CN113846124A - 一种用于糖代谢相关疾病基因治疗的核酸构建体 - Google Patents
一种用于糖代谢相关疾病基因治疗的核酸构建体 Download PDFInfo
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Abstract
本发明涉及基因治疗或生物医药技术领域,特别是涉及一种用于糖代谢相关疾病基因治疗的核酸构建体,所述核酸构建体包括编码GLP‑1或其类似物的多核苷酸,所述GLP‑1或其类似物的总数量为两个以上,GLP‑1与GLP‑1之间、GLP‑1与其类似物之间或GLP‑1类似物与GLP‑1与类似物之间通过编码连接肽的多核苷酸连接。本发明的核酸构建体在体内外均可用于高效表达具有活性的GLP‑1及其类似物,这为极具潜力的2型糖尿病、机体代谢紊乱、糖尿病相关并发症以及肥胖症等糖代谢相关疾病的药物开发了技术路线。
Description
技术领域
本发明涉及基因治疗或生物医药技术领域,特别是涉及一种用于糖代谢相关疾病基因治疗的核酸构建体。
背景技术
糖尿病是一种身体无法产生完全功能的胰岛素,或者无法正确利用及储存葡萄糖的疾病。葡萄糖存留于血液中引起血糖过高并引发一系列并发症为糖尿病的基础。2型糖尿病,或称成人型糖尿病、非胰岛素依赖型糖尿病,占糖尿病患者中的90%左右。良好的血糖控制对于逆转和减轻2型糖尿病并发症、降低致残率和死亡率、改善患者的生活质量至关重要。
基于胰岛素注射糖尿病治疗取得了一定进展,但是仍难以实现最佳血糖控制,在胰岛素注射治疗中伴随的低血糖和体重进一步增加,会加剧患者机体代谢紊乱以及糖尿病相关并发症,并导致相关的死亡风险。
基于胰高血糖素样肽1(glucagon-like peptide-1,GLP-1)的糖尿病治疗通过增加葡萄糖依赖性胰岛素分泌、减慢胃排空、降低餐后高血糖、减少食物摄入量等机制来安全有效的控制血糖,同时还能获得减肥和心血管保护等多方面的临床获益作用。基于GLP-1的糖尿病治疗有望代替胰岛素成为治疗糖尿病的一线药物。
目前主要的基于GLP-1的糖尿病治疗包括GLP-1受体激动剂和二肽基肽酶4(dipeptidyl peptidase-4,DPP-4)抑制剂,均以提高GLP-1有效浓度为目标。其中以Liraglutide(利拉鲁泰),Semaglutide(索马鲁泰)和Dulaglutide(度拉鲁肽)为代表的新的长效胰高血糖素样肽-1(GLP-1)受体激动剂在临床疗效及患者依从性等方面表现突出,已经作为一线用药进入欧美国家糖尿病指南。
但GLP-1及其类似物在体内很快被DPP-4降解,半衰期较短。虽然GLP-1类似物显著延长了GLP-1在体内的作用时间,但仍需频繁(1-7天)皮下给药。并且由于多肽药物的研制、生产、使用成本均远高于小分子糖尿病治疗药物,GLP-1类似物糖尿病治疗并不具有成本优势,长效以至于终身的GLP-1治疗对于部分中重度以及难控制血糖的糖尿病患者,具有现实及紧迫的临床需求。
病毒及非病毒载体递送的GLP-1类似物基因治疗技术路线,从理论上解决了GLP-1长期甚至终身表达的目标,可以一劳永逸提高GLP-1有效浓度,对于中重度及难控制糖尿病患者具有长期临床获益的巨大价值。然而多肽的基因表达载体的表达效率远低于大分子蛋白的基因表达载体,作为30个氨基酸左右的GLP-1及其类似物,过小的表达框架使其在任何病毒及非病毒载体上的有效表达均有不小的难度。同时较短血浆半衰期进一步增加了GLP-1及其类似物达到有效血液浓度的障碍。目前为止,尚未有任何病毒及非病毒载体递送的GLP-1类似物基因治疗技术路线获得有效的临床进展。
针对以上问题,基于病毒及非病毒载体递送的基因治疗技术路线的药物研发主要需要在以下两个方面取得突破:
如何以重组病毒或非病毒载体递送的基因载体,以基因组整合或非整合的方式在机体内长期稳定并高效的表达GLP-1及其类似物,使2型糖尿病及相关疾病患者一次治疗长期甚至终身获益,同时减少生产和使用成本。
发明内容
鉴于以上所述现有技术的缺点,本发明的目的在于提供一种用于糖代谢相关疾病基因治疗的核酸构建体,用于解决现有技术中的问题。
为实现上述目的及其他相关目的,本发明提供一种核酸构建体,所述核酸构建体包括编码GLP-1或其类似物的多核苷酸,所述GLP-1或其类似物的总数量为两个以上,GLP-1与GLP-1之间、GLP-1与其类似物之间或GLP-1类似物与GLP-1与类似物之间通过编码连接肽的多核苷酸连接。
本发明还提供一种慢病毒,所述慢病毒由所述核酸构建体经病毒包装而成。
本发明还提供一种慢病毒载体系统,所述慢病毒载体系统包括所述的核酸构建体以及辅助质粒。
本发明还提供一种腺相关病毒载体系统,所述腺相关病毒载体系统包括所述的核酸构建体以及辅助质粒。
本发明还提供一种腺相关病毒,所述腺相关病毒由所述腺相关病毒载体系统经病毒包装而成。
本发明还提供一种细胞系,所述细胞系为经所述慢病毒或腺相关病毒感染的细胞系。
本发明还提供所述核酸构建体、慢病毒、慢病毒载体系统、腺相关病毒、腺相关病毒载体系统、细胞系在制备预防、治疗糖代谢相关疾病的产品中的用途。
如上所述,本发明的用于糖代谢相关疾病基因治疗的核酸构建体,具有以下有益效果:本发明创造了含串联共价二聚体或多聚体形式的GLP-1或其类似物的表达框架的核酸构建体,该核酸构建体在体内外均可用于高效表达具有活性的GLP-1及其类似物,这为极具潜力的2型糖尿病、机体代谢紊乱、糖尿病相关并发症以及肥胖症等糖代谢相关疾病的基因治疗药物开发了技术路线。本发明的应用范围包括各种形式基于GLP-1及其类似物的糖代谢相关疾病的基因治疗,例如所述构建体能用于病毒载体或非病毒载体递送的糖代谢相关疾病的基因治疗药物的临床研究及新药研发及生产。
附图说明
图1A显示为本发明的pKL-kan-lenti-EF1α-WPRE质粒图谱。
图1B显示为本发明的pAAV-MCS-CMV-EGFP(反)质粒图谱。
图2显示为表达GLP1受体激动剂的核酸构建体的表达框架的设计。
图3A显示为表达GLP1受体激动剂的核酸构建体(慢病毒载体)的设计。
图3B显示为表达GLP1受体激动剂的核酸构建体(腺相关病毒载体)的设计。
图4A显示为表达GLP1受体激动剂的慢病毒转导细胞后对GLP1受体激动剂的表达(Westernblotting)还原凝胶电泳检测结果。
图4B显示为表达GLP1受体激动剂的慢病毒转导细胞后对GLP1受体激动剂的表达(Westernblotting)非还原凝胶电泳检测结果。
图5显示为表达GLP1受体激动剂的慢病毒转导的细胞培养上清对带有GLP1受体细胞的激活效果。A1-A4(即第一行)分别为GLP1(7-37)8nM,40nM,200nM,1000nM;B1-B4(即第二行)分别为KLDi02不同转导体积:2μL、5μL、10μL、20μL;C1-C4(即第三行)分别为KLDi01不同转导体积:2μL、5μL、10μL、20μL。
图6显示为DB/DB小鼠接受表达GLP1受体激动剂的腺相关病毒后的血糖浓度。
图7显示为C57BL/6小鼠接受表达GLP1受体激动剂的腺相关病毒后的糖耐量。
具体实施方式
除非下文另外定义,本发明所提及的所有技术和科学用语具有本发明所属领域的技术人员通常理解的意义。
术语“核酸构建体”是指可以被引入靶细胞或组织中的人工构建的核酸区段,所述核酸构建体可以为慢病毒载体或腺相关病毒载体,慢病毒载体或腺相关病毒载体包括载体骨架即空载体与表达框架。
术语“表达框架”是指具有编码蛋白质潜能的序列。
本发明提供一种核酸构建体,所述核酸构建体包括编码GLP-1或其类似物的多核苷酸,所述GLP-1或其类似物的总数量为两个以上,GLP-1与GLP-1之间、GLP-1与其类似物之间或GLP-1类似物与GLP-1与类似物之间通过编码连接肽的多核苷酸连接。
在一种实施方式中,GLP-1类似物为GLP-1N端的7-36或7-37个氨基酸肽链。这一部分氨基酸是GLP-1生物活性部分,因此药物研发等均集中在这些氨基酸序列。GLP-1类似物还可以为在GLP-1N端的7-36或7-37个氨基酸中的个别氨基酸的替换、删除、增加,或者个别氨基酸与不同化合物连接。
所述GLP-1或其类似物的总数量例如可以是两个、三个、四个、五个或更多个。
所述GLP-1或其类似物的总数量的意思选自以下任一:
1)当构建体中含GLP-1而不含其类似物时,GLP-1的总数量为两个以上;
2)当构建体中含GLP-1类似物而不含GLP-1时,GLP-1类似物的总数量为两个以上;
3)当构建体中既含GLP-1又含GLP-1类似物时,GLP-1和GLP-1类似物的数量为两个以上。
所述的核酸构建体中GLP-1或其类似物之间的连接方式如下:信号肽——GLP-1或GLP-1类似物——连接肽——GLP-1或GLP-1类似物——连接肽——GLP-1或GLP-1类似物,其中GLP-1或GLP-1类似物——连接肽——GLP-1或GLP-1类似物单元的个数为一个或多个,例如可以是两个、三个、四个、五个或更多个。
所述连接肽由侧链小的氨基酸组成。例如:(Gly)4,GSGGSG,GSGGSGG GSGGSGGG,GGGGSGGG,(GGGGS)3。
所述核酸构建体可以有效的在体内表达胰高血糖素样肽1(glucagon-likepeptide-1,GLP-1)或其类似物。所述核酸构建体表达的GLP1或其类似物作为GLP1受体的激动剂,所以所述核酸构建体亦可称为表达GLP1受体激动剂的构建体。
所述核酸构建体为用于糖代谢相关疾病基因治疗的核酸构建体。
具体的,糖代谢相关疾病包括糖尿病或其并发症、肥胖症。
在一种实施方式中,所述糖尿病为2型糖尿病。
所述核酸构建体包括如SEQ ID NO:4或SEQ ID NO:6所示的表达框架。所述表达框架表达的GLP1或其类似物作为GLP1受体的激动剂。或者包括跟SEQ ID NO.4或SEQ ID NO.6比较至少具有75%,80%,85%,90%,95%,96%,97%,98%,或99%同源性的核苷酸序列。
所述核酸构建体中不包括能够表达除GLP1或其类似物外的蛋白或多肽的多核苷酸。即,所述核酸构建体能够有效表达的蛋白或多肽仅有GLP1或其类似物。
所述核酸构建体编码的多肽氨基酸序列如SEQ ID NO:3或SEQ ID NO:5所示。
所述核酸构建体被用于生产基于病毒的基因治疗载体,优选的所述基因治疗载体为慢病毒载体或腺相关病毒载体。
所述核酸构建体为病毒载体或非病毒载体。
在一些实施方式中,所述核酸构建体为慢病毒载体。所述慢病毒载体中载体骨架可以现有技术中的载体骨架。
在一种实施方式中,所述慢病毒载体的核苷酸序列如SEQ ID NO.12或SEQ IDNO.13所示。或者包括跟SEQ ID NO.12或SEQ ID NO.13比较至少具有75%,80%,85%,90%,95%,96%,97%,98%,或99%同源性的核苷酸序列。
在一些实施方式中,所述核酸构建体为腺相关病毒载体。所述腺相关病毒载体中载体骨架可以现有技术中的载体骨架。
在一种实施方式中,所述腺相关病毒载体的核苷酸序列如SEQ ID NO.17或SEQ IDNO.18所示。或者包括跟SEQ ID NO.17或SEQ ID NO.18比较至少具有75%,80%,85%,90%,95%,96%,97%,98%,或99%同源性的核苷酸序列。
本发明还提供一种慢病毒载体系统,所述慢病毒载体系统包括所述的核酸构建体以及辅助质粒。
更进一步的,所述辅助质粒编码gag和pol蛋白的一个或者多个核苷酸序列以及其它必需的病毒包装组件核苷酸序列,所述辅助质粒可以包括包装质粒和包膜质粒。
进一步的,所述慢病毒载体系统还包括宿主细胞。所述宿主细胞携带慢病毒载体。所述宿主细胞可以选自本领域各种可适用的宿主细胞,只要不对本发明的发明目的产生限制即可。具体的可适用的细胞可以为生产慢病毒的细胞,例如可以为293T细胞。
本发明还提供一种慢病毒,所述慢病毒由所述慢病毒载体系统经病毒包装而成。
本发明还提供一种腺相关病毒载体系统,所述腺相关病毒载体系统包括所述的核酸构建体以及辅助质粒。
更进一步的,所述辅助质粒编码gag和pol蛋白的一个或者多个核苷酸序列以及其它必需的病毒包装组件核苷酸序列,所述辅助质粒可以包括包装质粒和包膜质粒。
进一步的,所述腺相关病毒载体系统还包括宿主细胞。所述宿主细胞携带腺相关病毒载体。所述宿主细胞可以选自本领域各种可适用的宿主细胞,只要不对本发明的发明目的产生限制即可。具体的可适用的细胞可以为生产腺相关病毒的细胞,例如可以为293T细胞。
本发明还提供一种腺相关病毒,所述腺相关病毒由所述腺相关病毒载体系统经病毒包装而成。
本发明还提供一种细胞系,所述细胞系为经所述慢病毒或腺相关病毒感染的细胞系。
该细胞系可以作为生物制剂用于制备预防或治疗神经退行性疾病的产品。
本发明还提供所述核酸构建体、慢病毒、慢病毒载体系统、腺相关病毒、腺相关病毒载体系统、细胞系在制备预防、治疗糖代谢相关疾病的产品中的用途。
具体的,糖代谢相关疾病包括糖尿病或其并发症、肥胖症。
在一种实施方式中,所述糖尿病为2型糖尿病。
以下通过特定的具体实例说明本发明的实施方式,本领域技术人员可由本说明书所揭露的内容轻易地了解本发明的其他优点与功效。本发明还可以通过另外不同的具体实施方式加以实施或应用,本说明书中的各项细节也可以基于不同观点与应用,在没有背离本发明的精神下进行各种修饰或改变。
在进一步描述本发明具体实施方式之前,应理解,本发明的保护范围不局限于下述特定的具体实施方案;还应当理解,本发明实施例中使用的术语是为了描述特定的具体实施方案,而不是为了限制本发明的保护范围;在本发明说明书和权利要求书中,除非文中另外明确指出,单数形式“一个”、“一”和“这个”包括复数形式。
当实施例给出数值范围时,应理解,除非本发明另有说明,每个数值范围的两个端点以及两个端点之间任何一个数值均可选用。除非另外定义,本发明中使用的所有技术和科学术语与本技术领域技术人员通常理解的意义相同。除实施例中使用的具体方法、设备、材料外,根据本技术领域的技术人员对现有技术的掌握及本发明的记载,还可以使用与本发明实施例中所述的方法、设备、材料相似或等同的现有技术的任何方法、设备和材料来实现本发明。
以下实施例包括:构建一系列能表达作为GLP1受体激动剂的多肽及融合多肽的核酸构建体,并将其克隆到重组腺相关病毒或重组慢病毒载体中。在293T细胞中,包装相应的腺相关病毒和慢病毒载体,以一定生物学滴度病毒感染293T细胞,以及分化及未分化肌肉细胞系。用细胞上清中产生的多肽及融合多肽定量、定性检测,同时在报告基因细胞系的感染活性分析中检验其对GLP-1受体(GLP-1R)的结合和中和活性。再将上述包含多肽及融合多肽表达框架的腺相关病毒和慢病毒载体经纯化后,经静脉或肌肉注射的方式递送至野生型或糖尿病模型小鼠体内,在不同时间点检测受试小鼠的GLP-1多肽及融合多肽的血浆浓度,抗GLP-1多肽抗体,以及餐后血糖。具体的各实验步骤如下:
实施例1:表达GLP1受体激动剂的核酸构建体表达框架的结构设计
如图2所示,GLP1受体激动剂携带有信号肽序列,图2所示的整体序列在细胞中以一个完整的前体分子的形式被翻译出来,并被分泌出细胞。前体分子包括GLP1、三个串联的柔性单元氨基酸组成的连接肽和辅助肽;其中辅助肽可以为GLP1或GLP1-连接肽-GLP1,也可以为人源抗体IgG1CH2和IgG1CH3或其他结构。本发明中应用的GLP1受体激动剂分子的结构为:
1.单体基因表达框架(GLP-1),由N端信号肽MALLTNLLPLCCLALLALPAQS(SEQ IDNO:30)以及单个GLP-1(7-37)基因HAEGTFTSDVSSYLE GQAAKEFIAWLVKGRG(SEQ ID NO:31)组成。构建的GLP1受体激动剂表达框架GLP1蛋白序列见SEQ ID NO:1,DNA序列见SEQ ID NO:2。
2.双GLP-1融合多肽基因表达框架(GLP1-GLP1),由N端信号肽MALLTNLLPLCCLALLALPAQS(SEQ ID NO:30),GLP-1(7-37)基因HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRG(SEQ ID NO:31),三个串联的柔性单元氨基酸组成的连接肽GGGGSGGGGSGGGGS(SEQ ID NO:32),GLP-1(7-37)基因HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRG(SEQ ID NO:31)组成。构建的GLP1受体激动剂表达框架KLDi02的蛋白序列见SEQ ID NO:3,DNA序列见SEQ ID NO:4。
3.叁GLP-1融合多肽基因表达框架(GLP1-GLP1-GLP1),由N端信号肽MALLTNLLPLCCLALLALPAQS(SEQ ID NO:30),GLP-1(7-37)基因HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRG(SEQ ID NO:31),连接肽GGGGSGGGGSGGGGS(SEQ IDNO:32),GLP-1(7-37)基因HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRG(SEQ ID NO:31),连接肽GGGGSGGGGSGGGGS(SEQ ID NO:32),GLP-1(7-37)基因HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRG(SEQ ID NO:31)组成。构建的GLP1受体激动剂基因表达框架KLDi03的蛋白序列见SEQ IDNO:5,DNA序列见SEQ ID NO:6。
4.GLP-1-Fc片段融合蛋白基因表达框架(GLP-1-Fc),由N端信号肽MXXXX,单个GLP-1(7-37)基因HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRG(SEQ ID NO:31)以及人源抗体IgG1CH2和IgG1CH3组成。构建的GLP1受体激动剂基因表达框架KLDi01的蛋白序列见SEQ IDNO:7,DNA序列见SEQ ID NO:8。此构建体将一个GLP-1类似物基因与抗体Fc片段编码序列相连,利用类抗体分子二聚体的形成,优化GLP-1类似物基因的表达效率以及增加大分子药物的血浆半衰期。此构建体为现有技术,在申请中作为对照组。
实施例2:表达GLP1受体激动剂的核酸构建体的构建
如图3A所示,将上述GLP1受体激动剂基因表达框架以多片段重组连接的方式克隆到当前应用的最新一代慢病毒载体骨架pKL-kan-lenti-EF1α-WPRE(图1A,核苷酸序列如SEQ ID NO:9)中。所述慢病毒载体骨架包括:5′LTR,其中用CMV启动子替换LTR的启动子区域;ψ包装信号;逆转录病毒输出元件RRE;cPPT;启动子CBH;编码HIV中和抗体片段的多肽的多核苷酸;转录后调节元件为wPRE;PPT;ΔU3 3’LTR;以及poly A信号。实施例1设计的基因表达框架GLP1、GLP1-Fc、GLP1-GLP1和GLP1-GLP1-GLP1经通用生物系统(安徽)有限公司合成后,通过本领域熟知的以同源重组的方法克隆至慢病毒载体骨架pKL-kan-lenti-EF1α-WPRE上的多克隆位点EcoRI/EcoRV之间,克隆完成后以测序确认序列信息,质粒分别命名为pKL-Kan-lenti-EF1α-GLP1(DNA序列见SEQ ID NO:10)、pKL-Kan-lenti-EF1α-KLDi01(DNA序列见SEQ ID NO:11)、pKL-Kan-lenti-EF1α-KLDi02(DNA序列见SEQ ID NO:12)和pKL-Kan-lenti-EF1α-KLDi03(DNA序列见SEQ ID NO:13)。
如图3B所示,将来源于pX261的CBH启动子(DNA序列见SEQ ID NO:14)和存在于pKL-Kan-lenti-EF1α-KLDi01、pKL-Kan-lenti-EF1α-KLDi02和pKL-Kan-lenti-EF1α-KLDi03的GLP1受体激动剂基因表达框架KLDi01、KLDi02、KLDi03以多片段重组连接的方式克隆到当前应用的最新一代腺相关病毒载体骨架pAAV-MCS-CMV-EGFP(反)(序列见SEQ IDNO:15)(见图1B)的多克隆位点MluI/SalI之间。所述腺相关病毒载体骨架包括:AAV2 ITR,启动子CBH;编码GLP1受体激动剂的多核苷酸;SV40 poly A信号,AAV2 ITR。质粒分别命名为pAAV-CBH-KLDi01(DNA序列见SEQ ID NO:16),pAAV-CBH-KLDi02(DNA序列见SEQ ID NO:17),pAAV-CBH-KLDi03(DNA序列见SEQ ID NO:18)。
实施例3:表达GLP1受体激动剂的病毒包装及纯化
以慢病毒载体(pKL-Kan-lenti-EF1α-GLP1、pKL-Kan-lenti-EF1α-KLDi01、pKL-Kan-lenti-EF1α-KLDi02和pKL-Kan-lenti-EF1α-KLDi03)在293T细胞系中进行抗体基因治疗慢病毒载体的包装。将实施2中构建的抗体基因慢病毒载体(pKL-Kan-lenti-CBH-GLP1、pKL-Kan-lenti-EF1α-KLDi01、pKL-Kan-lenti-EF1α-KLDi02和pKL-Kan-lenti-EF1α-KLDi03)、包膜质粒(pKL-Kan-Vsvg,其核苷酸序列如SEQ ID NO:19所示)和包装质粒(pKL-Kan-Rev,其核苷酸序列如SEQ ID NO:20所示;pKL-Kan-GagPol,其核苷酸序列如SEQ IDNO:21所示)混合后同时共转染293T细胞(购自美国模式菌种收集中心(ATCC),ATCC保藏号为CRL-3216),在该293T细胞系中进行HIV中和抗体基因治疗慢病毒的包装。转染方法为PEI阳离子聚合物介导的真核细胞瞬时转染,PEI阳离子聚合物为购自Polysciences的PEI-Max转染试剂(购自Polysciences,货号:24765-1),转染操作参照生产商推荐标准化操作进行,转染规模为15cm细胞培养皿。转染完成48小时后,收获慢病毒载体(转染细胞培养上清),首先在台式吊桶式机上,室温4000rpm离心5分钟去除细胞碎片,再4度10000g离心4小时获得病毒颗粒沉淀,去除离心上清后,以1mL DMEM完全培养基加入病毒颗粒沉淀中,以微量进样器重悬病毒颗粒,并将制备好的病毒重悬液分装冻存于-80度备用。
以AAV表达载体(pAAV-CBH-KLDi01,pAAV-CBH-KLDi02,pAAV-CBH-KLDi03)在293T细胞系中进行抗体基因治疗AAV载体的包装。将实施例2中构建的抗体基因AAV载体(pAAV-CBH-KLDi01,pAAV-CBH-KLDi02,pAAV-CBH-KLDi03)、包膜质粒(AAV2/8,其核苷酸序列如SEQID NO:22所示)和包装质粒(pHelper,其核苷酸序列如SEQ ID NO:23所示)混合后同时共转染293T细胞,在该293T细胞系中进行GLP1受体激动剂基因治疗载体AAV的包装。转染方法为PEI阳离子聚合物介导的真核细胞瞬时转染,PEI阳离子聚合物为购自Polysciences的PEI-Max转染试剂(购自Polysciences,货号:24765-1),转染操作参照生产商推荐标准化操作进行,转染规模为15cm细胞培养皿。转染后7h吸弃上清,更换为25ml产毒培养基。转染完成120小时后,收集上清和细胞,4200rpm离心10min,离心完成后分离上清跟细胞,细胞加裂解液跟核酶,裂解消化1h,10000g离心10min取裂解液上清。将裂解液上清及培养基上清经亲和层析纯化后分装冻存于-80℃备用。
实施例4:GLP1受体激动剂的细胞学表达
将包装好的慢病毒用于感染293T细胞。以慢病毒感染后细胞的培养上清载入SDS-PAGE,以GLP-1抗体(6F117):sc-71150作为1级抗体,以标记的羊抗鼠抗体作为2级抗体。Western blotting结果(图4A和图4B)显示,不同的GLP1受体激动剂慢病毒在体外转导细胞后均能有效地表达GLP1受体激动剂,并将成熟的激动剂蛋白分泌到细胞培养上清中。
实施例5:表达GLP1受体激动剂的慢病毒给药后上清表达的GLP1受体激动剂的功能验证
GLP1受体被GLP1激活后,从细胞膜表面移位到细胞内。EGFP标记蛋白对药物化合物或其他刺激的反应时的细胞易位。GLP1R-EGFP稳转U2OS细胞系可以辅助鉴定GLP1受体激动剂的活性。如果GLP1受体激动剂基因表达载体转导的细胞上清中,分泌表达的GLP1受体激动剂具有完全的功能,将可以观察到GLP1R-EGFP稳转U2OS细胞中绿色荧光集中在细胞核周围。具体的功能验证实验步骤如下:
1.收集慢病毒感染的293T细胞,用PBS清洗细胞后4200rpm,5min,离心收集细胞,重悬以20μL快速抽提溶液(QE DNAExtraction Solution)并用PCR仪运行以下程序以裂解细胞和抽提总DNA。
表1细胞QE裂解PCR程序
温度 | 时间 |
65℃ | 15min |
68℃ | 15min |
95℃ | 10min |
通过本领域熟知的方法,以定量PCR计算出293T细胞感染慢病毒拷贝数(VectorCopy Number,VCN)。
定量PCR使用的引物探针序列为:
LV Forward primer 5’-AGTAAGACCACCGCACAGCA-3’(SEQ ID NO:24)
LV Reverse primer 5’-CCTTGGTGGGTGCTACTCCT-3’(SEQ ID NO:25)
LV probe 5’-CCTCCAGGTCTGAAGATCAGCGGCCGC-3’(SEQ ID NO:26)
HK Forward primer 5’-GCTGTCATCTCTTGTGGGCTGT-3’(SEQ ID NO:27)
HK probe 5’-CCTGTCATGCCCACACAAATCTCTCC-3’(SEQ ID NO:28)
HK Reverse primer 5’-ACTCATGGGAGCTGCTGGTTC-3’(SEQ ID NO:29)
其中LV probe中5’端带有6FAM荧光基团,3’带有TAMRA荧光基团;
HK probe中5’端带有CY5荧光基团,3’带有DGB荧光基团。
定量PCR运行程序为:94℃5min;95℃10s,60℃30s,40cyclers。
VCN结果见表2。
表2慢病毒转导的细胞转导效率(单位:ng/mL)
转导体积 | 2μL | 5μL | 10μL | 20μL |
LV-KLDi02 | 11.96 | 26.54 | 43.11 | 90.51 |
LV-KLDi01 | 12.21 | 12.64 | 22.78 | 44.63 |
2.在这个实验中,使用商业化购买的GLP1(7-37)(GLPBIO,货号:GC30058)为阳性对照。将GLP1受体激动剂基因治疗载体转导的细胞培养上清与阳性对照,分别与GLP1R-EGFP稳转的U2OS细胞(北纳创联,货号:BNCC352040)孵育一段时间。荧光显微镜通过观察EGFP在细胞内的荧光来鉴别GLP1受体激动剂的活性。
结果显示(图5)在体外细胞试验中,GLP1受体激动剂慢病毒转导的细胞培养上清中GLP1受体激动剂与GLP1阳性对照(相对分子量为3355.67)有一样的活性,且生物活性高于8nM GLP1阳性对照。培养上清中的GLP1受体激动剂的活性相对于GLP1等效值可以超过26.85ng/mL。
实施例6:表达GLP1受体激动剂的腺相关病毒对糖尿病模型动物的高血糖药效学数据
由于慢病毒在小鼠肌肉注射的情况下表达框架不能整合到肌肉细胞,所以在小鼠体内几乎没有表达(数据未显示)。因此本申请采用腺相关病毒载体递送的GLP1受体激动剂表达框,通过肌肉注射方法给药。将不同类型的能表达GLP1受体激动剂的腺相关病毒通过肌肉注射给DB/DB小鼠给药后,每周剪尾用血糖仪(罗氏,卓越金)测血糖,比较不同给药组降血糖效果。
结果显示(图6)在体内试验中,pAAV-CBH-KLDi02包装的腺相关病毒作为GLP1受体激动剂给药组小鼠血糖显著低于生理盐水组小鼠。
在给药后第4周、第6周、第8周取血,分离血清,用人GLP1(7-36)ELISA试剂盒(abcam,ab184857)检测血清中GLP1受体激动剂的含量。试验组小鼠(表3)血清中GLP1受体激动剂的浓度显著达到正常人所需GLP1水平。
表3 DB/DB小鼠接受表达GLP1受体激动剂的腺相关病毒后的血清GLP1受体激动剂浓度
实施例7:多肽及融合多肽对糖尿病模型动物的高血糖药效学数据
将不同剂量的GLP1受体激动剂腺相关病毒通过肌肉注射给C57BL/6小鼠给药。在给药5周后进行糖耐量检测,评估GLP1受体激动剂腺相关病毒对正常餐食引起的血糖升高的影响。糖耐量检测:禁食12h,腹腔注射2.0g/kg体重的葡萄糖,在注射葡萄糖后0h,30min,60min,120min,剪尾取血测定血糖。
结果显示(图7)在体内试验中,不同剂量的pAAV-CBH-KLDi02包装的腺相关病毒作为GLP1受体激动剂给药组小鼠的血糖显著低于生理盐水组小鼠,且成剂量正相关。
综合以上实施例,本发明将两个以上的GLP-1或其类似物基因表达框架以柔性单元氨基酸组成的肽连接体(GGGGS)3连接后组成串联共价二聚体或多聚体。实验结果显示,二聚体或多聚体形式的GLP-1或其类似物表达框架经重组病毒载体递送后获得了远高于单体GLP-1类似物表达框架的表达效率。同时实验结果显示,无论是在体外细胞学实验或者体内动物学实验中,二聚体或多聚体形式的GLP-1类似物均具有完全的GLP-1受体结合和激动能力。基于上述表达框架的GLP-1类似物分子经腺相关病毒递送至小鼠体内后显示出有效的血药浓度,同时也显示出积极的动物体内血糖调节效果。
以上的实施例是为了说明本发明公开的实施方案,并不能理解为对本发明的限制。此外,本文所列出的各种修改以及发明中方法的变化,在不脱离本发明的范围和精神的前提下对本领域内的技术人员来说是显而易见的。虽然已结合本发明的多种具体优选实施例对本发明进行了具体的描述,但应当理解,本发明不应仅限于这些具体实施例。事实上,各种如上所述的对本领域内的技术人员来说显而易见的修改来获取发明都应包括在本发明的范围内。
序列表
<110> 康霖生物科技(杭州)有限公司
<120> 一种用于糖代谢相关疾病基因治疗的核酸构建体
<160> 32
<170> SIPOSequenceListing 1.0
<210> 1
<211> 50
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 1
Met Ala Leu Leu Thr Asn Leu Leu Pro Leu Cys Cys Leu Ala Leu Leu
1 5 10 15
Ala Leu Pro Ala Gln Ser His Ala Glu Gly Thr Phe Thr Ser Asp Val
20 25 30
Ser Ser Tyr Leu Glu Gly Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu
35 40 45
Val Lys
50
<210> 2
<211> 162
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 2
atggccctcc ttacgaacct cctgccgttg tgctgcctgg ccctcctcgc cttgccagcg 60
cagagccacg ccgaaggcac gttcacctcc gacgtgagca gctacctgga gggccaggcc 120
gcgaaagagt tcatcgcttg gctcgtcaag ggccggggct ga 162
<210> 3
<211> 99
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 3
Met Ala Leu Leu Thr Asn Leu Leu Pro Leu Cys Cys Leu Ala Leu Leu
1 5 10 15
Ala Leu Pro Ala Gln Ser His Ala Glu Gly Thr Phe Thr Ser Asp Val
20 25 30
Ser Ser Tyr Leu Glu Gly Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu
35 40 45
Val Lys Gly Arg Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
50 55 60
Gly Gly Gly Ser His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser
65 70 75 80
Tyr Leu Glu Gly Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Lys
85 90 95
Gly Arg Gly
<210> 4
<211> 300
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 4
atggccctcc ttacgaacct cctgccgttg tgctgcctgg ccctcctcgc cttgccagcg 60
cagagccacg ccgaaggcac gttcacctcc gacgtgagca gctacctgga gggccaggcc 120
gcgaaagagt tcatcgcttg gctcgtcaag ggccggggcg gaggaggagg aagcggagga 180
ggaggctcag gcggcggcgg ctctcacgcc gaaggcacgt tcacctccga cgtgagcagc 240
tacctggagg gccaggccgc gaaagagttc atcgcttggc tcgtcaaggg ccggggctga 300
<210> 5
<211> 145
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 5
Met Ala Leu Leu Thr Asn Leu Leu Pro Leu Cys Cys Leu Ala Leu Leu
1 5 10 15
Ala Leu Pro Ala Gln Ser His Ala Glu Gly Thr Phe Thr Ser Asp Val
20 25 30
Ser Ser Tyr Leu Glu Gly Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu
35 40 45
Val Lys Gly Arg Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
50 55 60
Gly Gly Gly Ser His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser
65 70 75 80
Tyr Leu Glu Gly Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Lys
85 90 95
Gly Arg Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly
100 105 110
Gly Ser His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu
115 120 125
Glu Gly Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg
130 135 140
Gly
145
<210> 6
<211> 438
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 6
atggccctgc tgaccaatct gctgcctctg tgctgtctgg ccctgctggc cctgcccgct 60
cagtctcacg ccgagggaac attcacttcc gatgtgagca gctacctgga gggccaggcc 120
