含人丙型肝炎病毒全长基因组的核酸构建物、核酸构建物转入其中的重组全长病毒基因组复制型细胞和生产丙型肝炎病毒颗粒的方法

摘要

本发明提供有效复制含全长HCV基因组序列的RNA的方法和使用细胞培养系统生产含全长HCV复制子RNA或全长HCV基因组RNA的HCV病毒颗粒的方法。此外，本发明涉及生产丙型肝炎病毒颗粒的方法，其包括培养其中导入复制子RNA的细胞，并在培养基中产生病毒颗粒，其中所述复制子RNA包含的核苷酸序列含有基因型2a丙型肝炎病毒的全长基因组RNA序列、至少一种选择标记基因和/或至少一种报告基因以及至少一种IRES序列，或含有基因型2a丙型肝炎病毒的全长基因组RNA。再进一步，本发明还涉及丙型肝炎疫苗和抗丙型肝炎病毒颗粒的抗体。

说明书

技术领域

本发明涉及含丙型肝炎病毒全长基因组的核酸构建物、生产丙型肝炎病毒颗粒的体外方法和所生产的丙型肝炎病毒颗粒的用途。

背景技术

丙型肝炎病毒(HCV)属于黄病毒科家族，是一种具有单链正义RNA基因组的病毒，已知引起丙型肝炎。

HCV通过持续感染引起慢性肝炎。目前，在世界范围内观察到的慢性肝炎的重要病因是持续HCV感染。实际上，大约50％的持续感染个体发展成慢性肝炎。这些患者约有20％在10-20年间由慢性肝炎转变成肝硬化，这些患者中有一些进一步发展成晚期致死性病症，例如肝癌。

丙型肝炎目前主要通过使用干扰素-α或干扰素-β的疗法或使用干扰素-α和利巴韦林(一种嘌呤-核苷衍生物)组合的疗法治疗。但是，即使在实施这些疗法时，在所有受治疗患者中也仅有约60％观察到疗效。当观察到疗效后终止治疗时，一半以上的患者疾病复发。

一个重要目标是开发有效对抗丙型肝炎的治疗剂或预防剂。在工业化国家丙型肝炎发病率高，最终引发严重后果，目前还没有可用的病因疗法。因此，需要开发HCV-特异性化学疗法和疫苗疗法。开发抗HCV药物的目标可为抑制HCV复制或抑制HCV感染细胞。

最近，已制备了HCV亚基因组RNA复制子系统，作为HCV衍生的自主复制RNA(参见特许文献1、2和3，非特许文献1-4)。在HCV亚基因组RNA复制子系统中，制备其中HCV基因组结构基因被去除并被药物选择性标记基因取代的HCV复制子RNA，并将其导入培养细胞中，由此使复制子RNA在细胞中自主复制。使用这些系统使得有可能分析HCV的复制机制。但是，这是一个其中仅可在HCV病毒增殖和复制过程中评价病毒RNA复制的实验系统，不能分析被感染细胞中HCV病毒颗粒形成和胞外释放或感染另一细胞的过程。

此时，只能在使用动物如黑猩猩的实验系统中评价HCV病毒颗粒形成和胞外释放以及感染另一细胞的过程(非特许文献5)。但是，使用活生物(例如动物)的实验系统很复杂，非常难分析。因此，为了分析HCV病毒颗粒形成和胞外释放以及感染另一细胞的过程，为了生产具有抑制该过程的作用机制的抗HCV药物，必须建立重现该过程的高度简单化的实验系统，即在细胞培养实验系统中的HCV病毒颗粒生产系统。

此外，一旦可使用细胞培养系统稳定提供HCV病毒颗粒，就有可能使用分子生物学技术对病毒减毒或生产非感染性HCV病毒，这可用于疫苗。

特许文献1：日本特许公开(公报)第2001-17187号

特许文献2：国际专利申请PCT/JP03/15038

特许文献3：JP特许申请第2003-329082号

非特许文献1：Lohmann等，Science，(1999)285，110-113页

非特许文献2：Blight等，Science，(2000)290，1972-1974页

非特许文献3：Friebe等，J.Virol.，(2001)75(24)：12047-12057页

非特许文献4：Ikeda等，J.Virol.，(2002)76(6)：2997-3006页

非特许文献5：Kolykhalov等，Science，(1997)277，570-574页

非特许文献6：Kato等，Gastroenterology，(2003)125，1808-1817页

非特许文献7：Yanagi等，Proc.Natl.Acad.Sci.，(1997)96(16)：8738-8743页

非特许文献8：Okamoto等，J.Gen.Virol.，(1991)73，2697-26704页

非特许文献9：Aoyagi等，J.Clin.Microbiol.，(1999)37(6)：1802-1808页

发明公开

本发明的目标是提供含全长HCV基因组序列的RNA的有效复制方法和在细胞培养系统中生产含全长HCV复制子RNA或全长HCV基因组RNA的HCV病毒颗粒的方法。本发明的目标迄今为止还从未实现过。

为实现以上目标，本发明的发明人深入研究，结果开发出一种在细胞培养系统中生产HCV病毒颗粒的方法。即，本发明如下所述。

[1]一种复制子RNA，其包含的核苷酸序列含有基因型2a丙型肝炎病毒基因组RNA的5′非翻译区、核心蛋白编码序列、E1蛋白编码序列、E2蛋白编码序列、NS2蛋白编码序列、NS3蛋白编码序列、NS4A蛋白编码序列、NS4B蛋白编码序列、NS5A蛋白编码序列、NS5B蛋白编码序列和3′非翻译区，至少一种选择标记基因和/或至少一种报告基因，以及至少一种IRES序列。

