat actaaataca aggactacca acaggtgggc 17760 atcctacacc aacacaacaa ctctggattt gttggctacc ttgcccccac catgcgcgaa 17820 ggacaggcct accctgctaa cttcccctat ccgcttatag gcaagaccgc agttgacagc 17880 attacccaga aaaagtttct ttgcgatcgc accctttggc gcatcccatt ctccagtaac 17940 tttatgtcca tgggcgcact cacagacctg ggccaaaacc ttctctacgc caactccgcc 18000 cacgcgctag acatgacttt tgaggtggat cccatggacg agcccaccct tctttatgtt 18060 ttgtttgaag tctttgacgt ggtccgtgtg caccggccgc accgcggcgt catcgaaacc 18120 gtgtacctgc gcacgccctt ctcggccggc aacgccacaa cataaagaag caagcaacat 18180 caacaacagc tgccgccatg ggctccagtg agcaggaact gaaagccatt gtcaaagatc 18240 ttggttgtgg gccatatttt ttgggcacct atgacaagcg ctttccaggc tttgtttctc 18300 cacacaagct cgcctgcgcc atagtcaata cggccggtcg cgagactggg ggcgtacact 18360 ggatggcctt tgcctggaac ccgcactcaa aaacatgcta cctctttgag ccctttggct 18420 tttctgacca gcgactcaag caggtttacc agtttgagta cgagtcactc ctgcgccgta 18480 gcgccattgc ttcttccccc gaccgctgta taacgctgga aaagtccacc caaagcgtac 18540 aggggcccaa ctcggccgcc tgtggactat tctgctgcat gtttctccac gcctttgcca 18600 actggcccca aactcccatg gatcacaacc ccaccatgaa ccttattacc ggggtaccca 18660 actccatgct caacagtccc caggtacagc ccaccctgcg tcgcaaccag gaacagctct 18720 acagcttcct ggagcgccac tcgccctact tccgcagcca cagtgcgcag attaggagcg 18780 ccacttcttt ttgtcacttg aaaaacatgt aaaaataatg tactagagac actttcaata 18840 aaggcaaatg cttttatttg tacactctcg ggtgattatt tacccccacc cttgccgtct 18900 gcgccgttta aaaatcaaag gggttctgcc gcgcatcgct atgcgccact ggcagggaca 18960 cgttgcgata ctggtgttta gtgctccact taaactcagg cacaaccatc cgcggcagct 19020 cggtgaagtt ttcactccac aggctgcgca ccatcaccaa cgcgtttagc aggtcgggcg 19080 ccgatatctt gaagtcgcag ttggggcctc cgccctgcgc gcgcgagttg cgatacacag 19140 ggttgcagca ctggaacact atcagcgccg ggtggtgcac gctggccagc acgctcttgt 19200 cggagatcag atccgcgtcc aggtcctccg cgttgctcag ggcgaacgga gtcaactttg 19260 gtagctgcct tcccaaaaag ggcgcgtgcc caggctttga gttgcactcg caccgtagtg 19320 gcatcaaaag gtgaccgtgc ccggtctggg cgttaggata cagcgcctgc ataaaagcct 19380 tgatctgctt aaaagccacc tgagcctttg cgccttcaga gaagaacatg ccgcaagact 19440 tgccggaaaa ctgattggcc ggacaggccg cgtcgtgcac gcagcacctt gcgtcggtgt 19500 tggagatctg caccacattt cggccccacc ggttcttcac gatcttggcc ttgctagact 19560 gctccttcag cgcgcgctgc ccgttttcgc tcgtcacatc catttcaatc acgtgctcct 19620 tatttatcat aatgcttccg tgtagacact taagctcgcc ttcgatctca gcgcagcggt 19680 gcagccacaa cgcgcagccc gtgggctcgt gatgcttgta ggtcacctct gcaaacgact 19740 gcaggtacgc ctgcaggaat cgccccatca tcgtcacaaa ggtcttgttg ctggtgaagg 19800 tcagctgcaa cccgcggtgc tcctcgttca gccaggtctt gcatacggcc gccagagctt 19860 ccacttggtc aggcagtagt ttgaagttcg cctttagatc gttatccacg tggtacttgt 19920 ccatcagcgc gcgcgcagcc tccatgccct tctcccacgc agacacgatc ggcacactca 19980 gcgggttcat caccgtaatt tcactttccg cttcgctggg ctcttcctct tcctcttgcg 20040 tccgcatacc acgcgccact gggtcgtctt cattcagccg ccgcactgtg cgcttacctc 20100 ctttgccatg cttgattagc accggtgggt tgctgaaacc caccatttgt agcgccacat 20160 cttctctttc ttcctcgctg tccacgatta cctctggtga tggcgggcgc tcgggcttgg 20220 gagaagggcg cttctttttc ttcttgggcg caatggccaa atccgccgcc gaggtcgatg 20280 gccgcgggct gggtgtgcgc ggcaccagcg cgtcttgtga tgagtcttcc tcgtcctcgg 20340 actcgatacg ccgcctcatc cgcttttttg ggggcgcccg gggaggcggc ggcgacgggg 20400 acggggacga cacgtcctcc atggttgggg gacgtcgcgc cgcaccgcgt ccgcgctcgg 20460 gggtggtttc gcgctgctcc tcttcccgac tggccatttc cttctcctat aggcagaaaa 20520 agatcatgga gtcagtcgag aagaaggaca gcctaaccgc cccctctgag ttcgccacca 20580 ccgcctccac cgatgccgcc aacgcgccta ccaccttccc cgtcgaggca cccccgcttg 20640 aggaggagga agtgattatc gagcaggacc caggttttgt aagcgaagac gacgaggacc 20700 gctcagtacc aacagaggat aaaaagcaag accaggacaa cgcagaggca aacgaggaac 20760 aagtcgggcg gggggacgaa aggcatggcg actacctaga tgtgggagac gacgtgctgt 20820 tgaagcatct gcagcgccag tgcgccatta tctgcgacgc gttgcaagag cgcagcgatg 20880 tgcccctcgc catagcggat gtcagccttg cctacgaacg ccacctattc tcaccgcgcg 20940 taccccccaa acgccaagaa aacggcacat gcgagcccaa cccgcgcctc aacttctacc 21000 ccgtatttgc cgtgccagag gtgcttgcca cctatcacat ctttttccaa aactgcaaga 21060 tacccctatc ctgccgtgcc aaccgcagcc gagcggacaa gcagctggcc ttgcggcagg 21120 gcgctgtcat acctgatatc gcctcgctca acgaagtgcc aaaaatcttt gagggtcttg 21180 gacgcgacga gaagcgcgcg gcaaacgctc tgcaacagga aaacagcgaa aatgaaagtc 21240 actctggagt gttggtggaa ctcgagggtg acaacgcgcg cctagccgta ctaaaacgca 21300 gcatcgaggt cacccacttt gcctacccgg cacttaacct accccccaag gtcatgagca 21360 cagtcatgag tgagctgatc gtgcgccgtg cgcagcccct ggagagggat gcaaatttgc 21420 aagaacaaac agaggagggc ctacccgcag ttggcgacga gcagctagcg cgctggcttc 21480 aaacgcgcga gcctgccgac ttggaggagc gacgcaaact aatgatggcc gcagtgctcg 21540 ttaccgtgga gcttgagtgc atgcagcggt tctttgctga cccggagatg cagcgcaagc 21600 tagaggaaac attgcactac acctttcgac agggctacgt acgccaggcc tgcaagatct 21660 ccaacgtgga gctctgcaac ctggtctcct accttggaat tttgcacgaa aaccgccttg 21720 ggcaaaacgt gcttcattcc acgctcaagg gcgaggcgcg ccgcgactac gtccgcgact 21780 gcgtttactt atttctatgc tacacctggc agacggccat gggcgtttgg cagcagtgct 21840 tggaggagtg caacctcaag gagctgcaga aactgctaaa gcaaaacttg aaggacctat 21900 ggacggcctt caacgagcgc tccgtggccg cgcacctggc ggacatcatt ttccccgaac 21960 gcctgcttaa aaccctgcaa cagggtctgc cagacttcac cagtcaaagc atgttgcaga 22020 actttaggaa ctttatccta gagcgctcag gaatcttgcc cgccacctgc tgtgcacttc 22080 ctagcgactt tgtgcccatt aagtaccgcg aatgccctcc gccgctttgg ggccactgct 22140 accttctgca gctagccaac taccttgcct accactctga cataatggaa gacgtgagcg 22200 gtgacggtct actggagtgt cactgtcgct gcaacctatg caccccgcac cgctccctgg 22260 tttgcaattc gcagctgctt aacgaaagtc aaattatcgg tacctttgag ctgcagggtc 22320 cctcgcctga cgaaaagtcc gcggctccgg ggttgaaact cactccgggg ctgtggacgt 22380 cggcttacct tcgcaaattt gtacctgagg actaccacgc ccacgagatt aggttctacg 22440 aagaccaatc ccgcccgcca aatgcggagc ttaccgcctg cgtcattacc cagggccaca 22500 ttcttggcca attgcaagcc atcaacaaag cccgccaaga gtttctgcta cgaaagggac 22560 ggggggttta cttggacccc cagtccggcg aggagctcaa cccaatcccc ccgccgccgc 22620 agccctatca gcagcagccg cgggcccttg cttcccagga tggcacccaa aaagaagctg 22680 cagctgccgc cgccacccac ggacgaggag gaatactggg acagtcaggc agaggaggtt 22740 ttggacgagg aggaggagga catgatggaa gactgggaga gcctagacga ggaagcttcc 22800 gaggtcgaag aggtgtcaga cgaaacaccg tcaccctcgg tcgcattccc ctcgccggcg 22860 ccccagaaat cggcaaccgg ttccagcatg gctacaacct ccgctcctca ggcgccgccg 22920 gcactgcccg ttcgccgacc caaccgtaga tgggacacca ctggaaccag ggccggtaag 22980 tccaagcagc cgccgccgtt agcccaagag caacaacagc gccaaggcta ccgctcatgg 23040 cgcgggcaca agaacgccat agttgcttgc ttgcaagact gtgggggcaa catctccttc 23100 gcccgccgct ttcttctcta ccatcacggc gtggccttcc cccgtaacat cctgcattac 23160 taccgtcatc tctacagccc atactgcacc ggcggcagcg gcagcggcag caacagcagc 23220 ggccacacag aagcaaaggc gaccggatag caagactctg acaaagccca agaaatccac 23280 agcggcggca gcagcaggag gaggagcgct gcgtctggcg cccaacgaac ccgtatcgac 23340 ccgcgagctt agaaacagga tttttcccac tctgtatgct atatttcaac agagcagggg 23400 ccaagaacaa gagctgaaaa taaaaaacag gtctctgcga tccctcaccc gcagctgcct 23460 gtatcacaaa agcgaagatc agcttcggcg cacgctggaa gacgcggagg ctctcttcag 23520 taaatactgc gcgctgactc ttaaggacta gtttcgcgcc ctttctcaaa tttaagcgcg 23580 aaaactacgt catctccagc ggccacaccc ggcgccagca cctgtcgtca gcgccattat 23640 gagcaaggaa attcccacgc cctacatgtg gagttaccag ccacaaatgg gacttgcggc 23700 tggagctgcc caagactact caacccgaat aaactacatg agcgcgggac cccacatgat 23760 atcccgggtc aacggaatcc gcgcccaccg aaaccgaatt ctcttggaac aggcggctat 23820 taccaccaca cctcgtaata accttaatcc ccgtagttgg cccgctgccc tggtgtacca 23880 ggaaagtccc gctcccacca ctgtggtact tcccagagac gcccaggccg aagttcagat 23940 gactaactca ggggcgcagc ttgcgggcgg ctttcgtcac agggtgcggt cgcccgggca 24000 gggtataact cacctgacaa tcagagggcg aggtattcag ctcaacgacg agtcggtgag 24060 ctcctcgctt ggtctccgtc cggacgggac atttcagatc ggcggcgccg gccgtccttc 24120 attcacgcct cgtcaggcaa tcctaactct gcagacctcg tcctctgagc cgcgctctgg 24180 aggcattgga actctgcaat ttattgagga gtttgtgcca tcggtctact ttaacccctt 24240 ctcgggacct cccggccact atccggatca atttattcct aactttgacg cggtaaagga 24300 ctcggcggac ggctacgact gaatgttaag tggagaggca gagcaactgc gcctgaaaca 24360 cctggtccac tgtcgccgcc acaagtgctt tgcccgcgac tccggtgagt tttgctactt 24420 tgaattgccc gaggatcata tcgagggccc ggcgcacggc gtccggctta ccgcccaggg 24480 agagcttgcc cgtagcctga ttcgggagtt tacccagcgc cccctgctag ttgagcggga 24540 caggggaccc tgtgttctca ctgtgatttg caactgtcct aaccttggat tacatcaaga 24600 tcctctagtt ataactagag tacccgggga tcttattccc tttaactaat aaaaaaaaat 24660 aataaagcat cacttactta aaatcagtta gcaaatttct gtccagttta ttcagcagca 