gccaaggagt tcatcgcctg gctggtgaag ggcagaggcg gcggaggcgg atccggagga 180
ggaggaagcg gcggaggcgg ttcccacgct gagggaacct tcacaagcga cgtgtcctcc 240
tacctggagg gacaggccgc caaagagttt atcgcctggc tcgtgaaggg ccggggcgga 300
ggaggaggtt ccggaggagg tggcagcggc ggaggaggta gccacgctga gggcaccttt 360
acctccgatg tgtcctccta tctggagggc caagccgcca aggaattcat cgcctggctg 420
gtcaagggca ggggctga 438
<210> 7
<211> 297
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 7
Met Ala Leu Leu Thr Asn Leu Leu Pro Leu Cys Cys Leu Ala Leu Leu
1 5 10 15
Ala Leu Pro Ala Gln Ser His Ala Glu Gly Thr Phe Thr Ser Asp Val
20 25 30
Ser Ser Tyr Leu Glu Gly Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu
35 40 45
Val Lys Gly Gly Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
50 55 60
Gly Gly Gly Ser Ala Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys
65 70 75 80
Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro
85 90 95
Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys
100 105 110
Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp
115 120 125
Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu
130 135 140
Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu
145 150 155 160
His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn
165 170 175
Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly
180 185 190
Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu
195 200 205
Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr
210 215 220
Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn
225 230 235 240
Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe
245 250 255
Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn
260 265 270
Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr
275 280 285
Gln Lys Ser Leu Ser Leu Ser Leu Gly
290 295
<210> 8
<211> 894
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 8
atggccctcc ttacgaacct cctgccgttg tgctgcctgg ccctcctcgc cttgccagcg 60
cagagccacg ccgaaggcac gttcacctcc gacgtgagca gctacctgga gggccaggcc 120
gcgaaagagt tcatcgcttg gctcgtcaag ggcggaggag gaggcggagg aagcggcgga 180
ggaggaagcg gaggcggagg tagcgccgag agcaagtacg gccctccttg tcctccctgc 240
cctgccccag aggccgctgg aggaccatcc gtgtttctgt ttccccccaa gcctaaggac 300
accctgatga tcagcaggac ccccgaggtg acctgcgtgg tggtggacgt gagccaggag 360
gaccctgagg tgcagttcaa ttggtacgtg gatggcgtgg aggtgcacaa cgccaagaca 420
aagcccagag aggagcagtt taatagcacc tacagagtgg tgtccgtgct gaccgtgctg 480
caccaggatt ggctgaatgg caaggagtac aagtgcaagg tgtccaataa gggcctgccc 540
agcagcatcg agaagaccat cagcaaggcc aagggccagc ccagagagcc tcaggtgtac 600
acactgcctc cctcccagga ggagatgacc aagaaccagg tgtccctgac ctgtctggtg 660
aagggcttct accccagcga tatcgccgtg gagtgggagt ccaacggcca gcccgagaat 720
aactacaaga ccacccctcc tgtgctggat agcgacggca gctttttcct gtactccaga 780
ctgaccgtgg acaagtctag atggcaggag ggcaacgtgt tttcttgtag cgtgatgcac 840
gaggccctgc acaatcacta cacccagaag tccctgtctc tgagcctggg ctga 894
<210> 9
<211> 7128
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 9
ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 60
acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 120
tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 180
ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc 240
ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 300
ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 360
actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 420
gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc 480
tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 540
caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 600
atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 660
acgttaaggg attttggtca tgaagcgctt ttgaagctcg gatccgaaca aacgacccaa 720
cacccgtgcg ttttattctg tctttttatt gccgatcccc tcagaagaac tcgtcaagaa 780
ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc 840
ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac gctatgtcct 900
gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag cggccatttt 960
ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc tcgccgtcgg 1020
gcatgctcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga tgctcttcgt 1080
ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc tcgatgcgat 1140
gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg 1200
catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg agatcctgcc 1260
ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagcacag 1320
ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcttgcagtt 1380
cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca 1440
gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata 1500
gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa 1560
acgatcctca tcctgtctct tgatcagagc ttgatcccct gcgccatcag atccttggcg 1620
gcaagaaagc catccagttt actttgcagg gcttcccaac cttaccagag gcctgcgccg 1680
cggccagctg gctagcaatt cccgggttaa ctctagagac attgattatt gactagttat 1740
taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca 1800
taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca 1860
ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg 1920
gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg 1980
ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc 2040
ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg 2100
atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca 2160
agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt 2220
ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg 2280
gaggtctata taagcagagc tcgtttagtg aaccggggtc tctctggtta gaccagatct 2340
gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc 2400
cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc 2460
tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa 2520
agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac 2580
ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta 2640
gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg 2700
ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg 2760
ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg 2820
ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag aagaacttag 2880
atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga 2940
caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca 3000
gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg 3060
aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa 3120
agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt 3180
tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca 3240
gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc 3300
aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg 3360
ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac 3420
tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga 3480
tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa 3540
tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg 3600
aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata 3660
taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac 3720
tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc 3780
caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca 3840
gagacagatc cattcgatta gtgaacggat ctcgacggta tcggttaact tttaaaagaa 3900
aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca 3960
tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttatc gatcacgaga 4020
ctagcctcga ggcatgcctg caggaattcg ctccggtgcc cgtcagtggg cagagcgcac 4080
atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgaaccg gtgcctagag 4140
aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga 4200
gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg 4260
gtttgccgcc agaacacagg taagtgccgt gtgtggttcc cgcgggcctg gcctctttac 4320
gggttatggc ccttgcgtgc cttgaattac ttccacgccc ctggctgcag tacgtgattc 4380
ttgatcccga gcttcgggtt ggaagtgggt gggagagttc gaggccttgc gcttaaggag 4440
ccccttcgcc tcgtgcttga gttgaggcct ggcttgggcg ctggggccgc cgcgtgcgaa 4500
tctggtggca ccttcgcgcc tgtctcgctg ctttcgataa gtctctagcc atttaaaatt 4560
tttgatgacc tgctgcgacg ctttttttct ggcaagatag tcttgtaaat gcgggccaag 4620
atctgcacac tggtatttcg gtttttgggg ccgcgggcgg cgacggggcc cgtgcgtccc 4680
agcgcacatg ttcggcgagg cggggcctgc gagcgcggcc accgagaatc ggacgggggt 4740
agtctcaagc tggccggcct gctctggtgc ctggcctcgc gccgccgtgt atcgccccgc 4800
cctgggcggc aaggctggcc cggtcggcac cagttgcgtg agcggaaaga tggccgcttc 4860
ccggccctgc tgcagggagc tcaaaatgga ggacgcggcg ctcgggagag cgggcgggtg 4920
agtcacccac acaaaggaaa agggcctttc cgtcctcagc cgtcgcttca tgtgactcca 4980
cggagtaccg ggcgccgtcc aggcacctcg attagttctc gagcttttgg agtacgtcgt 5040
ctttaggttg gggggagggg ttttatgcga tggagtttcc ccacactgag tgggtggaga 5100
ctgaagttag gccagcttgg cacttgatgt aattctcctt ggaatttgcc ctttttgagt 5160
ttggatcttg gttcattctc aagcctcaga cagtggttca aagttttttt cttccatttc 5220
aggtgtcgtg aggatctatt tccggtgaga cccaagctgg ctagctaaac ttacgcgtgc 5280
ctcggatcct ccagtgtggt gtgcagatat ccagcacagt cccgggccga gtctagacgt 5340
ttaaacccgc tgatcaggtc gacaatcaac ctctggatta caaaatttgt gaaagattga 5400
ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt 5460
tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt 5520
tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg 5580
tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg 5640
ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc 5700
gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc 5760
tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct 5820
tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg 5880
ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg 5940
ccgcctcccc gcggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca 6000
ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatct 6060
gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg 6120
ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt 6180
gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt 6240
gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc 6300
aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa 6360
ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg 6420
tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta 6480
actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga 6540
ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag 6600
tagtgaggag gcttttttgg aggccgctag cgtcgaccat tacttattgt tttagctgtc 6660
ctcatgaatg tcttttcact acccatttgc ttatcctgca tctctcagcc ttgactccac 6720
tcagttctct tgcttagaga taccaccttt cccctgaagt gttccttcca tgttttacgg 6780
cgagatggtt tctcctcgcc tggccactca gccttagttg tctctgttgt cttatagagg 6840
tctacttgaa gaaggaaaaa cagggggcat ggtttgactg tcctgtgagc ccttcttccc 6900
tgcctccccc actcacagtg acccggaatc cctcgacatg gcagtctagc actagtgcgg 6960
ccgcagatct gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 7020
ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 7080
aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaa 7128
<210> 10
<211> 7269
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 10
ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 60
acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 120
tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 180
ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc 240
ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 300
ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 360
actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 420
gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc 480
tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 540
caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 600
atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 660
acgttaaggg attttggtca tgaagcgctt ttgaagctcg gatccgaaca aacgacccaa 720
cacccgtgcg ttttattctg tctttttatt gccgatcccc tcagaagaac tcgtcaagaa 780
ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc 840
ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac gctatgtcct 900
gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag cggccatttt 960
ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc tcgccgtcgg 1020
gcatgctcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga tgctcttcgt 1080
ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc tcgatgcgat 1140
gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg 1200
catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg agatcctgcc 1260
ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagcacag 1320
ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcttgcagtt 1380
cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca 1440
gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata 1500
gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa 1560
acgatcctca tcctgtctct tgatcagagc ttgatcccct gcgccatcag atccttggcg 1620
gcaagaaagc catccagttt actttgcagg gcttcccaac cttaccagag gcctgcgccg 1680
cggccagctg gctagcaatt cccgggttaa ctctagagac attgattatt gactagttat 1740
taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca 1800
taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca 1860
ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg 1920
gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg 1980
ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc 2040
ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg 2100
atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca 2160
agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt 2220
ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg 2280
gaggtctata taagcagagc tcgtttagtg aaccggggtc tctctggtta gaccagatct 2340
gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc 2400
cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc 2460
tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa 2520
agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac 2580
ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta 2640
gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg 2700
ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg 2760
ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg 2820
ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag aagaacttag 2880
atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga 2940
caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca 3000
gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg 3060
aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa 3120
agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt 3180
tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca 3240
gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc 3300
aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg 3360
ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac 3420
tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga 3480
tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa 3540
tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg 3600
aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata 3660
taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac 3720
tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc 3780
caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca 3840
gagacagatc cattcgatta gtgaacggat ctcgacggta tcggttaact tttaaaagaa 3900
aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca 3960
tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttatc gatcacgaga 4020
ctagcctcga ggcatgcctg caggaattcg ctccggtgcc cgtcagtggg cagagcgcac 4080
atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgaaccg gtgcctagag 4140
aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga 4200
gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg 4260
gtttgccgcc agaacacagg taagtgccgt gtgtggttcc cgcgggcctg gcctctttac 4320
gggttatggc ccttgcgtgc cttgaattac ttccacgccc ctggctgcag tacgtgattc 4380
ttgatcccga gcttcgggtt ggaagtgggt gggagagttc gaggccttgc gcttaaggag 4440
ccccttcgcc tcgtgcttga gttgaggcct ggcttgggcg ctggggccgc cgcgtgcgaa 4500
tctggtggca ccttcgcgcc tgtctcgctg ctttcgataa gtctctagcc atttaaaatt 4560