在该复制子RNA中，优选核苷酸序列含在5′-3′方向为以下顺序的5′非翻译区、至少一种选择标记基因和/或至少一种报告基因，和至少一种IRES序列，以及核心蛋白编码序列、E1蛋白编码序列、E2蛋白编码序列、NS2蛋白编码序列、NS3蛋白编码序列、NS4A蛋白编码序列、NS4B蛋白编码序列、NS5A蛋白编码序列、NS5B蛋白编码序列和3′非翻译区。

在此复制子RNA的更优选实施方案中，基因型2a丙型肝炎病毒基因组RNA是含SEQ ID NO：12所示核苷酸序列的RNA。

在此复制子RNA的再更优选实施方案中，5′非翻译区含SEQ IDNO：1所示核苷酸序列，核心蛋白编码序列含SEQ ID NO：2所示核苷酸序列，E1蛋白编码序列含SEQ ID NO：3所示核苷酸序列，E2蛋白编码序列含SEQ ID NO：4所示核苷酸序列，NS2蛋白编码序列含SEQ ID NO：5所示核苷酸序列，NS3蛋白编码序列含SEQ ID NO：6所示核苷酸序列，NS4A蛋白编码序列含SEQ ID NO：7所示核苷酸序列，NS4B蛋白编码序列含SEQ ID NO：8所示核苷酸序列，NS5A蛋白编码序列含SEQ ID NO：9所示核苷酸序列，NS5B蛋白编码序列含SEQ ID NO：10所示核苷酸序列，而3′非翻译区含SEQ ID NO：11所示核苷酸序列。

[2]含以下(a)或(b)RNA的复制子RNA：

(a)含SEQ ID NO：13所示核苷酸序列的RNA；或

(b)含通过缺失、置换或添加1-100个核苷酸而衍生自SEQ ID NO：13所示核苷酸序列的核苷酸序列、并具有自主复制能力和病毒颗粒生产能力的RNA。

[3]一种生产细胞的方法，其中所述细胞复制复制子RNA并生产病毒颗粒，该方法包括将上述[1]或[2]中任一项的复制子RNA导入到细胞中。

用于该方法的细胞优选为增殖性细胞。同样或否则，用于该方法的细胞优选为真核细胞。

用于该方法的真核细胞优选为人肝源细胞、人子宫颈源细胞或人胎肾源细胞。所述真核细胞更优选为Huh7细胞、HepG2细胞、IMY-N9细胞、HeLa细胞或293细胞。

[4]利用上述[3]的方法可获得的细胞，其复制复制子RNA并生产病毒颗粒。

[5]生产丙型肝炎病毒颗粒的方法，包括培养上述[4]的细胞，以使该细胞生产病毒颗粒。

[6]利用上述[5]的方法可获得的丙型肝炎病毒颗粒。

[7]生产丙型肝炎病毒感染细胞的方法，包括培养上述[4]的细胞，并用培养物中的病毒颗粒感染其它细胞。

[8]通过上述[7]的方法可获得的丙型肝炎病毒感染细胞。

[9]筛选抗丙型肝炎病毒物质的方法，包括在测试物质存在下培养选自以下(a)、(b)和(c)中的至少一种：

(a)上述[4]的细胞，

(b)上述[8]的丙型肝炎病毒感染细胞，和

(c)上述[6]的丙型肝炎病毒颗粒和丙型肝炎病毒允许细胞；

并检测所获培养物中的复制子RNA或病毒颗粒。

[10]丙型肝炎疫苗，包含上述[6]的丙型肝炎病毒颗粒或其部分。

[11]使用上述[6]的丙型肝炎病毒颗粒或其部分作为抗原生产丙型肝炎疫苗的方法。

[12]通过使用上述[1]或[2]的复制子RNA生产用于基因治疗的嗜肝病毒载体的方法。

[13]利用上述[12]的方法可获得的嗜肝病毒载体。

[14]在细胞中复制和/或表达外源基因的方法，包括将编码外源基因的RNA插入到上述[1]或[2]中任一项的复制子RNA中，并将其导入到所述细胞中。

[15]一种细胞生产方法，其中所述细胞复制RNA并生产病毒颗粒，该方法包括将含SEQ ID NO：12所示核苷酸序列的RNA导入到该细胞中。

[16]生产丙型肝炎病毒颗粒的方法，包括将含SEQ ID NO：12所示核苷酸序列的RNA导入到细胞中，并培养细胞，使该细胞生产病毒颗粒。

[17]上述[15]或[16]的方法，其中所述细胞为增殖性细胞。

[18]生产含外源基因的病毒载体的方法，包括将编码外源基因的RNA插入到含SEQ ID NO：12所示核苷酸序列的RNA中，将其导入到细胞中，并培养细胞，使该细胞生产病毒颗粒。

[19]抗上述[6]的丙型肝炎病毒颗粒的抗体。

在本申请要求其优先权的日本特许申请第2004-045489号的说明书和附图中的内容结合到本文中。

附图简述

图1是一幅示意图，显示了构建模板DNA的方法，该模板DNA用于制备本发明的全长HCV复制子HCV或全长HCV基因组RNA。图1的上部显示了质粒克隆pJFH1的结构，其通过将全长HCV基因组插入到T7启动子下游产生。图1的下部显示了含全长HCV基因组序列的质粒克隆pFGREP-JFH1的结构，其中含新霉素抗性基因和EMCV IRES的DNA片段插入到pJFH1的T7启动子和5′非翻译区下游。该图中显示的术语如下。T7：T7RNA启动子，5′UTR：5′非翻译区，C：核心蛋白，E1、E2：包膜蛋白。NS2、NS3、NS4A、NS4B、4A、4B：非结构蛋白。3′UTR：3′非翻译区。AgeI、PmeI、XbaI：限制酶AgeI、PmeI和XbaI的限制性位点。GDD：对应于NS5B蛋白活性中心的氨基酸基序GDD的位点。neo：新霉素抗性基因。EMCVIRES：脑心肌炎病毒内部核糖体进入位点；