24720 cctccttgcc ctcctcccag ctctggtatt gcagcttcct cctggctgca aactttctcc 24780 acaatctaaa tggaatgtca gtttcctcct gttcctgtcc atccgcaccc actatcttca 24840 tgttgttgca gatgaagcgc gcaagaccgt ctgaagatac cttcaacccc gtgtatccat 24900 atgacacgga aaccggtcct ccaactgtgc cttttcttac tcctcccttt gtatccccca 24960 atgggtttca agagagtccc cctggggtac tctctttgcg cctatccgaa cctctagtta 25020 cctccaatgg catgcttgcg ctcaaaatgg gcaacggcct ctctctggac gaggccggca 25080 accttacctc ccaaaatgta accactgtga gcccacctct caaaaaaacc aagtcaaaca 25140 taaacctgga aatatctgca cccctcacag ttacctcaga agccctaact gtggctgccg 25200 ccgcacctct aatggtcgcg ggcaacacac tcaccatgca atcacaggcc ccgctaaccg 25260 tgcacgactc caaacttagc attgccaccc aaggacccct cacagtgtca gaaggaaagc 25320 tagccctgca aacatcaggc cccctcacca ccaccgatag cagtaccctt actatcactg 25380 cctcaccccc tctaactact gccactggta gcttgggcat tgacttgaaa gagcccattt 25440 atacacaaaa tggaaaacta ggactaaagt acggggctcc tttgcatgta acagacgacc 25500 taaacacttt gaccgtagca actggtccag gtgtgactat taataatact tccttgcaaa 25560 ctaaagttac tggagccttg ggttttgatt cacaaggcaa tatgcaactt aatgtagcag 25620 gaggactaag gattgattct caaaacagac gccttatact tgatgttagt tatccgtttg 25680 atgctcaaaa ccaactaaat ctaagactag gacagggccc tctttttata aactcagccc 25740 acaacttgga tattaactac aacaaaggcc tttacttgtt tacagcttca aacaattcca 25800 aaaagcttga ggttaaccta agcactgcca aggggttgat gtttgacgct acagccatag 25860 ccattaatgc aggagatggg cttgaatttg gttcacctaa tgcaccaaac acaaatcccc 25920 tcaaaacaaa aattggccat ggcctagaat ttgattcaaa caaggctatg gttcctaaac 25980 taggaactgg ccttagtttt gacagcacag gtgccattac agtaggaaac aaaaataatg 26040 ataagctaac tttgtggacc acaccagctc catctcctaa ctgtagacta aatgcagaga 26100 aagatgctaa actcactttg gtcttaacaa aatgtggcag tcaaatactt gctacagttt 26160 cagttttggc tgttaaaggc agtttggctc caatatctgg aacagttcaa agtgctcatc 26220 ttattataag atttgacgaa aatggagtgc tactaaacaa ttccttcctg gacccagaat 26280 attggaactt tagaaatgga gatcttactg aaggcacagc ctatacaaac gctgttggat 26340 ttatgcctaa cctatcagct tatccaaaat ctcacggtaa aactgccaaa agtaacattg 26400 tcagtcaagt ttacttaaac ggagacaaaa ctaaacctgt aacactaacc attacactaa 26460 acggtacaca ggaaacagga gacacaactc caagtgcata ctctatgtca ttttcatggg 26520 actggtctgg ccacaactac attaatgaaa tatttgccac atcctcttac actttttcat 26580 acattgccca agaataaaga atcgtttgtg ttatgtttca acgtgtttat ttttcaattg 26640 cagaaaattt caagtcattt ttcattcagt agtatagccc caccaccaca tagcttatac 26700 agatcaccgt accttaatca aactcacaga accctagtat tcaacctgcc acctccctcc 26760 caacacacag agtacacagt cctttctccc cggctggcct taaaaagcat catatcatgg 26820 gtaacagaca tattcttagg tgttatattc cacacggttt cctgtcgagc caaacgctca 26880 tcagtgatat taataaactc cccgggcagc tcacttaagt tcatgtcgct gtccagctgc 26940 tgagccacag gctgctgtcc aacttgcggt tgcttaacgg gcggcgaagg agaagtccac 27000 gcctacatgg gggtagagtc ataatcgtgc atcaggatag ggcggtggtg ctgcagcagc 27060 gcgcgaataa actgctgccg ccgccgctcc gtcctgcagg aatacaacat ggcagtggtc 27120 tcctcagcga tgattcgcac cgcccgcagc ataaggcgcc ttgtcctccg ggcacagcag 27180 cgcaccctga tctcacttaa atcagcacag taactgcagc acagcaccac aatattgttc 27240 aaaatcccac agtgcaaggc gctgtatcca aagctcatgg cggggaccac agaacccacg 27300 tggccatcat accacaagcg caggtagatt aagtggcgac ccctcataaa cacgctggac 27360 ataaacatta cctcttttgg catgttgtaa ttcaccacct cccggtacca tataaacctc 27420 tgattaaaca tggcgccatc caccaccatc ctaaaccagc tggccaaaac ctgcccgccg 27480 gctatacact gcagggaacc gggactggaa caatgacagt ggagagccca ggactcgtaa 27540 ccatggatca tcatgctcgt catgatatca atgttggcac aacacaggca cacgtgcata 27600 cacttcctca ggattacaag ctcctcccgc gttagaacca tatcccaggg aacaacccat 27660 tcctgaatca gcgtaaatcc cacactgcag ggaagacctc gcacgtaact cacgttgtgc 27720 attgtcaaag tgttacattc gggcagcagc ggatgatcct ccagtatggt agcgcgggtt 27780 tctgtctcaa aaggaggtag acgatcccta ctgtacggag tgcgccgaga caaccgagat 27840 cgtgttggtc gtagtgtcat gccaaatgga acgccggacg tagtcatatt tcctgaagca 27900 aaaccaggtg cgggcgtgac aaacagatct gcgtctccgg tctcgccgct tagatcgctc 27960 tgtgtagtag ttgtagtata tccactctct caaagcatcc aggcgccccc tggcttcggg 28020 ttctatgtaa actccttcat gcgccgctgc cctgataaca tccaccaccg cagaataagc 28080 cacacccagc caacctacac attcgttctg cgagtcacac acgggaggag cgggaagagc 28140 tggaagaacc atgttttttt ttttattcca aaagattatc caaaacctca aaatgaagat 28200 ctattaagtg aacgcgctcc cctccggtgg cgtggtcaaa ctctacagcc aaagaacaga 28260 taatggcatt tgtaagatgt tgcacaatgg cttccaaaag gcaaacggcc ctcacgtcca 28320 agtggacgta aaggctaaac ccttcagggt gaatctcctc tataaacatt ccagcacctt 28380 caaccatgcc caaataattc tcatctcgcc accttctcaa tatatctcta agcaaatccc 28440 gaatattaag tccggccatt gtaaaaatct gctccagagc gccctccacc ttcagcctca 28500 agcagcgaat catgattgca aaaattcagg ttcctcacag acctgtataa gattcaaaag 28560 cggaacatta acaaaaatac cgcgatcccg taggtccctt cgcagggcca gctgaacata 28620 atcgtgcagg tctgcacgga ccagcgcggc cacttccccg ccaggaacct tgacaaaaga 28680 acccacactg attatgacac gcatactcgg agctatgcta accagcgtag ccccgatgta 28740 agctttcaat agcttgttgc atgggcggcg atataaaatg caaggtgctg ctcaaaaaat 28800 caggcaaagc ctcgcgcaaa aaagaaagca catcgtagtc atgctcatgc agataaaggc 28860 aggtaagctc cggaaccacc acagaaaaag acaccatttt tctctcaaac atgtctgcgg 28920 gtttctgcat aaacacaaaa taaaataaca aaaaaacatt taaacattag aagcctgtct 28980 tacaacagga aaaacaaccc ttataagcat aagacggact acggccatgc cggcgtgacc 29040 gtaaaaaaac tggtcaccgt gattaaaaag caccaccgac agctcctcgg tcatgtccgg 29100 agtcataatg taagactcgg taaacacatc aggttgattc atcggtcagt gctaaaaagc 29160 gaccgaaata gcccggggga atacataccc gcaggcgtag agacaacatt acagccccca 29220 taggaggtat aacaaaatta ataggagaga aaaacacata aacacctgaa aaaccctcct 29280 gcctaggcaa aatagcaccc tcccgctcca gaacaacata cagcgcttca cagcggcagc 29340 ctaacagtca gccttaccag taaaaaagaa aacctattaa aaaaacacca ctcgacacgg 29400 caccagctca atcagtcaca gtgtaaaaaa gggccaagtg cagagcgagt atatatagga 29460 ctaaaaaatg acgtaacggt taaagtccac aaaaaacacc cagaaaaccg cacgcgaacc 29520 tacgcccaga aacgaaagcc aaaaaaccca caacttcctc aaatcgtcac ttccgttttc 29580 ccacgttacg taacttccca ttttaagaaa actacaattc ccaacacata caagttactc 29640 cgccctaaaa cctacgtcac ccgccccgtt cccacgcccc gcgccacgtc acaaactcca 29700 ccccctcatt atcatattgg cttcaatcca aaataaggta tattattgat gatgttaatt 29760 aacatgcatg gatccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc 29820 gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 29880 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 29940 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 30000 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 30060 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 30120 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 30180 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 30240 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 30300 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 30360 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 30420 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc 30480 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 30540 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 30600 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 30660 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 30720 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 30780 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 30840 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 30900 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 30960 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 31020 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 31080 ccattgctgc agccatgaga ttatcaaaaa ggatcttcac ctagatcctt ttcacgtaga 31140 aagccagtcc gcagaaacgg tgctgacccc ggatgaatgt cagctactgg gctatctgga 31200 caagggaaaa cgcaagcgca aagagaaagc aggtagcttg cagtgggctt acatggcgat 31260 agctagactg ggcggtttta tggacagcaa gcgaaccgga attgccagct ggggcgccct 31320 ctggtaaggt tgggaagccc tgcaaagtaa actggatggc tttctcgccg ccaaggatct 31380 gatggcgcag gggatcaagc tctgatcaag agacaggatg aggatcgttt cgcatgattg 31440 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg 31500 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg 31560 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg 31620 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg 31680 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc 31740 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc 31800 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc 31860 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc 31920 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg 31980 