tttgatgacc tgctgcgacg ctttttttct ggcaagatag tcttgtaaat gcgggccaag 4620
atctgcacac tggtatttcg gtttttgggg ccgcgggcgg cgacggggcc cgtgcgtccc 4680
agcgcacatg ttcggcgagg cggggcctgc gagcgcggcc accgagaatc ggacgggggt 4740
agtctcaagc tggccggcct gctctggtgc ctggcctcgc gccgccgtgt atcgccccgc 4800
cctgggcggc aaggctggcc cggtcggcac cagttgcgtg agcggaaaga tggccgcttc 4860
ccggccctgc tgcagggagc tcaaaatgga ggacgcggcg ctcgggagag cgggcgggtg 4920
agtcacccac acaaaggaaa agggcctttc cgtcctcagc cgtcgcttca tgtgactcca 4980
cggagtaccg ggcgccgtcc aggcacctcg attagttctc gagcttttgg agtacgtcgt 5040
ctttaggttg gggggagggg ttttatgcga tggagtttcc ccacactgag tgggtggaga 5100
ctgaagttag gccagcttgg cacttgatgt aattctcctt ggaatttgcc ctttttgagt 5160
ttggatcttg gttcattctc aagcctcaga cagtggttca aagttttttt cttccatttc 5220
aggtgtcgtg aggatctatt tccggtgaga cccaagctgg ctagctaaac ttacgcgtgc 5280
caccatggcc ctccttacga acctcctgcc gttgtgctgc ctggccctcc tcgccttgcc 5340
agcgcagagc cacgccgaag gcacgttcac ctccgacgtg agcagctacc tggagggcca 5400
ggccgcgaaa gagttcatcg cttggctcgt caagggccgg ggctgagata tccagcacag 5460
tcccgggccg agtctagacg tttaaacccg ctgatcaggt cgacaatcaa cctctggatt 5520
acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg 5580
gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct 5640
cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc 5700
aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca 5760
ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac 5820
tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt 5880
ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct 5940
ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc 6000
cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga 6060
cgagtcggat ctccctttgg gccgcctccc cgcggtacct ttaagaccaa tgacttacaa 6120
ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca 6180
ctcccaacga agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat 6240
ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt 6300
gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc 6360
cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat 6420
tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat 6480
tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt 6540
tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg 6600
gctctagcta tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc 6660
cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg 6720
gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggccgcta gcgtcgacca 6780
ttacttattg ttttagctgt cctcatgaat gtcttttcac tacccatttg cttatcctgc 6840
atctctcagc cttgactcca ctcagttctc ttgcttagag ataccacctt tcccctgaag 6900
tgttccttcc atgttttacg gcgagatggt ttctcctcgc ctggccactc agccttagtt 6960
gtctctgttg tcttatagag gtctacttga agaaggaaaa acagggggca tggtttgact 7020
gtcctgtgag cccttcttcc ctgcctcccc cactcacagt gacccggaat ccctcgacat 7080
ggcagtctag cactagtgcg gccgcagatc tgcttcctcg ctcactgact cgctgcgctc 7140
ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 7200
agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 7260
ccgtaaaaa 7269
<210> 11
<211> 8001
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 11
ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 60
acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 120
tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 180
ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc 240
ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 300
ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 360
actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 420
gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc 480
tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 540
caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 600
atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 660
acgttaaggg attttggtca tgaagcgctt ttgaagctcg gatccgaaca aacgacccaa 720
cacccgtgcg ttttattctg tctttttatt gccgatcccc tcagaagaac tcgtcaagaa 780
ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc 840
ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac gctatgtcct 900
gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag cggccatttt 960
ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc tcgccgtcgg 1020
gcatgctcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga tgctcttcgt 1080
ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc tcgatgcgat 1140
gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg 1200
catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg agatcctgcc 1260
ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagcacag 1320
ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcttgcagtt 1380
cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca 1440
gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata 1500
gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa 1560
acgatcctca tcctgtctct tgatcagagc ttgatcccct gcgccatcag atccttggcg 1620
gcaagaaagc catccagttt actttgcagg gcttcccaac cttaccagag gcctgcgccg 1680
cggccagctg gctagcaatt cccgggttaa ctctagagac attgattatt gactagttat 1740
taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca 1800
taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca 1860
ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg 1920
gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg 1980
ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc 2040
ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg 2100
atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca 2160
agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt 2220
ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg 2280
gaggtctata taagcagagc tcgtttagtg aaccggggtc tctctggtta gaccagatct 2340
gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc 2400
cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc 2460
tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa 2520
agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac 2580
ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta 2640
gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg 2700
ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg 2760
ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg 2820
ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag aagaacttag 2880
atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga 2940
caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca 3000
gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg 3060
aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa 3120
agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt 3180
tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca 3240
gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc 3300
aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg 3360
ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac 3420
tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga 3480
tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa 3540
tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg 3600
aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata 3660
taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac 3720
tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc 3780
caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca 3840
gagacagatc cattcgatta gtgaacggat ctcgacggta tcggttaact tttaaaagaa 3900
aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca 3960
tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttatc gatcacgaga 4020
ctagcctcga ggcatgcctg caggaattcg ctccggtgcc cgtcagtggg cagagcgcac 4080
atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgaaccg gtgcctagag 4140
aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga 4200
gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg 4260
gtttgccgcc agaacacagg taagtgccgt gtgtggttcc cgcgggcctg gcctctttac 4320
gggttatggc ccttgcgtgc cttgaattac ttccacgccc ctggctgcag tacgtgattc 4380
ttgatcccga gcttcgggtt ggaagtgggt gggagagttc gaggccttgc gcttaaggag 4440
ccccttcgcc tcgtgcttga gttgaggcct ggcttgggcg ctggggccgc cgcgtgcgaa 4500
tctggtggca ccttcgcgcc tgtctcgctg ctttcgataa gtctctagcc atttaaaatt 4560
tttgatgacc tgctgcgacg ctttttttct ggcaagatag tcttgtaaat gcgggccaag 4620
atctgcacac tggtatttcg gtttttgggg ccgcgggcgg cgacggggcc cgtgcgtccc 4680
agcgcacatg ttcggcgagg cggggcctgc gagcgcggcc accgagaatc ggacgggggt 4740
agtctcaagc tggccggcct gctctggtgc ctggcctcgc gccgccgtgt atcgccccgc 4800
cctgggcggc aaggctggcc cggtcggcac cagttgcgtg agcggaaaga tggccgcttc 4860
ccggccctgc tgcagggagc tcaaaatgga ggacgcggcg ctcgggagag cgggcgggtg 4920
agtcacccac acaaaggaaa agggcctttc cgtcctcagc cgtcgcttca tgtgactcca 4980
cggagtaccg ggcgccgtcc aggcacctcg attagttctc gagcttttgg agtacgtcgt 5040
ctttaggttg gggggagggg ttttatgcga tggagtttcc ccacactgag tgggtggaga 5100
ctgaagttag gccagcttgg cacttgatgt aattctcctt ggaatttgcc ctttttgagt 5160
ttggatcttg gttcattctc aagcctcaga cagtggttca aagttttttt cttccatttc 5220
aggtgtcgtg aggatctatt tccggtgaga cccaagctgg ctagctaaac ttacgcgtgc 5280
caccatggcc ctccttacga acctcctgcc gttgtgctgc ctggccctcc tcgccttgcc 5340
agcgcagagc cacgccgaag gcacgttcac ctccgacgtg agcagctacc tggagggcca 5400
ggccgcgaaa gagttcatcg cttggctcgt caagggcgga ggaggaggcg gaggaagcgg 5460
cggaggagga agcggaggcg gaggtagcgc cgagagcaag tacggccctc cttgtcctcc 5520
ctgccctgcc ccagaggccg ctggaggacc atccgtgttt ctgtttcccc ccaagcctaa 5580
ggacaccctg atgatcagca ggacccccga ggtgacctgc gtggtggtgg acgtgagcca 5640
ggaggaccct gaggtgcagt tcaattggta cgtggatggc gtggaggtgc acaacgccaa 5700
gacaaagccc agagaggagc agtttaatag cacctacaga gtggtgtccg tgctgaccgt 5760
gctgcaccag gattggctga atggcaagga gtacaagtgc aaggtgtcca ataagggcct 5820
gcccagcagc atcgagaaga ccatcagcaa ggccaagggc cagcccagag agcctcaggt 5880
gtacacactg cctccctccc aggaggagat gaccaagaac caggtgtccc tgacctgtct 5940
ggtgaagggc ttctacccca gcgatatcgc cgtggagtgg gagtccaacg gccagcccga 6000
gaataactac aagaccaccc ctcctgtgct ggatagcgac ggcagctttt tcctgtactc 6060
cagactgacc gtggacaagt ctagatggca ggagggcaac gtgttttctt gtagcgtgat 6120
gcacgaggcc ctgcacaatc actacaccca gaagtccctg tctctgagcc tgggctgaga 6180
tatccagcac agtcccgggc cgagtctaga cgtttaaacc cgctgatcag gtcgacaatc 6240
aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt 6300
ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg 6360
ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc 6420
ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt 6480
ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg 6540
ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg 6600
gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct 6660
gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc 6720
cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc 6780
ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcggtac ctttaagacc 6840
aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga 6900
agggctaatt cactcccaac gaagacaaga tctgcttttt gcttgtactg ggtctctctg 6960
gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc 7020
tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg 7080
taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc 7140
atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag 7200
gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac 7260
aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc 7320
ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccatcccgcc cctaactccg 7380
cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc 7440
gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccgc 7500
tagcgtcgac cattacttat tgttttagct gtcctcatga atgtcttttc actacccatt 7560
tgcttatcct gcatctctca gccttgactc cactcagttc tcttgcttag agataccacc 7620
tttcccctga agtgttcctt ccatgtttta cggcgagatg gtttctcctc gcctggccac 7680
tcagccttag ttgtctctgt tgtcttatag aggtctactt gaagaaggaa aaacaggggg 7740
catggtttga ctgtcctgtg agcccttctt ccctgcctcc cccactcaca gtgacccgga 7800
atccctcgac atggcagtct agcactagtg cggccgcaga tctgcttcct cgctcactga 7860
ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat 7920
acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca 7980
aaaggccagg aaccgtaaaa a 8001
<210> 12
<211> 7407
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 12
ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 60
acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 120
tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 180
ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc 240
ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 300
ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 360
actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 420
gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc 480
tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 540
caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 600
atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 660
acgttaaggg attttggtca tgaagcgctt ttgaagctcg gatccgaaca aacgacccaa 720
cacccgtgcg ttttattctg tctttttatt gccgatcccc tcagaagaac tcgtcaagaa 780
ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc 840
ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac gctatgtcct 900
gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag cggccatttt 960
ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc tcgccgtcgg 1020
gcatgctcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga tgctcttcgt 1080
ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc tcgatgcgat 1140
gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg 1200
catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg agatcctgcc 1260
ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagcacag 1320
ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcttgcagtt 1380
cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca 1440
gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata 1500
gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa 1560
acgatcctca tcctgtctct tgatcagagc ttgatcccct gcgccatcag atccttggcg 1620
gcaagaaagc catccagttt actttgcagg gcttcccaac cttaccagag gcctgcgccg 1680
cggccagctg gctagcaatt cccgggttaa ctctagagac attgattatt gactagttat 1740
taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca 1800
taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca 1860
ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg 1920
gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg 1980
ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc 2040
ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg 2100
atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca 2160
agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt 2220
ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg 2280
gaggtctata taagcagagc tcgtttagtg aaccggggtc tctctggtta gaccagatct 2340
gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc 2400
cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc 2460
tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa 2520
agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac 2580
ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta 2640
gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg 2700
ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg 2760
ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg 2820
ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag aagaacttag 2880
atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga 2940
caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca 3000
gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg 3060
aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa 3120
agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt 3180
tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca 3240
gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc 3300
aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg 3360
ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac 3420
tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga 3480
tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa 3540
tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg 3600
aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata 3660
taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac 3720
tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc 3780
caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca 3840
gagacagatc cattcgatta gtgaacggat ctcgacggta tcggttaact tttaaaagaa 3900
aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca 3960
tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttatc gatcacgaga 4020
ctagcctcga ggcatgcctg caggaattcg ctccggtgcc cgtcagtggg cagagcgcac 4080
atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgaaccg gtgcctagag 4140
aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga 4200
gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg 4260
gtttgccgcc agaacacagg taagtgccgt gtgtggttcc cgcgggcctg gcctctttac 4320
gggttatggc ccttgcgtgc cttgaattac ttccacgccc ctggctgcag tacgtgattc 4380
ttgatcccga gcttcgggtt ggaagtgggt gggagagttc gaggccttgc gcttaaggag 4440
ccccttcgcc tcgtgcttga gttgaggcct ggcttgggcg ctggggccgc cgcgtgcgaa 4500
tctggtggca ccttcgcgcc tgtctcgctg ctttcgataa gtctctagcc atttaaaatt 4560
tttgatgacc tgctgcgacg ctttttttct ggcaagatag tcttgtaaat gcgggccaag 4620
atctgcacac tggtatttcg gtttttgggg ccgcgggcgg cgacggggcc cgtgcgtccc 4680
agcgcacatg ttcggcgagg cggggcctgc gagcgcggcc accgagaatc ggacgggggt 4740
agtctcaagc tggccggcct gctctggtgc ctggcctcgc gccgccgtgt atcgccccgc 4800
cctgggcggc aaggctggcc cggtcggcac cagttgcgtg agcggaaaga tggccgcttc 4860
ccggccctgc tgcagggagc tcaaaatgga ggacgcggcg ctcgggagag cgggcgggtg 4920
agtcacccac acaaaggaaa agggcctttc cgtcctcagc cgtcgcttca tgtgactcca 4980
cggagtaccg ggcgccgtcc aggcacctcg attagttctc gagcttttgg agtacgtcgt 5040
ctttaggttg gggggagggg ttttatgcga tggagtttcc ccacactgag tgggtggaga 5100
ctgaagttag gccagcttgg cacttgatgt aattctcctt ggaatttgcc ctttttgagt 5160
ttggatcttg gttcattctc aagcctcaga cagtggttca aagttttttt cttccatttc 5220
aggtgtcgtg aggatctatt tccggtgaga cccaagctgg ctagctaaac ttacgcgtgc 5280
caccatggcc ctccttacga acctcctgcc gttgtgctgc ctggccctcc tcgccttgcc 5340
agcgcagagc cacgccgaag gcacgttcac ctccgacgtg agcagctacc tggagggcca 5400
ggccgcgaaa gagttcatcg cttggctcgt caagggccgg ggcggaggag gaggaagcgg 5460
aggaggaggc tcaggcggcg gcggctctca cgccgaaggc acgttcacct ccgacgtgag 5520
cagctacctg gagggccagg ccgcgaaaga gttcatcgct tggctcgtca agggccgggg 5580
ctgagatatc cagcacagtc ccgggccgag tctagacgtt taaacccgct gatcaggtcg 5640
acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg 5700
ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc 5760
gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt 5820
tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca 5880
ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc 5940
ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc 6000
tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc 6060
tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc 6120
tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc 6180
ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg cggtaccttt 6240
aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg 6300
actggaaggg ctaattcact cccaacgaag acaagatctg ctttttgctt gtactgggtc 6360
tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct 6420
taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga 6480
ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag 6540
tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag 6600
tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa 6660
tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa 6720
tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccat cccgccccta 6780
actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca 6840
gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga 6900
ggccgctagc gtcgaccatt acttattgtt ttagctgtcc tcatgaatgt cttttcacta 6960
cccatttgct tatcctgcat ctctcagcct tgactccact cagttctctt gcttagagat 7020
accacctttc ccctgaagtg ttccttccat gttttacggc gagatggttt ctcctcgcct 7080
ggccactcag ccttagttgt ctctgttgtc ttatagaggt ctacttgaag aaggaaaaac 7140
agggggcatg gtttgactgt cctgtgagcc cttcttccct gcctccccca ctcacagtga 7200
cccggaatcc ctcgacatgg cagtctagca ctagtgcggc cgcagatctg cttcctcgct 7260
cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc 7320
ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg 7380
ccagcaaaag gccaggaacc gtaaaaa 7407
<210> 13
<211> 7545
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 13
ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 60
acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 120
tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 180
ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc 240
ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 300
ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 360
actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 420
gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc 480
tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 540
caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 600
atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 660
acgttaaggg attttggtca tgaagcgctt ttgaagctcg gatccgaaca aacgacccaa 720
cacccgtgcg ttttattctg tctttttatt gccgatcccc tcagaagaac tcgtcaagaa 780
ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc 840
ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac gctatgtcct 900
gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag cggccatttt 960
ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc tcgccgtcgg 1020
gcatgctcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga tgctcttcgt 1080
ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc tcgatgcgat 1140
gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg 1200
catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg agatcctgcc 1260
ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagcacag 1320
ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcttgcagtt 1380
cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca 1440
gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata 1500
gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa 1560
acgatcctca tcctgtctct tgatcagagc ttgatcccct gcgccatcag atccttggcg 1620
gcaagaaagc catccagttt actttgcagg gcttcccaac cttaccagag gcctgcgccg 1680
cggccagctg gctagcaatt cccgggttaa ctctagagac attgattatt gactagttat 1740
taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca 1800
taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca 1860
ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg 1920
gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg 1980
ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc 2040
ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg 2100
atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca 2160
agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt 2220
ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg 2280
gaggtctata taagcagagc tcgtttagtg aaccggggtc tctctggtta gaccagatct 2340
gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc 2400
cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc 2460
tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa 2520
agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac 2580
ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta 2640
gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg 2700
ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg 2760
ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg 2820
ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag aagaacttag 2880
atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga 2940
caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca 3000
gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg 3060
aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa 3120
agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt 3180
tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca 3240
gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc 3300
aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg 3360
ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac 3420
tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga 3480
tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa 3540
tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg 3600
aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata 3660
taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac 3720
tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc 3780
caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca 3840
gagacagatc cattcgatta gtgaacggat ctcgacggta tcggttaact tttaaaagaa 3900
aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca 3960
tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttatc gatcacgaga 4020
ctagcctcga ggcatgcctg caggaattcg ctccggtgcc cgtcagtggg cagagcgcac 4080
atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgaaccg gtgcctagag 4140
aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga 4200
gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg 4260
gtttgccgcc agaacacagg taagtgccgt gtgtggttcc cgcgggcctg gcctctttac 4320
gggttatggc ccttgcgtgc cttgaattac ttccacgccc ctggctgcag tacgtgattc 4380
ttgatcccga gcttcgggtt ggaagtgggt gggagagttc gaggccttgc gcttaaggag 4440
ccccttcgcc tcgtgcttga gttgaggcct ggcttgggcg ctggggccgc cgcgtgcgaa 4500
tctggtggca ccttcgcgcc tgtctcgctg ctttcgataa gtctctagcc atttaaaatt 4560
tttgatgacc tgctgcgacg ctttttttct ggcaagatag tcttgtaaat gcgggccaag 4620
atctgcacac tggtatttcg gtttttgggg ccgcgggcgg cgacggggcc cgtgcgtccc 4680
agcgcacatg ttcggcgagg cggggcctgc gagcgcggcc accgagaatc ggacgggggt 4740
agtctcaagc tggccggcct gctctggtgc ctggcctcgc gccgccgtgt atcgccccgc 4800
cctgggcggc aaggctggcc cggtcggcac cagttgcgtg agcggaaaga tggccgcttc 4860
ccggccctgc tgcagggagc tcaaaatgga ggacgcggcg ctcgggagag cgggcgggtg 4920
agtcacccac acaaaggaaa agggcctttc cgtcctcagc cgtcgcttca tgtgactcca 4980
cggagtaccg ggcgccgtcc aggcacctcg attagttctc gagcttttgg agtacgtcgt 5040
ctttaggttg gggggagggg ttttatgcga tggagtttcc ccacactgag tgggtggaga 5100
ctgaagttag gccagcttgg cacttgatgt aattctcctt ggaatttgcc ctttttgagt 5160
ttggatcttg gttcattctc aagcctcaga cagtggttca aagttttttt cttccatttc 5220
aggtgtcgtg aggatctatt tccggtgaga cccaagctgg ctagctaaac ttacgcgtgc 5280
caccatggcc ctgctgacca atctgctgcc tctgtgctgt ctggccctgc tggccctgcc 5340
cgctcagtct cacgccgagg gaacattcac ttccgatgtg agcagctacc tggagggcca 5400
ggccgccaag gagttcatcg cctggctggt gaagggcaga ggcggcggag gcggatccgg 5460
aggaggagga agcggcggag gcggttccca cgctgaggga accttcacaa gcgacgtgtc 5520
ctcctacctg gagggacagg ccgccaaaga gtttatcgcc tggctcgtga agggccgggg 5580
cggaggagga ggttccggag gaggtggcag cggcggagga ggtagccacg ctgagggcac 5640
ctttacctcc gatgtgtcct cctatctgga gggccaagcc gccaaggaat tcatcgcctg 5700
gctggtcaag ggcaggggct gagatatcca gcacagtccc gggccgagtc tagacgttta 5760
aacccgctga tcaggtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg 5820
gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt 5880
atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc 5940
tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt 6000
ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga 6060
ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct 6120
gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga 6180
cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct 6240
gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc 6300
tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg 6360
cctccccgcg gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt 6420
tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac aagatctgct 6480
ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta 6540
actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg 6600
tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg 6660
gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa 6720
gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata 6780
aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg 6840
tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact 6900
ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta 6960
atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag 7020
tgaggaggct tttttggagg ccgctagcgt cgaccattac ttattgtttt agctgtcctc 7080
atgaatgtct tttcactacc catttgctta tcctgcatct ctcagccttg actccactca 7140
gttctcttgc ttagagatac cacctttccc ctgaagtgtt ccttccatgt tttacggcga 7200
gatggtttct cctcgcctgg ccactcagcc ttagttgtct ctgttgtctt atagaggtct 7260
acttgaagaa ggaaaaacag ggggcatggt ttgactgtcc tgtgagccct tcttccctgc 7320
ctcccccact cacagtgacc cggaatccct cgacatggca gtctagcact agtgcggccg 7380
cagatctgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 7440
atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 7500
gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaa 7545
<210> 14
<211> 814
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 14
cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 60
gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 120
atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 180
aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 240
catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 300
catggtcgag gtgagcccca cgttctgctt cactctcccc atctcccccc cctccccacc 360
cccaattttg tatttattta ttttttaatt attttgtgca gcgatggggg cggggggggg 420
gggggggcgc gcgccaggcg gggcggggcg gggcgagggg cggggcgggg cgaggcggag 480
aggtgcggcg gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg 540
gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgacgct 600
gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg ctctgactga 660
ccgcgttact cccacaggtg agcgggcggg acggcccttc tcctccgggc tgtaattagc 720
tgagcaagag gtaagggttt aagggatggt tggttggtgg ggtattaatg tttaattacc 780
tggagcacct gcctgaaatc actttttttc aggt 814
<210> 15
<211> 4400
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 15
acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 60
ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 120
ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 180
gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 240
gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 300
ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 360
actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 420
gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 480
ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta 540
ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 600
gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 660
tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 720
tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 780
aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 840
aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 900
tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 960
gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 1020
agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 1080
aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 1140
gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 1200
caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 1260
cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 1320
ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 1380
ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 1440
gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 1500
cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 1560
gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 1620
caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 1680
tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 1740
acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 1800
aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc 1860
gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca 1920
tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc 1980
gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag 2040
agcagattgt actgagagtg caccataaaa ttgtaaacgt taatattttg ttaaaattcg 2100
cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc 2160
cttataaatc aaaagaatag cccgagatag ggttgagtgt tgttccagtt tggaacaaga 2220
gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg 2280
atggcccact acgtgaacca tcacccaaat caagtttttt ggggtcgagg tgccgtaaag 2340
cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga 2400
acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg 2460
tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg 2520
cgtactatgg ttgctttgac gtatgcggtg tgaaataccg cacagatgcg taaggagaaa 2580
ataccgcatc aggcgcccct gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg 2640
ggcaaagccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg 2700
cagagaggga gtggccaact ccatcactag gggttcctac gcgttagtta ttaatagtaa 2760
tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg 2820
gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg 2880
tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta 2940
cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt 3000
gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac 3060
tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt 3120
tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac 3180
cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt 3240
cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat 3300
ataagcagag ctggtttagt gaaccgtcag atccgctagc gctaccggtc gccaccatgg 3360
tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg 3420
acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca 3480
agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg 3540
tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc 3600
acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca 3660
aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga 3720
accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc 3780
tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca 3840
tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc 3900
actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc 3960
tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc 4020
tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaggtcg 4080
acgtttattt gtgaaatttg tgatgctatt gctttatttg taaccatcta gctttatttg 4140
tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa 4200
caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggtttttta 4260
aagtttaaac aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg 4320
ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gcggcctcag tgagcgagcg 4380
agcgcgcagc tgcctgcagg 4400
<210> 16
<211> 4791
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 16
acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 60
ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 120
ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 180
gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 240
gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 300
ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 360
actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 420
gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 480
ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta 540
ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 600
gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 660
tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 720
tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 780
aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 840
aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 900
tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 960
gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 1020
agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 1080
aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 1140
gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 1200
caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 1260
cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 1320
ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 1380
ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 1440
gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 1500
cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 1560
gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 1620
caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 1680
tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 1740
acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 1800
aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc 1860
gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca 1920
tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc 1980
gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag 2040
agcagattgt actgagagtg caccataaaa ttgtaaacgt taatattttg ttaaaattcg 2100
cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc 2160
cttataaatc aaaagaatag cccgagatag ggttgagtgt tgttccagtt tggaacaaga 2220
gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg 2280
atggcccact acgtgaacca tcacccaaat caagtttttt ggggtcgagg tgccgtaaag 2340
cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga 2400
acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg 2460
tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg 2520
cgtactatgg ttgctttgac gtatgcggtg tgaaataccg cacagatgcg taaggagaaa 2580
ataccgcatc aggcgcccct gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg 2640
ggcaaagccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg 2700
cagagaggga gtggccaact ccatcactag gggttcctac gcgtcgttac ataacttacg 2760
gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg 2820
tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta 2880
cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt 2940
gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac 3000
tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc 3060
cccacgttct gcttcactct ccccatctcc cccccctccc cacccccaat tttgtattta 3120
tttatttttt aattattttg tgcagcgatg ggggcggggg gggggggggg gcgcgcgcca 3180
ggcggggcgg ggcggggcga ggggcggggc ggggcgaggc ggagaggtgc ggcggcagcc 3240
aatcagagcg gcgcgctccg aaagtttcct tttatggcga ggcggcggcg gcggcggccc 3300
tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcga cgctgccttc gccccgtgcc 3360
ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt tactcccaca 3420
ggtgagcggg cgggacggcc cttctcctcc gggctgtaat tagctgagca agaggtaagg 3480
gtttaaggga tggttggttg gtggggtatt aatgtttaat tacctggagc acctgcctga 3540
aatcactttt tttcaggttg gaccggtgcc accatggccc tccttacgaa cctcctgccg 3600
ttgtgctgcc tggccctcct cgccttgcca gcgcagagcc acgccgaagg cacgttcacc 3660
tccgacgtga gcagctacct ggagggccag gccgcgaaag agttcatcgc ttggctcgtc 3720
aagggcggag gaggaggcgg aggaagcggc ggaggaggaa gcggaggcgg aggtagcgcc 3780
gagagcaagt acggccctcc ttgtcctccc tgccctgccc cagaggccgc tggaggacca 3840
tccgtgtttc tgtttccccc caagcctaag gacaccctga tgatcagcag gacccccgag 3900
gtgacctgcg tggtggtgga cgtgagccag gaggaccctg aggtgcagtt caattggtac 3960
gtggatggcg tggaggtgca caacgccaag acaaagccca gagaggagca gtttaatagc 4020
acctacagag tggtgtccgt gctgaccgtg ctgcaccagg attggctgaa tggcaaggag 4080
tacaagtgca aggtgtccaa taagggcctg cccagcagca tcgagaagac catcagcaag 4140
gccaagggcc agcccagaga gcctcaggtg tacacactgc ctccctccca ggaggagatg 4200
accaagaacc aggtgtccct gacctgtctg gtgaagggct tctaccccag cgatatcgcc 4260
gtggagtggg agtccaacgg ccagcccgag aataactaca agaccacccc tcctgtgctg 4320
gatagcgacg gcagcttttt cctgtactcc agactgaccg tggacaagtc tagatggcag 4380
gagggcaacg tgttttcttg tagcgtgatg cacgaggccc tgcacaatca ctacacccag 4440
aagtccctgt ctctgagcct gggctgagtc gacgtttatt tgtgaaattt gtgatgctat 4500
tgctttattt gtaaccatct agctttattt gtgaaatttg tgatgctatt gctttatttg 4560
taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc 4620
aggttcaggg ggagatgtgg gaggtttttt aaagtttaaa caggaacccc tagtgatgga 4680
gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc 4740
ccgacgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag g 4791
<210> 17
<211> 4197
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 17
acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 60
ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 120
ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 180
gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 240
gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 300
ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 360
actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 420
gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 480
ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta 540
ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 600
gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 660
tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 720
tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 780
aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 840
aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 900
tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 960
gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 1020
agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 1080
aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 1140
gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 1200
caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 1260
cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 1320
ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 1380
ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 1440
gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 1500
cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 1560
gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 1620
caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 1680
tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 1740
acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 1800
aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc 1860
gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca 1920
tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc 1980
gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag 2040
agcagattgt actgagagtg caccataaaa ttgtaaacgt taatattttg ttaaaattcg 2100
cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc 2160
cttataaatc aaaagaatag cccgagatag ggttgagtgt tgttccagtt tggaacaaga 2220
gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg 2280
atggcccact acgtgaacca tcacccaaat caagtttttt ggggtcgagg tgccgtaaag 2340
cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga 2400
acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg 2460
tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg 2520
cgtactatgg ttgctttgac gtatgcggtg tgaaataccg cacagatgcg taaggagaaa 2580
ataccgcatc aggcgcccct gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg 2640
ggcaaagccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg 2700
cagagaggga gtggccaact ccatcactag gggttcctac gcgtcgttac ataacttacg 2760
gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg 2820
tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta 2880
cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt 2940
gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac 3000
tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc 3060
cccacgttct gcttcactct ccccatctcc cccccctccc cacccccaat tttgtattta 3120
tttatttttt aattattttg tgcagcgatg ggggcggggg gggggggggg gcgcgcgcca 3180
ggcggggcgg ggcggggcga ggggcggggc ggggcgaggc ggagaggtgc ggcggcagcc 3240
aatcagagcg gcgcgctccg aaagtttcct tttatggcga ggcggcggcg gcggcggccc 3300
tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcga cgctgccttc gccccgtgcc 3360
ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt tactcccaca 3420
ggtgagcggg cgggacggcc cttctcctcc gggctgtaat tagctgagca agaggtaagg 3480
gtttaaggga tggttggttg gtggggtatt aatgtttaat tacctggagc acctgcctga 3540
aatcactttt tttcaggttg gaccggtgcc accatggccc tccttacgaa cctcctgccg 3600
ttgtgctgcc tggccctcct cgccttgcca gcgcagagcc acgccgaagg cacgttcacc 3660
tccgacgtga gcagctacct ggagggccag gccgcgaaag agttcatcgc ttggctcgtc 3720
aagggccggg gcggaggagg aggaagcgga ggaggaggct caggcggcgg cggctctcac 3780
gccgaaggca cgttcacctc cgacgtgagc agctacctgg agggccaggc cgcgaaagag 3840
ttcatcgctt ggctcgtcaa gggccggggc tgagtcgacg tttatttgtg aaatttgtga 3900
tgctattgct ttatttgtaa ccatctagct ttatttgtga aatttgtgat gctattgctt 3960
tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta 4020
tgtttcaggt tcagggggag atgtgggagg ttttttaaag tttaaacagg aacccctagt 4080
gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa 4140
ggtcgcccga cgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcagg 4197
<210> 18
<211> 4335
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 18
acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 60
ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 120
ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 180
gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 240
gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 300
ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 360
actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 420
gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 480
ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta 540
ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 600
gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 660
tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 720
tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 780
aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 840
aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 900
tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 960
gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 1020
agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 1080
aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 1140
gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 1200
caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 1260
cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 1320
ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 1380
ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 1440
gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 1500
cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 1560
gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 1620
caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 1680
tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 1740
acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 1800
aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc 1860
gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca 1920
tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc 1980
gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag 2040
agcagattgt actgagagtg caccataaaa ttgtaaacgt taatattttg ttaaaattcg 2100
cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc 2160
cttataaatc aaaagaatag cccgagatag ggttgagtgt tgttccagtt tggaacaaga 2220
gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg 2280
atggcccact acgtgaacca tcacccaaat caagtttttt ggggtcgagg tgccgtaaag 2340
cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga 2400
acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg 2460
tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg 2520
cgtactatgg ttgctttgac gtatgcggtg tgaaataccg cacagatgcg taaggagaaa 2580
ataccgcatc aggcgcccct gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg 2640
ggcaaagccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg 2700
cagagaggga gtggccaact ccatcactag gggttcctac gcgtcgttac ataacttacg 2760
gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg 2820
tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta 2880
cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt 2940
gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac 3000
tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc 3060
cccacgttct gcttcactct ccccatctcc cccccctccc cacccccaat tttgtattta 3120
tttatttttt aattattttg tgcagcgatg ggggcggggg gggggggggg gcgcgcgcca 3180
ggcggggcgg ggcggggcga ggggcggggc ggggcgaggc ggagaggtgc ggcggcagcc 3240
aatcagagcg gcgcgctccg aaagtttcct tttatggcga ggcggcggcg gcggcggccc 3300
tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcga cgctgccttc gccccgtgcc 3360
ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt tactcccaca 3420
ggtgagcggg cgggacggcc cttctcctcc gggctgtaat tagctgagca agaggtaagg 3480
gtttaaggga tggttggttg gtggggtatt aatgtttaat tacctggagc acctgcctga 3540
aatcactttt tttcaggttg gaccggtgcc accatggccc tgctgaccaa tctgctgcct 3600
ctgtgctgtc tggccctgct ggccctgccc gctcagtctc acgccgaggg aacattcact 3660
tccgatgtga gcagctacct ggagggccag gccgccaagg agttcatcgc ctggctggtg 3720
aagggcagag gcggcggagg cggatccgga ggaggaggaa gcggcggagg cggttcccac 3780
gctgagggaa ccttcacaag cgacgtgtcc tcctacctgg agggacaggc cgccaaagag 3840
tttatcgcct ggctcgtgaa gggccggggc ggaggaggag gttccggagg aggtggcagc 3900
ggcggaggag gtagccacgc tgagggcacc tttacctccg atgtgtcctc ctatctggag 3960
ggccaagccg ccaaggaatt catcgcctgg ctggtcaagg gcaggggctg agtcgacgtt 4020
tatttgtgaa atttgtgatg ctattgcttt atttgtaacc atctagcttt atttgtgaaa 4080
tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca 4140
acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt ttttaaagtt 4200
taaacaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac 4260
tgaggccggg cgaccaaagg tcgcccgacg cccgggcggc ctcagtgagc gagcgagcgc 4320
gcagctgcct gcagg 4335
<210> 19
<211> 5612
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 19
ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 60
acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 120
tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 180
ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc 240
ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 300
ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 360
actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 420
gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc 480
tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 540
caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 600
atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 660
acgttaaggg attttggtca tgaagcgctt ttgaagctcg gatccgaaca aacgacccaa 720
cacccgtgcg ttttattctg tctttttatt gccgatcccc tcagaagaac tcgtcaagaa 780
ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc 840
ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac gctatgtcct 900
gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag cggccatttt 960
ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc tcgccgtcgg 1020
gcatgctcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga tgctcttcgt 1080
ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc tcgatgcgat 1140
gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg 1200
catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg agatcctgcc 1260
ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagcacag 1320
ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcttgcagtt 1380
cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca 1440
gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata 1500
gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa 1560
acgatcctca tcctgtctct tgatcagagc ttgatcccct gcgccatcag atccttggcg 1620
gcaagaaagc catccagttt actttgcagg gcttcccaac cttaccagag gcctgcgccg 1680
cggccagctg gctagcaatt cccgggttaa ctctagagac attgattatt gactagttat 1740
taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca 1800
taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca 1860
ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg 1920
gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg 1980
ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc 2040
ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg 2100
atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca 2160
agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt 2220
ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg 2280
gaggtctata taagcagagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca 2340
cgctgttttg acctccatag aagacaccgg gaccgatcca gcctcccctc gaagcttaca 2400
tgtggtaccg agctcggatc ctgagaactt cagggtgagt ctatgggacc cttgatgttt 2460
tctttcccct tcttttctat ggttaagttc atgtcatagg aaggggagaa gtaacagggt 2520
acacatattg accaaatcag ggtaattttg catttgtaat tttaaaaaat gctttcttct 2580
tttaatatac ttttttgttt atcttatttc taatactttc cctaatctct ttctttcagg 2640
gcaataatga tacaatgtat catgcctctt tgcaccattc taaagaataa cagtgataat 2700
ttctgggtta aggcaatagc aatatttctg catataaata tttctgcata taaattgtaa 2760
ctgatgtaag aggtttcata ttgctaatag cagctacaat ccagctacca ttctgctttt 2820
attttatggt tgggataagg ctggattatt ctgagtccaa gctaggccct tttgctaatc 2880
atgttcatac ctcttatctt cctcccacag ctcctgggca acgtgctggt ctgtgtgctg 2940
gcccatcact ttggcaaagc acgtgagatc tgaattctga cactatgaag tgccttttgt 3000
acttagcctt tttattcatt ggggtgaatt gcaagttcac catagttttt ccacacaacc 3060
aaaaaggaaa ctggaaaaat gttccttcta attaccatta ttgcccgtca agctcagatt 3120
taaattggca taatgactta ataggcacag ccttacaagt caaaatgccc aagagtcaca 3180
aggctattca agcagacggt tggatgtgtc atgcttccaa atgggtcact acttgtgatt 3240
tccgctggta tggaccgaag tatataacac attccatccg atccttcact ccatctgtag 3300
aacaatgcaa ggaaagcatt gaacaaacga aacaaggaac ttggctgaat ccaggcttcc 3360
ctcctcaaag ttgtggatat gcaactgtga cggatgccga agcagtgatt gtccaggtga 3420
ctcctcacca tgtgctggtt gatgaataca caggagaatg ggttgattca cagttcatca 3480
acggaaaatg cagcaattac atatgcccca ctgtccataa ctctacaacc tggcattctg 3540
actataaggt caaagggcta tgtgattcta acctcatttc catggacatc accttcttct 3600
cagaggacgg agagctatca tccctgggaa aggagggcac agggttcaga agtaactact 3660
ttgcttatga aactggaggc aaggcctgca aaatgcaata ctgcaagcat tggggagtca 3720
gactcccatc aggtgtctgg ttcgagatgg ctgataagga tctctttgct gcagccagat 3780
tccctgaatg cccagaaggg tcaagtatct ctgctccatc tcagacctca gtggatgtaa 3840
gtctaattca ggacgttgag aggatcttgg attattccct ctgccaagaa acctggagca 3900
aaatcagagc gggtcttcca atctctccag tggatctcag ctatcttgct cctaaaaacc 3960
caggaaccgg tcctgctttc accataatca atggtaccct aaaatacttt gagaccagat 4020
acatcagagt cgatattgct gctccaatcc tctcaagaat ggtcggaatg atcagtggaa 4080
ctaccacaga aagggaactg tgggatgact gggcaccata tgaagacgtg gaaattggac 4140
ccaatggagt tctgaggacc agttcaggat ataagtttcc tttatacatg attggacatg 4200
gtatgttgga ctccgatctt catcttagct caaaggctca ggtgttcgaa catcctcaca 4260
ttcaagacgc tgcttcgcaa cttcctgatg atgagagttt attttttggt gatactgggc 4320
tatccaaaaa tccaatcgag cttgtagaag gttggttcag tagttggaaa agctctattg 4380
cctctttttt ctttatcata gggttaatca ttggactatt cttggttctc cgagttggta 4440
tccatctttg cattaaatta aagcacacca agaaaagaca gatttataca gacatagaga 4500
tgaaccgact tggaaagtaa ctcaaatcct gcacaacaga ttcttcatgt ttggaccaaa 4560
tcaacttgtg ataccatgct caaagaggcc tcaattatat ttgagttttt aatttttatg 4620
aaaaaaaaaa aaaaaaacgg aattcacccc accagtgcag gctgcctatc agaaagtggt 4680
ggctggtgtg gctaatgccc tggcccacaa gtatcactaa gctcgctttc ttgctgtcca 4740
atttctatta aaggttcctt tgttccctaa gtccaactac taaactgggg gatattatga 4800
agggccttga gcatctggat tctgcctaat aaaaaacatt tattttcatt gcaatgatgt 4860
atttaaatta tttctgaata ttttactaaa aagggaatgt gggaggtcag tgcatttaaa 4920
acataaagaa atgaagagct agttcaaacc ttgggaaaat acactatatc ttaaactcca 4980
tgaaagaagg tgaggctgca aacagctaat gcacattggc aacagcccct gatgcctatg 5040
ccttattcat ccctcagaaa aggattcaag tagaggcttg atttggaggt taaagttttg 5100
ctatgctgta ttttagtcga ccattactta ttgttttagc tgtcctcatg aatgtctttt 5160
cactacccat ttgcttatcc tgcatctctc agccttgact ccactcagtt ctcttgctta 5220
gagataccac ctttcccctg aagtgttcct tccatgtttt acggcgagat ggtttctcct 5280
cgcctggcca ctcagcctta gttgtctctg ttgtcttata gaggtctact tgaagaagga 5340
aaaacagggg gcatggtttg actgtcctgt gagcccttct tccctgcctc ccccactcac 5400
agtgacccgg aatccctcga catggcagtc tagcactagt gcggccgcag atctgcttcc 5460
tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca 5520
aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca 5580
aaaggccagc aaaaggccag gaaccgtaaa aa 5612
<210> 20
<211> 3387
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 20
ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 60
acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 120
tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 180
ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc 240
ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 300
ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 360
actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 420
gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc 480
tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 540
caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 600
atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 660
acgttaaggg attttggtca tgaagcgctt ttgaagctcg gatccgaaca aacgacccaa 720
cacccgtgcg ttttattctg tctttttatt gccgatcccc tcagaagaac tcgtcaagaa 780
ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc 840
ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac gctatgtcct 900
gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag cggccatttt 960
ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc tcgccgtcgg 1020
gcatgctcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga tgctcttcgt 1080
ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc tcgatgcgat 1140
gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg 1200
catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg agatcctgcc 1260
ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagcacag 1320
ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcttgcagtt 1380
cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca 1440
gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata 1500
gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa 1560
acgatcctca tcctgtctct tgatcagagc ttgatcccct gcgccatcag atccttggcg 1620
gcaagaaagc catccagttt actttgcagg gcttcccaac cttaccagag gcctgcgccg 1680
cggccagctg gctagcaatt cccgggttaa ctctagagaa tgtagtctta tgcaatactc 1740
ttgtagtctt gcaacatggt aacgatgagt tagcaacatg ccttacaagg agagaaaaag 1800
caccgtgcat gccgattggt ggaagtaagg tggtacgatc gtgccttatt aggaaggcaa 1860
cagacgggtc tgacatggat tggacgaacc actgaattcc gcattgcaga gatattgtat 1920
ttaagtgcct agctcgatac aataaacgcc atttgaccat tcaccacatt ggtgtgcacc 1980
tccaagctcg agctcgttta gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt 2040
ttgacctcca tagaagacac cgggaccgat ccagcctccc ctcgaagcta gtcgattagg 2100
catctcctat ggcaggaaga agcggagaca gcgacgaaga cctcctcaag gcagtcagac 2160
tcatcaagtt tctctatcaa agcaacccac ctcccaatcc cgaggggacc cgacaggccc 2220
gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac 2280
ggatccttag cacttatctg ggacgatctg cggagcctgt gcctcttcag ctaccaccgc 2340
ttgagagact tactcttgat tgtaacgagg attgtggaac ttctgggacg cagggggtgg 2400
gaagccctca aatattggtg gaatctccta caatattgga gtcaggagct aaagaatagt 2460
gctgttagct tgctcaatgc cacagctata gcagtagctg aggggacaga tagggttata 2520
gaagtagtac aagaagcttg gcactggccg tcgttttaca acgtcgtgat ctgagcctgg 2580
gagatctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg 2640
cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc aggaaaaccc 2700
tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag 2760
cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg 2820
cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag 2880
caaccatagt gtcgaccatt acttattgtt ttagctgtcc tcatgaatgt cttttcacta 2940
cccatttgct tatcctgcat ctctcagcct tgactccact cagttctctt gcttagagat 3000
accacctttc ccctgaagtg ttccttccat gttttacggc gagatggttt ctcctcgcct 3060
ggccactcag ccttagttgt ctctgttgtc ttatagaggt ctacttgaag aaggaaaaac 3120
agggggcatg gtttgactgt cctgtgagcc cttcttccct gcctccccca ctcacagtga 3180
cccggaatcc ctcgacatgg cagtctagca ctagtgcggc cgcagatctg cttcctcgct 3240
cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc 3300
ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg 3360
ccagcaaaag gccaggaacc gtaaaaa 3387
<210> 21
<211> 9171
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 21
ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 60
acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 120
tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 180
ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc 240
ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 300
ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 360
actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 420
gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc 480
tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 540
caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 600
atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 660
acgttaaggg attttggtca tgaagcgctt ttgaagctcg gatccgaaca aacgacccaa 720
cacccgtgcg ttttattctg tctttttatt gccgatcccc tcagaagaac tcgtcaagaa 780
ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc 840
ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac gctatgtcct 900
gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag cggccatttt 960
ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc tcgccgtcgg 1020
gcatgctcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga tgctcttcgt 1080
ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc tcgatgcgat 1140
gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg 1200
catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg agatcctgcc 1260
ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagcacag 1320
ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcttgcagtt 1380
cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca 1440
gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata 1500
gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa 1560
acgatcctca tcctgtctct tgatcagagc ttgatcccct gcgccatcag atccttggcg 1620
gcaagaaagc catccagttt actttgcagg gcttcccaac cttaccagag gcctgcgccg 1680
cggccagctg gctagcaatt cccgggttaa ctctagagac attgattatt gactagttat 1740
taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca 1800
taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca 1860
ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg 1920
gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg 1980
ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc 2040
ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg 2100
atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca 2160
agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt 2220
ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg 2280
gaggtctata taagcagagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca 2340
cgctgttttg acctccatag aagacaccgg gaccgatcca gcctcccctc gaagcttaca 2400
tgtggtaccg agctcggatc ctgagaactt cagggtgagt ctatgggacc cttgatgttt 2460
tctttcccct tcttttctat ggttaagttc atgtcatagg aaggggagaa gtaacagggt 2520
acacatattg accaaatcag ggtaattttg catttgtaat tttaaaaaat gctttcttct 2580
tttaatatac ttttttgttt atcttatttc taatactttc cctaatctct ttctttcagg 2640
gcaataatga tacaatgtat catgcctctt tgcaccattc taaagaataa cagtgataat 2700
ttctgggtta aggcaatagc aatatttctg catataaata tttctgcata taaattgtaa 2760
ctgatgtaag aggtttcata ttgctaatag cagctacaat ccagctacca ttctgctttt 2820
attttatggt tgggataagg ctggattatt ctgagtccaa gctaggccct tttgctaatc 2880
atgttcatac ctcttatctt cctcccacag ctcctgggca acgtgctggt ctgtgtgctg 2940
gcccatcact ttggcaaagc acgtgagatc tgaattcgag atctgccgcc gccatgggtg 3000
cgagagcgtc agtattaagc gggggagaat tagatcgatg ggaaaaaatt cggttaaggc 3060
cagggggaaa gaaaaaatat aaattaaaac atatagtatg ggcaagcagg gagctagaac 3120
gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa atactgggac 3180
agctacaacc atcccttcag acaggatcag aagaacttag atcattatat aatacagtag 3240
caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa gctttagaca 3300