图2是一幅照片，显示了RNA印迹分析的结果，其表明rJFH-1在其中已导入全长HCV基因组RNA rJFH-1的Huh7细胞中复制；

图3显示了培养基中HCV核心蛋白定量的结果。空心圆代表其中已导入rJFH1的细胞，实心圆代表其中已导入rJFH1/GND的细胞；

图4图示了HCV核心蛋白和全长HCV基因组RNA的量，以及通过蔗糖密度梯度分级分离已导入rJFH-1的Huh7细胞的培养上清液所收集的各个级分的比重。实心圆、空心圆和阴影圆分别代表HCV核心蛋白、全长HCV基因组RNA和比重。

图5是一幅照片，显示了其中已转染入全长HCV复制子RNArFGREP-JFH1的Huh7细胞的集落形成；

图6是一幅照片，显示了全长HCV复制子RNA在全长HCV复制子RNA复制型细胞克隆中的复制，其中所述克隆已通过将rFGREP-JFH1转染入Huh7细胞中建立；

图7是一幅照片，显示了使用宿主细胞的基因组DNA作为模板和新霉素抗性基因特异性引物的PCR扩增的结果，用于证实新霉素抗性基因整合入基因组DNA中。M：DNA大小标记物，P：阳性对照，N：Huh7细胞；

图8是一幅照片，显示了蛋白质印迹分析的结果，其表明核心蛋白在其中已导入全长HCV复制子RNA rFGREP-JFH1的Huh7细胞中表达；

图9是一幅照片，显示了蛋白质印迹分析的结果，其表明NS3蛋白在其中已导入全长HCV复制子RNA rFGREP-JFH1的Huh7细胞中表达；

图10是一幅照片，显示了蛋白质印迹分析的结果，其表明NS5A蛋白在其中已导入全长HCV复制子RNA rFGREP-JFH1的Huh7细胞中表达；

图11图示了HCV核心蛋白和全长HCV复制子RNA的量，以及通过蔗糖密度梯度分级分离已导入rFGREP-JFH1的Huh7细胞的培养上清液所收集的各个级分的比重。实心圆、空心圆和阴影圆分别代表HCV核心蛋白、全长HCV复制子RNA和比重；

图12是一幅照片，显示了Huh7细胞的集落形成，其中已加入全长HCV复制子RNA复制型细胞培养上清液中的病毒颗粒。

实施本发明的最佳方式

本发明如下详细阐述。

1.全长HCV复制子RNA

丙型肝炎病毒(HCV)基因组是单股正链RNA，含约9600个核苷酸。此基因组RNA含5′非翻译区(也称为5′NTR或5′UTR)、由结构区和非结构区组成的翻译区以及3′非翻译区(也称为3′NTR或3′UTR)。HCV结构蛋白在结构区中编码，多种非结构蛋白在非结构区中编码。

这样的HCV结构蛋白(core、E1蛋白和E2蛋白)和非结构蛋白(NS2、NS3、NS4A、NS4B、NS5A和NS5B)如下产生：首先将翻译区翻译成单个连续的多蛋白，然后通过蛋白酶限制性切割多蛋白释放。在这些结构蛋白和非结构蛋白(即HCV的病毒蛋白)中，core是核心蛋白，E1和E2是包膜蛋白。非结构蛋白是参与病毒自身复制的蛋白，已知NS2具有金属蛋白酶活性，已知NS3具有丝氨酸蛋白酶活性(在N端侧的1/3)和解旋酶活性(在C端侧的2/3)。此外，NS4A是NS3蛋白酶活性的辅因子，已报道NS5B具有RNA依赖性RNA聚合酶活性。

本发明的发明人使用HVC基因组RNA构建了具有自主复制能力和病毒颗粒生产能力的复制子RNA。

已通过修饰HCV基因组RNA产生的具有自主复制能力的RNA在本文中叫做“复制子RNA”或“RNA复制子”。在本说明书中，源自HCV的复制子RNA还可叫做HCV-RNA复制子。含全长HCV基因组RNA的本发明复制子RNA在本文叫做“全长HCV复制子RNA”。本发明的全长HCV复制子RNA具有生产病毒颗粒的能力。

在本发明的全长HCV复制子RNA的优选实施方案中，丙型肝炎病毒优选为但不限于基因型2a丙型肝炎病毒。在本发明中，“基因型2a丙型肝炎病毒”或“基因型2a HCV”指按照Simmonds等(参见Simmonds，P.等，Hepatology，(1994)10，1321-1324页)的国际分类法鉴别为基因型2a的肝炎病毒。在本发明中，“基因型2a丙型肝炎病毒”或“基因型2a HCV”不仅包括具有天然HCV基因组RNA的病毒，而且包括具有其中天然HCV基因组序列已被人工修饰的基因组RNA的病毒。基因型2a HCV的具体实例包括JFH-1毒株(参见日本特许公开(公告)第2002-171978号)。

在本说明书中，“丙型肝炎病毒基因组RNA”指含丙型肝炎病毒单链正义RNA基因组整个区域的核苷酸序列的RNA。基因型2a丙型肝炎病毒的基因组RNA优选为但不限于含SEQ ID NO：12所示核苷酸序列的RNA。

本发明全长HCV复制子的一个实施方案是包含核苷酸序列的复制子RNA，该核苷酸序列包含5′非翻译区、核心蛋白编码序列、E1蛋白编码序列、E2蛋白编码序列、NS2蛋白编码序列、NS3蛋白编码序列、NS4A蛋白编码序列、NS4B蛋白编码序列、NS5A蛋白编码序列、NS5B蛋白编码序列和3′非翻译区、至少一种选择标记基因或报告基因以及至少一种IRES序列。