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct 32040 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt 32100 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc 32160 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt 32220 tcttctgaat tttgttaaaa tttttgttaa atcagctcat tttttaacca ataggccgaa 32280 atcggcaaca tcccttataa atcaaaagaa tagaccgcga tagggttgag tgttgttcca 32340 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc 32400 gtctatcagg gcgatggccc actacgtgaa ccatcaccca aatcaagttt tttgcggtcg 32460 aggtgccgta aagctctaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg 32520 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg 32580 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgcgcg cttaatgcgc 32640 cgctacaggg cgcgtccatt cgccattcag gatcgaatta attcttaatt aacatcatca 32700 ataatatacc ttattttgga ttgaagccaa tatgataatg agggggtgga gtttgtgacg 32760 tggcgcgggg cgtgggaacg gggcgggtga cgtagtagtg tggcggaagt gtgatgttgc 32820 aagtgtggcg gaacacatgt aagcgacgga tgtggcaaaa gtgacgtttt tggtgtgcgc 32880 cggtgtacac aggaagtgac aattttcgcg cggttttagg cggatgttgt agtaaatttg 32940 ggcgtaaccg agtaagattt ggccattttc gcgggaaaac tgaataagag gaagtgaaat 33000 ctgaataatt ttgtgttact catagcgcgt aatactgcga tctatacatt gaatcaatat 33060 tggcaattag ccatattagt cattggttat atagcataaa tcaatattgg ctattggcca 33120 ttgcatacgt tgtatctata tcataatatg tacatttata ttggctcatg tccaatatga 33180 ccgccatgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta 33240 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc 33300 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg 33360 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg 33420 gcagtacatc aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa 33480 tggcccgcct ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac 33540 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg 33600 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg 33660 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg 33720 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta 33780 gtgaaccgtc agatcctcac tctcttccgc atcgctgtct gcgagggcca gctgttgggc 33840 tcgcggttga ggacaaactc ttcgcggtct ttccagtact cttggatcgg aaacccgtcg 33900 gcctccgaac ggtactccgc caccgaggga cctgagccag tccgcatcga ccggatcgga 33960 aaacctctcg agaaaggcgt ctaaccagtc acagtcgcaa ggtaggctga gcaccgtggc 34020 gggcggcagc gggtggcggt cggggttgtt tctggcggag gtgctgctga tgatgtaatt 34080 aaagtaggcg gtcttgagcc ggcggatggt cgaggtgagg tgtggcaggc ttgagatcca 34140 gctgttgggg tgagtactcc ctctcaaaag cgggcatgac ttctgcgcta agattgtcag 34200 tttccaaaaa cgaggaggat ttgatattca cctggcccga tctggccata cacttgagtg 34260 acaatgacat ccactttgcc tttctctcca caggtgtcca ctcccaggtc caagtttgga 34320 agatccccat caattactac ttgcttcctg taggaatcct ctattgttgg atcatattcg 34380 tccacaaaat gattctgaat tagctgtatc gtcaaggcac tcttgcaagc ttcatcatgg 34440 ccgacagcaa aagttatcag gcatgcacct ggtagctagt ctttaaacca atagattgca 34500 tcggtttaaa aggcaagacc gtcaaattgc gggaaagggg tcaacagccg ttcagtacca 34560 agtctcaggg gaaactttga gatggccttg caaagggtat ggtaataagc tgacggacat 34620 ggtcctaacc acgcagccaa gtcctaagtc aacagatctt ctgttgatat ggatgcagtt 34680 cacagactaa atgtcggtcg gggaagatgt attcttctca taagatatag tcggacctct 34740 ccttaatggg agctagcgga tgaagtgatg caacactgga gccgctggga actaatttgt 34800 atgcgaaagt atattgatta gttttggagt actcgcgaca tgatcacgct tcgtacccct 34860 gccatcaaca cgcgtctgcg ttcgaccagg ctgcgcgttc tcgcggccat agcaaccgac 34920 gtacggcgtt gcgccctcgc cggcagcaag aagccacgga agtccgcctg gagcagaaaa 34980 tgcccacgct actgcgggtt tatatagacg gtcctcacgg gatggggaaa accaccacca 35040 cgcaactgct ggtggccctg ggttcgcgcg acgatatcgt ctacgtaccc gagccgatga 35100 cttactggca ggtgctgggg gcttccgaga caatcgcgaa catctacacc acacaacacc 35160 gcctcgacca gggtgagata tcggccgggg acgcggcggt ggtaatgaca agcgcccaga 35220 taacaatggg catgccttat gccgtgaccg acgccgttct ggctcctcat gtcggggggg 35280 aggctgggag ttcacatgcc ccgcccccgg ccctcaccct catcttcgac cgccatccca 35340 tcgccgccct cctgtgctac ccggccgcgc gataccttat gggcagcatg accccccagg 35400 ccgtgctggc gttcgtggcc ctcatcccgc cgaccttgcc cggcacaaac atcgtgttgg 35460 gggcccttcc ggaggacaga cacatcgacc gcctggccaa acgccagcgc cccggcgagc 35520 ggcttgacct ggctatgctg gccgcgattc gccgcgttta cgggctgctt gccaatacgg 35580 tgcggtatct gcagggcggc gggtcgtggt gggaggattg gggacagctt tcggggacgg 35640 ccgtgccgcc ccagggtgcc gagccccaga gcaacgcggg cccacgaccc catatcgggg 35700 acacgttatt taccctgttt cgggcccccg agttgctggc ccccaacggc gacctgtata 35760 acgtgtttgc ctgggccttg gacgtcttgg ccaaacgcct ccgtcccatg cacgtcttta 35820 tcctggatta cgaccaatcg cccgccggct gccgggacgc cctgctgcaa cttacctccg 35880 ggatggtcca gacccacgtc accaccccag gctccatacc gacgatctgc gacctggcgc 35940 gcacgtttgc ccgggagatg ggggaggcta ataattcgaa ggcgaaagat cccaacgaaa 36000 agagagacca catggtcctt cttgagtttg taacagctgc tgggattaca catggcatgg 36060 atgaactgta caactgagga tcccccgacc tcgacctctg gctaataaag gaaatttatt 36120 ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg 36180 gcaaatcatt tggtcgagat ccctcggaga tcggatctgg gcgtgg 36226 <210> 44 <211> 38160 <212> DNA <213> Artificial Sequence <220> <223> Ad-AS100Rib2-LacZ <400> 44 ttaagggtgg gaaagaatat ataaggtggg ggtcttatgt agttttgtat ctgttttgca 60 gcagccgccg ccgccatgag caccaactcg tttgatggaa gcattgtgag ctcatatttg 120 acaacgcgca tgcccccatg ggccggggtg cgtcagaatg tgatgggctc cagcattgat 180 ggtcgccccg tcctgcccgc aaactctact accttgacct acgagaccgt gtctggaacg 240 ccgttggaga ctgcagcctc cgccgccgct tcagccgctg cagccaccgc ccgcgggatt 300 gtgactgact ttgctttcct gagcccgctt gcaagcagtg cagcttcccg ttcatccgcc 360 cgcgatgaca agttgacggc tcttttggca caattggatt ctttgacccg ggaacttaat 420 gtcgtttctc agcagctgtt ggatctgcgc cagcaggttt ctgccctgaa ggcttcctcc 480 cctcccaatg cggtttaaaa cataaataaa aaaccagact ctgtttggat ttggatcaag 540 caagtgtctt gctgtcttta tttaggggtt ttgcgcgcgc ggtaggcccg ggaccagcgg 600 tctcggtcgt tgagggtcct gtgtattttt tccaggacgt ggtaaaggtg actctggatg 660 ttcagataca tgggcataag cccgtctctg gggtggaggt agcaccactg cagagcttca 720 tgctgcgggg tggtgttgta gatgatccag tcgtagcagg agcgctgggc gtggtgccta 780 aaaatgtctt tcagtagcaa gctgattgcc aggggcaggc ccttggtgta agtgtttaca 840 aagcggttaa gctgggatgg gtgcatacgt ggggatatga gatgcatctt ggactgtatt 900 tttaggttgg ctatgttccc agccatatcc ctccggggat tcatgttgtg cagaaccacc 960 agcacagtgt atccggtgca cttgggaaat ttgtcatgta gcttagaagg aaatgcgtgg 1020 aagaacttgg agacgccctt gtgacctcca agattttcca tgcattcgtc cataatgatg 1080 gcaatgggcc cacgggcggc ggcctgggcg aagatatttc tgggatcact aacgtcatag 1140 ttgtgttcca ggatgagatc gtcataggcc atttttacaa agcgcgggcg gagggtgcca 1200 gactgcggta taatggttcc atccggccca ggggcgtagt taccctcaca gatttgcatt 1260 tcccacgctt tgagttcaga tggggggatc atgtctacct gcggggcgat gaagaaaacg 1320 gtttccgggg taggggagat cagctgggaa gaaagcaggt tcctgagcag ctgcgactta 1380 ccgcagccgg tgggcccgta aatcacacct attaccgggt gcaactggta gttaagagag 1440 ctgcagctgc cgtcatccct gagcaggggg gccacttcgt taagcatgtc cctgactcgc 1500 atgttttccc tgaccaaatc cgccagaagg cgctcgccgc ccagcgatag cagttcttgc 1560 aaggaagcaa agtttttcaa cggtttgaga ccgtccgccg taggcatgct tttgagcgtt 1620 tgaccaagca gttccaggcg gtcccacagc tcggtcacct gctctacggc atctcgatcc 1680 agcatatctc ctcgtttcgc gggttggggc ggctttcgct gtacggcagt agtcggtgct 1740 cgtccagacg ggccagggtc atgtctttcc acgggcgcag ggtcctcgtc agcgtagtct 1800 gggtcacggt gaaggggtgc gctccgggct gcgcgctggc cagggtgcgc ttgaggctgg 1860 tcctgctggt gctgaagcgc tgccggtctt cgccctgcgc gtcggccagg tagcatttga 1920 ccatggtgtc atagtccagc ccctccgcgg cgtggccctt ggcgcgcagc ttgcccttgg 1980 aggaggcgcc gcacgagggg cagtgcagac ttttgagggc gtagagcttg ggcgcgagaa 2040 ataccgattc cggggagtag gcatccgcgc cgcaggcccc gcagacggtc tcgcattcca 2100 cgagccaggt gagctctggc cgttcggggt caaaaaccag gtttccccca tgctttttga 2160 tgcgtttctt acctctggtt tccatgagcc ggtgtccacg ctcggtgacg aaaaggctgt 2220 ccgtgtcccc gtatacagac ttgagaggcc tgtcctcgag cggtgttccg cggtcctcct 2280 cgtatagaaa ctcggaccac tctgagacaa aggctcgcgt ccaggccagc acgaaggagg 2340 ctaagtggga ggggtagcgg tcgttgtcca ctagggggtc cactcgctcc agggtgtgaa 2400 gacacatgtc gccctcttcg gcatcaagga aggtgattgg tttgtaggtg taggccacgt 2460 gaccgggtgt tcctgaaggg gggctataaa agggggtggg ggcgcgttcg tcctcactct 2520 cttccgcatc gctgtctgcg agggccagct gttggggtga gtactccctc tgaaaagcgg 2580 gcatgacttc tgcgctaaga ttgtcagttt ccaaaaacga ggaggatttg atattcacct 2640 ggcccgcggt gatgcctttg agggtggccg catccatctg gtcagaaaag acaatctttt 2700 tgttgtcaag cttggtggca aacgacccgt agagggcgtt ggacagcaac ttggcgatgg 2760 agcgcagggt ttggtttttg tcgcgatcgg cgcgctcctt ggccgcgatg tttagctgca 2820 cgtattcgcg cgcaacgcac