CN107177625B - 一种定点突变的人工载体系统及定点突变方法 - Google Patents
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Abstract
本申请涉及一套用于水稻基因组碱基定点替换的人工系统及定点突变方法。该人工系统包括：第I调节元件，其包括能够编码如氨基酸序列I的核苷酸序列；其中所述氨基酸序列I选自SEQ ID Nos.1‑6中的一种；第II调节元件，其包括依次从5’端到3’端的第II‑1核苷酸序列和第II‑2核苷酸序列；所述第II‑1核苷酸序列包括靶核苷酸序列，所述第II‑2核苷酸序列包括来源于化脓链球菌的sgRNA核酸序列，第II‑1核苷酸序列与第II‑2核苷酸序列转录融合，其产物能引导第I调控元件编码的蛋白至目标生物基因组中待突变的靶位点处，并将靶位点处的C诱导突变为T、A和G中的一种；或G突变为A、T和C中的一种。
Description
技术领域
本发明涉及一套植物基因组碱基定点突变的人工系统及定点突变方法，特别是用于水稻基因组单碱基置换的人工系统。
背景技术
水稻(Oryza sativa L.)是世界主要粮食作物之一，世界上近一半人口，包括几乎整个东亚和东南亚的人口，都以稻米为食。在中国，水稻播种面积占全国粮食作物的1/4，而产量则占一半以上。提高产量，改善稻米品质，增加水稻植株抗病、抗逆性等研究以保证粮食的稳定供给是人类社会可持续性发展的重大课题。水稻也是单子叶植物的模式系统，其研究技术、方法、理论和成果对其它禾本科植物，如小麦、玉米、高粱等具有重要指导作用。
进入二十一世纪以来，全基因组测序，基因挖掘和基因编辑等技术手段的不断涌现让现代生物技术在农业上发挥了前所未有的作用。其中，基因功能的鉴定以及对有重大应用价值的基因的转化应用是最为关键的步骤。随着基因组大数据工程的快速发展，海量的植物基因组序列等待着解读，基因的挖掘和应用严重滞后，由此显得更加迫切。值得注意的是，利用现代生物技术所获得的转基因材料，在应用上涉及田间释放的转基因生物安全问题，近些年来引发了公众的普遍顾虑。
目前在植物研究领域，CRISPR/Cas9技术已在拟南芥、烟草、水稻、大豆、番茄、玉米、大麦、小麦、马铃薯、蘑菇等模式植物和农作物上展现出骄人的成绩，获得了大量基因突变材料和基因编辑产品。但是这些编辑事件基本上都是利用植物细胞非同源末端连接修复机制获得的，属于基因功能丧失性突变体。
获得基因功能获得性突变体，对于基础研究和应用研究的意义更为重大。然而，传统的基因功能获得性突变体是通过同源重组机制而获得的。在大部分复杂生物体内，非同源末端连接修复机制在外源DNA片段的插入过程中占主导地位，同源重组发生的概率非常低，从而定制突变体的效率只有千分之一到百分之二。最近出现的利用胞嘧啶去氨酶介导的单碱基替换技术，为人们高效获得基因功能获得性突变体提供了全新思路。然而，初步结果发现，现有的单碱基替换技术，如rBE3系统，是基于来源于小鼠的APOBEC1蛋白，其对靶位点TC和CC具有偏好性，而对靶位点GC和AC编辑效率较低。
发明内容
由于水稻基因组中GC和AC位点含量极高，因此，为了克服现有技术对GC和AC位点不敏感的缺陷，本申请提供了一套人工系统，所述人工系统包括
第I调节元件，其包括能够编码如氨基酸序列I的核苷酸序列；其中所述氨基酸序列I选自SEQ ID No.1、SEQ ID No.2、SEQ ID No.3、SEQ ID No.4、SEQ ID No.5和SEQ IDNo.6中的一种；
第II调节元件，其包括依次从5’端到3’端的第II-1核苷酸序列和第II-2核苷酸序列；所述第II-1核苷酸序列包括靶核苷酸序列；所述第II-2核苷酸序列包括来源于化脓链球菌(Streptococcus pyogenes)的sgRNA核酸序列；所述第II-1核苷酸序列和所述第II-2核苷酸序列转录融合，其产物能引导第I调控元件编码的蛋白至目标生物基因组中待突变的靶位点处，并将所述靶位点处的C突变为T、A和G中的一种，或将所述靶位点处的G突变为A、T和C中的一种；
当所述第II调节元件为多个时，包含在其中的多个第II-1核苷酸序列两两不相同。另外，当所述第II调节元件为多个时，它们可以串联的形成连接在一起。
通过利用本申请的人工系统，可以将水稻基因组内源位点的特定的C定点突变为T、A和G中的一种，或G定点突变为A、T和C中的一种，筛选得到水稻基因功能获得性突变体。但是，使用的靶核苷酸序列为靶位点处为C那条链上的核苷酸序列。通过本申请SEQ IDNo.2的验证，加之生物信息学的分析，以及结合本领域的常规技术知识，可以合理的得出SEQ ID No.1、SEQ ID No.3、SEQ ID No.4、SEQ ID No.5和SEQ ID No.6也均适用于本申请的人工系统。
在一个具体实施方式中，所述第I调节元件的核苷酸序列为能够适于在水稻中表达的核苷酸序列，所述第II调节元件的核苷酸序列为能够适于在水稻中发生转录的核苷酸序列。
在一个具体实施方式中，能够编码如SEQ ID No.1所示蛋白的核苷酸编码序列如SEQ ID No.7所示；能够编码如SEQ ID No.2所示蛋白的核苷酸编码序列如SEQ ID No.8所示；能够编码如SEQ ID No.3所示蛋白的核苷酸编码序列如SEQ ID No.9所示；能够编码如SEQ ID No.4所示蛋白的核苷酸编码序列如SEQ ID No.10所示；能够编码如SEQ ID No.5所示蛋白的核苷酸编码序列如SEQ ID No.11所示；能够编码如SEQ ID No.6所示蛋白的核苷酸编码序列如SEQ ID No.12所示。经过密码子优化分析，在本申请中随机挑选出适合在水稻中表达如SEQ ID No.2所示氨基酸的核苷酸序列其中之一如SEQ ID No.8所示。相应地，根据如SEQ ID No.8所示的核苷酸序列信息，根据生物信息学的分析，以及结合本领域的常规技术知识，可以合理的得出适合在水稻中表达如SEQ ID No.1、SEQ ID No.3、SEQ IDNo.4、SEQ ID No.5和SEQ ID No.6所示氨基酸的核苷酸序列。
在一个具体实施方式中，所述第II-2核苷酸序列如SEQ ID No.13所示。
在一个具体实施方式中，所述第II-1核苷酸序列包括IIS型限制性内切酶的酶切位点，所述靶核苷酸序列通过所述IIS型限制性内切酶的酶切位点的切割被克隆，以使所述第II-1核苷酸序列与第II-2序列转录融合；当所述第II调节元件为多个时，用于克隆不同靶核苷酸序列的所述IIS型限制性内切酶的酶切位点两两不相同。
其中，由于所述靶核苷酸序列是根据基因编辑位点而变化的，因此可以将其他元件构建好，包括事先克隆到相关位置的限制性内切酶的酶切位点。在使用的之前，再根据基因编辑目的将所述靶核苷酸序列通过限制性内切酶的酶切位点的切割而被克隆。当所述第II调节元件为多个时，包含在其中的多个第II-1核苷酸序列中的限制性内切酶的酶切位点两两不相同，如此，可以有效的保障不同的靶核苷酸顺利的被克隆到目标位置。多个靶核苷酸序列可用于目标生物基因组上的多个待突变的靶位点的碱基替换。
在一个具体实施方式中，优选所述克隆位点的核苷酸序列包括SEQ ID No.14和/或SEQ ID No.15。
在一个具体实施方式中，通过如下方式确定所述靶核苷酸序列：
1)确定水稻基因组上需要被改造的核苷酸序列；
2)判断步骤1)中确定的需要被改造的核苷酸序列或其反向互补序列中是否携带有待突变的核苷酸C，并判断所述待突变的核苷酸C突变为T、A和G中的一种、或所述待突变的核苷酸G突变为A、T和C中的一种后引起的改变是否符合预期；
3)在需要被改造的核苷酸序列或其反向互补序列中筛选靶标序列：向所述待突变的核苷酸C的3′端方向搜索以确认存在能够被氨基酸序列I识别的识别模序。且所述待突变的核苷酸C处在所述识别模序5′端上游的-19至-13的位置，由此确定的所述识别模序5′端上游17至21个核苷酸序列(不含识别模序)为所述靶核苷酸序列。
在一个具体实施方式中，当第I调节元件中含有能够编码SEQ ID No.1、SEQ IDNo.2和SEQ ID No.3所示中的一种的氨基酸序列时，所识别模序为5′-NGG-3′、5′-NGA-3′、5′-GAGN-3′、5′-AAGN-3′中的一种，所述靶核苷酸序列上游的17至21个核苷酸序列，淘汰含有连续五个T的核苷酸序列；其中，所述N为A、G、C和T中的一种。
在一个具体实施方式中，当第I调节元件中含有能够编码SEQ ID No.4、SEQ IDNo.5和SEQ ID No.6所示中的一种氨基酸序列时，所述模序为5′-NGA-3′、5′-TGCG-3′、5′-TGTG-3′、5′-GAAG-3′、5′-CGCG-3′中的一种，所述靶核苷酸序列为所述识别模序5′端上游的17至21个核苷酸序列，淘汰含有连续五个T的核苷酸序列；其中，所述N为A、G、C和T中的一种。
在一个具体实施方式中，所述靶核苷酸序列如SEQ ID No.16所示。
在一个具体实施方式中，所述人工系统还包括在所述第I调节元件的5’端的能够用于水稻中的，且能够启动所述第I调节元件转录的第一启动子；和/或所述人工系统还包括在所述第II调节元件的5’端的能够用于水稻中的，且能够启动所述第II调节元件转录的第二启动子。
在一个具体实施方式中，所述第一启动子为RNA聚合酶II型启动子；和/或第二启动子为RNA聚合酶III型启动子。
在一个具体实施方式中，第一启动子为SEQ ID No.17；和/或第二启动子为SEQ IDNo.18和/或SEQ ID No.19。
在一个具体实施方式中，所述人工系统还包括在所述第I调节元件的3’端的能够终止所述第I调节元件转录的第一终止子；和/或所述人工系统还包括在所述第II调节元件的3’端的能够终止所述第II调节元件转录的第二终止子。
在一个具体实施方式中，第一终止子为SEQ ID No.20；和/或第二终止子为SEQ IDNo.21。
在一个具体实施方式中，所述第I调节元件和所述第II元件能够被克隆到至少一个载体上。例如，所述第I调节元件表达框架和所述第II调节元件表达框架能够被克隆或整合到同一个载体上，或第I调节元件表达框架和第II调节元件表达框架混合或分别位于不同的载体上时，可以采用基因枪法、农杆菌侵染法或PEG介导转化法将两个表达框架或含有两个表达框架的载体导入到水稻的愈伤组织或水稻原生质体中。
在一个具体实施方式中，所述第I调节元件能够被克隆到pUbi-ccdB上；所述第II调节元件被克隆到入门载体pENTR4上。pUbi-ccdB为基于Gateway反应并用于水稻遗传转化的双元载体。
在一个具体实施方式中，所述第一启动子、第I调节元件和第一终止子能够被克隆到pUbi-ccdB载体上。
在一个具体实施方式中，第二启动子、第II调节元件和第二终止子被克隆到pENTR4载体上。
本申请之二提供了一种如本申请之一中任意一人工系统在用于将水稻基因组中的C定点突变为T、A和G中的一种，或将水稻基因组中的G定点突变为A、T和C中的一种的应用。
本申请之三提供了一种将水稻基因组上的C定点突变为T、A或G的方法，其包括如下步骤：
1)将本申请之一中任意一人工系统通过农杆菌介导的方法导入到水稻愈伤组织，然后培养以获得水稻植株；
2)筛选获得含有定点突变的水稻植株。
进一步地，所述水稻植株能够产生含有定点替换碱基的种子，水稻获得了新的经济性状。
在进行所述的人工系统导入时，可以采用农杆菌侵染法，也可以采用基因枪法或PEG介导转化的方法中的一种将所述的人工系统导入到水稻愈伤组织或水稻原生质体中，这是本领域技术人员容易理解的。
本领域的技术人员公知，水稻基因组DNA由两条链组成，因此，所述靶核苷酸序列可以在其中的任意一条链上，并且靶位点处为C。例如，当所述靶核苷酸序列位于某一功能基因的一条链中时，该功能基因的特定位点上的C被定点突变为T、A和G中的一种后，如果其中的一种突变能够获得预期的其对应的功能蛋白中的氨基酸，则可以采用此系统来实现，即可以通过直接的碱基替换三联体密码子中的C为T、A和G中的一种，或通过间接的碱基替换三联体密码子中的G为A、T和C中的一种来改变所述三联体密码子编码氨基酸为所需氨基酸的核苷酸序列，得到水稻基因功能获得性突变体；或当所述靶核苷酸序列位于某一功能基因的另一条链中时，该功能基因的特定位点上的G被定点突变为A、T和C中的一种后，如果其中的一种突变能够获得预期的其对应的功能蛋白中的氨基酸，也可以采用此系统来实现，即可以通过将该链中的G替换为A、T和C中的一种来改变该链中的所述三联体密码子编码氨基酸，或通过间接的碱基替换三联体密码子中的G为A、T和C中的一种，得到水稻基因功能获得性突变体。
本申请的有益效果在于：
a)第II调节元件可以为多个，这样可以同时编辑水稻细胞内多个基因靶位点。
b)本申请的人工系统的碱基编辑效率高，特别是GC和AC处的靶位点碱基编辑效率高，可达26.9％，而现有的rBE3系统不能实现此位点相应的编辑。因此，本申请的人工系统在水稻基因功能研究和分子育种中具有广泛的应用价值。
附图说明
图1显示了本申请实施例带有pi-d2基因的靶向序列相对应pi-d2基因的位置，以及利用rBE5系统获得的Pi-d2基因功能获得性突变体核苷酸突变信息。其中，识别模序序列由下划实线表示；靶核苷酸序列为加粗显示部分，氨基酸替换位点由下划虚线表示。
具体实施方式
pUbi-ccdB载体的来源：pUbi-ccdB载体由本实验室通过改造pCAMBIA1300而来，其中插入了attR1-ccdB-attR2模块，用于gateway反应接受来自于入门载体的attL1-靶向序列转录模块-attL2模块。
pENTR4载体的来源：购自美国Invitrogen公司。
实施例1
1.载体的构建
通过DNA克隆的常规操作，将玉米的组成型启动子Ubi-p(SEQ ID No.17)、SEQ IDNo.8、Nos终止子(SEQ ID No.20)按照从5’到3’的顺序克隆到pUbi-ccdB载体上，命名为pUbi:rBE5，用于水稻的转基因植株研究。
将OsU6-p启动子(SEQ ID No.18)、两个BsaI酶切位点(SEQ ID No.14)、sgRNA序列(SEQ ID No.13)、(T)8终止子(SEQ ID No.21)、粳稻U6snRNA启动子(SEQ ID No.19)、两个BtgZI酶切位点(SEQ ID No.15)、sgRNA序列(SEQ ID No.13)、(T)8终止子(SEQ ID No.21)按照从5’到3’的顺序克隆到pENTR4载体的多克隆位点上，命名为pENTR4:sgRNA。其中的两个BtgZI或两个BsaI酶切位点可以用于分别克隆以下实施例2中特定基因的靶向序列。
2.针对Pi-d2基因的识别序列设计和克隆。
每个基因的转录本序列和基因组序列从MSU水稻基因组数据库中获得(http://rice.plantbiology.msu.edu/)。1)判断编辑位点的正链或负链是否携带有核酸碱基C，相应碱基C突变为T、A或G后引起的氨基酸改变是否符合预期；2)在3′端方向搜索NGG、NGA、GAGN、AAGN等模序，所述N是A或G或C或T，使突变碱基C处在所述模序上游的的-19至-13位置；3)合成19-20bp的靶向序列，进行载体构建。