本发明的全长HCV复制子RNA不限于但优选包含：在5′-3′方向为以下顺序的5′非翻译区、至少一种选择标记基因或报告基因、至少一种IRES序列、核心蛋白编码序列、E1蛋白编码序列、E2蛋白编码序列、NS2蛋白编码序列、NS3蛋白编码序列、NS4A蛋白编码序列、NS4B蛋白编码序列、NS5A蛋白编码序列、NS5B蛋白编码序列和3′非翻译区。

在本申请的说明书中，“5′非翻译区”(5′NTR或5′UTR)、“核心蛋白编码序列”(核心区或C区)、“E1蛋白编码序列”(E1区)、“E2蛋白编码序列”(E2区)、“NS2蛋白编码序列”(NS2区)、“NS3蛋白编码序列”(NS3区)、“NS4A蛋白编码序列”(NS4A区)、“NS4B蛋白编码序列”(NS4B区)、“NS5A蛋白编码序列”(NS5A区)、“NS5B蛋白编码序列”(NS5B区)和“3′非翻译区”(3′NTR或3′UTR)，以及其它特定区域或位点，以包含基因型2a HCV病毒JFH-1毒株(日本特许公开(公报)第2002-171978号)基因组完整区域的全长基因组RNA(SEQ ID NO：12)为基准定义。

另外，本发明丙型肝炎病毒(HCV)基因组中的部分区域或位点可以SEQ ID NO：1-11所示序列为基准定义，SEQ ID NO：1-11是JFH-1毒株基因组RNA(SEQ ID NO：12)的部分核苷酸序列。JFH-1毒株全长基因组RNA(源自JFH-1克隆；SEQ ID NO：12)的“5′非翻译区”包含SEQ ID NO：1所示核苷酸序列。“核心蛋白编码序列”包含SEQ IDNO：2所示核苷酸序列。“E1蛋白编码序列”包含SEQ ID NO：3所示核苷酸序列。“E2蛋白编码序列”包含SEQ ID NO：4所示核苷酸序列。“NS2蛋白编码序列”包含SEQ ID NO：5所示核苷酸序列。“NS3蛋白编码序列”包含SEQ ID NO：6所示核苷酸序列。“NS4A蛋白编码序列”包含SEQ ID NO：7所示核苷酸序列。“NS4B蛋白编码序列”包含SEQ ID NO：8所示核苷酸序列。“NS5A蛋白编码序列”包含SEQ IDNO：9所示核苷酸序列。“NS5B蛋白编码序列”包含SEQ ID NO：10所示核苷酸序列。“3′非翻译区”包含SEQ ID NO：11所示核苷酸序列。

例如，HCV源RNA序列中的区域或位点可由SEQ ID NO：1-12的核苷酸序列中的核苷酸编号限定，核苷酸编号通过比对RNA序列和SEQ ID NO：1-12所示核苷酸序列来确定。在比对中，可存在空位、添加、缺失、置换等。

在本发明的更优选实施方案中，包含在全长HCV复制子RNA中的5′非翻译区、核心蛋白编码序列、E1蛋白编码序列、E2蛋白编码序列、NS2蛋白编码序列、NS3蛋白编码序列、NS4A蛋白编码序列、NS4B蛋白编码序列、NS5A蛋白编码序列、NS5B蛋白编码序列和3′非翻译区优选分别包含SEQ ID NO：1-11所示核苷酸序列。

本发明的全长HCV复制子RNA的优选实施方案为含SEQ ID NO：1-11所示核苷酸序列、至少一种标记基因和/或报告基因和至少一种IRES序列的复制子RNA。

本发明中的“选择标记基因”指赋予细胞选择能力的基因，使得只有表达该基因的细胞才可被选择。选择标记基因的一般性实例包括抗生素抗性基因。本发明中优选的选择标记基因实例包括新霉素抗性基因、胸苷激酶基因、卡那霉素抗性基因、抗硫胺素抗性基因、腺苷酰转移酶基因、Zeocin抗性基因和嘌呤霉素抗性基因。优选新霉素抗性基因和胸苷激酶基因，更优选新霉素抗性基因。但是，本发明中的选择标记基因不限于这些基因。

此外，在本发明中，“报告基因”指其编码基因产物可作为该基因表达指示剂的标记基因。报告基因的一般性实例包括催化光发射反应或显色反应的酶的结构基因。本发明中的报告基因的优选实例包括源自转座子Tn9的氯霉素乙酰转移酶基因、大肠杆菌源的β-葡糖醛酸酶基因或β-半乳糖苷酶基因、荧光素酶基因、绿色荧光蛋白基因、得自水母的水母发光蛋白基因和分泌型胎盘碱性磷酸酶(SEAP)基因。但是，本发明中的报告基因不限于这些基因。

在全长复制子RNA中可包含以上选择标记基因和报告基因中的仅一种，或者可同时包含二者。一种或多种选择标记基因或报告基因可存在于一个全长HCV复制子RNA中。

在本发明中，“IRES序列”指内部核糖体进入位点，其允许核糖体通过结合RNA内部启动翻译。本发明中的IRES序列的优选实例包括但不限于EMCV IRES(脑心肌炎病毒的内部核糖体进入位点)、FMDV IRES和HCV IRES。更优选EMCV IRES和HCV IRES，EMCVIRES是最优选的序列。

本发明的全长HCV复制子RNA的再更优选实施方案是含SEQID NO：13所示核苷酸序列的RNA。此外，含在SEQ ID NO：13所示核苷酸序列中通过缺失、置换或添加1-100个核苷酸(优选1-30个，更优选1-10个，再更优选1-6个，最优选1至若干(2-5)个核苷酸)而衍生自SEQ ID NO：13所示核苷酸序列的核苷酸序列、具有自主复制能力和病毒颗粒生产能力的复制子RNA是全长HCV复制子RNA的优选实施方案，也包括在本发明范围中。