cgccattcgg gaaagacggt ggtgcgctcg tcgggcacca 2880 ggtgcacgcg ccaaccgcgg ttgtgcaggg tgacaaggtc aacgctggtg gctacctctc 2940 cgcgtaggcg ctcgttggtc cagcagaggc ggccgccctt gcgcgagcag aatggcggta 3000 gggggtctag ctgcgtctcg tccggggggt ctgcgtccac ggtaaagacc ccgggcagca 3060 ggcgcgcgtc gaagtagtct atcttgcatc cttgcaagtc tagcgcctgc tgccatgcgc 3120 gggcggcaag cgcgcgctcg tatgggttga gtgggggacc ccatggcatg gggtgggtga 3180 gcgcggaggc gtacatgccg caaatgtcgt aaacgtagag gggctctctg agtattccaa 3240 gatatgtagg gtagcatctt ccaccgcgga tgctggcgcg cacgtaatcg tatagttcgt 3300 gcgagggagc gaggaggtcg ggaccgaggt tgctacgggc gggctgctct gctcggaaga 3360 ctatctgcct gaagatggca tgtgagttgg atgatatggt tggacgctgg aagacgttga 3420 agctggcgtc tgtgagacct accgcgtcac gcacgaagga ggcgtaggag tcgcgcagct 3480 tgttgaccag ctcggcggtg acctgcacgt ctagggcgca gtagtccagg gtttccttga 3540 tgatgtcata cttatcctgt cccttttttt tccacagctc gcggttgagg acaaactctt 3600 cgcggtcttt ccagtactct tggatcggaa acccgtcggc ctccgaacgg taagagccta 3660 gcatgtagaa ctggttgacg gcctggtagg cgcagcatcc cttttctacg ggtagcgcgt 3720 atgcctgcgc ggccttccgg agcgaggtgt gggtgagcgc aaaggtgtcc ctgaccatga 3780 ctttgaggta ctggtatttg aagtcagtgt cgtcgcatcc gccctgctcc cagagcaaaa 3840 agtccgtgcg ctttttggaa cgcggatttg gcagggcgaa ggtgacatcg ttgaagagta 3900 tctttcccgc gcgaggcata aagttgcgtg tgatgcggaa gggtcccggc acctcggaac 3960 ggttgttaat tacctgggcg gcgagcacga tctcgtcaaa gccgttgatg ttgtggccca 4020 caatgtaaag ttccaagaag cgcgggatgc ccttgatgga aggcaatttt ttaagttcct 4080 cgtaggtgag ctcttcaggg gagctgagcc cgtgctctga aagggcccag tctgcaagat 4140 gagggttgga agcgacgaat gagctccaca ggtcacgggc cattagcatt tgcaggtggt 4200 cgcgaaaggt cctaaactgg cgacctatgg ccattttttc tggggtgatg cagtagaagg 4260 taagcgggtc ttgttcccag cggtcccatc caaggttcgc ggctaggtct cgcgcggcag 4320 tcactagagg ctcatctccg ccgaacttca tgaccagcat gaagggcacg agctgcttcc 4380 caaaggcccc catccaagta taggtctcta catcgtaggt gacaaagaga cgctcggtgc 4440 gaggatgcga gccgatcggg aagaactgga tctcccgcca ccaattggag gagtggctat 4500 tgatgtggtg aaagtagaag tccctgcgac gggccgaaca ctcgtgctgg cttttgtaaa 4560 aacgtgcgca gtactggcag cggtgcacgg gctgtacatc ctgcacgagg ttgacctgac 4620 gaccgcgcac aaggaagcag agtgggaatt tgagcccctc gcctggcggg tttggctggt 4680 ggtcttctac ttcggctgct tgtccttgac cgtctggctg ctcgagggga gttacggtgg 4740 atcggaccac cacgccgcgc gagcccaaag tccagatgtc cgcgcgcggc ggtcggagct 4800 tgatgacaac atcgcgcaga tgggagctgt ccatggtctg gagctcccgc ggcgtcaggt 4860 caggcgggag ctcctgcagg tttacctcgc atagacgggt cagggcgcgg gctagatcca 4920 ggtgatacct aatttccagg ggctggttgg tggcggcgtc gatggcttgc aagaggccgc 4980 atccccgcgg cgcgactacg gtaccgcgcg gcgggcggtg ggccgcgggg gtgtccttgg 5040 atgatgcatc taaaagcggt gacgcgggcg agcccccgga ggtagggggg gctccggacc 5100 cgccgggaga gggggcaggg gcacgtcggc gccgcgcgcg ggcaggagct ggtgctgcgc 5160 gcgtaggttg ctggcgaacg cgacgacgcg gcggttgatc tcctgaatct ggcgcctctg 5220 cgtgaagacg acgggcccgg tgagcttgag cctgaaagag agttcgacag aatcaatttc 5280 ggtgtcgttg acggcggcct ggcgcaaaat ctcctgcacg tctcctgagt tgtcttgata 5340 ggcgatctcg gccatgaact gctcgatctc ttcctcctgg agatctccgc gtccggctcg 5400 ctccacggtg gcggcgaggt cgttggaaat gcgggccatg agctgcgaga aggcgttgag 5460 gcctccctcg ttccagacgc ggctgtagac cacgccccct tcggcatcgc gggcgcgcat 5520 gaccacctgc gcgagattga gctccacgtg ccgggcgaag acggcgtagt ttcgcaggcg 5580 ctgaaagagg tagttgaggg tggtggcggt gtgttctgcc acgaagaagt acataaccca 5640 gcgtcgcaac gtggattcgt tgatatcccc caaggcctca aggcgctcca tggcctcgta 5700 gaagtccacg gcgaagttga aaaactggga gttgcgcgcc gacacggtta actcctcctc 5760 cagaagacgg atgagctcgg cgacagtgtc gcgcacctcg cgctcaaagg ctacaggggc 5820 ctcttcttct tcttcaatct cctcttccat aagggcctcc ccttcttctt cttctggcgg 5880 cggtggggga ggggggacac ggcggcgacg acggcgcacc gggaggcggt cgacaaagcg 5940 ctcgatcatc tccccgcggc gacggcgcat ggtctcggtg acggcgcggc cgttctcgcg 6000 ggggcgcagt tggaagacgc cgcccgtcat gtcccggtta tgggttggcg gggggctgcc 6060 atgcggcagg gatacggcgc taacgatgca tctcaacaat tgttgtgtag gtactccgcc 6120 gccgagggac ctgagcgagt ccgcatcgac cggatcggaa aacctctcga gaaaggcgtc 6180 taaccagtca cagtcgcaag gtaggctgag caccgtggcg ggcggcagcg ggcggcggtc 6240 ggggttgttt ctggcggagg tgctgctgat gatgtaatta aagtaggcgg tcttgagacg 6300 gcggatggtc gacagaagca ccatgtcctt gggtccggcc tgctgaatgc gcaggcggtc 6360 ggccatgccc caggcttcgt tttgacatcg gcgcaggtct ttgtagtagt cttgcatgag 6420 cctttctacc ggcacttctt cttctccttc ctcttgtcct gcatctcttg catctatcgc 6480 tgcggcggcg gcggagtttg gccgtaggtg gcgccctctt cctcccatgc gtgtgacccc 6540 gaagcccctc atcggctgaa gcagggctag gtcggcgaca acgcgctcgg ctaatatggc 6600 ctgctgcacc tgcgtgaggg tagactggaa gtcatccatg tccacaaagc ggtggtatgc 6660 gcccgtgttg atggtgtaag tgcagttggc cataacggac cagttaacgg tctggtgacc 6720 cggctgcgag agctcggtgt acctgagacg cgagtaagcc ctcgagtcaa atacgtagtc 6780 gttgcaagtc cgcaccaggt actggtatcc caccaaaaag tgcggcggcg gctggcggta 6840 gaggggccag cgtagggtgg ccggggctcc gggggcgaga tcttccaaca taaggcgatg 6900 atatccgtag atgtacctgg acatccaggt gatgccggcg gcggtggtgg aggcgcgcgg 6960 aaagtcgcgg acgcggttcc agatgttgcg cagcggcaaa aagtgctcca tggtcgggac 7020 gctctggccg gtcaggcgcg cgcaatcgtt gacgctctac cgtgcaaaag gagagcctgt 7080 aagcgggcac tcttccgtgg tctggtggat aaattcgcaa gggtatcatg gcggacgacc 7140 ggggttcgag ccccgtatcc ggccgtccgc cgtgatccat gcggttaccg cccgcgtgtc 7200 gaacccaggt gtgcgacgtc agacaacggg ggagtgctcc ttttggcttc cttccaggcg 7260 cggcggctgc tgcgctagct tttttggcca ctggccgcgc gcagcgtaag cggttaggct 7320 ggaaagcgaa agcattaagt ggctcgctcc ctgtagccgg agggttattt tccaagggtt 7380 gagtcgcggg acccccggtt cgagtctcgg accggccgga ctgcggcgaa cgggggtttg 7440 cctccccgtc atgcaagacc ccgcttgcaa attcctccgg aaacagggac gagccccttt 7500 tttgcttttc ccagatgcat ccggtgctgc ggcagatgcg cccccctcct cagcagcggc 7560 aagagcaaga gcagcggcag acatgcaggg caccctcccc tcctcctacc gcgtcaggag 7620 gggcgacatc cgcggttgac gcggcagcag atggtgatta cgaacccccg cggcgccggg 7680 cccggcacta cctggacttg gaggagggcg agggcctggc gcggctagga gcgccctctc 7740 ctgagcggta cccaagggtg cagctgaagc gtgatacgcg tgaggcgtac gtgccgcggc 7800 agaacctgtt tcgcgaccgc gagggagagg agcccgagga gatgcgggat cgaaagttcc 7860 acgcagggcg cgagctgcgg catggcctga atcgcgagcg gttgctgcgc gaggaggact 7920 ttgagcccga cgcgcgaacc gggattagtc ccgcgcgcgc acacgtggcg gccgccgacc 7980 tggtaaccgc atacgagcag acggtgaacc aggagattaa ctttcaaaaa agctttaaca 8040 accacgtgcg tacgcttgtg gcgcgcgagg aggtggctat aggactgatg catctgtggg 8100 actttgtaag cgcgctggag caaaacccaa atagcaagcc gctcatggcg cagctgttcc 8160 ttatagtgca gcacagcagg gacaacgagg cattcaggga tgcgctgcta aacatagtag 8220 agcccgaggg ccgctggctg ctcgatttga taaacatcct gcagagcata gtggtgcagg 8280 agcgcagctt gagcctggct gacaaggtgg ccgccatcaa ctattccatg cttagcctgg 8340 gcaagtttta cgcccgcaag atataccata ccccttacgt tcccatagac aaggaggtaa 8400 agatcgaggg gttctacatg cgcatggcgc tgaaggtgct taccttgagc gacgacctgg 8460 gcgtttatcg caacgagcgc atccacaagg ccgtgagcgt gagccggcgg cgcgagctca 8520 gcgaccgcga gctgatgcac agcctgcaaa gggccctggc tggcacgggc agcggcgata 8580 gagaggccga gtcctacttt gacgcgggcg ctgacctgcg ctgggcccca agccgacgcg 8640 ccctggaggc agctggggcc ggacctgggc tggcggtggc acccgcgcgc gctggcaacg 8700 tcggcggcgt ggaggaatat gacgaggacg atgagtacga gccagaggac ggcgagtact 8760 aagcggtgat gtttctgatc agatgatgca agacgcaacg gacccggcgg tgcgggcggc 8820 gctgcagagc cagccgtccg gccttaactc cacggacgac tggcgccagg tcatggaccg 8880 catcatgtcg ctgactgcgc gcaatcctga cgcgttccgg cagcagccgc aggccaaccg 8940 gctctccgca attctggaag cggtggtccc ggcgcgcgca aaccccacgc acgagaaggt 9000 gctggcgatc gtaaacgcgc tggccgaaaa cagggccatc cggcccgacg aggccggcct 9060 ggtctacgac gcgctgcttc agcgcgtggc tcgttacaac agcggcaacg tgcagaccaa 9120 cctggaccgg ctggtggggg atgtgcgcga ggccgtggcg cagcgtgagc gcgcgcagca 9180 gcagggcaac ctgggctcca tggttgcact aaacgccttc ctgagtacac agcccgccaa 9240 cgtgccgcgg ggacaggagg actacaccaa ctttgtgagc gcactgcggc taatggtgac 9300 tgagacaccg caaagtgagg tgtaccagtc tgggccagac tattttttcc agaccagtag 9360 acaaggcctg cagaccgtaa acctgagcca ggctttcaaa aacttgcagg ggctgtgggg 9420 ggtgcgggct cccacaggcg accgcgcgac cgtgtctagc ttgctgacgc ccaactcgcg 9480 cctgttgctg ctgctaatag cgcccttcac ggacagtggc agcgtgtccc gggacacata 9540 cctaggtcac ttgctgacac tgtaccgcga ggccataggt caggcgcatg tggacgagca 9600 tactttccag gagattacaa gtgtcagccg cgcgctgggg caggaggaca cgggcagcct 9660 ggaggcaacc ctaaactacc tgctgaccaa ccggcggcag aagatcccct cgttgcacag 9720 tttaaacagc gaggaggagc gcattttgcg ctacgtgcag cagagcgtga gccttaacct 9780 gatgcgcgac ggggtaacgc ccagcgtggc gctggacatg accgcgcgca acatggaacc 9840 gggcatgtat gcctcaaacc ggccgtttat caaccgccta atggactact tgcatcgcgc 9900 ggccgccgtg aaccccgagt atttcaccaa