agatagagga agagcaaaac aaaagtaaga aaaaagcaca gcaagcagca gctgacacag 3360
gacacagcaa tcaggtcagc caaaattacc ctatagtgca gaacatccag gggcaaatgg 3420
tacatcaggc catatcacct agaactttaa atgcatgggt aaaagtagta gaagagaagg 3480
ctttcagccc agaagtgata cccatgtttt cagcattatc agaaggagcc accccacaag 3540
atttaaacac catgctaaac acagtggggg gacatcaagc agccatgcaa atgttaaaag 3600
agaccatcaa tgaggaagct gcagaatggg atagagtgca tccagtgcat gcagggccta 3660
ttgcaccagg ccagatgaga gaaccaaggg gatcagacat cgctggaact actagtaccc 3720
ttcaggaaca aataggatgg atgacacata atccacctat cccagtagga gaaatctata 3780
aaagatggat aatcctggga ttaaataaaa tagtaagaat gtatagccct accagcattc 3840
tggacataag acaaggacca aaggaaccct ttagagacta tgtagaccga ttctataaaa 3900
ctctaagagc cgagcaagct tcacaagagg taaaaaattg gatgacagaa accttgttgg 3960
tccaaaatgc gaacccagat tgtaagacta ttttaaaagc attgggacca ggagcgacac 4020
tagaagaaat gatgacagca tgtcagggag tggggggacc cggccataaa gcaagagttt 4080
tggctgaagc aatgagccaa gtaacaaatc cagctaccat aatgatacag aaaggcaatt 4140
ttaggaacca aagaaagact gttaagtgtt tcaattgtgg caaagaaggg cacatagcca 4200
aaaattgcag ggcccctagg aaaaagggct gttggaaatg tggaaaggaa ggacaccaaa 4260
tgaaagattg tactgagaga caggctaatt ttttagggaa gatctggcct tcccacaagg 4320
gaaggccagg gaattttctt cagagcagac cagagccaac agccccacca gaagagagct 4380
tcaggtttgg ggaagagaca acaactccct ctcagaagca ggagccgata gacaaggaac 4440
tgtatccttt agcttccctc agatcactct ttggcagcga cccctcgtca caataaagat 4500
aggggggcaa ttaaaggaag ctctattaga tactggtgct gacgacacag tattagaaga 4560
aatgaatttg ccaggaagat ggaaaccaaa aatgataggg ggaattggag gttttatcaa 4620
agtaagacag tatgatcaga tactcataga aatctgcgga cataaagcta taggtacagt 4680
attagtagga cctacacctg tcaacataat tggaagaaat ctgttgactc agattggctg 4740
cactttaaat tttcccatta gtcctattga gactgtacca gtaaaattaa agccaggaat 4800
ggatggccca aaagttaaac aatggccatt gacagaagaa aaaataaaag cattagtaga 4860
aatttgtaca gaaatggaaa aggaaggaaa aatttcaaaa attgggcctg aaaatccata 4920
caatactcca gtatttgcca taaagaaaaa agacagtact aaatggagaa aattagtaga 4980
tttcagagaa cttaataaga gaactcaaga tttctgggaa gttcaattag gaataccaca 5040
tcctgcaggg ttaaaacaga aaaaatcagt aacagtactg gatgtgggcg atgcatattt 5100
ttcagttccc ttagataaag acttcaggaa gtatactgca tttaccatac ctagtataaa 5160
caatgagaca ccagggatta gatatcagta caatgtgctt ccacagggat ggaaaggatc 5220
accagcaata ttccagtgta gcatgacaaa aatcttagag ccttttagaa aacaaaatcc 5280
agacatagtc atctatcaat acatggatga tttgtatgta ggatctgact tagaaatagg 5340
gcagcataga acaaaaatag aggaactgag acaacatctg ttgaggtggg gatttaccac 5400
accagacaaa aaacatcaga aagaacctcc attcctttgg atgggttatg aactccatcc 5460
tgataaatgg acagtacagc ctatagtgct gccagaaaag gacagctgga ctgtcaatga 5520
catacagaaa ttagtgggaa aattgaattg ggcaagtcag atttatgcag ggattaaagt 5580
aaggcaatta tgtaaacttc ttaggggaac caaagcacta acagaagtag taccactaac 5640
agaagaagca gagctagaac tggcagaaaa cagggagatt ctaaaagaac cggtacatgg 5700
agtgtattat gacccatcaa aagacttaat agcagaaata cagaagcagg ggcaaggcca 5760
atggacatat caaatttatc aagagccatt taaaaatctg aaaacaggaa agtatgcaag 5820
aatgaagggt gcccacacta atgatgtgaa acaattaaca gaggcagtac aaaaaatagc 5880
cacagaaagc atagtaatat ggggaaagac tcctaaattt aaattaccca tacaaaagga 5940
aacatgggaa gcatggtgga cagagtattg gcaagccacc tggattcctg agtgggagtt 6000
tgtcaatacc cctcccttag tgaagttatg gtaccagtta gagaaagaac ccataatagg 6060
agcagaaact ttctatgtag atggggcagc caatagggaa actaaattag gaaaagcagg 6120
atatgtaact gacagaggaa gacaaaaagt tgtcccccta acggacacaa caaatcagaa 6180
gactgagtta caagcaattc atctagcttt gcaggattcg ggattagaag taaacatagt 6240
gacagactca caatatgcat tgggaatcat tcaagcacaa ccagataaga gtgaatcaga 6300
gttagtcagt caaataatag agcagttaat aaaaaaggaa aaagtctacc tggcatgggt 6360
accagcacac aaaggaattg gaggaaatga acaagtagat aaattggtca gtgctggaat 6420
caggaaagta ctatttttag atggaataga taaggcccaa gaagaacatg agaaatatca 6480
cagtaattgg agagcaatgg ctagtgattt taacctacca cctgtagtag caaaagaaat 6540
agtagccagc tgtgataaat gtcagctaaa aggggaagcc atgcatggac aagtagactg 6600
tagcccagga atatggcagc tagattgtac acatttagaa ggaaaagtta tcttggtagc 6660
agttcatgta gccagtggat atatagaagc agaagtaatt ccagcagaga cagggcaaga 6720
aacagcatac ttcctcttaa aattagcagg aagatggcca gtaaaaacag tacatacaga 6780
caatggcagc aatttcacca gtactacagt taaggccgcc tgttggtggg cggggatcaa 6840
gcaggaattt ggcattccct acaatccgca gtcacaagga gtaatagaat ctatgaataa 6900
agaattaaag aaaattatag gacaggtaag agatcaggct gaacatctta aaacagcagt 6960
acaaatggca gtattcatcc acaattttaa aagaaaaggg gggattgggg ggtacagtgc 7020
aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca 7080
aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagagatc cagtttggaa 7140
aggaccagca aagctcctct ggaaaggtga aggggcagta gtaatacaag ataatagtga 7200
cataaaagta gtgccaagaa gaaaagcaaa gatcatcagg gattatggaa aacagatggc 7260
aggtgatgat tgtgtggcaa gtagacagga tgaggattaa cacatggaat tccggagcgg 7320
ccgcaggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt 7380
caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca 7440
atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca 7500
agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg 7560
ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt 7620
ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag 7680
aaattaacaa ttacacaagc ttccgcggaa ttcaccccac cagtgcaggc tgcctatcag 7740
aaagtggtgg ctggtgtggc taatgccctg gcccacaagt atcactaagc tcgctttctt 7800
gctgtccaat ttctattaaa ggttcctttg ttccctaagt ccaactacta aactggggga 7860
tattatgaag ggccttgagc atctggattc tgcctaataa aaaacattta ttttcattgc 7920
aatgatgtat ttaaattatt tctgaatatt ttactaaaaa gggaatgtgg gaggtcagtg 7980
catttaaaac ataaagaaat gaagagctag ttcaaacctt gggaaaatac actatatctt 8040
aaactccatg aaagaaggtg aggctgcaaa cagctaatgc acattggcaa cagcccctga 8100
tgcctatgcc ttattcatcc ctcagaaaag gattcaagta gaggcttgat ttggaggtta 8160
aagttttgct atgctgtatt ttacattact tattgtttta gctgtcctca tgaatgtctt 8220
ttcactaccc atttgcttat cctgcatctc tcagccttga ctccactcag ttctcttgct 8280
tagagatacc acctttcccc tgaagtgttc cttccatgtt ttacggcgag atggtttctc 8340
ctcgcctggc cactcagcct tagttgtctc tgttgtctta tagaggtcta cttgaagaag 8400
gaaaaacagg gggcatggtt tgactgtcct gtgagccctt cttccctgcc tcccccactc 8460
acagtgaccc ggaatccctc gacatggcag tctagcacta gtgcggccgc agatctgctt 8520
cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact 8580
caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag 8640
caaaaggcca gcaaaaggcc aggaaccgta aaaagtcgac cattacttat tgttttagct 8700
gtcctcatga atgtcttttc actacccatt tgcttatcct gcatctctca gccttgactc 8760
cactcagttc tcttgcttag agataccacc tttcccctga agtgttcctt ccatgtttta 8820
cggcgagatg gtttctcctc gcctggccac tcagccttag ttgtctctgt tgtcttatag 8880
aggtctactt gaagaaggaa aaacaggggg catggtttga ctgtcctgtg agcccttctt 8940
ccctgcctcc cccactcaca gtgacccgga atccctcgac atggcagtct agcactagtg 9000
cggccgcaga tctgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg 9060
agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 9120
aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa a 9171
<210> 22
<211> 11635
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 22
ggtacccaac tccatgctta acagtcccca ggtacagccc accctgcgtc gcaaccagga 60
acagctctac agcttcctgg agcgccactc gccctacttc cgcagccaca gtgcgcagat 120
taggagcgcc acttcttttt gtcacttgaa aaacatgtaa aaataatgta ctaggagaca 180
ctttcaataa aggcaaatgt ttttatttgt acactctcgg gtgattattt accccccacc 240
cttgccgtct gcgccgttta aaaatcaaag gggttctgcc gcgcatcgct atgcgccact 300
ggcagggaca cgttgcgata ctggtgttta gtgctccact taaactcagg cacaaccatc 360
cgcggcagct cggtgaagtt ttcactccac aggctgcgca ccatcaccaa cgcgtttagc 420
aggtcgggcg ccgatatctt gaagtcgcag ttggggcctc cgccctgcgc gcgcgagttg 480
cgatacacag ggttgcagca ctggaacact atcagcgccg ggtggtgcac gctggccagc 540
acgctcttgt cggagatcag atccgcgtcc aggtcctccg cgttgctcag ggcgaacgga 600
gtcaactttg gtagctgcct tcccaaaaag ggtgcatgcc caggctttga gttgcactcg 660
caccgtagtg gcatcagaag gtgaccgtgc ccggtctggg cgttaggata cagcgcctgc 720
atgaaagcct tgatctgctt aaaagccacc tgagcctttg cgccttcaga gaagaacatg 780
ccgcaagact tgccggaaaa ctgattggcc ggacaggccg cgtcatgcac gcagcacctt 840
gcgtcggtgt tggagatctg caccacattt cggccccacc ggttcttcac gatcttggcc 900
ttgctagact gctccttcag cgcgcgctgc ccgttttcgc tcgtcacatc catttcaatc 960
acgtgctcct tatttatcat aatgctcccg tgtagacact taagctcgcc ttcgatctca 1020
gcgcagcggt gcagccacaa cgcgcagccc gtgggctcgt ggtgcttgta ggttacctct 1080
gcaaacgact gcaggtacgc ctgcaggaat cgccccatca tcgtcacaaa ggtcttgttg 1140
ctggtgaagg tcagctgcaa cccgcggtgc tcctcgttta gccaggtctt gcatacggcc 1200
gccagagctt ccacttggtc aggcagtagc ttgaagtttg cctttagatc gttatccacg 1260
tggtacttgt ccatcaacgc gcgcgcagcc tccatgccct tctcccacgc agacacgatc 1320
ggcaggctca gcgggtttat caccgtgctt tcactttccg cttcactgga ctcttccttt 1380
tcctcttgcg tccgcatacc ccgcgccact gggtcgtctt cattcagccg ccgcaccgtg 1440
cgcttacctc ccttgccgtg cttgattagc accggtgggt tgctgaaacc caccatttgt 1500
agcgccacat cttctctttc ttcctcgctg tccacgatca cctctgggga tggcgggcgc 1560
tcgggcttgg gagaggggcg cttctttttc tttttggacg caatggccaa atccgccgtc 1620
gaggtcgatg gccgcgggct gggtgtgcgc ggcaccagcg catcttgtga cgagtcttct 1680
tcgtcctcgg actcgagacg ccgcctcagc cgcttttttg ggggcgcgcg gggaggcggc 1740
ggcgacggcg acggggacga cacgtcctcc atggttggtg gacgtcgcgc cgcaccgcgt 1800
ccgcgctcgg gggtggtttc gcgctgctcc tcttcccgac tggccatttc cttctcctat 1860
aggcagaaaa agatcatgga gtcagtcgag aaggaggaca gcctaaccgc cccctttgag 1920
ttcgccacca ccgcctccac cgatgccgcc aacgcgccta ccaccttccc cgtcgaggca 1980
cccccgcttg aggaggagga agtgattatc gagcaggacc caggttttgt aagcgaagac 2040
gacgaggatc gctcagtacc aacagaggat aaaaagcaag accaggacga cgcagaggca 2100
aacgaggaac aagtcgggcg gggggaccaa aggcatggcg actacctaga tgtgggagac 2160
gacgtgctgt tgaagcatct gcagcgccag tgcgccatta tctgcgacgc gttgcaagag 2220
cgcagcgatg tgcccctcgc catagcggat gtcagccttg cctacgaacg ccacctgttc 2280
tcaccgcgcg taccccccaa acgccaagaa aacggcacat gcgagcccaa cccgcgcctc 2340
aacttctacc ccgtatttgc cgtgccagag gtgcttgcca cctatcacat ctttttccaa 2400
aactgcaaga tacccctatc ctgccgtgcc aaccgcagcc gagcggacaa gcagctggcc 2460
ttgcggcagg gcgctgtcat acctgatatc gcctcgctcg acgaagtgcc aaaaatcttt 2520
gagggtcttg gacgcgacga gaaacgcgcg gcaaacgctc tgcaacaaga aaacagcgaa 2580
aatgaaagtc actgtggagt gctggtggaa cttgagggtg acaacgcgcg cctagccgtg 2640
ctgaaacgca gcatcgaggt cacccacttt gcctacccgg cacttaacct accccccaag 2700
gttatgagca cagtcatgag cgagctgatc gtgcgccgtg cacgacccct ggagagggat 2760
gcaaacttgc aagaacaaac cgaggagggc ctacccgcag ttggcgatga gcagctggcg 2820
cgctggcttg agacgcgcga gcctgccgac ttggaggagc gacgcaagct aatgatggcc 2880
gcagtgcttg ttaccgtgga gcttgagtgc atgcagcggt tctttgctga cccggagatg 2940
cagcgcaagc tagaggaaac gttgcactac acctttcgcc agggctacgt gcgccaggcc 3000
tgcaaaattt ccaacgtgga gctctgcaac ctggtctcct accttggaat tttgcacgaa 3060
aaccgcctcg ggcaaaacgt gcttcattcc acgctcaagg gcgaggcgcg ccgcgactac 3120
gtccgcgact gcgtttactt atttctgtgc tacacctggc aaacggccat gggcgtgtgg 3180
cagcaatgcc tggaggagcg caacctaaag gagctgcaga agctgctaaa gcaaaacttg 3240
aaggacctat ggacggcctt caacgagcgc tccgtggccg cgcacctggc ggacattatc 3300
ttccccgaac gcctgcttaa aaccctgcaa cagggtctgc cagacttcac cagtcaaagc 3360
atgttgcaaa actttaggaa ctttatccta gagcgttcag gaattctgcc cgccacctgc 3420
tgtgcgcttc ctagcgactt tgtgcccatt aagtaccgtg aatgccctcc gccgctttgg 3480
ggtcactgct accttctgca gctagccaac taccttgcct accactccga catcatggaa 3540
gacgtgagcg gtgacggcct actggagtgt cactgtcgct gcaacctatg caccccgcac 3600
cgctccctgg tctgcaattc gcaactgctt agcgaaagtc aaattatcgg tacctttgag 3660
ctgcagggtc cctcgcctga cgaaaagtcc gcggctccgg ggttgaaact cactccgggg 3720
ctgtggacgt cggcttacct tcgcaaattt gtacctgagg actaccacgc ccacgagatt 3780
aggttctacg aagaccaatc ccgcccgcca aatgcggagc ttaccgcctg cgtcattacc 3840
cagggccaca tccttggcca attgcaagcc atcaacaaag cccgccaaga gtttctgcta 3900
cgaaagggac ggggggttta cctggacccc cagtccggcg aggagctcaa cccaatcccc 3960
ccgccgccgc agccctatca gcagccgcgg gcccttgctt cccaggatgg cacccaaaaa 4020
gaagctgcag ctgccgccgc cgccacccac ggacgaggag gaatactggg acagtcaggc 4080
agaggaggtt ttggacgagg aggaggagat gatggaagac tgggacagcc tagacgaagc 4140
ttccgaggcc gaagaggtgt cagacgaaac accgtcaccc tcggtcgcat tcccctcgcc 4200
ggcgccccag aaattggcaa ccgttcccag catcgctaca acctccgctc ctcaggcgcc 4260
gccggcactg cctgttcgcc gacccaaccg tagatgggac accactggaa ccagggccgg 4320
taagtctaag cagccgccgc cgttagccca agagcaacaa cagcgccaag gctaccgctc 4380
gtggcgcggg cacaagaacg ccatagttgc ttgcttgcaa gactgtgggg gcaacatctc 4440
cttcgcccgc cgctttcttc tctaccatca cggcgtggcc ttcccccgta acatcctgca 4500
ttactaccgt catctctaca gcccctactg caccggcggc agcggcagcg gcagcaacag 4560
cagcggtcac acagaagcaa aggcgaccgg atagcaagac tctgacaaag cccaagaaat 4620
ccacagcggc ggcagcagca ggaggaggag cgctgcgtct ggcgcccaac gaacccgtat 4680
cgacccgcga gcttagaaat aggatttttc ccactctgta tgctatattt caacaaagca 4740
ggggccaaga acaagagctg aaaataaaaa acaggtctct gcgctccctc acccgcagct 4800
gcctgtatca caaaagcgaa gatcagcttc ggcgcacgct ggaagacgcg gaggctctct 4860
tcagcaaata ctgcgcgctg actcttaagg actagtttcg cgccctttct caaatttaag 4920
cgcgaaaact acgtcatctc cagcggccac acccggcgcc agcacctgtc gtcagcgcca 4980
ttatgagcaa ggaaattccc acgccctaca tgtggagtta ccagccacaa atgggacttg 5040
cggctggagc tgcccaagac tactcaaccc gaataaacta catgagcgcg ggaccccaca 5100
tgatatcccg ggtcaacgga atccgcgccc accgaaaccg aattctcctc gaacaggcgg 5160
ctattaccac cacacctcgt aataacctta atccccgtag ttggcccgct gccctggtgt 5220
accaggaaag tcccgctccc accactgtgg tacttcccag agacgcccag gccgaagttc 5280
agatgactaa ctcaggggcg cagcttgcgg gcggctttcg tcacagggtg cggtcgcccg 5340
ggcgttttag ggcggagtaa cttgcatgta ttgggaattg tagttttttt aaaatgggaa 5400
gtgacgtatc gtgggaaaac ggaagtgaag atttgaggaa gttgtgggtt ttttggcttt 5460
cgtttctggg cgtaggttcg cgtgcggttt tctgggtgtt ttttgtggac tttaaccgtt 5520
acgtcatttt ttagtcctat atatactcgc tctgtacttg gcccttttta cactgtgact 5580
gattgagctg gtgccgtgtc gagtggtgtt ttttaatagg tttttttact ggtaaggctg 5640
actgttatgg ctgccgctgt ggaagcgctg tatgttgttc tggagcggga gggtgctatt 5700
ttgcctaggc aggagggttt ttcaggtgtt tatgtgtttt tctctcctat taattttgtt 5760
atacctccta tgggggctgt aatgttgtct ctacgcctgc gggtatgtat tcccccgggc 5820
tatttcggtc gctttttagc actgaccgat gttaaccaac ctgatgtgtt taccgagtct 5880
tacattatga ctccggacat gaccgaggaa ctgtcggtgg tgctttttaa tcacggtgac 5940
cagttttttt acggtcacgc cggcatggcc gtagtccgtc ttatgcttat aagggttgtt 6000
tttcctgttg taagacaggc ttctaatgtt taaatgtttt tttttttgtt attttatttt 6060
gtgtttaatg caggaacccg cagacatgtt tgagagaaaa atggtgtctt tttctgtggt 6120
ggttccggaa cttacctgcc tttatctgca tgagcatgac tacgatgtgc ttgctttttt 6180
gcgcgaggct ttgcctgatt ttttgagcag caccttgcat tttatatcgc cgcccatgca 6240
acaagcttac ataggggcta cgctggttag catagctccg agtatgcgtg tcataatcag 6300
tgtgggttct tttgtcatgg ttcctggcgg ggaagtggcc gcgctggtcc gtgcagacct 6360
gcacgattat gttcagctgg ccctgcgaag ggacctacgg gatcgcggta tttttgttaa 6420
tgttccgctt ttgaatctta tacaggtctg tgaggaacct gaatttttgc aatcatgatt 6480
cgctgcttga ggctgaaggt ggagggcgct ctggagcaga tttttacaat ggccggactt 6540
aatattcggg atttgcttag agacatattg ataaggtggc gagatgaaaa ttatttgggc 6600
atggttgaag gtgctggaat gtttatagag gagattcacc ctgaagggtt tagcctttac 6660
gtccacttgg acgtgagggc agtttgcctt ttggaagcca ttgtgcaaca tcttacaaat 6720
gccattatct gttctttggc tgtagagttt gaccacgcca ccggagggga gcgcgttcac 6780
ttaatagatc ttcattttga ggttttggat aatcttttgg aataaaaaaa aaaaaacatg 6840
gttcttccag ctcttcccgc tcctcccgtg tgtgactcgc agaacgaatg tgtaggttgg 6900
ctgggtgtgg cttattctgc ggtggtggat gttatcaggg cagcggcgca tgaaggagtt 6960
tacatagaac ccgaagccag ggggcgcctg gatgctttga gagagtggat atactacaac 7020
tactacacag agcgagctaa gcgacgagac cggagacgca gatctgtttg tcacgcccgc 7080
acctggtttt gcttcaggaa atatgactac gtccggcgtt ccatttggca tgacactacg 7140
accaacacga tctcggttgt ctcggcgcac tccgtacagt agggatcgcc tacctccttt 7200
tgagacagag acccgcgcta ccatactgga ggatcatccg ctgctgcccg aatgtaacac 7260
tttgacaatg cacaacgtga gttacgtgcg aggtcttccc tgcagtgtgg gatttacgct 7320