本发明的全长HCV复制子RNA还可包含编码任选外源基因的RNA，其中所述外源基因要在全长复制子RNA已导入其中的细胞中表达。编码外源基因的RNA还可连接在5′非翻译区下游，或连接在选择标记基因或报告基因的上游或下游，或连接在3′非翻译区上游。编码外源基因的RNA可插入在核心蛋白编码序列、E1蛋白编码序列、E2蛋白编码序列、NS2蛋白编码序列、NS3蛋白编码序列、NS4A蛋白编码序列、NS4B蛋白编码序列、NS5A蛋白编码序列和NS5B蛋白编码序列之间的任意位点。

当含外源基因编码RNA的全长HCV复制子RNA在已导入其的细胞中翻译时，其可表达外源基因编码的基因产物。因此，含外源基因编码RNA的全长HCV复制子RNA还可适用于在细胞中生产外源基因的基因产物。

本发明的全长HCV复制子RNA可进一步包含核酶。核酶连接在选择标记基因和/或报告基因下游，使得可通过核酶的自切割活性将选择标记基因和/或报告基因由IRES序列、核心蛋白编码序列、E1蛋白编码序列、E2蛋白编码序列、NS2蛋白编码序列、NS3蛋白编码序列、NS4A蛋白编码序列、NS4B蛋白编码序列、NS5A蛋白编码序列和NS5B蛋白编码序列以及3′非翻译区上切下。

在本发明的全长HCV复制子RNA中，上述选择标记基因和/或报告基因、病毒蛋白编码序列和外源基因、核酶等被连接在一起，使得它们由全长HCV复制子RNA以正确的读框翻译。在这些序列中，全长复制子RNA编码的蛋白优选通过蛋白酶切割位点等互相连接，使得蛋白被翻译或表达为多蛋白，接着由蛋白酶切割成各个蛋白。

本发明还涉及编码本发明复制子RNA的DNA载体，优选表达载体。

在本发明中，RNA的“自主复制能力”指RNA在导入细胞中时能够自主生长。RNA的自主复制能力可通过以下方法证实，但该方法不是限制性的。用目的RNA转染Huh7细胞并培养。由所获得的培养细胞提取RNA，并使用能够特异性检测导入RNA的探针对RNA进行RNA印迹杂交。目的RNA检测证实自主复制。在本说明书实施例中有关集落形成能力实验、HCV蛋白表达验证、复制子RNA检测等的描述中阐述了证实自主复制能力的具体方法的实例。

此外，在本发明中，RNA的“病毒颗粒生产能力”指当RNA导入到细胞(例如培养细胞，如Huh7细胞)中时在细胞中产生病毒颗粒。病毒颗粒生产能力可通过例如将使用RNA特异性引物的RT-PCR法用于检测导入该RNA的细胞的培养上清液来证实。其还可通过对培养上清液进行蔗糖密度梯度法以分离病毒颗粒和通过检测HCV蛋白来证实。在本说明书实施例中有关集落形成能力实验、HCV蛋白表达验证、复制子RNA检测等的描述中阐述了具体方法的实例。

2.全长HCV复制子RNA的制备

可使用本领域技术人员已知的基因工程技术制备本发明的全长HCV复制子RNA。可使用例如但不限于JFH-1毒株作为基因型2a丙型肝炎病毒，利用以下的方法制备全长HCV复制子RNA。

首先，常规地再构建对应于JFH-1毒株基因组RNA完整区域(SEQ ID NO：12；该序列在国际DNA数据库中在检索号AB047639下)的DNA，并将其插入到RNA启动子下游，以便制备DNA克隆。本文使用的“对应于RNA的DNA”指具有通过用T(胸腺嘧啶)取代U(尿嘧啶)而来源于RNA核苷酸序列的核苷酸序列的DNA。以上RNA启动子优选包含在质粒克隆中。优选RNA启动子的实例不限于但包括T7RNA启动子、SP6 RNA启动子和SP3 RNA启动子，特别优选T7 RNA启动子。

接着，将选择标记基因和/或报告基因以及IRES序列编码DNA插入到上述DNA克隆中。优选将选择标记基因和/或报告基因插入到5′非翻译区下游，将IRES序列插入到更下游。

随后，使用如上制备的DNA克隆作为模板，使用RNA聚合酶合成RNA。RNA合成可通过标准方法由5′非翻译区开始。当DNA克隆为质粒克隆时，可用使用限制酶由质粒克隆切下的DNA片段作为模板合成RNA。另外，优选要合成的RNA的3′端与病毒基因组RNA的3′非翻译区的末端具有相同序列，未添加或缺失其它序列。由此合成的RNA是本发明的全长HCV复制子RNA。

3.HCV颗粒的制备

可通过将如上所述制备的全长HCV复制子RNA导入细胞中，获得可复制全长HCV复制子RNA(优选持续复制(即具有复制子RNA复制能力))的重组细胞。在本说明书中，复制全长HCV复制子RNA的重组细胞被称为“全长HCV复制子RNA复制型细胞”。

全长HCV复制子RNA复制型细胞可生产病毒颗粒。生产的病毒颗粒在由HCV病毒蛋白组成的外壳中含全长HCV复制子RNA。因此，本发明的全长HCV复制子RNA复制型细胞生产的病毒颗粒是HCV颗粒。也就是说，在本发明中，可通过培养全长HCV复制子RNA复制型细胞，在细胞培养系统中制备HCV颗粒。优选地，可通过培养全长HCV复制子RNA复制型细胞，并收集在培养物(优选培养上清液)中产生的病毒颗粒，获得HCV颗粒。