tgccatcttg aacccgcact ggctaccgcc 9960 ccctggtttc tacaccgggg gattcgaggt gcccgagggt aacgatggat tcctctggga 10020 cgacatagac gacagcgtgt tttccccgca accgcagacc ctgctagagt tgcaacagcg 10080 cgagcaggca gaggcggcgc tgcgaaagga aagcttccgc aggccaagca gcttgtccga 10140 tctaggcgct gcggccccgc ggtcagatgc tagtagccca tttccaagct tgatagggtc 10200 tcttaccagc actcgcacca cccgcccgcg cctgctgggc gaggaggagt acctaaacaa 10260 ctcgctgctg cagccgcagc gcgaaaaaaa cctgcctccg gcatttccca acaacgggat 10320 agagagccta gtggacaaga tgagtagatg gaagacgtac gcgcaggagc acagggacgt 10380 gccaggcccg cgcccgccca cccgtcgtca aaggcacgac cgtcagcggg gtctggtgtg 10440 ggaggacgat gactcggcag acgacagcag cgtcctggat ttgggaggga gtggcaaccc 10500 gtttgcgcac cttcgcccca ggctggggag aatgttttaa aaaaaaaaaa gcatgatgca 10560 aaataaaaaa ctcaccaagg ccatggcacc gagcgttggt tttcttgtat tccccttagt 10620 atgcggcgcg cggcgatgta tgaggaaggt cctcctccct cctacgagag tgtggtgagc 10680 gcggcgccag tggcggcggc gctgggttct cccttcgatg ctcccctgga cccgccgttt 10740 gtgcctccgc ggtacctgcg gcctaccggg gggagaaaca gcatccgtta ctctgagttg 10800 gcacccctat tcgacaccac ccgtgtgtac ctggtggaca acaagtcaac ggatgtggca 10860 tccctgaact accagaacga ccacagcaac tttctgacca cggtcattca aaacaatgac 10920 tacagcccgg gggaggcaag cacacagacc atcaatcttg acgaccggtc gcactggggc 10980 ggcgacctga aaaccatcct gcataccaac atgccaaatg tgaacgagtt catgtttacc 11040 aataagttta aggcgcgggt gatggtgtcg cgcttgccta ctaaggacaa tcaggtggag 11100 ctgaaatacg agtgggtgga gttcacgctg cccgagggca actactccga gaccatgacc 11160 atagacctta tgaacaacgc gatcgtggag cactacttga aagtgggcag acagaacggg 11220 gttctggaaa gcgacatcgg ggtaaagttt gacacccgca acttcagact ggggtttgac 11280 cccgtcactg gtcttgtcat gcctggggta tatacaaacg aagccttcca tccagacatc 11340 attttgctgc caggatgcgg ggtggacttc acccacagcc gcctgagcaa cttgttgggc 11400 atccgcaagc ggcaaccctt ccaggagggc tttaggatca cctacgatga tctggagggt 11460 ggtaacattc ccgcactgtt ggatgtggac gcctaccagg cgagcttgaa agatgacacc 11520 gaacagggcg ggggtggcgc aggcggcagc aacagcagtg gcagcggcgc ggaagagaac 11580 tccaacgcgg cagccgcggc aatgcagccg gtggaggaca tgaacgatca tgccattcgc 11640 ggcgacacct ttgccacacg ggctgaggag aagcgcgctg aggccgaagc agcggccgaa 11700 gctgccgccc ccgctgcgca acccgaggtc gagaagcctc agaagaaacc ggtgatcaaa 11760 cccctgacag aggacagcaa gaaacgcagt tacaacctaa taagcaatga cagcaccttc 11820 acccagtacc gcagctggta ccttgcatac aactacggcg accctcagac cggaatccgc 11880 tcatggaccc tgctttgcac tcctgacgta acctgcggct cggagcaggt ctactggtcg 11940 ttgccagaca tgatgcaaga ccccgtgacc ttccgctcca cgcgccagat cagcaacttt 12000 ccggtggtgg gcgccgagct gttgcccgtg cactccaaga gcttctacaa cgaccaggcc 12060 gtctactccc aactcatccg ccagtttacc tctctgaccc acgtgttcaa tcgctttccc 12120 gagaaccaga ttttggcgcg cccgccagcc cccaccatca ccaccgtcag tgaaaacgtt 12180 cctgctctca cagatcacgg gacgctaccg ctgcgcaaca gcatcggagg agtccagcga 12240 gtgaccatta ctgacgccag acgccgcacc tgcccctacg tttacaaggc cctgggcata 12300 gtctcgccgc gcgtcctatc gagccgcact ttttgagcaa gcatgtccat ccttatatcg 12360 cccagcaata acacaggctg gggcctgcgc ttcccaagca agatgtttgg cggggccaag 12420 aagcgctccg accaacaccc agtgcgcgtg cgcgggcact accgcgcgcc ctggggcgcg 12480 cacaaacgcg gccgcactgg gcgcaccacc gtcgatgacg ccatcgacgc ggtggtggag 12540 gaggcgcgca actacacgcc cacgccgcca ccagtgtcca cagtggacgc ggccattcag 12600 accgtggtgc gcggagcccg gcgctatgct aaaatgaaga gacggcggag gcgcgtagca 12660 cgtcgccacc gccgccgacc cggcactgcc gcccaacgcg cggcggcggc cctgcttaac 12720 cgcgcacgtc gcaccggccg acgggcggcc atgcgggccg ctcgaaggct ggccgcgggt 12780 attgtcactg tgccccccag gtccaggcga cgagcggccg ccgcagcagc cgcggccatt 12840 agtgctatga ctcagggtcg caggggcaac gtgtattggg tgcgcgactc ggttagcggc 12900 ctgcgcgtgc ccgtgcgcac ccgccccccg cgcaactaga ttgcaagaaa aaactactta 12960 gactcgtact gttgtatgta tccagcggcg gcggcgcgca acgaagctat gtccaagcgc 13020 aaaatcaaag aagagatgct ccaggtcatc gcgccggaga tctatggccc cccgaagaag 13080 gaagagcagg attacaagcc ccgaaagcta aagcgggtca aaaagaaaaa gaaagatgat 13140 gatgatgaac ttgacgacga ggtggaactg ctgcacgcta ccgcgcccag gcgacgggta 13200 cagtggaaag gtcgacgcgt aaaacgtgtt ttgcgacccg gcaccaccgt agtctttacg 13260 cccggtgagc gctccacccg cacctacaag cgcgtgtatg atgaggtgta cggcgacgag 13320 gacctgcttg agcaggccaa cgagcgcctc ggggagtttg cctacggaaa gcggcataag 13380 gacatgctgg cgttgccgct ggacgagggc aacccaacac ctagcctaaa gcccgtaaca 13440 ctgcagcagg tgctgcccgc gcttgcaccg tccgaagaaa agcgcggcct aaagcgcgag 13500 tctggtgact tggcacccac cgtgcagctg atggtaccca agcgccagcg actggaagat 13560 gtcttggaaa aaatgaccgt ggaacctggg ctggagcccg aggtccgcgt gcggccaatc 13620 aagcaggtgg cgccgggact gggcgtgcag accgtggacg ttcagatacc cactaccagt 13680 agcaccagta ttgccaccgc cacagagggc atggagacac aaacgtcccc ggttgcctca 13740 gcggtggcgg atgccgcggt gcaggcggtc gctgcggccg cgtccaagac ctctacggag 13800 gtgcaaacgg acccgtggat gtttcgcgtt tcagcccccc ggcgcccgcg cggttcgagg 13860 aagtacggcg ccgccagcgc gctactgccc gaatatgccc tacatccttc cattgcgcct 13920 acccccggct atcgtggcta cacctaccgc cccagaagac gagcaactac ccgacgccga 13980 accaccactg gaacccgccg ccgccgtcgc cgtcgccagc ccgtgctggc cccgatttcc 14040 gtgcgcaggg tggctcgcga aggaggcagg accctggtgc tgccaacagc gcgctaccac 14100 cccagcatcg tttaaaagcc ggtctttgtg gttcttgcag atatggccct cacctgccgc 14160 ctccgtttcc cggtgccggg attccgagga agaatgcacc gtaggagggg catggccggc 14220 cacggcctga cgggcggcat gcgtcgtgcg caccaccggc ggcggcgcgc gtcgcaccgt 14280 cgcatgcgcg gcggtatcct gcccctcctt attccactga tcgccgcggc gattggcgcc 14340 gtgcccggaa ttgcatccgt ggccttgcag gcgcagagac actgattaaa aacaagttgc 14400 atgtggaaaa atcaaaataa aaagtctgga ctctcacgct cgcttggtcc tgtaactatt 14460 ttgtagaatg gaagacatca actttgcgtc tctggccccg cgacacggct cgcgcccgtt 14520 catgggaaac tggcaagata tcggcaccag caatatgagc ggtggcgcct tcagctgggg 14580 ctcgctgtgg agcggcatta aaaatttcgg ttccaccgtt aagaactatg gcagcaaggc 14640 ctggaacagc agcacaggcc agatgctgag ggataagttg aaagagcaaa atttccaaca 14700 aaaggtggta gatggcctgg cctctggcat tagcggggtg gtggacctgg ccaaccaggc 14760 agtgcaaaat aagattaaca gtaagcttga tccccgccct cccgtagagg agcctccacc 14820 ggccgtggag acagtgtctc cagaggggcg tggcgaaaag cgtccgcgcc ccgacaggga 14880 agaaactctg gtgacgcaaa tagacgagcc tccctcgtac gaggaggcac taaagcaagg 14940 cctgcccacc acccgtccca tcgcgcccat ggctaccgga gtgctgggcc agcacacacc 15000 cgtaacgctg gacctgcctc cccccgccga cacccagcag aaacctgtgc tgccaggccc 15060 gaccgccgtt gttgtaaccc gtcctagccg cgcgtccctg cgccgcgccg ccagcggtcc 15120 gcgatcgttg cggcccgtag ccagtggcaa ctggcaaagc acactgaaca gcatcgtggg 15180 tctgggggtg caatccctga agcgccgacg atgcttctga atagctaacg tgtcgtatgt 15240 gtgtcatgta tgcgtccatg tcgccgccag aggagctgct gagccgccgc gcgcccgctt 15300 tccaagatgg ctaccccttc gatgatgccg cagtggtctt acatgcacat ctcgggccag 15360 gacgcctcgg agtacctgag ccccgggctg gtgcagtttg cccgcgccac cgagacgtac 15420 ttcagcctga ataacaagtt tagaaacccc acggtggcgc ctacgcacga cgtgaccaca 15480 gaccggtccc agcgtttgac gctgcggttc atccctgtgg accgtgagga tactgcgtac 15540 tcgtacaagg cgcggttcac cctagctgtg ggtgataacc gtgtgctgga catggcttcc 15600 acgtactttg acatccgcgg cgtgctggac aggggcccta cttttaagcc ctactctggc 15660 actgcctaca acgccctggc tcccaagggt gccccaaatc cttgcgaatg ggatgaagct 15720 gctactgctc ttgaaataaa cctagaagaa gaggacgatg acaacgaaga cgaagtagac 15780 gagcaagctg agcagcaaaa aactcacgta tttgggcagg cgccttattc tggtataaat 15840 attacaaagg agggtattca aataggtgtc gaaggtcaaa cacctaaata tgccgataaa 15900 acatttcaac ctgaacctca aataggagaa tctcagtggt acgaaactga aattaatcat 15960 gcagctggga gagtccttaa aaagactacc ccaatgaaac catgttacgg ttcatatgca 16020 aaacccacaa atgaaaatgg agggcaaggc attcttgtaa agcaacaaaa tggaaagcta 16080 gaaagtcaag tggaaatgca atttttctca actactgagg cgaccgcagg caatggtgat 16140 aacttgactc ctaaagtggt attgtacagt gaagatgtag atatagaaac cccagacact 16200 catatttctt acatgcccac tattaaggaa ggtaactcac gagaactaat gggccaacaa 16260 tctatgccca acaggcctaa ttacattgct tttagggaca attttattgg tctaatgtat 16320 tacaacagca cgggtaatat gggtgttctg gcgggccaag catcgcagtt gaatgctgtt 16380 gtagatttgc aagacagaaa cacagagctt tcataccagc ttttgcttga ttccattggt 16440 gatagaacca ggtacttttc tatgtggaat caggctgttg acagctatga tccagatgtt 16500 agaattattg aaaatcatgg aactgaagat gaacttccaa attactgctt tccactggga 16560 ggtgtgatta atacagagac tcttaccaag gtaaaaccta aaacaggtca ggaaaatgga 16620 tgggaaaaag atgctacaga attttcagat aaaaatgaaa taagagttgg aaataatttt 16680 gccatggaaa tcaatctaaa tgccaacctg tggagaaatt tcctgtactc caacatagcg 16740 ctgtatttgc ccgacaagct aaagtacagt ccttccaacg taaaaatttc tgataaccca 16800 aacacctacg actacatgaa caagcgagtg gtggctcccg ggttagtgga ctgctacatt 16860 aaccttggag cacgctggtc ccttgactat atggacaacg tcaacccatt taaccaccac 16920 