针对Pi-d2基因的靶核苷酸序列为5′-GAGCATAATGACAATAATAA-3′(SEQ IDNo.16)。分别合成gPi-d2-F1(5′-GTGTGAGCATAATGACAATAATAA-3′，SEQ ID No.22)和gPi-d2-R1(5′-AAACTTATTATTGTCATTATGCTC-3′，SEQ ID No.23)引物后，使用T4多聚核苷酸激酶进行磷酸化处理，退火形成双链，克隆到pENTR4:sgRNA载体的BsaI酶切位点中，测序确认插入片段(靶核苷酸序列)完全正确，获得pENTR4:gPi-d2。将pENTR4:gPi-d2用AatII酶切进行线性化，再通过Gateway反应将第II核苷酸序列转录元件转移到pUbi:rBE5中，获得为pUbi:rBE5-gPi-d2最终载体，其对于Pi-d2基因上的改造位点序列为5′-TTATTATTGTCATTATGCTC-3′(SEQ ID No.24)，该序列与SEQ ID No.16互补；基因编辑后序列为5′-TTATTATTGTCATTATACTC-3′(SEQ ID No.25)。
3.rBE系统转化粳稻品种Kitaake。
1)水稻愈伤诱导：
去壳的未成熟的水稻种子用50％的市售消毒液处理25分钟；无菌水清洗3-5次，然后将种子转移至无菌的培养皿中，吸出多余的水份；将种子放置于MSD平板上(4.43g/L MS粉；30g/L蔗糖；2ml/L 2,4-D；8g/L植物凝胶；pH5.7)，于光照培养室培养10天，诱导愈伤组织形成；去除种子的胚和芽，将愈伤组织转移至一个新的MSD培养皿上，培养5天，直至能够用于农杆菌的转化。
2)农杆菌转化：
将pUbi:rBE5-gPi-d2通过电击法转入到农杆菌菌株EHA105中，在LB培养基中过夜培养12小时；收集农杆菌，用MSD溶液重悬，使其OD600＝0.1，待用。
3)水稻愈伤的农杆菌侵染：
将愈伤组织置于上述农杆菌悬浮液中30分钟；除去农杆菌悬浮液，将愈伤组织转移至无菌的吸水纸上；将愈伤组织转移至新的含有100μM乙酰丁香酮的MSD培养基上，室温避光培养3天。
4)水稻抗性愈伤筛选：
将暗培养后的愈伤组织转移至MSD培养基(200mg/L特美汀；50mg/L潮霉素B)上，光照培养2周至1个月，直至愈伤组织的表面出现抗性愈伤，转移抗性愈伤到新的MSD培养基(200mg/L特美汀，50mg/L潮霉素B)上，每2周换一次培养基。
5)抗性愈伤分化与生根
将抗性愈伤组织转移至再生培养基上(4.43g/L MS粉；30g/L蔗糖；25g/L山梨醇；0.5mg/L NAA；3mg/L BA；100mg/L特美汀；50mg/L潮霉素B；12g/L琼脂粉；pH＝5.7)，直至长成植株幼苗，每7-10天转移一次；转移幼苗至1/2MS培养基(2.21g/L MS粉；15g/L蔗糖；8g/L植物凝胶；pH5.7)中生根。
4.对T0代转基因水稻中Pi-d2基因靶位点进行鉴定。
用CTAB法提取抗性愈伤和转基因植株的基因组DNA。根据Pi-d2基因的靶位点DNA序列设计特异性的PCR引物：Pi-d2-F1：5′-CGGGTTGTAAGAGTGCCTGT-3′(SEQ ID No.26)，Pi-d2-R1：5′-CTCCAGCTTCTTCACAGCAA-3′(SEQ ID No.27)，利用I-5高保真酶混合液(MACLAB)进行PCR扩增目的片段491bp，PCR产物直接测序或连接pGEM-T载体后测序，基因编辑效果如图1所示，pUbi:rBE5系统对Pi-d2的靶位点编辑效率高达26.9％。
实施例2
通过DNA克隆的常规操作，将玉米的组成型启动子Ubi-p(SEQ ID No.17)、SEQ IDNo.28、Nos终止子(SEQ ID No.20)按照从5’到3’的顺序克隆到pUbi-ccdB载体上，命名为pUbi:rBE3。将pENTR4:gPi-d2用AatII酶切进行线性化，再通过Gateway反应将靶向序列转录元件转移到pUbi:rBE3中，获得为pUbi:rBE3-gPi-d2最终载体，用于水稻的转基因植株研究。其对于Pi-d2在基因组靶位点序列与实施例1相同，为5′-TTATTATTGTCATTATGCTC-3′(SEQ ID No.24)。SEQ ID No.28所示的核苷酸序列编码如SEQ ID No.29所示的氨基酸序列。
其他操作同实施例1。
转基因植株群体中未筛选获得到Pi-d2的编辑植株。
本申请通过转化水稻愈伤，获得含有rBE3(pUbi:rBE3-gPi-d2)和rRE5(pUbi:rBE5-gPi-d2)系统的转基因水稻，通过靶位点测序发现，对于GC靶位点，rBE5系统成功地在转基因水稻植株中对Pi-d2位点进行了单碱基定点替换。我们从26株独立的转基因水稻株系中筛选获得到7株目标突变体，基因编辑效率为26.9％，而已报道的单碱基替换载体rBE3系统对其的编辑效率为零。综上所述，我们构建的这套新载体进行针对GC和AC靶位点的定点碱基突变方法，对于用基因编辑技术获得指定基因功能获得性水稻突变体具有重要价值。
LHA1760261 核苷酸序和氨基酸列表
<110> 中国农业科学院植物保护研究所
<120>一种定点突变的人工载体系统及定点突变方法
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<223> SEQ ID No. 2
<400> 2
MDSLLMNRREFLYQFKNVRWAKGRRETYLCYVVKRRDSATSFSLDFGYLRNKNGCHVELLFLRYISDWDLDPGRCYRVTWFISWSPCYDCARHVADFLRGNPNLSLRIFTARLYFCEDRKAEPEGLRRLHRAGVQIAIMTFKDYFYCWNTFVENHGRTFKAWEGLHENSVRLSRQLRRILLPLYEVDDLRDAFRTSGSETPGTSESATPESDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDANAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDRPKKKRKVGG；
<210>3
<211> 1676
<212> PRT
<213> 人工序列
<223> SEQ ID No. 3
<400> 3
MDSLLMNRREFLYQFKNVRWAKGRRETYLCYVVKRRDSATSFSLDFGYLRNKNGCHVELLFLRYISDWDLDPGRCYRVTWFISWSPCYDCARHVADFLRGNPNLSLRIFTARLYFCEDRKAEPEGLRRLHRAGVQIAIMTFKDYFYCWNTFVENHGRTFKAWEGLHENSVRLSRQLRRILLPLYEVDDLRDAFRTSGSETPGTSESATPESDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDANAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTAYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKMLSGGSPKKKRKV；
<210> 4
<211> 1572
<212> PRT
<213> 人工序列
<223> SEQ ID No. 4
<400> 4
MDSLLMNRREFLYQFKNVRWAKGRRETYLCYVVKRRDSATSFSLDFGYLRNKNGCHVELLFLRYISDWDLDPGRCYRVTWFISWSPCYDCARHVADFLRGNPNLSLRIFTARLYFCEDRKAEPEGLRRLHRAGVQIAIMTFKDYFYCWNTFVENHGRTFKAWEGLHENSVRLSRQLRRILLPLYEVDDLRDAFRTDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDANAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFVSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDRPKKKRKVGG；
<210> 5
<211> 1588
<212> PRT
<213>人工序列
<223> SEQ ID No. 5
<400> 5
MDSLLMNRREFLYQFKNVRWAKGRRETYLCYVVKRRDSATSFSLDFGYLRNKNGCHVELLFLRYISDWDLDPGRCYRVTWFISWSPCYDCARHVADFLRGNPNLSLRIFTARLYFCEDRKAEPEGLRRLHRAGVQIAIMTFKDYFYCWNTFVENHGRTFKAWEGLHENSVRLSRQLRRILLPLYEVDDLRDAFRTSGSETPGTSESATPESDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDANAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFVSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDRPKKKRKVGG；
<210> 6
<211> 1676
<212> PRT
<213>人工序列
<223> SEQ ID No. 6
<400> 6
MDSLLMNRREFLYQFKNVRWAKGRRETYLCYVVKRRDSATSFSLDFGYLRNKNGCHVELLFLRYISDWDLDPGRCYRVTWFISWSPCYDCARHVADFLRGNPNLSLRIFTARLYFCEDRKAEPEGLRRLHRAGVQIAIMTFKDYFYCWNTFVENHGRTFKAWEGLHENSVRLSRQLRRILLPLYEVDDLRDAFRTSGSETPGTSESATPESDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDANAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFVSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTAYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKMLSGGSPKKKRKV；
<210> 7
<211> 4719
<212> DNA
<213>人工序列
<223> SEQ ID No. 7
<400> 7
ATGGATAGCCTTCTCATGAACAGAAGAGAGTTTCTCTATCAGTTTAAAAATGTTCGGTGGGCGAAGGGGAGGAGAGAGACATATCTCTGCTATGTTGTTAAGCGGAGAGATTCTGCGACCTCATTCTCACTCGATTTTGGTTATTTGAGGAACAAGAATGGATGTCATGTCGAATTGTTGTTTCTCCGGTATATTTCCGACTGGGATTTGGACCCAGGGCGGTGTTACCGGGTCACATGGTTTATTTCCTGGAGTCCATGTTACGACTGTGCGCGCCATGTCGCCGACTTCCTCAGGGGTAATCCTAACTTGTCCTTGCGGATTTTTACAGCCAGACTCTATTTCTGTGAGGATCGGAAGGCGGAACCCGAGGGGCTGAGAAGACTGCACCGCGCTGGCGTCCAAATCGCCATCATGACTTTTAAGGATTATTTCTACTGTTGGAACACGTTCGTCGAGAACCACGGTCGGACCTTCAAAGCCTGGGAAGGGCTGCATGAAAATTCCGTGAGGTTGTCCCGGCAACTCCGCAGAATACTCCTGCCCCTTTATGAGGTCGACGATCTCAGAGACGCCTTTAGAACTGATAAAAAGTATTCAATCGGACTTGCTATTGGGACAAACTCTGTGGGCTGGGCGGTAATTACCGACGAGTACAAGGTGCCTAGTAAGAAATTTAAAGTGCTCGGAAACACTGACAGGCACTCTATAAAGAAGAACCTGATCGGGGCACTGCTTTTCGACTCCGGAGAGACGGCGGAGGCGACGCGTCTCAAGCGTACCGCGCGCCGCAGGTACACAAGAAGGAAGAATAGGATCTGCTACTTGCAGGAAATCTTCAGTAACGAGATGGCGAAGGTCGACGATAGTTTCTTTCATCGGTTGGAAGAATCGTTCCTCGTAGAGGAGGACAAAAAGCACGAGCGTCACCCAATATTCGGGAATATTGTTGACGAGGTTGCCTACCATGAGAAATATCCTACAATATATCACCTCCGTAAGAAGCTTGTCGATTCAACTGATAAGGCTGATCTCAGACTCATCTATCTTGCCCTCGCACATATGATTAAGTTTCGTGGCCACTTCTTGATTGAAGGCGACCTCAACCCGGACAACTCAGATGTTGACAAGCTTTTTATACAGCTCGTCCAGACATATAACCAGCTGTTTGAAGAGAATCCCATCAATGCGAGTGGGGTTGATGCTAACGCCATTTTGTCCGCCAGGTTGTCCAAATCTCGCAGACTGGAAAACCTGATCGCACAGCTTCCCGGTGAAAAGAAAAACGGGCTCTTCGGCAATCTCATCGCACTGTCCCTCGGCCTCACCCCAAACTTCAAGTCTAACTTCGACCTGGCCGAGGATGCGAAGCTCCAGCTGTCAAAAGATACATACGACGACGATTTGGACAATCTGCTTGCGCAAATAGGCGACCAGTATGCGGACCTGTTCCTGGCTGCCAAAAATCTGTCAGATGCAATCCTCCTGTCCGATATATTGCGTGTGAACACCGAAATCACGAAGGCACCGCTTAGCGCATCCATGATCAAGAGATACGACGAGCACCATCAGGACCTCACACTCCTCAAGGCGCTTGTTCGTCAGCAGCTTCCCGAGAAATATAAGGAAATTTTTTTCGATCAAAGCAAGAATGGATATGCTGGCTATATTGACGGTGGCGCTTCGCAGGAGGAGTTCTATAAATTCATTAAGCCGATTCTGGAGAAGATGGACGGAACGGAGGAGCTCCTCGTCAAGCTTAACCGGGAAGACCTGTTGCGGAAGCAGAGGACTTTTGATAACGGCTCTATTCCGCACCAAATCCATCTGGGTGAGTTGCACGCAATCTTGAGAAGACAAGAGGATTTCTACCCGTTCCTTAAGGATAACAGAGAGAAGATAGAAAAAATACTGACCTTCAGGATACCATACTATGTGGGCCCACTGGCGCGCGGAAATAGTCGTTTCGCATGGATGACTAGAAAGTCCGAAGAAACGATCACGCCATGGAATTTTGAGGAAGTGGTCGACAAGGGCGCCTCTGCCCAGAGCTTCATCGAAAGGATGACCAATTTTGACAAAAATCTGCCTAACGAAAAGGTGCTTCCGAAGCACAGCCTGTTGTATGAATACTTCACAGTTTATAACGAGCTCACTAAGGTCAAGTACGTCACGGAGGGCATGCGTAAGCCTGCTTTCCTGTCTGGTGAACAAAAAAAGGCGATTGTGGACCTCCTTTTCAAGACGAACCGTAAAGTTACTGTGAAGCAACTGAAAGAGGATTACTTTAAGAAAATTGAGTGCTTCGACAGTGTGGAGATTTCCGGTGTCGAGGACCGGTTTAACGCCAGCCTGGGTACGTATCATGACCTGCTTAAAATTATCAAGGATAAAGATTTCCTGGATAATGAAGAGAACGAAGATATACTGGAGGACATTGTGTTGACTTTGACCCTCTTCGAGGACAGAGAGATGATTGAGGAAAGACTGAAGACCTACGCACACCTTTTTGATGACAAGGTCATGAAACAACTCAAGCGCCGGCGCTATACTGGCTGGGGCCGGCTTTCTCGCAAGCTCATCAATGGGATTCGGGATAAGCAATCAGGCAAGACAATTTTGGACTTCCTCAAATCCGACGGATTCGCAAATAGGAATTTTATGCAGCTGATACATGACGACTCTTTGACATTCAAAGAAGACATACAGAAGGCTCAGGTCTCCGGCCAAGGAGATTCTTTGCACGAGCATATCGCTAACTTGGCAGGTAGCCCCGCCATAAAAAAGGGCATTCTTCAAACGGTAAAAGTTGTTGACGAACTCGTGAAGGTTATGGGCCGTCATAAGCCGGAAAACATTGTTATTGAAATGGCTAGGGAAAATCAGACGACCCAGAAGGGACAGAAAAATAGCAGGGAGCGGATGAAGAGAATTGAAGAGGGAATTAAGGAGCTTGGATCTCAGATTCTTAAGGAGCACCCTGTGGAGAACACCCAACTTCAGAATGAAAAGCTCTACCTTTACTACCTTCAAAACGGCCGGGATATGTACGTCGATCAGGAACTTGACATTAACCGGTTGAGCGATTATGACGTTGACCATATTGTGCCCCAATCTTTCCTTAAAGACGACTCTATCGACAATAAAGTGCTGACGCGCAGCGATAAAAATCGCGGTAAGTCGGATAATGTCCCGTCGGAAGAGGTGGTTAAAAAAATGAAGAACTATTGGAGGCAACTCCTGAATGCCAAGCTGATCACTCAGAGGAAATTCGACAATCTCACCAAGGCAGAAAGGGGTGGACTTAGCGAGCTCGACAAGGCCGGTTTTATCAAAAGACAGCTGGTGGAGACACGCCAAATCACCAAACACGTTGCCCAGATCCTGGATTCGAGGATGAACACGAAGTATGACGAGAACGACAAGTTGATTAGGGAAGTCAAGGTCATCACTTTGAAGTCCAAGCTGGTGAGCGACTTTCGCAAAGACTTCCAGTTTTACAAAGTCAGGGAAATTAATAACTACCACCACGCCCACGACGCCTACCTTAACGCCGTGGTTGGCACAGCACTCATCAAGAAATACCCTAAGCTCGAATCTGAGTTCGTCTATGGCGACTATAAGGTCTACGACGTTAGAAAAATGATCGCGAAATCTGAGCAGGAAATAGGCAAGGCAACTGCCAAGTACTTCTTCTATTCCAATATCATGAACTTTTTTAAGACGGAGATTACCCTGGCGAATGGTGAGATCCGCAAGCGCCCTTTGATTGAGACAAACGGAGAAACAGGAGAGATCGTATGGGACAAAGGGCGGGACTTTGCTACTGTTAGGAAGGTGCTCTCTATGCCACAAGTTAACATTGTCAAAAAAACTGAAGTGCAGACAGGTGGGTTTAGCAAGGAATCTATCCTGCCGAAGAGGAACTCTGACAAGCTGATCGCCCGCAAGAAAGATTGGGATCCGAAAAAGTACGGAGGATTCGACTCCCCCACAGTTGCGTACTCCGTGCTTGTCGTGGCCAAAGTGGAGAAGGGCAAGTCTAAGAAGCTCAAGAGCGTCAAAGAGTTGTTGGGGATCACGATTATGGAGCGGTCGTCTTTCGAAAAGAATCCGATAGATTTTCTCGAGGCCAAGGGTTATAAAGAAGTCAAGAAGGATCTTATCATCAAGCTCCCTAAGTACTCCCTCTTTGAGCTTGAAAACGGACGGAAAAGAATGCTGGCTTCAGCGGGTGAACTTCAGAAGGGTAATGAACTCGCTCTGCCCTCAAAATATGTGAATTTCCTTTACCTGGCATCACACTATGAGAAGCTTAAGGGGTCTCCAGAGGACAACGAGCAGAAGCAACTGTTCGTTGAACAACACAAGCACTACCTTGACGAGATTATCGAGCAAATCAGCGAGTTTAGCAAGCGCGTTATACTGGCAGACGCAAATCTTGATAAGGTCCTTAGCGCCTACAACAAGCATAGAGACAAACCCATCCGGGAGCAGGCCGAGAACATTATTCATCTCTTCACCTTGACGAATCTTGGGGCCCCGGCCGCGTTCAAGTACTTCGATACTACCATAGACAGAAAGCGCTATACATCGACAAAGGAAGTTCTTGACGCCACGCTGATCCACCAAAGTATAACAGGCCTCTATGAGACACGCATCGACCTTTCGCAGTTGGGCGGTGACCGCCCCAAAAAGAAGAGGAAAGTTGGCGGGTGA；
<210> 8
<211> 4767
<212> DNA
<213>人工序列
<223> SEQ ID No. 8
<400> 8
ATGGATAGCCTTCTCATGAACAGAAGAGAGTTTCTCTATCAGTTTAAAAATGTTCGGTGGGCGAAGGGGAGGAGAGAGACATATCTCTGCTATGTTGTTAAGCGGAGAGATTCTGCGACCTCATTCTCACTCGATTTTGGTTATTTGAGGAACAAGAATGGATGTCATGTCGAATTGTTGTTTCTCCGGTATATTTCCGACTGGGATTTGGACCCAGGGCGGTGTTACCGGGTCACATGGTTTATTTCCTGGAGTCCATGTTACGACTGTGCGCGCCATGTCGCCGACTTCCTCAGGGGTAATCCTAACTTGTCCTTGCGGATTTTTACAGCCAGACTCTATTTCTGTGAGGATCGGAAGGCGGAACCCGAGGGGCTGAGAAGACTGCACCGCGCTGGCGTCCAAATCGCCATCATGACTTTTAAGGATTATTTCTACTGTTGGAACACGTTCGTCGAGAACCACGGTCGGACCTTCAAAGCCTGGGAAGGGCTGCATGAAAATTCCGTGAGGTTGTCCCGGCAACTCCGCAGAATACTCCTGCCCCTTTATGAGGTCGACGATCTCAGAGACGCCTTTAGAACTAGCGGAAGCGAGACGCCAGGGACTTCTGAATCGGCCACCCCCGAGAGCGATAAAAAGTATTCAATCGGACTTGCTATTGGGACAAACTCTGTGGGCTGGGCGGTAATTACCGACGAGTACAAGGTGCCTAGTAAGAAATTTAAAGTGCTCGGAAACACTGACAGGCACTCTATAAAGAAGAACCTGATCGGGGCACTGCTTTTCGACTCCGGAGAGACGGCGGAGGCGACGCGTCTCAAGCGTACCGCGCGCCGCAGGTACACAAGAAGGAAGAATAGGATCTGCTACTTGCAGGAAATCTTCAGTAACGAGATGGCGAAGGTCGACGATAGTTTCTTTCATCGGTTGGAAGAATCGTTCCTCGTAGAGGAGGACAAAAAGCACGAGCGTCACCCAATATTCGGGAATATTGTTGACGAGGTTGCCTACCATGAGAAATATCCTACAATATATCACCTCCGTAAGAAGCTTGTCGATTCAACTGATAAGGCTGATCTCAGACTCATCTATCTTGCCCTCGCACATATGATTAAGTTTCGTGGCCACTTCTTGATTGAAGGCGACCTCAACCCGGACAACTCAGATGTTGACAAGCTTTTTATACAGCTCGTCCAGACATATAACCAGCTGTTTGAAGAGAATCCCATCAATGCGAGTGGGGTTGATGCTAACGCCATTTTGTCCGCCAGGTTGTCCAAATCTCGCAGACTGGAAAACCTGATCGCACAGCTTCCCGGTGAAAAGAAAAACGGGCTCTTCGGCAATCTCATCGCACTGTCCCTCGGCCTCACCCCAAACTTCAAGTCTAACTTCGACCTGGCCGAGGATGCGAAGCTCCAGCTGTCAAAAGATACATACGACGACGATTTGGACAATCTGCTTGCGCAAATAGGCGACCAGTATGCGGACCTGTTCCTGGCTGCCAAAAATCTGTCAGATGCAATCCTCCTGTCCGATATATTGCGTGTGAACACCGAAATCACGAAGGCACCGCTTAGCGCATCCATGATCAAGAGATACGACGAGCACCATCAGGACCTCACACTCCTCAAGGCGCTTGTTCGTCAGCAGCTTCCCGAGAAATATAAGGAAATTTTTTTCGATCAAAGCAAGAATGGATATGCTGGCTATATTGACGGTGGCGCTTCGCAGGAGGAGTTCTATAAATTCATTAAGCCGATTCTGGAGAAGATGGACGGAACGGAGGAGCTCCTCGTCAAGCTTAACCGGGAAGACCTGTTGCGGAAGCAGAGGACTTTTGATAACGGCTCTATTCCGCACCAAATCCATCTGGGTGAGTTGCACGCAATCTTGAGAAGACAAGAGGATTTCTACCCGTTCCTTAAGGATAACAGAGAGAAGATAGAAAAAATACTGACCTTCAGGATACCATACTATGTGGGCCCACTGGCGCGCGGAAATAGTCGTTTCGCATGGATGACTAGAAAGTCCGAAGAAACGATCACGCCATGGAATTTTGAGGAAGTGGTCGACAAGGGCGCCTCTGCCCAGAGCTTCATCGAAAGGATGACCAATTTTGACAAAAATCTGCCTAACGAAAAGGTGCTTCCGAAGCACAGCCTGTTGTATGAATACTTCACAGTTTATAACGAGCTCACTAAGGTCAAGTACGTCACGGAGGGCATGCGTAAGCCTGCTTTCCTGTCTGGTGAACAAAAAAAGGCGATTGTGGACCTCCTTTTCAAGACGAACCGTAAAGTTACTGTGAAGCAACTGAAAGAGGATTACTTTAAGAAAATTGAGTGCTTCGACAGTGTGGAGATTTCCGGTGTCGAGGACCGGTTTAACGCCAGCCTGGGTACGTATCATGACCTGCTTAAAATTATCAAGGATAAAGATTTCCTGGATAATGAAGAGAACGAAGATATACTGGAGGACATTGTGTTGACTTTGACCCTCTTCGAGGACAGAGAGATGATTGAGGAAAGACTGAAGACCTACGCACACCTTTTTGATGACAAGGTCATGAAACAACTCAAGCGCCGGCGCTATACTGGCTGGGGCCGGCTTTCTCGCAAGCTCATCAATGGGATTCGGGATAAGCAATCAGGCAAGACAATTTTGGACTTCCTCAAATCCGACGGATTCGCAAATAGGAATTTTATGCAGCTGATACATGACGACTCTTTGACATTCAAAGAAGACATACAGAAGGCTCAGGTCTCCGGCCAAGGAGATTCTTTGCACGAGCATATCGCTAACTTGGCAGGTAGCCCCGCCATAAAAAAGGGCATTCTTCAAACGGTAAAAGTTGTTGACGAACTCGTGAAGGTTATGGGCCGTCATAAGCCGGAAAACATTGTTATTGAAATGGCTAGGGAAAATCAGACGACCCAGAAGGGACAGAAAAATAGCAGGGAGCGGATGAAGAGAATTGAAGAGGGAATTAAGGAGCTTGGATCTCAGATTCTTAAGGAGCACCCTGTGGAGAACACCCAACTTCAGAATGAAAAGCTCTACCTTTACTACCTTCAAAACGGCCGGGATATGTACGTCGATCAGGAACTTGACATTAACCGGTTGAGCGATTATGACGTTGACCATATTGTGCCCCAATCTTTCCTTAAAGACGACTCTATCGACAATAAAGTGCTGACGCGCAGCGATAAAAATCGCGGTAAGTCGGATAATGTCCCGTCGGAAGAGGTGGTTAAAAAAATGAAGAACTATTGGAGGCAACTCCTGAATGCCAAGCTGATCACTCAGAGGAAATTCGACAATCTCACCAAGGCAGAAAGGGGTGGACTTAGCGAGCTCGACAAGGCCGGTTTTATCAAAAGACAGCTGGTGGAGACACGCCAAATCACCAAACACGTTGCCCAGATCCTGGATTCGAGGATGAACACGAAGTATGACGAGAACGACAAGTTGATTAGGGAAGTCAAGGTCATCACTTTGAAGTCCAAGCTGGTGAGCGACTTTCGCAAAGACTTCCAGTTTTACAAAGTCAGGGAAATTAATAACTACCACCACGCCCACGACGCCTACCTTAACGCCGTGGTTGGCACAGCACTCATCAAGAAATACCCTAAGCTCGAATCTGAGTTCGTCTATGGCGACTATAAGGTCTACGACGTTAGAAAAATGATCGCGAAATCTGAGCAGGAAATAGGCAAGGCAACTGCCAAGTACTTCTTCTATTCCAATATCATGAACTTTTTTAAGACGGAGATTACCCTGGCGAATGGTGAGATCCGCAAGCGCCCTTTGATTGAGACAAACGGAGAAACAGGAGAGATCGTATGGGACAAAGGGCGGGACTTTGCTACTGTTAGGAAGGTGCTCTCTATGCCACAAGTTAACATTGTCAAAAAAACTGAAGTGCAGACAGGTGGGTTTAGCAAGGAATCTATCCTGCCGAAGAGGAACTCTGACAAGCTGATCGCCCGCAAGAAAGATTGGGATCCGAAAAAGTACGGAGGATTCGACTCCCCCACAGTTGCGTACTCCGTGCTTGTCGTGGCCAAAGTGGAGAAGGGCAAGTCTAAGAAGCTCAAGAGCGTCAAAGAGTTGTTGGGGATCACGATTATGGAGCGGTCGTCTTTCGAAAAGAATCCGATAGATTTTCTCGAGGCCAAGGGTTATAAAGAAGTCAAGAAGGATCTTATCATCAAGCTCCCTAAGTACTCCCTCTTTGAGCTTGAAAACGGACGGAAAAGAATGCTGGCTTCAGCGGGTGAACTTCAGAAGGGTAATGAACTCGCTCTGCCCTCAAAATATGTGAATTTCCTTTACCTGGCATCACACTATGAGAAGCTTAAGGGGTCTCCAGAGGACAACGAGCAGAAGCAACTGTTCGTTGAACAACACAAGCACTACCTTGACGAGATTATCGAGCAAATCAGCGAGTTTAGCAAGCGCGTTATACTGGCAGACGCAAATCTTGATAAGGTCCTTAGCGCCTACAACAAGCATAGAGACAAACCCATCCGGGAGCAGGCCGAGAACATTATTCATCTCTTCACCTTGACGAATCTTGGGGCCCCGGCCGCGTTCAAGTACTTCGATACTACCATAGACAGAAAGCGCTATACATCGACAAAGGAAGTTCTTGACGCCACGCTGATCCACCAAAGTATAACAGGCCTCTATGAGACACGCATCGACCTTTCGCAGTTGGGCGGTGACCGCCCCAAAAAGAAGAGGAAAGTTGGCGGGTGA；