或者，可利用通过将全长HCV基因组RNA导入到细胞中获得的重组细胞生产HCV颗粒。全长HCV基因组RNA在其中导入本发明的全长HCV基因组RNA(优选来源于JFH-1克隆的全长HCV基因组RNA，更优选具有SEQ ID NO：12所示核苷酸序列的RNA)的细胞中高效复制。在本说明书中，复制全长HCV基因组RNA的细胞称为“全长HCV基因组RNA复制型细胞”。全长HCV基因组RNA复制型细胞可生产病毒颗粒。全长HCV基因组RNA复制型细胞生产的病毒颗粒在由HCV病毒蛋白组成的壳中包含全长HCV基因组RNA。因此，由其中导入本发明的全长HCV基因组RNA的细胞生产的病毒颗粒是HCV颗粒。不限于但优选地可通过培养其中导入来源于JFH-1克隆的全长HCV基因组RNA(例如具有SEQ ID NO：12所示核苷酸序列的RNA)的细胞，在细胞培养系统中制备HCV颗粒。例如，可通过培养其中导入全长HCV基因组RNA(例如具有SEQ IDNO：12所示核苷酸序列的RNA)的细胞，并收集在培养物(优选培养上清液)中产生的病毒颗粒，获得HCV颗粒。

对于要导入上述全长HCV复制子RNA或全长HCV基因组RNA的细胞，可使用任何细胞，只要其可被传代培养。这样的细胞优选为真核细胞，更优选为人细胞，再更优选为人肝源细胞、人子宫颈源细胞或人胎肾源细胞。优选可使用增殖性细胞，包括癌细胞系、干细胞系等细胞，更优选使用Huh7细胞、HepG2细胞、IMY-N9细胞、HeLa细胞和293细胞等。对于这些细胞，可使用市售的细胞，这些细胞可得自细胞保藏机构，或者也可使用由任意细胞(例如癌细胞或干细胞)建立的细胞系。

可使用本领域技术人员已知的任何技术，实现将全长HCV复制子RNA或全长HCV基因组RNA导入到细胞中。这样的导入方法的实例包括电穿孔、粒子枪法、脂转染法、磷酸钙法、微注射法、DEAESepharose法等。特别优选使用电穿孔的方法。

全长HCV复制子RNA或全长HCV基因组RNA可单独导入，或者可在与其它核酸混合后导入。为改变全长HCV复制子RNA或全长HCV基因组RNA的量，同时将要导入的RNA量保持在一定水平，将需要量的要导入的全长HCV复制子RNA或全长HCV基因组RNA与由要导入RNA的细胞提取的总细胞RNA混合，以使总RNA量达到一定水平，然后混合物用于导入到细胞中。可按照使用的导入方法确定用于导入到细胞中的复制子RNA的量，其优选为1pg-100μg，更优选为10pg-10μg。

可利用在全长HCV复制子RNA中的选择标记基因或报告基因的表达，选择全长HCV复制子RNA复制型细胞。具体地说，例如，可在培养基中培养这种进行了全长HCV复制子RNA细胞导入处理的细胞，其中细胞可由于表达选择标记基因而被选择。或者，在进行全长HCV复制子RNA细胞导入处理的细胞培养后，可检测报告基因(例如荧光蛋白)的表达。

作为实例，当全长HCV复制子RNA包含新霉素抗性基因作为选择标记基因时，将用全长HCV复制子RNA进行电穿孔法的细胞接种入培养皿中。在培养12-72小时(优选16-48小时)后，以0.05mg/ml-3.0mg/ml的浓度将G418(新霉素)加入到培养皿中。优选在接种后将细胞连续培养10-40天，更优选14-28天，同时一周更换培养基两次，可通过用结晶紫染色活细胞，选择复制导入的全长HCV复制子RNA的细胞的集落。

可通过标准方法由形成的集落克隆细胞。由此获得的复制全长HCV复制子RNA的细胞克隆在本说明书中被称为“全长HCV复制子RNA复制型细胞克隆”。本发明的全长HCV复制子RNA复制型细胞包括全长HCV复制子RNA复制型细胞克隆。

对于全长HCV复制子RNA复制型细胞，可通过检测复制的全长HCV复制子RNA，确认全长HCV复制子RNA的选择标记基因或报告基因没有整合在宿主基因组DNA中，并进一步检测HCV蛋白，证实全长HCV复制子RNA在细胞或细胞克隆中的实际复制。

可按照本领域技术人员已知的任意RNA检测方法，检测已复制的全长HCV复制子RNA。例如，可通过使用全长HCV复制子RNA特异性DNA片段作为探针的RNA杂交法，检测由细胞提取的总RNA中的全长HCV复制子RNA。

而且，可通过例如但不限于对由细胞提取出的基因组DNA进行PCR，以扩增至少一部分选择标记基因或报告基因，然后确认没有扩增产物，来证实全长HCV复制子RNA中的选择标记基因或报告基因没有整合在宿主基因组DNA中。因为一般认为在证实扩增产物的细胞中选择标记基因或报告基因可能已整合入宿主基因组中，所以有可能全长HCV复制子RNA自身不复制。在此情况下，可如下所述通过检测HCV蛋白进一步证实全长HCV复制子RNA的复制。

可通过例如使抗HCV蛋白(其由导入的全长HCV复制子RNA表达)的抗体与提取的细胞蛋白反应，检测HCV蛋白。该方法可通过本领域技术人员已知的任意蛋白检测方法进行。具体地说，例如，可通过将由细胞提取出的蛋白样品印迹在硝酸纤维素膜上，使抗HCV蛋白抗体(例如抗-NS3特异性抗体或由丙型肝炎患者收集的抗血清)与硝酸纤维素膜反应，并检测抗HCV蛋白抗体，检测HCV蛋白。如果在提取的细胞蛋白中检测出HCV蛋白，则可推断该细胞复制全长HCV复制子RNA，并表达HCV蛋白。