cgcaatgctg gcctgcgcta ccgctcaatg ttgctgggca atggtcgcta tgtgcccttc 16980 cacatccagg tgcctcagaa gttctttgcc attaaaaacc tccttctcct gccgggctca 17040 tacacctacg agtggaactt caggaaggat gttaacatgg ttctgcagag ctccctagga 17100 aatgacctaa gggttgacgg agccagcatt aagtttgata gcatttgcct ttacgccacc 17160 ttcttcccca tggcccacaa caccgcctcc acgcttgagg ccatgcttag aaacgacacc 17220 aacgaccagt cctttaacga ctatctctcc gccgccaaca tgctctaccc tatacccgcc 17280 aacgctacca acgtgcccat atccatcccc tcccgcaact gggcggcttt ccgcggctgg 17340 gccttcacgc gccttaagac taaggaaacc ccatcactgg gctcgggcta cgacccttat 17400 tacacctact ctggctctat accctaccta gatggaacct tttacctcaa ccacaccttt 17460 aagaaggtgg ccattacctt tgactcttct gtcagctggc ctggcaatga ccgcctgctt 17520 acccccaacg agtttgaaat taagcgctca gttgacgggg agggttacaa cgttgcccag 17580 tgtaacatga ccaaagactg gttcctggta caaatgctag ctaactacaa cattggctac 17640 cagggcttct atatcccaga gagctacaag gaccgcatgt actccttctt tagaaacttc 17700 cagcccatga gccgtcaggt ggtggatgat actaaataca aggactacca acaggtgggc 17760 atcctacacc aacacaacaa ctctggattt gttggctacc ttgcccccac catgcgcgaa 17820 ggacaggcct accctgctaa cttcccctat ccgcttatag gcaagaccgc agttgacagc 17880 attacccaga aaaagtttct ttgcgatcgc accctttggc gcatcccatt ctccagtaac 17940 tttatgtcca tgggcgcact cacagacctg ggccaaaacc ttctctacgc caactccgcc 18000 cacgcgctag acatgacttt tgaggtggat cccatggacg agcccaccct tctttatgtt 18060 ttgtttgaag tctttgacgt ggtccgtgtg caccggccgc accgcggcgt catcgaaacc 18120 gtgtacctgc gcacgccctt ctcggccggc aacgccacaa cataaagaag caagcaacat 18180 caacaacagc tgccgccatg ggctccagtg agcaggaact gaaagccatt gtcaaagatc 18240 ttggttgtgg gccatatttt ttgggcacct atgacaagcg ctttccaggc tttgtttctc 18300 cacacaagct cgcctgcgcc atagtcaata cggccggtcg cgagactggg ggcgtacact 18360 ggatggcctt tgcctggaac ccgcactcaa aaacatgcta cctctttgag ccctttggct 18420 tttctgacca gcgactcaag caggtttacc agtttgagta cgagtcactc ctgcgccgta 18480 gcgccattgc ttcttccccc gaccgctgta taacgctgga aaagtccacc caaagcgtac 18540 aggggcccaa ctcggccgcc tgtggactat tctgctgcat gtttctccac gcctttgcca 18600 actggcccca aactcccatg gatcacaacc ccaccatgaa ccttattacc ggggtaccca 18660 actccatgct caacagtccc caggtacagc ccaccctgcg tcgcaaccag gaacagctct 18720 acagcttcct ggagcgccac tcgccctact tccgcagcca cagtgcgcag attaggagcg 18780 ccacttcttt ttgtcacttg aaaaacatgt aaaaataatg tactagagac actttcaata 18840 aaggcaaatg cttttatttg tacactctcg ggtgattatt tacccccacc cttgccgtct 18900 gcgccgttta aaaatcaaag gggttctgcc gcgcatcgct atgcgccact ggcagggaca 18960 cgttgcgata ctggtgttta gtgctccact taaactcagg cacaaccatc cgcggcagct 19020 cggtgaagtt ttcactccac aggctgcgca ccatcaccaa cgcgtttagc aggtcgggcg 19080 ccgatatctt gaagtcgcag ttggggcctc cgccctgcgc gcgcgagttg cgatacacag 19140 ggttgcagca ctggaacact atcagcgccg ggtggtgcac gctggccagc acgctcttgt 19200 cggagatcag atccgcgtcc aggtcctccg cgttgctcag ggcgaacgga gtcaactttg 19260 gtagctgcct tcccaaaaag ggcgcgtgcc caggctttga gttgcactcg caccgtagtg 19320 gcatcaaaag gtgaccgtgc ccggtctggg cgttaggata cagcgcctgc ataaaagcct 19380 tgatctgctt aaaagccacc tgagcctttg cgccttcaga gaagaacatg ccgcaagact 19440 tgccggaaaa ctgattggcc ggacaggccg cgtcgtgcac gcagcacctt gcgtcggtgt 19500 tggagatctg caccacattt cggccccacc ggttcttcac gatcttggcc ttgctagact 19560 gctccttcag cgcgcgctgc ccgttttcgc tcgtcacatc catttcaatc acgtgctcct 19620 tatttatcat aatgcttccg tgtagacact taagctcgcc ttcgatctca gcgcagcggt 19680 gcagccacaa cgcgcagccc gtgggctcgt gatgcttgta ggtcacctct gcaaacgact 19740 gcaggtacgc ctgcaggaat cgccccatca tcgtcacaaa ggtcttgttg ctggtgaagg 19800 tcagctgcaa cccgcggtgc tcctcgttca gccaggtctt gcatacggcc gccagagctt 19860 ccacttggtc aggcagtagt ttgaagttcg cctttagatc gttatccacg tggtacttgt 19920 ccatcagcgc gcgcgcagcc tccatgccct tctcccacgc agacacgatc ggcacactca 19980 gcgggttcat caccgtaatt tcactttccg cttcgctggg ctcttcctct tcctcttgcg 20040 tccgcatacc acgcgccact gggtcgtctt cattcagccg ccgcactgtg cgcttacctc 20100 ctttgccatg cttgattagc accggtgggt tgctgaaacc caccatttgt agcgccacat 20160 cttctctttc ttcctcgctg tccacgatta cctctggtga tggcgggcgc tcgggcttgg 20220 gagaagggcg cttctttttc ttcttgggcg caatggccaa atccgccgcc gaggtcgatg 20280 gccgcgggct gggtgtgcgc ggcaccagcg cgtcttgtga tgagtcttcc tcgtcctcgg 20340 actcgatacg ccgcctcatc cgcttttttg ggggcgcccg gggaggcggc ggcgacgggg 20400 acggggacga cacgtcctcc atggttgggg gacgtcgcgc cgcaccgcgt ccgcgctcgg 20460 gggtggtttc gcgctgctcc tcttcccgac tggccatttc cttctcctat aggcagaaaa 20520 agatcatgga gtcagtcgag aagaaggaca gcctaaccgc cccctctgag ttcgccacca 20580 ccgcctccac cgatgccgcc aacgcgccta ccaccttccc cgtcgaggca cccccgcttg 20640 aggaggagga agtgattatc gagcaggacc caggttttgt aagcgaagac gacgaggacc 20700 gctcagtacc aacagaggat aaaaagcaag accaggacaa cgcagaggca aacgaggaac 20760 aagtcgggcg gggggacgaa aggcatggcg actacctaga tgtgggagac gacgtgctgt 20820 tgaagcatct gcagcgccag tgcgccatta tctgcgacgc gttgcaagag cgcagcgatg 20880 tgcccctcgc catagcggat gtcagccttg cctacgaacg ccacctattc tcaccgcgcg 20940 taccccccaa acgccaagaa aacggcacat gcgagcccaa cccgcgcctc aacttctacc 21000 ccgtatttgc cgtgccagag gtgcttgcca cctatcacat ctttttccaa aactgcaaga 21060 tacccctatc ctgccgtgcc aaccgcagcc gagcggacaa gcagctggcc ttgcggcagg 21120 gcgctgtcat acctgatatc gcctcgctca acgaagtgcc aaaaatcttt gagggtcttg 21180 gacgcgacga gaagcgcgcg gcaaacgctc tgcaacagga aaacagcgaa aatgaaagtc 21240 actctggagt gttggtggaa ctcgagggtg acaacgcgcg cctagccgta ctaaaacgca 21300 gcatcgaggt cacccacttt gcctacccgg cacttaacct accccccaag gtcatgagca 21360 cagtcatgag tgagctgatc gtgcgccgtg cgcagcccct ggagagggat gcaaatttgc 21420 aagaacaaac agaggagggc ctacccgcag ttggcgacga gcagctagcg cgctggcttc 21480 aaacgcgcga gcctgccgac ttggaggagc gacgcaaact aatgatggcc gcagtgctcg 21540 ttaccgtgga gcttgagtgc atgcagcggt tctttgctga cccggagatg cagcgcaagc 21600 tagaggaaac attgcactac acctttcgac agggctacgt acgccaggcc tgcaagatct 21660 ccaacgtgga gctctgcaac ctggtctcct accttggaat tttgcacgaa aaccgccttg 21720 ggcaaaacgt gcttcattcc acgctcaagg gcgaggcgcg ccgcgactac gtccgcgact 21780 gcgtttactt atttctatgc tacacctggc agacggccat gggcgtttgg cagcagtgct 21840 tggaggagtg caacctcaag gagctgcaga aactgctaaa gcaaaacttg aaggacctat 21900 ggacggcctt caacgagcgc tccgtggccg cgcacctggc ggacatcatt ttccccgaac 21960 gcctgcttaa aaccctgcaa cagggtctgc cagacttcac cagtcaaagc atgttgcaga 22020 actttaggaa ctttatccta gagcgctcag gaatcttgcc cgccacctgc tgtgcacttc 22080 ctagcgactt tgtgcccatt aagtaccgcg aatgccctcc gccgctttgg ggccactgct 22140 accttctgca gctagccaac taccttgcct accactctga cataatggaa gacgtgagcg 22200 gtgacggtct actggagtgt cactgtcgct gcaacctatg caccccgcac cgctccctgg 22260 tttgcaattc gcagctgctt aacgaaagtc aaattatcgg tacctttgag ctgcagggtc 22320 cctcgcctga cgaaaagtcc gcggctccgg ggttgaaact cactccgggg ctgtggacgt 22380 cggcttacct tcgcaaattt gtacctgagg actaccacgc ccacgagatt aggttctacg 22440 aagaccaatc ccgcccgcca aatgcggagc ttaccgcctg cgtcattacc cagggccaca 22500 ttcttggcca attgcaagcc atcaacaaag cccgccaaga gtttctgcta cgaaagggac 22560 ggggggttta cttggacccc cagtccggcg aggagctcaa cccaatcccc ccgccgccgc 22620 agccctatca gcagcagccg cgggcccttg cttcccagga tggcacccaa aaagaagctg 22680 cagctgccgc cgccacccac ggacgaggag gaatactggg acagtcaggc agaggaggtt 22740 ttggacgagg aggaggagga catgatggaa gactgggaga gcctagacga ggaagcttcc 22800 gaggtcgaag aggtgtcaga cgaaacaccg tcaccctcgg tcgcattccc ctcgccggcg 22860 ccccagaaat cggcaaccgg ttccagcatg gctacaacct ccgctcctca ggcgccgccg 22920 gcactgcccg ttcgccgacc caaccgtaga tgggacacca ctggaaccag ggccggtaag 22980 tccaagcagc cgccgccgtt agcccaagag caacaacagc gccaaggcta ccgctcatgg 23040 cgcgggcaca agaacgccat agttgcttgc ttgcaagact gtgggggcaa catctccttc 23100 gcccgccgct ttcttctcta ccatcacggc gtggccttcc cccgtaacat cctgcattac 23160 taccgtcatc tctacagccc atactgcacc ggcggcagcg gcagcggcag caacagcagc 23220 ggccacacag aagcaaaggc gaccggatag caagactctg acaaagccca agaaatccac 23280 agcggcggca gcagcaggag gaggagcgct gcgtctggcg cccaacgaac ccgtatcgac 23340 ccgcgagctt agaaacagga tttttcccac tctgtatgct atatttcaac agagcagggg 23400 ccaagaacaa gagctgaaaa taaaaaacag gtctctgcga tccctcaccc gcagctgcct 23460 gtatcacaaa agcgaagatc agcttcggcg cacgctggaa gacgcggagg ctctcttcag 23520 taaatactgc gcgctgactc ttaaggacta gtttcgcgcc ctttctcaaa tttaagcgcg 23580 aaaactacgt catctccagc ggccacaccc ggcgccagca cctgtcgtca gcgccattat 23640 gagcaaggaa attcccacgc cctacatgtg gagttaccag ccacaaatgg gacttgcggc 23700 tggagctgcc caagactact caacccgaat aaactacatg agcgcgggac cccacatgat 23760 atcccgggtc aacggaatcc gcgcccaccg aaaccgaatt ctcttggaac aggcggctat 23820 taccaccaca cctcgtaata accttaatcc ccgtagttgg cccgctgccc tggtgtacca 23880 ggaaagtccc gctcccacca ctgtggtact