gattcaggaa tgggttgttc cctgggatat ggttctgacg cgggaggagc ttgtaatcct 7380
gaggaagtgt atgcacgtgt gcctgtgttg tgccaacatt gatatcatga cgagcatgat 7440
gatccatggt tacgagtcct gggctctcca ctgtcattgt tccagtcccg gttccctgca 7500
gtgcatagcc ggcgggcagg ttttggccag ctggtttagg atggtggtgg atggcgccat 7560
gtttaatcag aggtttatat ggtaccggga ggtggtgaat tacaacatgc caaaagaggt 7620
aatgtttatg tccagcgtgt ttatgagggg tcgccactta atctacctgc gcttgtggta 7680
tgatggccac gtgggttctg tggtccccgc catgagcttt ggatacagcg ccttgcactg 7740
tgggattttg aacaatattg tggtgctgtg ctgcagttac tgtgctgatt taagtgagat 7800
cagggtgcgc tgctgtgccc ggaggacaag gcgtctcatg ctgcgggcgg tgcgaatcat 7860
cgctgaggag accactgcca tgttgtattc ctgcaggacg gagcggcggc ggcagcagtt 7920
tattcgcgcg ctgctgcagc accaccgccc tatcctgatg cacgattatg actctacccc 7980
catgtaggcg tggacttccc cttcgccgcc cgttgagcaa ccgcaagttg gacagcagcc 8040
tgtggctcag cagctggaca gcgacatgaa cttaagcgag ctgcccgggg agtttattaa 8100
tatcactgat gagcgtttgg ctcgacagga aaccgtgtgg aatataacac ctaagaatat 8160
gtctgttacc catgatatga tgctttttaa ggccagccgg ggagaaagga ctgtgtactc 8220
tgtgtgttgg gagggaggtg gcaggttgaa tactagggtt ctgtgagttt gattaaggta 8280
cggtgatcaa tataagctat gtggtggtgg ggctatacta ctgaatgaaa aatgacttga 8340
aattttctgc aattgaaaaa taaacacgtt gaaacataac atgcaacagg ttcacgattc 8400
tttattcctg ggcaatgtag gagaaggtgt aagagttggt agcaaaagtt tcagtggtgt 8460
attttccact ttcccaggac catgtaaaag acatagagta agtgcttacc tcgctagttt 8520
ctgtggattc actagaatcg atgtaggatg ttgcccctcc tgacgcggta ggagaagggg 8580
agggtgccct gcatgtctgc cgctgctctt gctcttgccg ctgctgagga ggggggcgca 8640
tctgccgcag caccggatgc atctgggaaa agcaaaaaag gggctcgtcc ctgtttccgg 8700
aggaatttgc aagcggggtc ttgcatgacg gggaggcaaa cccccgttcg ccgcagtccg 8760
gccggcccga gactcgaacc gggggtcctg cgactcaacc cttggaaaat aaccctccgg 8820
ctacagggag cgagccactt aatgctttcg ctttccagcc taaccgctta cgccgcgcgc 8880
ggccagtggc caaaaaagct agcgcagcag ccgccgcgcc tggaaggaag ccaaaaggag 8940
cgctcccccg ttgtctgacg tcgcacacct gggttcgaca cgcgggcggt aaccgcatgg 9000
atcacggcgg acggccggat ccggggttcg aaccccggtc gtccgccatg atacccttgc 9060
gaatttatcc accagaccac ggaagagtgc ccgcttacag gctctccttt tgcacggtct 9120
agagcgtcaa cgactgcgca cgcctcaccg gccagagcgt cccgaccatg gagcactttt 9180
tgccgctgcg caacatctgg aaccgcgtcc gcgactttcc gcgcgcctcc accaccgccg 9240
ccggcatcac ctggatgtcc aggtacatct acggattacg tcgacgttta aaccatatga 9300
tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag 9360
aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg 9420
tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg 9480
tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg 9540
cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga 9600
agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc 9660
tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt 9720
aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact 9780
ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg 9840
cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt 9900
accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt 9960
ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct 10020
ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg 10080
gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt 10140
aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt 10200
gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc 10260
gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg 10320
cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc 10380
gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg 10440
gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca 10500
ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga 10560
tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct 10620
ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg 10680
cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca 10740
accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata 10800
cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct 10860
tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact 10920
cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa 10980
acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc 11040
atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga 11100
tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga 11160
aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt 11220
aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag 11280
aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga 11340
acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg 11400
aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc 11460
ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg 11520
aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc 11580
gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgatg gatcc 11635
<210> 23
<211> 7336
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 23
atgccggggt tttacgagat tgtgattaag gtccccagcg accttgacga gcatctgccc 60
ggcatttctg acagctttgt gaactgggtg gccgagaagg aatgggagtt gccgccagat 120
tctgacatgg atctgaatct gattgagcag gcacccctga ccgtggccga gaagctgcag 180
cgcgactttc tgacggaatg gcgccgtgtg agtaaggccc cggaggctct tttctttgtg 240
caatttgaga agggagagag ctacttccac atgcacgtgc tcgtggaaac caccggggtg 300
aaatccatgg ttttgggacg tttcctgagt cagattcgcg aaaaactgat tcagagaatt 360
taccgcggga tcgagccgac tttgccaaac tggttcgcgg tcacaaagac cagaaatggc 420
gccggaggcg ggaacaaggt ggtggatgag tgctacatcc ccaattactt gctccccaaa 480
acccagcctg agctccagtg ggcgtggact aatatggaac agtatttaag cgcctgtttg 540
aatctcacgg agcgtaaacg gttggtggcg cagcatctga cgcacgtgtc gcagacgcag 600
gagcagaaca aagagaatca gaatcccaat tctgatgcgc cggtgatcag atcaaaaact 660
tcagccaggt acatggagct ggtcgggtgg ctcgtggaca aggggattac ctcggagaag 720
cagtggatcc aggaggacca ggcctcatac atctccttca atgcggcctc caactcgcgg 780
tcccaaatca aggctgcctt ggacaatgcg ggaaagatta tgagcctgac taaaaccgcc 840
cccgactacc tggtgggcca gcagcccgtg gaggacattt ccagcaatcg gatttataaa 900
attttggaac taaacgggta cgatccccaa tatgcggctt ccgtctttct gggatgggcc 960
acgaaaaagt tcggcaagag gaacaccatc tggctgtttg ggcctgcaac taccgggaag 1020
accaacatcg cggaggccat agcccacact gtgcccttct acgggtgcgt aaactggacc 1080
aatgagaact ttcccttcaa cgactgtgtc gacaagatgg tgatctggtg ggaggagggg 1140
aagatgaccg ccaaggtcgt ggagtcggcc aaagccattc tcggaggaag caaggtgcgc 1200
gtggaccaga aatgcaagtc ctcggcccag atagacccga ctcccgtgat cgtcacctcc 1260
aacaccaaca tgtgcgccgt gattgacggg aactcaacga ccttcgaaca ccagcagccg 1320
ttgcaagacc ggatgttcaa atttgaactc acccgccgtc tggatcatga ctttgggaag 1380
gtcaccaagc aggaagtcaa agactttttc cggtgggcaa aggatcacgt ggttgaggtg 1440
gagcatgaat tctacgtcaa aaagggtgga gccaagaaaa gacccgcccc cagtgacgca 1500
gatataagtg agcccaaacg ggtgcgcgag tcagttgcgc agccatcgac gtcagacgcg 1560
gaagcttcga tcaactacgc agacaggtac caaaacaaat gttctcgtca cgtgggcatg 1620
aatctgatgc tgtttccctg cagacaatgc gagagaatga atcagaattc aaatatctgc 1680
ttcactcacg gacagaaaga ctgtttagag tgctttcccg tgtcagaatc tcaacccgtt 1740
tctgtcgtca aaaaggcgta tcagaaactg tgctacattc atcatatcat gggaaaggtg 1800
ccagacgctt gcactgcctg cgatctggtc aatgtggatt tggatgactg catctttgaa 1860
caataaatga tttaaatcag gtatggctgc cgatggttat cttccagatt ggctcgagga 1920
caacctctct gagggcattc gcgagtggtg ggcgctgaaa cctggagccc cgaagcccaa 1980
agccaaccag caaaagcagg acgacggccg gggtctggtg cttcctggct acaagtacct 2040
cggacccttc aacggactcg acaaggggga gcccgtcaac gcggcggacg cagcggccct 2100
cgagcacgac aaggcctacg accagcagct gcaggcgggt gacaatccgt acctgcggta 2160
taaccacgcc gacgccgagt ttcaggagcg tctgcaagaa gatacgtctt ttgggggcaa 2220
cctcgggcga gcagtcttcc aggccaagaa gcgggttctc gaacctctcg gtctggttga 2280
ggaaggcgct aagacggctc ctggaaagaa gagaccggta gagccatcac cccagcgttc 2340
tccagactcc tctacgggca tcggcaagaa aggccaacag cccgccagaa aaagactcaa 2400
ttttggtcag actggcgact cagagtcagt tccagaccct caacctctcg gagaacctcc 2460
agcagcgccc tctggtgtgg gacctaatac aatggctgca ggcggtggcg caccaatggc 2520
agacaataac gaaggcgccg acggagtggg tagttcctcg ggaaattggc attgcgattc 2580
cacatggctg ggcgacagag tcatcaccac cagcacccga acctgggccc tgcccaccta 2640
caacaaccac ctctacaagc aaatctccaa cgggacatcg ggaggagcca ccaacgacaa 2700
cacctacttc ggctacagca ccccctgggg gtattttgac tttaacagat tccactgcca 2760
cttttcacca cgtgactggc agcgactcat caacaacaac tggggattcc ggcccaagag 2820
actcagcttc aagctcttca acatccaggt caaggaggtc acgcagaatg aaggcaccaa 2880
gaccatcgcc aataacctca ccagcaccat ccaggtgttt acggactcgg agtaccagct 2940
gccgtacgtt ctcggctctg cccaccaggg ctgcctgcct ccgttcccgg cggacgtgtt 3000
catgattccc cagtacggct acctaacact caacaacggt agtcaggccg tgggacgctc 3060
ctccttctac tgcctggaat actttccttc gcagatgctg agaaccggca acaacttcca 3120
gtttacttac accttcgagg acgtgccttt ccacagcagc tacgcccaca gccagagctt 3180
ggaccggctg atgaatcctc tgattgacca gtacctgtac tacttgtctc ggactcaaac 3240
aacaggaggc acggcaaata cgcagactct gggcttcagc caaggtgggc ctaatacaat 3300
ggccaatcag gcaaagaact ggctgccagg accctgttac cgccaacaac gcgtctcaac 3360
gacaaccggg caaaacaaca atagcaactt tgcctggact gctgggacca aataccatct 3420
gaatggaaga aattcattgg ctaatcctgg catcgctatg gcaacacaca aagacgacga 3480
ggagcgtttt tttcccagta acgggatcct gatttttggc aaacaaaatg ctgccagaga 3540
caatgcggat tacagcgatg tcatgctcac cagcgaggaa gaaatcaaaa ccactaaccc 3600
tgtggctaca gaggaatacg gtatcgtggc agataacttg cagcagcaaa acacggctcc 3660
tcaaattgga actgtcaaca gccagggggc cttacccggt atggtctggc agaaccggga 3720
cgtgtacctg cagggtccca tctgggccaa gattcctcac acggacggca acttccaccc 3780
gtctccgctg atgggcggct ttggcctgaa acatcctccg cctcagatcc tgatcaagaa 3840
cacgcctgta cctgcggatc ctccgaccac cttcaaccag tcaaagctga actctttcat 3900
cacgcaatac agcaccggac aggtcagcgt ggaaattgaa tgggagctgc agaaggaaaa 3960
cagcaagcgc tggaaccccg agatccagta cacctccaac tactacaaat ctacaagtgt 4020
ggactttgct gttaatacag aaggcgtgta ctctgaaccc cgccccattg gcacccgtta 4080
cctcacccgt aatctgtaat tgcctgttaa tcaataaacc ggttgattcg tttcagttga 4140
actttggtct ctgcgaaggg cgaattcgtt taaacctgca ggactagagg tcctgtatta 4200
gaggtcacgt gagtgttttg cgacattttg cgacaccatg tggtcacgct gggtatttaa 4260
gcccgagtga gcacgcaggg tctccatttt gaagcgggag gtttgaacgc gcagccgcca 4320
agccgaattc tgcagatatc catcacactg gcggccgctc gactagagcg gccgccaccg 4380
cggtggagct ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca 4440
tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga 4500
gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt 4560
gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga 4620
atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc 4680
actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg 4740
gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc 4800
cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc 4860
ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga 4920
ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc 4980
ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat 5040
agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg 5100
cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc 5160
aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga 5220
gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact 5280
agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt 5340
ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag 5400
cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg 5460
tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa 5520
aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata 5580
tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg 5640
atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata 5700
cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg 5760
gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct 5820
gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt 5880
tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc 5940
tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga 6000
tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt 6060
aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc 6120
atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa 6180
tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca 6240
catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca 6300
aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct 6360
tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc 6420
gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa 6480
tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt 6540
tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg 6600
taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta 6660
accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt 6720
tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca 6780
aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa 6840
gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat 6900
ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag 6960
gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg 7020
ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt 7080
gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg 7140
ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga 7200
cggccagtga gcgcgcgtaa tacgactcac tatagggcga attgggtacc gggccccccc 7260
tcgatcgagg tcgacggtat cgggggagct cgcagggtct ccattttgaa gcgggaggtt 7320
tgaacgcgca gccgcc 7336
<210> 24
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 24
agtaagacca ccgcacagca 20
<210> 25
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 25
ccttggtggg tgctactcct 20
<210> 26
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 26
cctccaggtc tgaagatcag cggccgc 27
<210> 27
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 27
gctgtcatct cttgtgggct gt 22
<210> 28
<211> 26
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 28
cctgtcatgc ccacacaaat ctctcc 26
<210> 29
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 29
actcatggga gctgctggtt c 21
<210> 30
<211> 22
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 30
Met Ala Leu Leu Thr Asn Leu Leu Pro Leu Cys Cys Leu Ala Leu Leu
1 5 10 15
Ala Leu Pro Ala Gln Ser
20
<210> 31
<211> 31
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 31
His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly
1 5 10 15
Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg Gly
20 25 30
<210> 32
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 32
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
1 5 10 15
Claims (17)
1.一种核酸构建体,其特征在于,所述核酸构建体包括编码GLP-1或其类似物的多核苷酸,所述GLP-1或其类似物的总数量为两个以上,GLP-1与GLP-1之间、GLP-1与其类似物之间或GLP-1类似物与GLP-1与类似物之间通过编码连接肽的多核苷酸连接。
2.根据权利要求1所述的核酸构建体,其特征在于,所述核酸构建体为用于糖代谢相关疾病基因治疗的核酸构建体。
3.根据权利要求1所述的核酸构建体,其特征在于,糖代谢相关疾病包括糖尿病或其并发症、肥胖症;优选的,所述糖尿病为2型糖尿病。
4.根据权利要求1所述的核酸构建体,其中GLP-1或其类似物之间的连接方式如下:信号肽——GLP-1或GLP-1类似物——连接肽——GLP-1或GLP-1类似物——连接肽——GLP-1或GLP-1类似物,其中GLP-1或GLP-1类似物——连接肽——GLP-1或GLP-1类似物单元的个数为一个或多个。
5.根据权利要求1所述的核酸构建体,其特征在于,所述核酸构建体包括如SEQ ID NO:4或SEQ ID NO:6所示的表达框架。
6.根据权利要求1所述的核酸构建体,其特征在于,所述核酸构建体编码的多肽氨基酸序列如SEQ ID NO:3或SEQ ID NO:5所示。
7.根据权利要求1所述的核酸构建体,其特征在于,所述核酸构建体被用于生产基于病毒的基因治疗载体;优选的,所述基因治疗载体为慢病毒载体或腺相关病毒载体。
8.根据权利要求7所述的核酸构建体,其特征在于,还包括以下任一特征:
1)所述慢病毒载体的核苷酸序列如SEQ ID NO.12或SEQ ID NO.13所示;
2)所述腺相关病毒载体的核苷酸序列如SEQ ID NO.17或SEQ ID NO.18所示。
9.一种慢病毒载体系统,其特征在于,所述慢病毒载体系统包括权利要求1-8任一所述的核酸构建体以及辅助质粒。
10.根据权利要求9所述的慢病毒载体系统,其特征在于,所述慢病毒载体系统还包括携带慢病毒载体的宿主细胞。
11.一种慢病毒,其特征在于,所述慢病毒由权利要求9或10所述的慢病毒载体系统经病毒包装而成。
12.一种腺相关病毒载体系统,其特征在于,所述腺相关病毒载体系统包括权利要求1-8任一所述的核酸构建体以及辅助质粒。
13.根据权利要求12所述的腺相关病毒载体系统,其特征在于,所述腺相关病毒载体系统还包括携带腺相关病毒载体的宿主细胞。
14.一种腺相关病毒,其特征在于,所述腺相关病毒由权利要求12或13所述的腺相关病毒载体系统经病毒包装而成。
15.一种细胞系,其特征在于,所述细胞系为经权利要求11所述的慢病毒或权利要求14所述的腺相关病毒感染的细胞系。
16.权利要求1-8任一所述的核酸构建体、权利要求9所述的慢病毒载体系统、权利要求11所述的慢病毒、权利要求12所述的腺相关病毒载体系统、权利要求14所述的腺相关病毒、权利要求15所述的细胞系在制备预防、治疗糖代谢相关疾病的产品中的用途。
17.根据权利要求16所述的用途,其特征在于,糖代谢相关疾病包括糖尿病或其并发症、肥胖症;优选的,所述糖尿病为2型糖尿病。
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WO2023045996A1 (zh) * | 2021-09-26 | 2023-03-30 | 康霖生物科技(杭州)有限公司 | 一种用于糖代谢相关疾病基因治疗的核酸构建体 |
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