可通过本领域技术人员已知的任意病毒检测方法，证实全长HCV复制子RNA复制型细胞或全长HCV基因组RNA复制型细胞的病毒颗粒生产能力。例如，通过蔗糖密度梯度分级分离怀疑生产病毒颗粒的细胞的培养上清液，并测定各个级分的级分密度、HCV核心蛋白浓度以及全长HCV复制子RNA或全长HCV基因组RNA的量。结果，如果核心蛋白的峰与全长HCV复制子RNA或全长HCV基因组RNA的峰一致，显示检测峰的级分的密度(例如1.18-1.20mg)小于分级分离25％NP40(聚氧乙烯(9)辛基苯基醚)处理的培养上清液所获得的等同级分的密度，则可认为细胞具有病毒颗粒生产能力。

例如可使用核心蛋白、E1蛋白或E2蛋白的抗体检测培养上清液中释放的HCV病毒颗粒。另外，可通过使用特异性引物的RT-PCR法扩增和检测培养上清液中的全长HCV复制子RNA，间接检测HCV病毒颗粒的存在。

4.用本发明的HCV颗粒感染另一细胞

本发明的HCV病毒颗粒具有感染细胞(优选HCV允许细胞)的能力。本发明还涉及生产丙型肝炎病毒感染细胞的方法，包括培养全长HCV复制子RNA复制型细胞或全长HCV基因组RNA复制型细胞，并用在由此获得的培养物(优选培养上清液)中的病毒颗粒感染另一细胞(优选HCV允许细胞)。在本发明中，HCV允许细胞指对HCV敏感的细胞，优选但不限于肝细胞或淋巴系细胞。具体地说，肝细胞包括原代肝细胞、Huh7细胞、HepG2细胞、IMY-N9细胞、HeLa细胞、203细胞等。淋巴系细胞包括但不限于Molt4细胞、HPB-Ma细胞、Daudi细胞等。

当用本发明的全长HCV复制子RNA复制型细胞生产的HCV颗粒感染细胞(例如HCV允许细胞)时，全长HCV复制子RNA在感染细胞中复制，病毒颗粒也在感染细胞中形成。因为用在全长HCV复制子RNA复制型细胞中产生的病毒颗粒感染的细胞表达选择标记基因和/或报告基因，所以可利用表达选择和/或检测感染细胞。通过用在本发明的全长HCV复制子RNA复制型细胞中产生的病毒颗粒感染细胞，全长HCV复制子RNA在细胞中复制，可进一步生产病毒颗粒。

更进一步，通过用在本发明的全长HCV基因组RNA复制型细胞中产生的HCV颗粒感染细胞(例如HCV允许细胞)，全长HCV基因组RNA在感染细胞中复制，病毒颗粒也在感染细胞中形成。通过用在本发明的全长HCV基因组RNA复制型细胞中产生的病毒颗粒感染细胞，全长HCV基因组RNA在细胞中复制，而且可生产病毒颗粒。

在全长HCV复制子RNA复制型细胞或全长HCV基因组RNA复制型细胞中产生的HCV病毒颗粒可感染HCV允许动物，例如黑猩猩等，并在其中诱发由HCV引起的肝炎。

5.本发明的其它实施方案

全长HCV复制子RNA在本发明的全长HCV复制子RNA复制型细胞中高效复制。另外，全长HCV基因组RNA在本发明的全长HCV基因组RNA复制型细胞中高效复制。因此，可使用本发明的全长HCV复制子RNA复制型细胞或全长HCV基因组RNA复制型细胞高效生产全长HCV复制子RNA或全长HCV基因组RNA。

在本发明中，可通过培养全长HCV复制子RNA复制型细胞，由培养物(培养细胞和/或培养基)中提取RNA，对RNA进行电泳法，并分离和纯化全长HCV复制子RNA，生产全长HCV复制子RNA。还可通过相似的方法，使用全长HCV基因组RNA复制型细胞生产全长HCV基因组RNA。通过这样的方法生产的RNA包含丙型肝炎病毒的全长基因组序列。在此情况下，可通过选择标记基因和/或报告基因和IRES序列中断丙型肝炎病毒的全长基因组序列。借助于包含所提供的丙型肝炎病毒全长基因组序列的RNA的生产方法，使得有可能更详细地分析丙型肝炎病毒基因组。

再者，本发明的全长HCV复制子RNA复制型细胞或全长HCV基因组RNA复制型细胞可适用于生产HCV蛋白。HCV蛋白可利用本领域技术人员已知的任何方法生产。例如，可通过将全长HCV复制子RNA或全长HCV基因组RNA导入到细胞中，培养重组细胞，并利用已知方法由如此获得的培养物(培养细胞和/或培养基)中收集蛋白，生产HCV蛋白。

再者，本发明的HCV病毒颗粒可具有嗜肝性。因此，可使用本发明的全长HCV复制子RNA生产嗜肝病毒载体。该病毒载体适用于基因治疗。在本发明中，通过将外源基因编码RNA整合入全长HCV复制子RNA或全长HCV基因组RNA中并将整合的RNA导入细胞中表达，外源基因可被导入到细胞中，并在细胞中复制。此外，通过制备其中全长HCV复制子RNA或全长HCV基因组RNA的E1蛋白编码序列和/或E2蛋白编码序列被其它生物物种来源病毒的外壳蛋白编码序列置换的RNA，使该RNA有可能感染不同的生物物种。还是在此情况下，外源基因整合入全长HCV复制子RNA或全长HCV基因组RNA中，这可用作在肝细胞中表达外源基因的嗜肝病毒载体。