tcccagagac gcccaggccg aagttcagat 23940 gactaactca ggggcgcagc ttgcgggcgg ctttcgtcac agggtgcggt cgcccgggca 24000 gggtataact cacctgacaa tcagagggcg aggtattcag ctcaacgacg agtcggtgag 24060 ctcctcgctt ggtctccgtc cggacgggac atttcagatc ggcggcgccg gccgtccttc 24120 attcacgcct cgtcaggcaa tcctaactct gcagacctcg tcctctgagc cgcgctctgg 24180 aggcattgga actctgcaat ttattgagga gtttgtgcca tcggtctact ttaacccctt 24240 ctcgggacct cccggccact atccggatca atttattcct aactttgacg cggtaaagga 24300 ctcggcggac ggctacgact gaatgttaag tggagaggca gagcaactgc gcctgaaaca 24360 cctggtccac tgtcgccgcc acaagtgctt tgcccgcgac tccggtgagt tttgctactt 24420 tgaattgccc gaggatcata tcgagggccc ggcgcacggc gtccggctta ccgcccaggg 24480 agagcttgcc cgtagcctga ttcgggagtt tacccagcgc cccctgctag ttgagcggga 24540 caggggaccc tgtgttctca ctgtgatttg caactgtcct aaccttggat tacatcaaga 24600 tcctctagtt ataactagag tacccgggga tcttattccc tttaactaat aaaaaaaaat 24660 aataaagcat cacttactta aaatcagtta gcaaatttct gtccagttta ttcagcagca 24720 cctccttgcc ctcctcccag ctctggtatt gcagcttcct cctggctgca aactttctcc 24780 acaatctaaa tggaatgtca gtttcctcct gttcctgtcc atccgcaccc actatcttca 24840 tgttgttgca gatgaagcgc gcaagaccgt ctgaagatac cttcaacccc gtgtatccat 24900 atgacacgga aaccggtcct ccaactgtgc cttttcttac tcctcccttt gtatccccca 24960 atgggtttca agagagtccc cctggggtac tctctttgcg cctatccgaa cctctagtta 25020 cctccaatgg catgcttgcg ctcaaaatgg gcaacggcct ctctctggac gaggccggca 25080 accttacctc ccaaaatgta accactgtga gcccacctct caaaaaaacc aagtcaaaca 25140 taaacctgga aatatctgca cccctcacag ttacctcaga agccctaact gtggctgccg 25200 ccgcacctct aatggtcgcg ggcaacacac tcaccatgca atcacaggcc ccgctaaccg 25260 tgcacgactc caaacttagc attgccaccc aaggacccct cacagtgtca gaaggaaagc 25320 tagccctgca aacatcaggc cccctcacca ccaccgatag cagtaccctt actatcactg 25380 cctcaccccc tctaactact gccactggta gcttgggcat tgacttgaaa gagcccattt 25440 atacacaaaa tggaaaacta ggactaaagt acggggctcc tttgcatgta acagacgacc 25500 taaacacttt gaccgtagca actggtccag gtgtgactat taataatact tccttgcaaa 25560 ctaaagttac tggagccttg ggttttgatt cacaaggcaa tatgcaactt aatgtagcag 25620 gaggactaag gattgattct caaaacagac gccttatact tgatgttagt tatccgtttg 25680 atgctcaaaa ccaactaaat ctaagactag gacagggccc tctttttata aactcagccc 25740 acaacttgga tattaactac aacaaaggcc tttacttgtt tacagcttca aacaattcca 25800 aaaagcttga ggttaaccta agcactgcca aggggttgat gtttgacgct acagccatag 25860 ccattaatgc aggagatggg cttgaatttg gttcacctaa tgcaccaaac acaaatcccc 25920 tcaaaacaaa aattggccat ggcctagaat ttgattcaaa caaggctatg gttcctaaac 25980 taggaactgg ccttagtttt gacagcacag gtgccattac agtaggaaac aaaaataatg 26040 ataagctaac tttgtggacc acaccagctc catctcctaa ctgtagacta aatgcagaga 26100 aagatgctaa actcactttg gtcttaacaa aatgtggcag tcaaatactt gctacagttt 26160 cagttttggc tgttaaaggc agtttggctc caatatctgg aacagttcaa agtgctcatc 26220 ttattataag atttgacgaa aatggagtgc tactaaacaa ttccttcctg gacccagaat 26280 attggaactt tagaaatgga gatcttactg aaggcacagc ctatacaaac gctgttggat 26340 ttatgcctaa cctatcagct tatccaaaat ctcacggtaa aactgccaaa agtaacattg 26400 tcagtcaagt ttacttaaac ggagacaaaa ctaaacctgt aacactaacc attacactaa 26460 acggtacaca ggaaacagga gacacaactc caagtgcata ctctatgtca ttttcatggg 26520 actggtctgg ccacaactac attaatgaaa tatttgccac atcctcttac actttttcat 26580 acattgccca agaataaaga atcgtttgtg ttatgtttca acgtgtttat ttttcaattg 26640 cagaaaattt caagtcattt ttcattcagt agtatagccc caccaccaca tagcttatac 26700 agatcaccgt accttaatca aactcacaga accctagtat tcaacctgcc acctccctcc 26760 caacacacag agtacacagt cctttctccc cggctggcct taaaaagcat catatcatgg 26820 gtaacagaca tattcttagg tgttatattc cacacggttt cctgtcgagc caaacgctca 26880 tcagtgatat taataaactc cccgggcagc tcacttaagt tcatgtcgct gtccagctgc 26940 tgagccacag gctgctgtcc aacttgcggt tgcttaacgg gcggcgaagg agaagtccac 27000 gcctacatgg gggtagagtc ataatcgtgc atcaggatag ggcggtggtg ctgcagcagc 27060 gcgcgaataa actgctgccg ccgccgctcc gtcctgcagg aatacaacat ggcagtggtc 27120 tcctcagcga tgattcgcac cgcccgcagc ataaggcgcc ttgtcctccg ggcacagcag 27180 cgcaccctga tctcacttaa atcagcacag taactgcagc acagcaccac aatattgttc 27240 aaaatcccac agtgcaaggc gctgtatcca aagctcatgg cggggaccac agaacccacg 27300 tggccatcat accacaagcg caggtagatt aagtggcgac ccctcataaa cacgctggac 27360 ataaacatta cctcttttgg catgttgtaa ttcaccacct cccggtacca tataaacctc 27420 tgattaaaca tggcgccatc caccaccatc ctaaaccagc tggccaaaac ctgcccgccg 27480 gctatacact gcagggaacc gggactggaa caatgacagt ggagagccca ggactcgtaa 27540 ccatggatca tcatgctcgt catgatatca atgttggcac aacacaggca cacgtgcata 27600 cacttcctca ggattacaag ctcctcccgc gttagaacca tatcccaggg aacaacccat 27660 tcctgaatca gcgtaaatcc cacactgcag ggaagacctc gcacgtaact cacgttgtgc 27720 attgtcaaag tgttacattc gggcagcagc ggatgatcct ccagtatggt agcgcgggtt 27780 tctgtctcaa aaggaggtag acgatcccta ctgtacggag tgcgccgaga caaccgagat 27840 cgtgttggtc gtagtgtcat gccaaatgga acgccggacg tagtcatatt tcctgaagca 27900 aaaccaggtg cgggcgtgac aaacagatct gcgtctccgg tctcgccgct tagatcgctc 27960 tgtgtagtag ttgtagtata tccactctct caaagcatcc aggcgccccc tggcttcggg 28020 ttctatgtaa actccttcat gcgccgctgc cctgataaca tccaccaccg cagaataagc 28080 cacacccagc caacctacac attcgttctg cgagtcacac acgggaggag cgggaagagc 28140 tggaagaacc atgttttttt ttttattcca aaagattatc caaaacctca aaatgaagat 28200 ctattaagtg aacgcgctcc cctccggtgg cgtggtcaaa ctctacagcc aaagaacaga 28260 taatggcatt tgtaagatgt tgcacaatgg cttccaaaag gcaaacggcc ctcacgtcca 28320 agtggacgta aaggctaaac ccttcagggt gaatctcctc tataaacatt ccagcacctt 28380 caaccatgcc caaataattc tcatctcgcc accttctcaa tatatctcta agcaaatccc 28440 gaatattaag tccggccatt gtaaaaatct gctccagagc gccctccacc ttcagcctca 28500 agcagcgaat catgattgca aaaattcagg ttcctcacag acctgtataa gattcaaaag 28560 cggaacatta acaaaaatac cgcgatcccg taggtccctt cgcagggcca gctgaacata 28620 atcgtgcagg tctgcacgga ccagcgcggc cacttccccg ccaggaacct tgacaaaaga 28680 acccacactg attatgacac gcatactcgg agctatgcta accagcgtag ccccgatgta 28740 agctttcaat agcttgttgc atgggcggcg atataaaatg caaggtgctg ctcaaaaaat 28800 caggcaaagc ctcgcgcaaa aaagaaagca catcgtagtc atgctcatgc agataaaggc 28860 aggtaagctc cggaaccacc acagaaaaag acaccatttt tctctcaaac atgtctgcgg 28920 gtttctgcat aaacacaaaa taaaataaca aaaaaacatt taaacattag aagcctgtct 28980 tacaacagga aaaacaaccc ttataagcat aagacggact acggccatgc cggcgtgacc 29040 gtaaaaaaac tggtcaccgt gattaaaaag caccaccgac agctcctcgg tcatgtccgg 29100 agtcataatg taagactcgg taaacacatc aggttgattc atcggtcagt gctaaaaagc 29160 gaccgaaata gcccggggga atacataccc gcaggcgtag agacaacatt acagccccca 29220 taggaggtat aacaaaatta ataggagaga aaaacacata aacacctgaa aaaccctcct 29280 gcctaggcaa aatagcaccc tcccgctcca gaacaacata cagcgcttca cagcggcagc 29340 ctaacagtca gccttaccag taaaaaagaa aacctattaa aaaaacacca ctcgacacgg 29400 caccagctca atcagtcaca gtgtaaaaaa gggccaagtg cagagcgagt atatatagga 29460 ctaaaaaatg acgtaacggt taaagtccac aaaaaacacc cagaaaaccg cacgcgaacc 29520 tacgcccaga aacgaaagcc aaaaaaccca caacttcctc aaatcgtcac ttccgttttc 29580 ccacgttacg taacttccca ttttaagaaa actacaattc ccaacacata caagttactc 29640 cgccctaaaa cctacgtcac ccgccccgtt cccacgcccc gcgccacgtc acaaactcca 29700 ccccctcatt atcatattgg cttcaatcca aaataaggta tattattgat gatgttaatt 29760 aacatgcatg gatccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc 29820 gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 29880 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 29940 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 30000 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 30060 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 30120 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 30180 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 30240 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 30300 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 30360 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 30420 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc 30480 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 30540 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 30600 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 30660 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 30720 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 30780 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 30840 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 30900 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 30960 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 31020 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 31080 ccattgctgc agccatgaga