<210> 9
<211> 5031
<212> DNA
<213>人工序列
<223> SEQ ID No. 9
<400> 9
ATGGATAGCCTTCTCATGAACAGAAGAGAGTTTCTCTATCAGTTTAAAAATGTTCGGTGGGCGAAGGGGAGGAGAGAGACATATCTCTGCTATGTTGTTAAGCGGAGAGATTCTGCGACCTCATTCTCACTCGATTTTGGTTATTTGAGGAACAAGAATGGATGTCATGTCGAATTGTTGTTTCTCCGGTATATTTCCGACTGGGATTTGGACCCAGGGCGGTGTTACCGGGTCACATGGTTTATTTCCTGGAGTCCATGTTACGACTGTGCGCGCCATGTCGCCGACTTCCTCAGGGGTAATCCTAACTTGTCCTTGCGGATTTTTACAGCCAGACTCTATTTCTGTGAGGATCGGAAGGCGGAACCCGAGGGGCTGAGAAGACTGCACCGCGCTGGCGTCCAAATCGCCATCATGACTTTTAAGGATTATTTCTACTGTTGGAACACGTTCGTCGAGAACCACGGTCGGACCTTCAAAGCCTGGGAAGGGCTGCATGAAAATTCCGTGAGGTTGTCCCGGCAACTCCGCAGAATACTCCTGCCCCTTTATGAGGTCGACGATCTCAGAGACGCCTTTAGAACTAGCGGAAGCGAGACGCCAGGGACTTCTGAATCGGCCACCCCCGAGAGCGATAAAAAGTATTCAATCGGACTTGCTATTGGGACAAACTCTGTGGGCTGGGCGGTAATTACCGACGAGTACAAGGTGCCTAGTAAGAAATTTAAAGTGCTCGGAAACACTGACAGGCACTCTATAAAGAAGAACCTGATCGGGGCACTGCTTTTCGACTCCGGAGAGACGGCGGAGGCGACGCGTCTCAAGCGTACCGCGCGCCGCAGGTACACAAGAAGGAAGAATAGGATCTGCTACTTGCAGGAAATCTTCAGTAACGAGATGGCGAAGGTCGACGATAGTTTCTTTCATCGGTTGGAAGAATCGTTCCTCGTAGAGGAGGACAAAAAGCACGAGCGTCACCCAATATTCGGGAATATTGTTGACGAGGTTGCCTACCATGAGAAATATCCTACAATATATCACCTCCGTAAGAAGCTTGTCGATTCAACTGATAAGGCTGATCTCAGACTCATCTATCTTGCCCTCGCACATATGATTAAGTTTCGTGGCCACTTCTTGATTGAAGGCGACCTCAACCCGGACAACTCAGATGTTGACAAGCTTTTTATACAGCTCGTCCAGACATATAACCAGCTGTTTGAAGAGAATCCCATCAATGCGAGTGGGGTTGATGCTAACGCCATTTTGTCCGCCAGGTTGTCCAAATCTCGCAGACTGGAAAACCTGATCGCACAGCTTCCCGGTGAAAAGAAAAACGGGCTCTTCGGCAATCTCATCGCACTGTCCCTCGGCCTCACCCCAAACTTCAAGTCTAACTTCGACCTGGCCGAGGATGCGAAGCTCCAGCTGTCAAAAGATACATACGACGACGATTTGGACAATCTGCTTGCGCAAATAGGCGACCAGTATGCGGACCTGTTCCTGGCTGCCAAAAATCTGTCAGATGCAATCCTCCTGTCCGATATATTGCGTGTGAACACCGAAATCACGAAGGCACCGCTTAGCGCATCCATGATCAAGAGATACGACGAGCACCATCAGGACCTCACACTCCTCAAGGCGCTTGTTCGTCAGCAGCTTCCCGAGAAATATAAGGAAATTTTTTTCGATCAAAGCAAGAATGGATATGCTGGCTATATTGACGGTGGCGCTTCGCAGGAGGAGTTCTATAAATTCATTAAGCCGATTCTGGAGAAGATGGACGGAACGGAGGAGCTCCTCGTCAAGCTTAACCGGGAAGACCTGTTGCGGAAGCAGAGGACTTTTGATAACGGCTCTATTCCGCACCAAATCCATCTGGGTGAGTTGCACGCAATCTTGAGAAGACAAGAGGATTTCTACCCGTTCCTTAAGGATAACAGAGAGAAGATAGAAAAAATACTGACCTTCAGGATACCATACTATGTGGGCCCACTGGCGCGCGGAAATAGTCGTTTCGCATGGATGACTAGAAAGTCCGAAGAAACGATCACGCCATGGAATTTTGAGGAAGTGGTCGACAAGGGCGCCTCTGCCCAGAGCTTCATCGAAAGGATGACCAATTTTGACAAAAATCTGCCTAACGAAAAGGTGCTTCCGAAGCACAGCCTGTTGTATGAATACTTCACAGTTTATAACGAGCTCACTAAGGTCAAGTACGTCACGGAGGGCATGCGTAAGCCTGCTTTCCTGTCTGGTGAACAAAAAAAGGCGATTGTGGACCTCCTTTTCAAGACGAACCGTAAAGTTACTGTGAAGCAACTGAAAGAGGATTACTTTAAGAAAATTGAGTGCTTCGACAGTGTGGAGATTTCCGGTGTCGAGGACCGGTTTAACGCCAGCCTGGGTACGTATCATGACCTGCTTAAAATTATCAAGGATAAAGATTTCCTGGATAATGAAGAGAACGAAGATATACTGGAGGACATTGTGTTGACTTTGACCCTCTTCGAGGACAGAGAGATGATTGAGGAAAGACTGAAGACCTACGCACACCTTTTTGATGACAAGGTCATGAAACAACTCAAGCGCCGGCGCTATACTGGCTGGGGCCGGCTTTCTCGCAAGCTCATCAATGGGATTCGGGATAAGCAATCAGGCAAGACAATTTTGGACTTCCTCAAATCCGACGGATTCGCAAATAGGAATTTTATGCAGCTGATACATGACGACTCTTTGACATTCAAAGAAGACATACAGAAGGCTCAGGTCTCCGGCCAAGGAGATTCTTTGCACGAGCATATCGCTAACTTGGCAGGTAGCCCCGCCATAAAAAAGGGCATTCTTCAAACGGTAAAAGTTGTTGACGAACTCGTGAAGGTTATGGGCCGTCATAAGCCGGAAAACATTGTTATTGAAATGGCTAGGGAAAATCAGACGACCCAGAAGGGACAGAAAAATAGCAGGGAGCGGATGAAGAGAATTGAAGAGGGAATTAAGGAGCTTGGATCTCAGATTCTTAAGGAGCACCCTGTGGAGAACACCCAACTTCAGAATGAAAAGCTCTACCTTTACTACCTTCAAAACGGCCGGGATATGTACGTCGATCAGGAACTTGACATTAACCGGTTGAGCGATTATGACGTTGACCATATTGTGCCCCAATCTTTCCTTAAAGACGACTCTATCGACAATAAAGTGCTGACGCGCAGCGATAAAAATCGCGGTAAGTCGGATAATGTCCCGTCGGAAGAGGTGGTTAAAAAAATGAAGAACTATTGGAGGCAACTCCTGAATGCCAAGCTGATCACTCAGAGGAAATTCGACAATCTCACCAAGGCAGAAAGGGGTGGACTTAGCGAGCTCGACAAGGCCGGTTTTATCAAAAGACAGCTGGTGGAGACACGCCAAATCACCAAACACGTTGCCCAGATCCTGGATTCGAGGATGAACACGAAGTATGACGAGAACGACAAGTTGATTAGGGAAGTCAAGGTCATCACTTTGAAGTCCAAGCTGGTGAGCGACTTTCGCAAAGACTTCCAGTTTTACAAAGTCAGGGAAATTAATAACTACCACCACGCCCACGACGCCTACCTTAACGCCGTGGTTGGCACAGCACTCATCAAGAAATACCCTAAGCTCGAATCTGAGTTCGTCTATGGCGACTATAAGGTCTACGACGTTAGAAAAATGATCGCGAAATCTGAGCAGGAAATAGGCAAGGCAACTGCCAAGTACTTCTTCTATTCCAATATCATGAACTTTTTTAAGACGGAGATTACCCTGGCGAATGGTGAGATCCGCAAGCGCCCTTTGATTGAGACAAACGGAGAAACAGGAGAGATCGTATGGGACAAAGGGCGGGACTTTGCTACTGTTAGGAAGGTGCTCTCTATGCCACAAGTTAACATTGTCAAAAAAACTGAAGTGCAGACAGGTGGGTTTAGCAAGGAATCTATCCTGCCGAAGAGGAACTCTGACAAGCTGATCGCCCGCAAGAAAGATTGGGATCCGAAAAAGTACGGAGGATTCGACTCCCCCACAGTTGCGTACTCCGTGCTTGTCGTGGCCAAAGTGGAGAAGGGCAAGTCTAAGAAGCTCAAGAGCGTCAAAGAGTTGTTGGGGATCACGATTATGGAGCGGTCGTCTTTCGAAAAGAATCCGATAGATTTTCTCGAGGCCAAGGGTTATAAAGAAGTCAAGAAGGATCTTATCATCAAGCTCCCTAAGTACTCCCTCTTTGAGCTTGAAAACGGACGGAAAAGAATGCTGGCTTCAGCGGGTGAACTTCAGAAGGGTAATGAACTCGCTCTGCCCTCAAAATATGTGAATTTCCTTTACCTGGCATCACACTATGAGAAGCTTAAGGGGTCTCCAGAGGACAACGAGCAGAAGCAACTGTTCGTTGAACAACACAAGCACTACCTTGACGAGATTATCGAGCAAATCAGCGAGTTTAGCAAGCGCGTTATACTGGCAGACGCAAATCTTGATAAGGTCCTTAGCGCCTACAACAAGCATAGAGACAAACCCATCCGGGAGCAGGCCGAGAACATTATTCATCTCTTCACCTTGACGAATCTTGGGGCCCCGGCCGCGTTCAAGTACTTCGATACTACCATAGACAGAAAGCGCTATACATCGACAAAGGAAGTTCTTGACGCCACGCTGATCCACCAAAGTATAACAGGCCTCTATGAGACACGCATCGACCTTTCGCAGTTGGGCGGTGACTCCGGCGGAAGTACAAACCTTTCAGACATTATAGAAAAGGAAACCGGCAAGCAACTCGTCATCCAGGAATCCATACTTATGCTCCCTGAAGAGGTGGAAGAAGTGATCGGTAATAAACCAGAGAGCGACATACTTGTCCACACCGCTTATGACGAAAGTACAGACGAAAACGTCATGCTTCTGACGAGTGATGCCCCCGAATACAAACCTTGGGCGCTCGTCATCCAGGATTCCAATGGGGAGAATAAAATAAAGATGCTCTCTGGAGGCAGCCCAAAGAAGAAGAGAAAGGTCTGA；
<210> 10
<211> 4719
<212> DNA
<213>人工序列
<223> SEQ ID No. 10
<400> 10