ttatcaaaaa ggatcttcac ctagatcctt ttcacgtaga 31140 aagccagtcc gcagaaacgg tgctgacccc ggatgaatgt cagctactgg gctatctgga 31200 caagggaaaa cgcaagcgca aagagaaagc aggtagcttg cagtgggctt acatggcgat 31260 agctagactg ggcggtttta tggacagcaa gcgaaccgga attgccagct ggggcgccct 31320 ctggtaaggt tgggaagccc tgcaaagtaa actggatggc tttctcgccg ccaaggatct 31380 gatggcgcag gggatcaagc tctgatcaag agacaggatg aggatcgttt cgcatgattg 31440 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg 31500 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg 31560 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg 31620 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg 31680 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc 31740 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc 31800 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc 31860 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc 31920 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg 31980 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct 32040 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt 32100 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc 32160 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt 32220 tcttctgaat tttgttaaaa tttttgttaa atcagctcat tttttaacca ataggccgaa 32280 atcggcaaca tcccttataa atcaaaagaa tagaccgcga tagggttgag tgttgttcca 32340 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc 32400 gtctatcagg gcgatggccc actacgtgaa ccatcaccca aatcaagttt tttgcggtcg 32460 aggtgccgta aagctctaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg 32520 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg 32580 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgcgcg cttaatgcgc 32640 cgctacaggg cgcgtccatt cgccattcag gatcgaatta attcttaatt aacatcatca 32700 ataatatacc ttattttgga ttgaagccaa tatgataatg agggggtgga gtttgtgacg 32760 tggcgcgggg cgtgggaacg gggcgggtga cgtagtagtg tggcggaagt gtgatgttgc 32820 aagtgtggcg gaacacatgt aagcgacgga tgtggcaaaa gtgacgtttt tggtgtgcgc 32880 cggtgtacac aggaagtgac aattttcgcg cggttttagg cggatgttgt agtaaatttg 32940 ggcgtaaccg agtaagattt ggccattttc gcgggaaaac tgaataagag gaagtgaaat 33000 ctgaataatt ttgtgttact catagcgcgt aatactgcga tctatacatt gaatcaatat 33060 tggcaattag ccatattagt cattggttat atagcataaa tcaatattgg ctattggcca 33120 ttgcatacgt tgtatctata tcataatatg tacatttata ttggctcatg tccaatatga 33180 ccgccatgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta 33240 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc 33300 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg 33360 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg 33420 gcagtacatc aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa 33480 tggcccgcct ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac 33540 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg 33600 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg 33660 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg 33720 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta 33780 gtgaaccgtc agatcctcac tctcttccgc atcgctgtct gcgagggcca gctgttgggc 33840 tcgcggttga ggacaaactc ttcgcggtct ttccagtact cttggatcgg aaacccgtcg 33900 gcctccgaac ggtactccgc caccgaggga cctgagccag tccgcatcga ccggatcgga 33960 aaacctctcg agaaaggcgt ctaaccagtc acagtcgcaa ggtaggctga gcaccgtggc 34020 gggcggcagc gggtggcggt cggggttgtt tctggcggag gtgctgctga tgatgtaatt 34080 aaagtaggcg gtcttgagcc ggcggatggt cgaggtgagg tgtggcaggc ttgagatcca 34140 gctgttgggg tgagtactcc ctctcaaaag cgggcatgac ttctgcgcta agattgtcag 34200 tttccaaaaa cgaggaggat ttgatattca cctggcccga tctggccata cacttgagtg 34260 acaatgacat ccactttgcc tttctctcca caggtgtcca ctcccaggtc caagtttgga 34320 agatccccat caattactac ttgcttcctg taggaatcct ctattgttgg atcatattcg 34380 tccacaaaat gattctgaat tagctgtatc gtcaaggcac tcttgcaagc ttcatcatgg 34440 ccgacagcaa aagttatcag gcatgcacct ggtagctagt ctttaaacca atagattgca 34500 tcggtttaaa aggcaagacc gtcaaattgc gggaaagggg tcaacagccg ttcagtacca 34560 agtctcaggg gaaactttga gatggccttg caaagggtat ggtaataagc tgacggacat 34620 ggtcctaacc acgcagccaa gtcctaagtc aacagatctt ctgttgatat ggatgcagtt 34680 cacagactaa atgtcggtcg gggaagatgt attcttctca taagatatag tcggacctct 34740 ccttaatggg agctagcgga tgaagtgatg caacactgga gccgctggga actaatttgt 34800 atgcgaaagt atattgatta gttttggagt actcgcgaca tcatcacgtc gttttacaac 34860 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca 34920 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc 34980 caacagttgc gcagcctgaa tggcgaatgg cgctttgcct ggtttccggc accagaagcg 35040 gtgccggaaa gctggctgga gtgcgatctt cctgaggccg atactgtcgt cgtcccctca 35100 aactggcaga tgcacggtta cgatgcgccc atctacacca acgtgaccta tcccattacg 35160 gtcaatccgc cgtttgttcc cacggagaat ccgacgggtt gttactcgct cacatttaat 35220 gttgatgaaa gctggctaca ggaaggccag acgcgaatta tttttgatgg cgttaactcg 35280 gcgtttcatc tgtggtgcaa cgggcgctgg gtcggttacg gccaggacag tcgtttgccg 35340 tctgaatttg acctgagcgc atttttacgc gccggagaaa accgcctcgc ggtgatggtg 35400 ctgcgttgga gtgacggcag ttatctggaa gatcaggata tgtggcggat gagcggcatt 35460 ttccgtgacg tctcgttgct gcataaaccg actacacaaa tcagcgattt ccatgttgcc 35520 actcgcttta atgatgattt cagccgcgct gtactggagg ctgaagttca gatgtgcggc 35580 gagttgcgtg actacctacg ggtaacagtt tctttatggc agggtgaaac gcaggtcgcc 35640 agcggcaccg cgcctttcgg cggtgaaatt atcgatgagc gtggtggtta tgccgatcgc 35700 gtcacactac gtctgaacgt cgaaaacccg aaactgtgga gcgccgaaat cccgaatctc 35760 tatcgtgcgg tggttgaact gcacaccgcc gacggcacgc tgattgaagc agaagcctgc 35820 gatgtcggtt tccgcgaggt gcggattgaa aatggtctgc tgctgctgaa cggcaagccg 35880 ttgctgattc gaggcgttaa ccgtcacgag catcatcctc tgcatggtca ggtcatggat 35940 gagcagacga tggtgcagga tatcctgctg atgaagcaga acaactttaa cgccgtgcgc 36000 tgttcgcatt atccgaacca tccgctgtgg tacacgctgt gcgaccgcta cggcctgtat 36060 gtggtggatg aagccaatat tgaaacccac ggcatggtgc caatgaatcg tctgaccgat 36120 gatccgcgct ggctaccggc gatgagcgaa cgcgtaacgc gaatggtgca gcgcgatcgt 36180 aatcacccga gtgtgatcat ctggtcgctg gggaatgaat caggccacgg cgctaatcac 36240 gacgcgctgt atcgctggat caaatctgtc gatccttccc gcccggtgca gtatgaaggc 36300 ggcggagccg acaccacggc caccgatatt atttgcccga tgtacgcgcg cgtggatgaa 36360 gaccagccct tcccggctgt gccgaaatgg tccatcaaaa aatggctttc gctacctgga 36420 gagacgcgcc cgctgatcct ttgcgaatac gcccacgcga tgggtaacag tcttggcggt 36480 ttcgctaaat actggcaggc gtttcgtcag tatccccgtt tacagggcgg cttcgtctgg 36540 gactgggtgg atcagtcgct gattaaatat gatgaaaacg gcaacccgtg gtcggcttac 36600 ggcggtgatt ttggcgatac gccgaacgat cgccagttct gtatgaacgg tctggtcttt 36660 gccgaccgca cgccgcatcc agcgctgacg gaagcaaaac accagcagca gtttttccag 36720 ttccgtttat ccgggcaaac catcgaagtg accagcgaat acctgttccg tcatagcgat 36780 aacgagctcc tgcactggat ggtggcgctg gatggtaagc cgctggcaag cggtgaagtg 36840 cctctggatg tcgctccaca aggtaaacag ttgattgaac tgcctgaact accgcagccg 36900 gagagcgccg ggcaactctg gctcacagta cgcgtagtgc aaccgaacgc gaccgcatgg 36960 tcagaagccg ggcacatcag cgcctggcag cagtggcgtc tggcggaaaa cctcagtgtg 37020 acgctccccg ccgcgtccca cgccatcccg catctgacca ccagcgaaat ggatttttgc 37080 atcgagctgg gtaataagcg ttggcaattt aaccgccagt caggctttct ttcacagatg 37140 tggattggcg ataaaaaaca actgctgacg ccgctgcgcg atcagttcac ccgtgcaccg 37200 ctggataacg acattggcgt aagtgaagcg acccgcattg accctaacgc ctgggtcgaa 37260 cgctggaagg cggcgggcca ttaccaggcc gaagcagcgt tgttgcagtg cacggcagat 37320 acacttgctg atgcggtgct gattacgacc gctcacgcgt ggcagcatca ggggaaaacc 37380 ttatttatca gccggaaaac ctaccggatt gatggtagtg gtcaaatggc gattaccgtt 37440 gatgttgaag tggcgagcga tacaccgcat ccggcgcgga ttggcctgaa ctgccagctg 37500 gcgcaggtag cagagcgggt aaactggctc ggattagggc cgcaagaaaa ctatcccgac 37560 cgccttactg ccgcctgttt tgaccgctgg gatctgccat tgtcagacat gtataccccg 37620 tacgtcttcc cgagcgaaaa cggtctgcgc tgcgggacgc gcgaattgaa ttatggccca 37680 caccagtggc gcggcgactt ccagttcaac atcagccgct acagtcaaca gcaactgatg 37740 gaaaccagcc atcgccatct gctgcacgcg gaagaaggca catggctgaa tatcgacggt 37800 ttccatatgg ggattggtgg cgacgactcc tggagcccgt cagtatcggc ggaattccag 37860 ctgagcgccg gtcgctacca ttaccagttg gtctggtgtc aaaaataatt cgaaggcgaa 37920 agatcccaac gaaaagagag accacatggt ccttcttgag tttgtaacag ctgctgggat 37980 tacacatggc atggatgaac tgtacaactg aggatccccc gacctcgacc tctggctaat 38040 aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa 38100 ggacatatgg gagggcaaat catttggtcg agatccctcg gagatcggat ctgggcgtgg 38160 38160

Claims

서열번호 26으로 표시되는 AS300KRib1인 Kras G12V RNA를 표적으로 하여 특이적으로 인지할 수 있는 트랜스-스플라이싱 그룹 Ｉ 리보자임.
삭제
삭제
삭제
삭제
삭제
삭제
삭제
삭제
삭제
삭제
삭제
삭제
제1항에 따른 리보자임을 유효성분으로 포함하는 항암용 약학 조성물.
삭제