ATGGATAGCCTTCTCATGAACAGAAGAGAGTTTCTCTATCAGTTTAAAAATGTTCGGTGGGCGAAGGGGAGGAGAGAGACATATCTCTGCTATGTTGTTAAGCGGAGAGATTCTGCGACCTCATTCTCACTCGATTTTGGTTATTTGAGGAACAAGAATGGATGTCATGTCGAATTGTTGTTTCTCCGGTATATTTCCGACTGGGATTTGGACCCAGGGCGGTGTTACCGGGTCACATGGTTTATTTCCTGGAGTCCATGTTACGACTGTGCGCGCCATGTCGCCGACTTCCTCAGGGGTAATCCTAACTTGTCCTTGCGGATTTTTACAGCCAGACTCTATTTCTGTGAGGATCGGAAGGCGGAACCCGAGGGGCTGAGAAGACTGCACCGCGCTGGCGTCCAAATCGCCATCATGACTTTTAAGGATTATTTCTACTGTTGGAACACGTTCGTCGAGAACCACGGTCGGACCTTCAAAGCCTGGGAAGGGCTGCATGAAAATTCCGTGAGGTTGTCCCGGCAACTCCGCAGAATACTCCTGCCCCTTTATGAGGTCGACGATCTCAGAGACGCCTTTAGAACTGATAAAAAGTATTCAATCGGACTTGCTATTGGGACAAACTCTGTGGGCTGGGCGGTAATTACCGACGAGTACAAGGTGCCTAGTAAGAAATTTAAAGTGCTCGGAAACACTGACAGGCACTCTATAAAGAAGAACCTGATCGGGGCACTGCTTTTCGACTCCGGAGAGACGGCGGAGGCGACGCGTCTCAAGCGTACCGCGCGCCGCAGGTACACAAGAAGGAAGAATAGGATCTGCTACTTGCAGGAAATCTTCAGTAACGAGATGGCGAAGGTCGACGATAGTTTCTTTCATCGGTTGGAAGAATCGTTCCTCGTAGAGGAGGACAAAAAGCACGAGCGTCACCCAATATTCGGGAATATTGTTGACGAGGTTGCCTACCATGAGAAATATCCTACAATATATCACCTCCGTAAGAAGCTTGTCGATTCAACTGATAAGGCTGATCTCAGACTCATCTATCTTGCCCTCGCACATATGATTAAGTTTCGTGGCCACTTCTTGATTGAAGGCGACCTCAACCCGGACAACTCAGATGTTGACAAGCTTTTTATACAGCTCGTCCAGACATATAACCAGCTGTTTGAAGAGAATCCCATCAATGCGAGTGGGGTTGATGCTAACGCCATTTTGTCCGCCAGGTTGTCCAAATCTCGCAGACTGGAAAACCTGATCGCACAGCTTCCCGGTGAAAAGAAAAACGGGCTCTTCGGCAATCTCATCGCACTGTCCCTCGGCCTCACCCCAAACTTCAAGTCTAACTTCGACCTGGCCGAGGATGCGAAGCTCCAGCTGTCAAAAGATACATACGACGACGATTTGGACAATCTGCTTGCGCAAATAGGCGACCAGTATGCGGACCTGTTCCTGGCTGCCAAAAATCTGTCAGATGCAATCCTCCTGTCCGATATATTGCGTGTGAACACCGAAATCACGAAGGCACCGCTTAGCGCATCCATGATCAAGAGATACGACGAGCACCATCAGGACCTCACACTCCTCAAGGCGCTTGTTCGTCAGCAGCTTCCCGAGAAATATAAGGAAATTTTTTTCGATCAAAGCAAGAATGGATATGCTGGCTATATTGACGGTGGCGCTTCGCAGGAGGAGTTCTATAAATTCATTAAGCCGATTCTGGAGAAGATGGACGGAACGGAGGAGCTCCTCGTCAAGCTTAACCGGGAAGACCTGTTGCGGAAGCAGAGGACTTTTGATAACGGCTCTATTCCGCACCAAATCCATCTGGGTGAGTTGCACGCAATCTTGAGAAGACAAGAGGATTTCTACCCGTTCCTTAAGGATAACAGAGAGAAGATAGAAAAAATACTGACCTTCAGGATACCATACTATGTGGGCCCACTGGCGCGCGGAAATAGTCGTTTCGCATGGATGACTAGAAAGTCCGAAGAAACGATCACGCCATGGAATTTTGAGGAAGTGGTCGACAAGGGCGCCTCTGCCCAGAGCTTCATCGAAAGGATGACCAATTTTGACAAAAATCTGCCTAACGAAAAGGTGCTTCCGAAGCACAGCCTGTTGTATGAATACTTCACAGTTTATAACGAGCTCACTAAGGTCAAGTACGTCACGGAGGGCATGCGTAAGCCTGCTTTCCTGTCTGGTGAACAAAAAAAGGCGATTGTGGACCTCCTTTTCAAGACGAACCGTAAAGTTACTGTGAAGCAACTGAAAGAGGATTACTTTAAGAAAATTGAGTGCTTCGACAGTGTGGAGATTTCCGGTGTCGAGGACCGGTTTAACGCCAGCCTGGGTACGTATCATGACCTGCTTAAAATTATCAAGGATAAAGATTTCCTGGATAATGAAGAGAACGAAGATATACTGGAGGACATTGTGTTGACTTTGACCCTCTTCGAGGACAGAGAGATGATTGAGGAAAGACTGAAGACCTACGCACACCTTTTTGATGACAAGGTCATGAAACAACTCAAGCGCCGGCGCTATACTGGCTGGGGCCGGCTTTCTCGCAAGCTCATCAATGGGATTCGGGATAAGCAATCAGGCAAGACAATTTTGGACTTCCTCAAATCCGACGGATTCGCAAATAGGAATTTTATGCAGCTGATACATGACGACTCTTTGACATTCAAAGAAGACATACAGAAGGCTCAGGTCTCCGGCCAAGGAGATTCTTTGCACGAGCATATCGCTAACTTGGCAGGTAGCCCCGCCATAAAAAAGGGCATTCTTCAAACGGTAAAAGTTGTTGACGAACTCGTGAAGGTTATGGGCCGTCATAAGCCGGAAAACATTGTTATTGAAATGGCTAGGGAAAATCAGACGACCCAGAAGGGACAGAAAAATAGCAGGGAGCGGATGAAGAGAATTGAAGAGGGAATTAAGGAGCTTGGATCTCAGATTCTTAAGGAGCACCCTGTGGAGAACACCCAACTTCAGAATGAAAAGCTCTACCTTTACTACCTTCAAAACGGCCGGGATATGTACGTCGATCAGGAACTTGACATTAACCGGTTGAGCGATTATGACGTTGACCATATTGTGCCCCAATCTTTCCTTAAAGACGACTCTATCGACAATAAAGTGCTGACGCGCAGCGATAAAAATCGCGGTAAGTCGGATAATGTCCCGTCGGAAGAGGTGGTTAAAAAAATGAAGAACTATTGGAGGCAACTCCTGAATGCCAAGCTGATCACTCAGAGGAAATTCGACAATCTCACCAAGGCAGAAAGGGGTGGACTTAGCGAGCTCGACAAGGCCGGTTTTATCAAAAGACAGCTGGTGGAGACACGCCAAATCACCAAACACGTTGCCCAGATCCTGGATTCGAGGATGAACACGAAGTATGACGAGAACGACAAGTTGATTAGGGAAGTCAAGGTCATCACTTTGAAGTCCAAGCTGGTGAGCGACTTTCGCAAAGACTTCCAGTTTTACAAAGTCAGGGAAATTAATAACTACCACCACGCCCACGACGCCTACCTTAACGCCGTGGTTGGCACAGCACTCATCAAGAAATACCCTAAGCTCGAATCTGAGTTCGTCTATGGCGACTATAAGGTCTACGACGTTAGAAAAATGATCGCGAAATCTGAGCAGGAAATAGGCAAGGCAACTGCCAAGTACTTCTTCTATTCCAATATCATGAACTTTTTTAAGACGGAGATTACCCTGGCGAATGGTGAGATCCGCAAGCGCCCTTTGATTGAGACAAACGGAGAAACAGGAGAGATCGTATGGGACAAAGGGCGGGACTTTGCTACTGTTAGGAAGGTGCTCTCTATGCCACAAGTTAACATTGTCAAAAAAACTGAAGTGCAGACAGGTGGGTTTAGCAAGGAATCTATCCTGCCGAAGAGGAACTCTGACAAGCTGATCGCCCGCAAGAAAGATTGGGATCCGAAAAAGTACGGAGGATTCGtCTCCCCCACAGTTGCGTACTCCGTGCTTGTCGTGGCCAAAGTGGAGAAGGGCAAGTCTAAGAAGCTCAAGAGCGTCAAAGAGTTGTTGGGGATCACGATTATGGAGCGGTCGTCTTTCGAAAAGAATCCGATAGATTTTCTCGAGGCCAAGGGTTATAAAGAAGTCAAGAAGGATCTTATCATCAAGCTCCCTAAGTACTCCCTCTTTGAGCTTGAAAACGGACGGAAAAGAATGCTGGCTTCAGCGGGTGAACTTCAGAAGGGTAATGAACTCGCTCTGCCCTCAAAATATGTGAATTTCCTTTACCTGGCATCACACTATGAGAAGCTTAAGGGGTCTCCAGAGGACAACGAGCAGAAGCAACTGTTCGTTGAACAACACAAGCACTACCTTGACGAGATTATCGAGCAAATCAGCGAGTTTAGCAAGCGCGTTATACTGGCAGACGCAAATCTTGATAAGGTCCTTAGCGCCTACAACAAGCATAGAGACAAACCCATCCGGGAGCAGGCCGAGAACATTATTCATCTCTTCACCTTGACGAATCTTGGGGCCCCGGCCGCGTTCAAGTACTTCGATACTACCATAGACAGAAAGCGCTATACATCGACAAAGGAAGTTCTTGACGCCACGCTGATCCACCAAAGTATAACAGGCCTCTATGAGACACGCATCGACCTTTCGCAGTTGGGCGGTGACCGCCCCAAAAAGAAGAGGAAAGTTGGCGGGTGA；
<210> 11
<211> 4767
<212> DNA
<213>人工序列
<223> SEQ ID No. 11
<400> 11
ATGGATAGCCTTCTCATGAACAGAAGAGAGTTTCTCTATCAGTTTAAAAATGTTCGGTGGGCGAAGGGGAGGAGAGAGACATATCTCTGCTATGTTGTTAAGCGGAGAGATTCTGCGACCTCATTCTCACTCGATTTTGGTTATTTGAGGAACAAGAATGGATGTCATGTCGAATTGTTGTTTCTCCGGTATATTTCCGACTGGGATTTGGACCCAGGGCGGTGTTACCGGGTCACATGGTTTATTTCCTGGAGTCCATGTTACGACTGTGCGCGCCATGTCGCCGACTTCCTCAGGGGTAATCCTAACTTGTCCTTGCGGATTTTTACAGCCAGACTCTATTTCTGTGAGGATCGGAAGGCGGAACCCGAGGGGCTGAGAAGACTGCACCGCGCTGGCGTCCAAATCGCCATCATGACTTTTAAGGATTATTTCTACTGTTGGAACACGTTCGTCGAGAACCACGGTCGGACCTTCAAAGCCTGGGAAGGGCTGCATGAAAATTCCGTGAGGTTGTCCCGGCAACTCCGCAGAATACTCCTGCCCCTTTATGAGGTCGACGATCTCAGAGACGCCTTTAGAACTAGCGGAAGCGAGACGCCAGGGACTTCTGAATCGGCCACCCCCGAGAGCGATAAAAAGTATTCAATCGGACTTGCTATTGGGACAAACTCTGTGGGCTGGGCGGTAATTACCGACGAGTACAAGGTGCCTAGTAAGAAATTTAAAGTGCTCGGAAACACTGACAGGCACTCTATAAAGAAGAACCTGATCGGGGCACTGCTTTTCGACTCCGGAGAGACGGCGGAGGCGACGCGTCTCAAGCGTACCGCGCGCCGCAGGTACACAAGAAGGAAGAATAGGATCTGCTACTTGCAGGAAATCTTCAGTAACGAGATGGCGAAGGTCGACGATAGTTTCTTTCATCGGTTGGAAGAATCGTTCCTCGTAGAGGAGGACAAAAAGCACGAGCGTCACCCAATATTCGGGAATATTGTTGACGAGGTTGCCTACCATGAGAAATATCCTACAATATATCACCTCCGTAAGAAGCTTGTCGATTCAACTGATAAGGCTGATCTCAGACTCATCTATCTTGCCCTCGCACATATGATTAAGTTTCGTGGCCACTTCTTGATTGAAGGCGACCTCAACCCGGACAACTCAGATGTTGACAAGCTTTTTATACAGCTCGTCCAGACATATAACCAGCTGTTTGAAGAGAATCCCATCAATGCGAGTGGGGTTGATGCTAACGCCATTTTGTCCGCCAGGTTGTCCAAATCTCGCAGACTGGAAAACCTGATCGCACAGCTTCCCGGTGAAAAGAAAAACGGGCTCTTCGGCAATCTCATCGCACTGTCCCTCGGCCTCACCCCAAACTTCAAGTCTAACTTCGACCTGGCCGAGGATGCGAAGCTCCAGCTGTCAAAAGATACATACGACGACGATTTGGACAATCTGCTTGCGCAAATAGGCGACCAGTATGCGGACCTGTTCCTGGCTGCCAAAAATCTGTCAGATGCAATCCTCCTGTCCGATATATTGCGTGTGAACACCGAAATCACGAAGGCACCGCTTAGCGCATCCATGATCAAGAGATACGACGAGCACCATCAGGACCTCACACTCCTCAAGGCGCTTGTTCGTCAGCAGCTTCCCGAGAAATATAAGGAAATTTTTTTCGATCAAAGCAAGAATGGATATGCTGGCTATATTGACGGTGGCGCTTCGCAGGAGGAGTTCTATAAATTCATTAAGCCGATTCTGGAGAAGATGGACGGAACGGAGGAGCTCCTCGTCAAGCTTAACCGGGAAGACCTGTTGCGGAAGCAGAGGACTTTTGATAACGGCTCTATTCCGCACCAAATCCATCTGGGTGAGTTGCACGCAATCTTGAGAAGACAAGAGGATTTCTACCCGTTCCTTAAGGATAACAGAGAGAAGATAGAAAAAATACTGACCTTCAGGATACCATACTATGTGGGCCCACTGGCGCGCGGAAATAGTCGTTTCGCATGGATGACTAGAAAGTCCGAAGAAACGATCACGCCATGGAATTTTGAGGAAGTGGTCGACAAGGGCGCCTCTGCCCAGAGCTTCATCGAAAGGATGACCAATTTTGACAAAAATCTGCCTAACGAAAAGGTGCTTCCGAAGCACAGCCTGTTGTATGAATACTTCACAGTTTATAACGAGCTCACTAAGGTCAAGTACGTCACGGAGGGCATGCGTAAGCCTGCTTTCCTGTCTGGTGAACAAAAAAAGGCGATTGTGGACCTCCTTTTCAAGACGAACCGTAAAGTTACTGTGAAGCAACTGAAAGAGGATTACTTTAAGAAAATTGAGTGCTTCGACAGTGTGGAGATTTCCGGTGTCGAGGACCGGTTTAACGCCAGCCTGGGTACGTATCATGACCTGCTTAAAATTATCAAGGATAAAGATTTCCTGGATAATGAAGAGAACGAAGATATACTGGAGGACATTGTGTTGACTTTGACCCTCTTCGAGGACAGAGAGATGATTGAGGAAAGACTGAAGACCTACGCACACCTTTTTGATGACAAGGTCATGAAACAACTCAAGCGCCGGCGCTATACTGGCTGGGGCCGGCTTTCTCGCAAGCTCATCAATGGGATTCGGGATAAGCAATCAGGCAAGACAATTTTGGACTTCCTCAAATCCGACGGATTCGCAAATAGGAATTTTATGCAGCTGATACATGACGACTCTTTGACATTCAAAGAAGACATACAGAAGGCTCAGGTCTCCGGCCAAGGAGATTCTTTGCACGAGCATATCGCTAACTTGGCAGGTAGCCCCGCCATAAAAAAGGGCATTCTTCAAACGGTAAAAGTTGTTGACGAACTCGTGAAGGTTATGGGCCGTCATAAGCCGGAAAACATTGTTATTGAAATGGCTAGGGAAAATCAGACGACCCAGAAGGGACAGAAAAATAGCAGGGAGCGGATGAAGAGAATTGAAGAGGGAATTAAGGAGCTTGGATCTCAGATTCTTAAGGAGCACCCTGTGGAGAACACCCAACTTCAGAATGAAAAGCTCTACCTTTACTACCTTCAAAACGGCCGGGATATGTACGTCGATCAGGAACTTGACATTAACCGGTTGAGCGATTATGACGTTGACCATATTGTGCCCCAATCTTTCCTTAAAGACGACTCTATCGACAATAAAGTGCTGACGCGCAGCGATAAAAATCGCGGTAAGTCGGATAATGTCCCGTCGGAAGAGGTGGTTAAAAAAATGAAGAACTATTGGAGGCAACTCCTGAATGCCAAGCTGATCACTCAGAGGAAATTCGACAATCTCACCAAGGCAGAAAGGGGTGGACTTAGCGAGCTCGACAAGGCCGGTTTTATCAAAAGACAGCTGGTGGAGACACGCCAAATCACCAAACACGTTGCCCAGATCCTGGATTCGAGGATGAACACGAAGTATGACGAGAACGACAAGTTGATTAGGGAAGTCAAGGTCATCACTTTGAAGTCCAAGCTGGTGAGCGACTTTCGCAAAGACTTCCAGTTTTACAAAGTCAGGGAAATTAATAACTACCACCACGCCCACGACGCCTACCTTAACGCCGTGGTTGGCACAGCACTCATCAAGAAATACCCTAAGCTCGAATCTGAGTTCGTCTATGGCGACTATAAGGTCTACGACGTTAGAAAAATGATCGCGAAATCTGAGCAGGAAATAGGCAAGGCAACTGCCAAGTACTTCTTCTATTCCAATATCATGAACTTTTTTAAGACGGAGATTACCCTGGCGAATGGTGAGATCCGCAAGCGCCCTTTGATTGAGACAAACGGAGAAACAGGAGAGATCGTATGGGACAAAGGGCGGGACTTTGCTACTGTTAGGAAGGTGCTCTCTATGCCACAAGTTAACATTGTCAAAAAAACTGAAGTGCAGACAGGTGGGTTTAGCAAGGAATCTATCCTGCCGAAGAGGAACTCTGACAAGCTGATCGCCCGCAAGAAAGATTGGGATCCGAAAAAGTACGGAGGATTCGtCTCCCCCACAGTTGCGTACTCCGTGCTTGTCGTGGCCAAAGTGGAGAAGGGCAAGTCTAAGAAGCTCAAGAGCGTCAAAGAGTTGTTGGGGATCACGATTATGGAGCGGTCGTCTTTCGAAAAGAATCCGATAGATTTTCTCGAGGCCAAGGGTTATAAAGAAGTCAAGAAGGATCTTATCATCAAGCTCCCTAAGTACTCCCTCTTTGAGCTTGAAAACGGACGGAAAAGAATGCTGGCTTCAGCGGGTGAACTTCAGAAGGGTAATGAACTCGCTCTGCCCTCAAAATATGTGAATTTCCTTTACCTGGCATCACACTATGAGAAGCTTAAGGGGTCTCCAGAGGACAACGAGCAGAAGCAACTGTTCGTTGAACAACACAAGCACTACCTTGACGAGATTATCGAGCAAATCAGCGAGTTTAGCAAGCGCGTTATACTGGCAGACGCAAATCTTGATAAGGTCCTTAGCGCCTACAACAAGCATAGAGACAAACCCATCCGGGAGCAGGCCGAGAACATTATTCATCTCTTCACCTTGACGAATCTTGGGGCCCCGGCCGCGTTCAAGTACTTCGATACTACCATAGACAGAAAGCGCTATACATCGACAAAGGAAGTTCTTGACGCCACGCTGATCCACCAAAGTATAACAGGCCTCTATGAGACACGCATCGACCTTTCGCAGTTGGGCGGTGACCGCCCCAAAAAGAAGAGGAAAGTTGGCGGGTGA；
<210> 12
<211> 5031
<212> DNA
<213>人工序列
<223> SEQ ID No. 12
<400> 12
ATGGATAGCCTTCTCATGAACAGAAGAGAGTTTCTCTATCAGTTTAAAAATGTTCGGTGGGCGAAGGGGAGGAGAGAGACATATCTCTGCTATGTTGTTAAGCGGAGAGATTCTGCGACCTCATTCTCACTCGATTTTGGTTATTTGAGGAACAAGAATGGATGTCATGTCGAATTGTTGTTTCTCCGGTATATTTCCGACTGGGATTTGGACCCAGGGCGGTGTTACCGGGTCACATGGTTTATTTCCTGGAGTCCATGTTACGACTGTGCGCGCCATGTCGCCGACTTCCTCAGGGGTAATCCTAACTTGTCCTTGCGGATTTTTACAGCCAGACTCTATTTCTGTGAGGATCGGAAGGCGGAACCCGAGGGGCTGAGAAGACTGCACCGCGCTGGCGTCCAAATCGCCATCATGACTTTTAAGGATTATTTCTACTGTTGGAACACGTTCGTCGAGAACCACGGTCGGACCTTCAAAGCCTGGGAAGGGCTGCATGAAAATTCCGTGAGGTTGTCCCGGCAACTCCGCAGAATACTCCTGCCCCTTTATGAGGTCGACGATCTCAGAGACGCCTTTAGAACTAGCGGAAGCGAGACGCCAGGGACTTCTGAATCGGCCACCCCCGAGAGCGATAAAAAGTATTCAATCGGACTTGCTATTGGGACAAACTCTGTGGGCTGGGCGGTAATTACCGACGAGTACAAGGTGCCTAGTAAGAAATTTAAAGTGCTCGGAAACACTGACAGGCACTCTATAAAGAAGAACCTGATCGGGGCACTGCTTTTCGACTCCGGAGAGACGGCGGAGGCGACGCGTCTCAAGCGTACCGCGCGCCGCAGGTACACAAGAAGGAAGAATAGGATCTGCTACTTGCAGGAAATCTTCAGTAACGAGATGGCGAAGGTCGACGATAGTTTCTTTCATCGGTTGGAAGAATCGTTCCTCGTAGAGGAGGACAAAAAGCACGAGCGTCACCCAATATTCGGGAATATTGTTGACGAGGTTGCCTACCATGAGAAATATCCTACAATATATCACCTCCGTAAGAAGCTTGTCGATTCAACTGATAAGGCTGATCTCAGACTCATCTATCTTGCCCTCGCACATATGATTAAGTTTCGTGGCCACTTCTTGATTGAAGGCGACCTCAACCCGGACAACTCAGATGTTGACAAGCTTTTTATACAGCTCGTCCAGACATATAACCAGCTGTTTGAAGAGAATCCCATCAATGCGAGTGGGGTTGATGCTAACGCCATTTTGTCCGCCAGGTTGTCCAAATCTCGCAGACTGGAAAACCTGATCGCACAGCTTCCCGGTGAAAAGAAAAACGGGCTCTTCGGCAATCTCATCGCACTGTCCCTCGGCCTCACCCCAAACTTCAAGTCTAACTTCGACCTGGCCGAGGATGCGAAGCTCCAGCTGTCAAAAGATACATACGACGACGATTTGGACAATCTGCTTGCGCAAATAGGCGACCAGTATGCGGACCTGTTCCTGGCTGCCAAAAATCTGTCAGATGCAATCCTCCTGTCCGATATATTGCGTGTGAACACCGAAATCACGAAGGCACCGCTTAGCGCATCCATGATCAAGAGATACGACGAGCACCATCAGGACCTCACACTCCTCAAGGCGCTTGTTCGTCAGCAGCTTCCCGAGAAATATAAGGAAATTTTTTTCGATCAAAGCAAGAATGGATATGCTGGCTATATTGACGGTGGCGCTTCGCAGGAGGAGTTCTATAAATTCATTAAGCCGATTCTGGAGAAGATGGACGGAACGGAGGAGCTCCTCGTCAAGCTTAACCGGGAAGACCTGTTGCGGAAGCAGAGGACTTTTGATAACGGCTCTATTCCGCACCAAATCCATCTGGGTGAGTTGCACGCAATCTTGAGAAGACAAGAGGATTTCTACCCGTTCCTTAAGGATAACAGAGAGAAGATAGAAAAAATACTGACCTTCAGGATACCATACTATGTGGGCCCACTGGCGCGCGGAAATAGTCGTTTCGCATGGATGACTAGAAAGTCCGAAGAAACGATCACGCCATGGAATTTTGAGGAAGTGGTCGACAAGGGCGCCTCTGCCCAGAGCTTCATCGAAAGGATGACCAATTTTGACAAAAATCTGCCTAACGAAAAGGTGCTTCCGAAGCACAGCCTGTTGTATGAATACTTCACAGTTTATAACGAGCTCACTAAGGTCAAGTACGTCACGGAGGGCATGCGTAAGCCTGCTTTCCTGTCTGGTGAACAAAAAAAGGCGATTGTGGACCTCCTTTTCAAGACGAACCGTAAAGTTACTGTGAAGCAACTGAAAGAGGATTACTTTAAGAAAATTGAGTGCTTCGACAGTGTGGAGATTTCCGGTGTCGAGGACCGGTTTAACGCCAGCCTGGGTACGTATCATGACCTGCTTAAAATTATCAAGGATAAAGATTTCCTGGATAATGAAGAGAACGAAGATATACTGGAGGACATTGTGTTGACTTTGACCCTCTTCGAGGACAGAGAGATGATTGAGGAAAGACTGAAGACCTACGCACACCTTTTTGATGACAAGGTCATGAAACAACTCAAGCGCCGGCGCTATACTGGCTGGGGCCGGCTTTCTCGCAAGCTCATCAATGGGATTCGGGATAAGCAATCAGGCAAGACAATTTTGGACTTCCTCAAATCCGACGGATTCGCAAATAGGAATTTTATGCAGCTGATACATGACGACTCTTTGACATTCAAAGAAGACATACAGAAGGCTCAGGTCTCCGGCCAAGGAGATTCTTTGCACGAGCATATCGCTAACTTGGCAGGTAGCCCCGCCATAAAAAAGGGCATTCTTCAAACGGTAAAAGTTGTTGACGAACTCGTGAAGGTTATGGGCCGTCATAAGCCGGAAAACATTGTTATTGAAATGGCTAGGGAAAATCAGACGACCCAGAAGGGACAGAAAAATAGCAGGGAGCGGATGAAGAGAATTGAAGAGGGAATTAAGGAGCTTGGATCTCAGATTCTTAAGGAGCACCCTGTGGAGAACACCCAACTTCAGAATGAAAAGCTCTACCTTTACTACCTTCAAAACGGCCGGGATATGTACGTCGATCAGGAACTTGACATTAACCGGTTGAGCGATTATGACGTTGACCATATTGTGCCCCAATCTTTCCTTAAAGACGACTCTATCGACAATAAAGTGCTGACGCGCAGCGATAAAAATCGCGGTAAGTCGGATAATGTCCCGTCGGAAGAGGTGGTTAAAAAAATGAAGAACTATTGGAGGCAACTCCTGAATGCCAAGCTGATCACTCAGAGGAAATTCGACAATCTCACCAAGGCAGAAAGGGGTGGACTTAGCGAGCTCGACAAGGCCGGTTTTATCAAAAGACAGCTGGTGGAGACACGCCAAATCACCAAACACGTTGCCCAGATCCTGGATTCGAGGATGAACACGAAGTATGACGAGAACGACAAGTTGATTAGGGAAGTCAAGGTCATCACTTTGAAGTCCAAGCTGGTGAGCGACTTTCGCAAAGACTTCCAGTTTTACAAAGTCAGGGAAATTAATAACTACCACCACGCCCACGACGCCTACCTTAACGCCGTGGTTGGCACAGCACTCATCAAGAAATACCCTAAGCTCGAATCTGAGTTCGTCTATGGCGACTATAAGGTCTACGACGTTAGAAAAATGATCGCGAAATCTGAGCAGGAAATAGGCAAGGCAACTGCCAAGTACTTCTTCTATTCCAATATCATGAACTTTTTTAAGACGGAGATTACCCTGGCGAATGGTGAGATCCGCAAGCGCCCTTTGATTGAGACAAACGGAGAAACAGGAGAGATCGTATGGGACAAAGGGCGGGACTTTGCTACTGTTAGGAAGGTGCTCTCTATGCCACAAGTTAACATTGTCAAAAAAACTGAAGTGCAGACAGGTGGGTTTAGCAAGGAATCTATCCTGCCGAAGAGGAACTCTGACAAGCTGATCGCCCGCAAGAAAGATTGGGATCCGAAAAAGTACGGAGGATTCGtCTCCCCCACAGTTGCGTACTCCGTGCTTGTCGTGGCCAAAGTGGAGAAGGGCAAGTCTAAGAAGCTCAAGAGCGTCAAAGAGTTGTTGGGGATCACGATTATGGAGCGGTCGTCTTTCGAAAAGAATCCGATAGATTTTCTCGAGGCCAAGGGTTATAAAGAAGTCAAGAAGGATCTTATCATCAAGCTCCCTAAGTACTCCCTCTTTGAGCTTGAAAACGGACGGAAAAGAATGCTGGCTTCAGCGGGTGAACTTCAGAAGGGTAATGAACTCGCTCTGCCCTCAAAATATGTGAATTTCCTTTACCTGGCATCACACTATGAGAAGCTTAAGGGGTCTCCAGAGGACAACGAGCAGAAGCAACTGTTCGTTGAACAACACAAGCACTACCTTGACGAGATTATCGAGCAAATCAGCGAGTTTAGCAAGCGCGTTATACTGGCAGACGCAAATCTTGATAAGGTCCTTAGCGCCTACAACAAGCATAGAGACAAACCCATCCGGGAGCAGGCCGAGAACATTATTCATCTCTTCACCTTGACGAATCTTGGGGCCCCGGCCGCGTTCAAGTACTTCGATACTACCATAGACAGAAAGCGCTATACATCGACAAAGGAAGTTCTTGACGCCACGCTGATCCACCAAAGTATAACAGGCCTCTATGAGACACGCATCGACCTTTCGCAGTTGGGCGGTGACTCCGGCGGAAGTACAAACCTTTCAGACATTATAGAAAAGGAAACCGGCAAGCAACTCGTCATCCAGGAATCCATACTTATGCTCCCTGAAGAGGTGGAAGAAGTGATCGGTAATAAACCAGAGAGCGACATACTTGTCCACACCGCTTATGACGAAAGTACAGACGAAAACGTCATGCTTCTGACGAGTGATGCCCCCGAATACAAACCTTGGGCGCTCGTCATCCAGGATTCCAATGGGGAGAATAAAATAAAGATGCTCTCTGGAGGCAGCCCAAAGAAGAAGAGAAAGGTCTGA；
<210> 13
<211> 76
<212> DNA
<213>化脓链球菌（Streptococcus pyogenes）
<223> SEQ ID No. 13
<400> 13
GTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTCCGTTATCAACTTGAAAAAGTGGCACCGAGTCGGTGC；
<210> 14
<211> 20
<212> DNA
<213> 人工序列
<223> SEQ ID No. 14
<400> 14
AGAGACCAAAGGAGGTCTCA；
<210> 15
<211> 37
<212> DNA
<213>人工序列
<223> SEQ ID No. 15
<400> 15
GGCTAGGATCCATCGCAGTCAGCGATGAGTACAGCAA；
<210> 16
<211> 20
<212> DNA
<213> 水稻（Oryza sativa L）
<223>针对Pi-d2的靶核苷酸序列
<400> 16
GAGCATAATGACAATAATAA；
<210> 17
<211> 1765
<212> DNA
<213>玉米（Zea mays L.）
<223> SEQ ID No. 17
<400> 17
GCAGCGTGACCCGGTCGTGCCCCTCTCTAGAGATAATGAGCATTGCATGTCTAAGTTATAAAAAATTACCACATATTTTTTTTGTCACACTTGTTTGAAGTGCAGTTTATCTATCTTTATACATATATTTAAACTTTACTCTACGAATAATATAATCTATAGTACTACAATAATATCAGTGTTTTAGAGAATCATATAAATGAACAGTTAGACATGGTCTAAAGGACAATTGAGTATTTTGACAACAGGACTCTACAGTTTTATCTTTTTAGTGTGCATGTGTTCTCCTTTTTTTTTGCAAATAGCTTCACCTATATAATACTTCATCCATTTTATTAGTACATCCATTTAGGGTTTAGGGTTAATGGTTTTTATAGACTAATTTTTTTAGTACATCTATTTTATTCTATTTTAGCCTCTAAATTAAGAAAACTAAAACTCTATTTTAGTTTTTTTATTTAATAATTTAGATATAAAATAGAATAAAATAAAGTGACTAAAAATTAAACAAATACCCTTTAAGAAATTAAAAAAACTAAGGAAACATTTTTCTTGTTTCGAGTAGATAATGCCAGCCTGTTAAACGCCGTCGACGAGTCTAACGGACACCAACCAGCGAACCAGCAGCGTCGCGTCGGGCCAAGCGAAGCAGACGGCACGGCATCTCTGTCGCTGCCTCTGGACCCCTCTCGAGAGTTCCGCTCCACCGTTGGACTTGCTCCGCTGTCGGCATCCAGAAATTGCGTGGCGGAGCGGCAGACGTGAGCCGGCACGGCAGGCGGCCTCCTCCTCCTCTCACGGCACGGCAGCTACGGGGGATTCCTTTCCCACCGCTCCTTCGCTTTCCCTTCCTCGCCCGCCGTAATAAATAGACACCCCCTCCACACCCTCTTTCCCCAACCTCGTGTTGTTCGGAGCGCACACACACACAACCAGATCTCCCCCAAATCCACCCGTCGGCACCTCCGCTTCAAGGTACGCCGCTCGTCCTCCCCCCCCCCCCCTCTCTACCTTCTCTAGATCGGCGTTCCGGTCCATGGTTAGGGCCCGGTAGTTCTACTTCTGTTCATGTTTGTGTTAGATCCGTGTTTGTGTTAGATCCGTGCTGCTAGCGTTCGTACACGGATGCGACCTGTACGTCAGACACGTTCTGATTGCTAACTTGCCAGTGTTTCTCTTTGGGGAATCCTGGGATGGCTCTAGCCGTTCCGCAGACGGGATCGATTTCATGATTTTTTTTGTTTCGTTGCATAGGGTTTGGTTTGCCCTTTTCCTTTATTTCAATATATGCCGTGCACTTGTTTGTCGGGTCATCTTTTCATGCTTTTTTTTTGTCTTGGTTGTGATGATGTGGTGTGGTTGGGCGGTCGTTCATTCGTTCTAGATCGGAGTAGAATACTGTTTCAAACTACCTGGTGTATTTATTAATTTTGGAACTGTATGTGTGTGTCATACATCTTCATAGTTACGAGTTTAAGATGGATGGAAATATCGATCTAGGATAGGTATACATGTTGATGTGGGTTTTACTGATGCATATACATGATGGCATATGCAGCATCTATTCATATGCTCTAACCTTGAGTACCTATCTATTATAATAAACAAGTATGTTTTATAATTATTTTGATCTTGATATACTTGGATGATGGCATATGCAGCAGCTATATGTGGATTTTTTTAGCCCTGCCTTCATACGCTATTTATTTGCTTGGTACTGTTTCTTTTGTCGATGCTCACCCTGTTGTTTGGTGTTACTTCTGCA；
<210> 18
<211> 326
<212> DNA
<213>水稻（Oryza sativa L）
<223> SEQ ID No. 18
<400> 18
AAGAACGAACTAAGCCGGACAAAAAAAGGAGCACATATACAAACCGGTTTTATTCATGAATGGTCACGATGGATGATGGGGCTCAGACTTGAGCTACGAGGCCGCAGGCGAGAGAAGCCTAGTGTGCTCTCTGCTTGTTTGGGCCGTAACGGAGGATACGGCCGACGAGCGTGTACTACCGCGCGGGATGCCGCTGGGCGCTGCGGGGGCCGTTGGATGGGGATCGGTGGGTCGCGGGAGCGTTGAGGGGAGACAGGTTTAGTACCACCTCGCCTACCGAACAATGAAGAACCCACCTTATAACCCCGCGCGCTGCCGCTTGTGTT；
<210> 19
<211> 245
<212> DNA
<213>水稻（Oryza sativa L）
<223> SEQ ID No. 19
<400> 19
GGATCATGAACCAACGGCCTGGCTGTATTTGGTGGTTGTGTAGGGAGATGGGGAGAAGAAAAGCCCGATTCTCTTCGCTGTGATGGGCTGGATGCATGCGGGGGAGCGGGAGGCCCAAGTACGTGCACGGTGAGCGGCCCACAGGGCGAGTGTGAGCGCGAGAGGCGGGAGGAACAGTTTAGTACCACATTGCCCAGCTAACTCGAACGCGACCAACTTATAAACCCGCGCGCTGTCGCTTGTGT；
<210> 20
<211> 253
<212> DNA
<213>CaMV (Califlower mosaic virus)
<223> SEQ ID No. 20
<400> 20
GATCGTTCAAACATTTGGCAATAAAGTTTCTTAAGATTGAATCCTGTTGCCGGTCTTGCGATGATTATCATATAATTTCTGTTGAATTACGTTAAGCATGTAATAATTAACATGTAATGCATGACGTTATTTATGAGATGGGTTTTTATGATTAGAGTCCCGCAATTATACATTTAATACGCGATAGAAAACAAAATATAGCGCGCAAACTAGGATAAATTATCGCGCGCGGTGTCATCTATGTTACTAGATC；
<210> 21
<211> 8
<212> DNA
<213>人工序列
<223> SEQ ID No. 21
<400> 21
TTTTTTTT；
<210> 22
<211> 24
<212> DNA
<213>人工序列
<223> gPi-d2-F1
<400> 22
GTGTGAGCATAATGACAATAATAA；
<210> 23
<211> 24
<212> DNA
<213>人工序列
<223> gPi-d2-R1
<400> 23
AAACTTATTATTGTCATTATGCTC；
<210> 24
<211> 20
<212>DNA
<213>人工序列
<223> Pi-d2基因上的编辑位点序列
<400> 24
TTATTATTGTCATTATGCTC；
<210> 25
<211> 20
<212> DNA
<213>人工序列
<223> Pi-d2基因被编辑后的部分核苷酸序列
<400> 25
TTATTATTGTCATTATACTC；
<210> 26
<211> 20
<212> DNA
<213>人工序列
<223> Pi-d2-F1
<400> 26
CGGGTTGTAAGAGTGCCTGT；
<210> 27
<211> 20
<212> DNA
<213>人工序列
<223> Pi-d2-R1
<400> 27
CTCCAGCTTCTTCACAGCAA；
<210> 28
<211> 5133
<212> DNA
<213>人工序列
<223> SEQ ID No. 28
<400> 28
ATGAGTAGCGAGACAGGTCCTGTTGCAGTTGACCCGACCCTTCGGAGAAGGATAGAGCCACACGAATTTGAAGTGTTTTTCGACCCTAGAGAACTGAGGAAGGAGACGTGCCTTCTGTACGAGATAAACTGGGGTGGTCGCCACTCTATTTGGAGGCACACTTCGCAAAACACGAACAAGCATGTGGAGGTGAACTTTATAGAAAAATTTACGACTGAGAGATACTTCTGCCCTAATACCCGGTGCTCCATCACCTGGTTCCTTAGCTGGAGCCCTTGTGGCGAATGCTCGAGGGCAATCACCGAGTTTCTGTCCAGATACCCACATGTGACGCTTTTTATATATATTGCCCGCTTGTATCACCACGCTGACCCTAGAAACCGCCAGGGTCTTCGCGATCTGATATCTTCAGGAGTTACCATCCAAATAATGACGGAACAAGAATCCGGTTACTGTTGGCGCAATTTCGTCAACTATAGCCCTTCCAATGAAGCTCATTGGCCTAGATATCCGCACCTCTGGGTCCGGCTGTATGTTCTCGAGCTTTACTGCATTATACTTGGACTTCCCCCCTGCTTGAATATTCTCCGCAGAAAGCAGCCTCAGCTTACGTTTTTTACGATTGCACTCCAAAGTTGTCATTATCAGAGACTGCCACCCCATATCTTGTGGGCTACGGGACTGAAGAGCGGAAGCGAGACGCCAGGGACTTCTGAATCGGCCACCCCCGAGAGCGATAAAAAGTATTCAATCGGACTTGCTATTGGGACAAACTCTGTGGGCTGGGCGGTAATTACCGACGAGTACAAGGTGCCTAGTAAGAAATTTAAAGTGCTCGGAAACACTGACAGGCACTCTATAAAGAAGAACCTGATCGGGGCACTGCTTTTCGACTCCGGAGAGACGGCGGAGGCGACGCGTCTCAAGCGTACCGCGCGCCGCAGGTACACAAGAAGGAAGAATAGGATCTGCTACTTGCAGGAAATCTTCAGTAACGAGATGGCGAAGGTCGACGATAGTTTCTTTCATCGGTTGGAAGAATCGTTCCTCGTAGAGGAGGACAAAAAGCACGAGCGTCACCCAATATTCGGGAATATTGTTGACGAGGTTGCCTACCATGAGAAATATCCTACAATATATCACCTCCGTAAGAAGCTTGTCGATTCAACTGATAAGGCTGATCTCAGACTCATCTATCTTGCCCTCGCACATATGATTAAGTTTCGTGGCCACTTCTTGATTGAAGGCGACCTCAACCCGGACAACTCAGATGTTGACAAGCTTTTTATACAGCTCGTCCAGACATATAACCAGCTGTTTGAAGAGAATCCCATCAATGCGAGTGGGGTTGATGCTAAGGCCATTTTGTCCGCCAGGTTGTCCAAATCTCGCAGACTGGAAAACCTGATCGCACAGCTTCCCGGTGAAAAGAAAAACGGGCTCTTCGGCAATCTCATCGCACTGTCCCTCGGCCTCACCCCAAACTTCAAGTCTAACTTCGACCTGGCCGAGGATGCGAAGCTCCAGCTGTCAAAAGATACATACGACGACGATTTGGACAATCTGCTTGCGCAAATAGGCGACCAGTATGCGGACCTGTTCCTGGCTGCCAAAAATCTGTCAGATGCAATCCTCCTGTCCGATATATTGCGTGTGAACACCGAAATCACGAAGGCACCGCTTAGCGCATCCATGATCAAGAGATACGACGAGCACCATCAGGACCTCACACTCCTCAAGGCGCTTGTTCGTCAGCAGCTTCCCGAGAAATATAAGGAAATTTTTTTCGATCAAAGCAAGAATGGATATGCTGGCTATATTGACGGTGGCGCTTCGCAGGAGGAGTTCTATAAATTCATTAAGCCGATTCTGGAGAAGATGGACGGAACGGAGGAGCTCCTCGTCAAGCTTAACCGGGAAGACCTGTTGCGGAAGCAGAGGACTTTTGATAACGGCTCTATTCCGCACCAAATCCATCTGGGTGAGTTGCACGCAATCTTGAGAAGACAAGAGGATTTCTACCCGTTCCTTAAGGATAACAGAGAGAAGATAGAAAAAATACTGACCTTCAGGATACCATACTATGTGGGCCCACTGGCGCGCGGAAATAGTCGTTTCGCATGGATGACTAGAAAGTCCGAAGAAACGATCACGCCATGGAATTTTGAGGAAGTGGTCGACAAGGGCGCCTCTGCCCAGAGCTTCATCGAAAGGATGACCAATTTTGACAAAAATCTGCCTAACGAAAAGGTGCTTCCGAAGCACAGCCTGTTGTATGAATACTTCACAGTTTATAACGAGCTCACTAAGGTCAAGTACGTCACGGAGGGCATGCGTAAGCCTGCTTTCCTGTCTGGTGAACAAAAAAAGGCGATTGTGGACCTCCTTTTCAAGACGAACCGTAAAGTTACTGTGAAGCAACTGAAAGAGGATTACTTTAAGAAAATTGAGTGCTTCGACAGTGTGGAGATTTCCGGTGTCGAGGACCGGTTTAACGCCAGCCTGGGTACGTATCATGACCTGCTTAAAATTATCAAGGATAAAGATTTCCTGGATAATGAAGAGAACGAAGATATACTGGAGGACATTGTGTTGACTTTGACCCTCTTCGAGGACAGAGAGATGATTGAGGAAAGACTGAAGACCTACGCACACCTTTTTGATGACAAGGTCATGAAACAACTCAAGCGCCGGCGCTATACTGGCTGGGGCCGGCTTTCTCGCAAGCTCATCAATGGGATTCGGGATAAGCAATCAGGCAAGACAATTTTGGACTTCCTCAAATCCGACGGATTCGCAAATAGGAATTTTATGCAGCTGATACATGACGACTCTTTGACATTCAAAGAAGACATACAGAAGGCTCAGGTCTCCGGCCAAGGAGATTCTTTGCACGAGCATATCGCTAACTTGGCAGGTAGCCCCGCCATAAAAAAGGGCATTCTTCAAACGGTAAAAGTTGTTGACGAACTCGTGAAGGTTATGGGCCGTCATAAGCCGGAAAACATTGTTATTGAAATGGCTAGGGAAAATCAGACGACCCAGAAGGGACAGAAAAATAGCAGGGAGCGGATGAAGAGAATTGAAGAGGGAATTAAGGAGCTTGGATCTCAGATTCTTAAGGAGCACCCTGTGGAGAACACCCAACTTCAGAATGAAAAGCTCTACCTTTACTACCTTCAAAACGGCCGGGATATGTACGTCGATCAGGAACTTGACATTAACCGGTTGAGCGATTATGACGTTGACCATATTGTGCCCCAATCTTTCCTTAAAGACGACTCTATCGACAATAAAGTGCTGACGCGCAGCGATAAAAATCGCGGTAAGTCGGATAATGTCCCGTCGGAAGAGGTGGTTAAAAAAATGAAGAACTATTGGAGGCAACTCCTGAATGCCAAGCTGATCACTCAGAGGAAATTCGACAATCTCACCAAGGCAGAAAGGGGTGGACTTAGCGAGCTCGACAAGGCCGGTTTTATCAAAAGACAGCTGGTGGAGACACGCCAAATCACCAAACACGTTGCCCAGATCCTGGATTCGAGGATGAACACGAAGTATGACGAGAACGACAAGTTGATTAGGGAAGTCAAGGTCATCACTTTGAAGTCCAAGCTGGTGAGCGACTTTCGCAAAGACTTCCAGTTTTACAAAGTCAGGGAAATTAATAACTACCACCACGCCCACGACGCCTACCTTAACGCCGTGGTTGGCACAGCACTCATCAAGAAATACCCTAAGCTCGAATCTGAGTTCGTCTATGGCGACTATAAGGTCTACGACGTTAGAAAAATGATCGCGAAATCTGAGCAGGAAATAGGCAAGGCAACTGCCAAGTACTTCTTCTATTCCAATATCATGAACTTTTTTAAGACGGAGATTACCCTGGCGAATGGTGAGATCCGCAAGCGCCCTTTGATTGAGACAAACGGAGAAACAGGAGAGATCGTATGGGACAAAGGGCGGGACTTTGCTACTGTTAGGAAGGTGCTCTCTATGCCACAAGTTAACATTGTCAAAAAAACTGAAGTGCAGACAGGTGGGTTTAGCAAGGAATCTATCCTGCCGAAGAGGAACTCTGACAAGCTGATCGCCCGCAAGAAAGATTGGGATCCGAAAAAGTACGGAGGATTCGACTCCCCCACAGTTGCGTACTCCGTGCTTGTCGTGGCCAAAGTGGAGAAGGGCAAGTCTAAGAAGCTCAAGAGCGTCAAAGAGTTGTTGGGGATCACGATTATGGAGCGGTCGTCTTTCGAAAAGAATCCGATAGATTTTCTCGAGGCCAAGGGTTATAAAGAAGTCAAGAAGGATCTTATCATCAAGCTCCCTAAGTACTCCCTCTTTGAGCTTGAAAACGGACGGAAAAGAATGCTGGCTTCAGCGGGTGAACTTCAGAAGGGTAATGAACTCGCTCTGCCCTCAAAATATGTGAATTTCCTTTACCTGGCATCACACTATGAGAAGCTTAAGGGGTCTCCAGAGGACAACGAGCAGAAGCAACTGTTCGTTGAACAACACAAGCACTACCTTGACGAGATTATCGAGCAAATCAGCGAGTTTAGCAAGCGCGTTATACTGGCAGACGCAAATCTTGATAAGGTCCTTAGCGCCTACAACAAGCATAGAGACAAACCCATCCGGGAGCAGGCCGAGAACATTATTCATCTCTTCACCTTGACGAATCTTGGGGCCCCGGCCGCGTTCAAGTACTTCGATACTACCATAGACAGAAAGCGCTATACATCGACAAAGGAAGTTCTTGACGCCACGCTGATCCACCAAAGTATAACAGGCCTCTATGAGACACGCATCGACCTTTCGCAGTTGGGCGGTGACTCCGGCGGAAGTACAAACCTTTCAGACATTATAGAAAAGGAAACCGGCAAGCAACTCGTCATCCAGGAATCCATACTTATGCTCCCTGAAGAGGTGGAAGAAGTGATCGGTAATAAACCAGAGAGCGACATACTTGTCCACACCGCTTATGACGAAAGTACAGACGAAAACGTCATGCTTCTGACGAGTGATGCCCCCGAATACAAACCTTGGGCGCTCGTCATCCAGGATTCCAATGGGGAGAATAAAATAAAGATGCTCTCTGGAGGCAGCCCAAAGAAGAAGAGAAAGGTCTGA；
<210> 29
<211> 1710
<212> PRT
<213>人工序列
<223> SEQ ID No. 29
<400> 29
MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGGRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLSRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSNEAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHILWATGLKSGSETPGTSESATPESDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTAYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKMLSGGSPKKKRKV。
Claims (14)
1.一套基因编辑人工系统，所述人工系统包括：
第I调节元件，其包括能够编码如氨基酸序列I的核苷酸序列；其中所述氨基酸序列I如SEQ ID No. 2所示；
第II调节元件，其包括依次从5’端到3’端的第II-1核苷酸序列和第II-2核苷酸序列；所述第II-1核苷酸序列包括靶核苷酸序列；所述第II-2核苷酸序列包括来源于化脓链球菌（Streptococcus pyogenes）的sgRNA核酸序列；所述第II-1核苷酸序列和所述第II-2核苷酸序列转录融合，其产物能引导第I调控元件编码的蛋白至目标生物基因组中待突变的靶位点处，并将所述靶位点处的C突变为T、A和G中的一种；或将所述靶位点处的G突变为A、T和C中的一种；
当所述第II调节元件为多个时，包含在其中的多个第II-1核苷酸序列两两不相同；
所述第I调节元件的核苷酸序列为能够适于在水稻中表达的核苷酸序列，所述第II调节元件的核苷酸序列为能够适于在水稻中发生转录的核苷酸序列；
能够编码如SEQ ID No. 2所示蛋白的核苷酸编码序列如SEQ ID No. 8所示；
所述第II-2核苷酸序列如SEQ ID No. 13所示；
通过如下方式确定所述靶核苷酸序列：
1）确定水稻基因组上需要被改造的核苷酸序列；
2）判断步骤1）中确定的需要被改造的核苷酸序列或其反向互补序列中是否携带有待突变的核苷酸C，并判断所述待突变的核苷酸C突变为T、A和G中的一种，或所述待突变的核苷酸G突变为A、T和C中的一种后引起的改变是否符合预期；
3）在需要被改造的核苷酸序列或其反向互补序列中筛选靶标序列：向所述待突变的核苷酸C的3ʹ端方向搜索以确认存在能够被氨基酸序列I识别的识别模序，且所述待突变的核苷酸C处在所述识别模序5ʹ端上游的-19至-13的位置，由此确定的所述识别模序5ʹ端上游17至21个核苷酸序列为所述靶核苷酸序列；
所述识别模序为5ʹ-NGG-3ʹ、5ʹ-NGA-3ʹ、5ʹ- GAGN-3ʹ、5ʹ-AAGN-3ʹ中的一种，所述靶核苷酸序列为所述识别模序5ʹ端上游的17至21个核苷酸序列，淘汰含有连续五个T的核苷酸序列；
其中，所述N为A、G、C和T中的一种。
2.根据权利要求1所述的人工系统，其特征在于，所述第II-1核苷酸序列包括IIS型限制性内切酶的酶切位点，所述靶核苷酸序列通过所述IIS型限制性内切酶的酶切位点而被克隆，以使所述第II-1核苷酸序列与第II-2序列转录融合；
当所述第II调节元件为多个时，用于克隆不同靶核苷酸序列的所述IIS型限制性内切酶的酶切位点两两不相同。
3.根据权利要求1所述的人工系统，其特征在于，所述靶核苷酸序列如SEQ ID No. 16所示。
4.根据权利要求1-3中任意一项所述的人工系统，其特征在于，所述人工系统还包括在所述第I调节元件的5ʹ端的能够用于水稻中的，且能够启动所述第I调节元件转录的第一启动子；和/或所述人工系统还包括在所述第II调节元件的5ʹ端的能够用于水稻中的，且能够启动所述第II调节元件转录的第二启动子；
所述人工系统还包括在所述第I调节元件的3’端的能够终止所述第I调节元件转录的第一终止子；和/或所述人工系统还包括在所述第II调节元件的3’端的能够终止所述第II调节元件转录的第二终止子。
5.根据权利要求4所述的人工系统，其特征在于，所述第一启动子为RNA聚合酶II型启动子；和/或第二启动子为RNA聚合酶III型启动子。
6.根据权利要求5所述的人工系统，其特征在于，第一启动子为SEQ ID No. 17；和/或第二启动子为SEQ ID No. 18和/或SEQ ID No. 19。
7.根据权利要求4所述的人工系统，其特征在于，第一终止子为SEQ ID No. 20；和/或第二终止子为SEQ ID No. 21。
8.根据权利要求4所述的人工系统，其特征在于，所述第I调节元件和所述第II元件能够被克隆到至少一个载体上。
9.根据权利要求8所述的人工系统，其特征在于，所述第I调节元件能够被克隆到pUbi-ccdB上，所述第II调节元件被克隆到入门载体pENTR4上。
10.根据权利要求9所述的人工系统，其特征在于，所述第一启动子、第I调节元件和第一终止子能够被克隆到pUbi-ccdB载体上。
11.根据权利要求9所述的人工系统，其特征在于，所述第二启动子、第II调节元件和第二终止子被克隆到pENTR4载体上。
12.根据权利要求8所述的人工系统，其特征在于，所述第I调节元件和所述第II调节元件能够被整合到同一个载体上，或被分布在多个载体上一起使用。
13.如权利要求1-12中任意一项所述的人工系统在用于将水稻基因组中的C定点突变为T、A和G中的一种、或将水稻基因组中的G定点突变为A、T和C中的一种的应用。
14.一种将水稻基因组上的C定点突变为T、A和G中的一种，或将水稻基因组中的G定点突变为A、T和C中的一种的方法，其包括如下步骤：
1）将如权利要求1-12中任意一项所述的人工系统通过农杆菌介导、基因枪轰击或PEG介导转化的方法中的一种导入到水稻愈伤组织或水稻原生质体中，然后培养获得水稻植株；
2）筛选获得含有定点突变的水稻植株；进一步地，所述水稻植株能够产生含有定点替换碱基的水稻种子。
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