本发明还涉及生产携带外源基因的病毒载体的方法，包括将外源基因编码RNA插入到含SEQ ID NO：12所示核苷酸序列的RNA中，将其导入到细胞中，并培养细胞，以生产病毒颗粒。

本发明提供含本发明的HCV颗粒或其部分的丙型肝炎疫苗，生产含本发明的HCV颗粒或其部分的丙型肝炎疫苗的方法。

具体地说，如上制备的HCV颗粒可直接用作疫苗，或者可在利用本领域已知方法减毒或灭活后使用。例如，可通过使用柱层析、过滤、离心等纯化HCV颗粒，获得HCV疫苗母液。可由此HCV疫苗母液制备减毒活HCV疫苗或灭活HCV疫苗。可通过例如将灭活剂(如福尔马林、β-丙内酯、戊二醛等)加入到病毒悬浮液中并混合，使其与病毒反应(Appaiahgari等，Vaccine，(2004)22(27-28)，3669-3675页)，进行病毒灭活。

为生产本发明的疫苗，有可能使用其中通过使用公开已知的技术引入突变减弱或消除病原性的HCV复制子RNA。

本发明的疫苗制备成溶液或悬浮液给予。还有可能制备为适于溶解或悬浮在液体中的固体物质形式。制品可在脂质体中乳化或囊化。活性免疫原性组分如HCV颗粒经常与药物可接受的并适于活性成分的赋形剂混合。合适的赋形剂包括例如水、生理盐水、葡萄糖、甘油、乙醇及其混合物。此外，如果有需要，疫苗可包含少量的助剂(例如增湿剂或乳化剂)、pH缓冲剂和/或用于增强疫苗效力的佐剂。有效佐剂的实例包括但不限于以下物质：氢氧化铝、N-乙酰-胞壁酰-L-苏氨酰-D-异谷氨酰胺(thr-MDP)、N-乙酰-正-胞壁酰-L-丙氨酰-D-异谷氨酰胺(CGP 11637，nor-MDP)、N-乙酰胞壁酰-L-丙氨酰-D-异谷氨酰胺-L-丙氨酸-2-(1′-2′-二棕榈酰-sn-甘油-3-羟基磷酰氧基)-乙胺(CGP19835A，称为MTP-PE)和RIBI。RIBI在2％鲨烯/Tween 80乳浊液中包含三种提取自细菌的组分，即单磷酰脂质A、二分枝菌酸海藻糖和细胞壁骨架(HPL+TDM+CWS)。可通过检测抗免疫原性HCV颗粒的抗体的量测定佐剂效力，其中所述抗体通过给予由HCV颗粒组成的疫苗产生。

本发明的疫苗一般胃肠外给予，例如通过注射，如皮下或肌内注射。适于其它给予途径的其它剂型包括栓剂，在某些情况下包括口服制剂。

如果有需要，可将一种或多种具有佐剂活性的上述化合物加入到HCV疫苗中。佐剂为此免疫系统的非特异性刺激因子，在宿主中增强对HCV疫苗的免疫应答。该技术领域已知佐剂的具体实例包括完全弗氏佐剂、不完全弗氏佐剂、维生素E、非离子型嵌段共聚物、胞壁酰二肽、皂苷、矿物油、植物油和Carbopol。特别适用于粘膜的佐剂包括例如大肠杆菌不耐热毒素(LT)和霍乱毒素(CT)。其它合适的佐剂包括例如氢氧化铝、磷酸铝或氧化铝、油乳浊液(例如Bayol(注册商标)或Marcol 52(注册商标))、皂苷或维生素E增溶物。在优选的实施方案中，本发明的疫苗包含佐剂。

例如，对于皮下、皮内、肌内和静脉内给予的注射，本发明的HCV疫苗中可包含的药物可接受的载体和稀释剂的具体实例包括稳定剂、糖类(例如山梨醇、甘露醇、淀粉、蔗糖、葡萄糖、葡聚糖)、蛋白(例如白蛋白或酪蛋白)、含蛋白的物质(例如牛血清白蛋白或脱脂奶)和缓冲液(例如磷酸盐缓冲液)。

用于栓剂的常规粘合剂和载体包括例如聚亚烷基二醇或甘油三酯。栓剂可由含0.5％-50％(优选1％-20％)活性成分的混合物配制。口服制剂可包含通常使用的赋形剂。这样的赋形剂包括例如药用级的甘露醇、乳糖、淀粉、硬脂酸镁、糖精钠、纤维素、碳酸镁等。

本发明的疫苗可以溶液、悬浮液、片剂、丸剂、胶囊、延迟释放制剂或粉末剂型生产，并可包含10-95％(优选25-70％)的活性成分(病毒颗粒或其部分)。

本发明的疫苗通过适于剂型的方法并以预防和/或治疗有效量给予。剂量为0.01μg-100,000μg，这取决于要治疗的患者、患者免疫系统中的抗体形成能力和需要的保护水平。其还依赖于给予途径，例如口服、皮下、皮内、肌内、静脉内等。

该疫苗可通过单次给予程序给予，或优选通过复合给予程序给予。在复合给予程序中，在给予开始时进行1-10次单独给予，接着以维持和/或增强免疫反应所需要的间隔给予。例如，可在1-4个月后进行另一类给予，作为第二次给予。必要时，可在几个月后持续给予。给予方案至少部分地按照个体患者的需要来确定，取决于主治医师的判断。

此外，含免疫原性HCV颗粒的疫苗可与其它免疫控制剂(例如免疫球蛋白)共同给予。

HCV颗粒疫苗可通过给予健康个体，以诱发对HCV的免疫应答，用于预防性地对抗可能的新HCV感染。HCV颗粒疫苗还可通过给予感染HCV的患者并在体内诱发针对HCV的强免疫应答，用作治疗性疫苗，以清除HCV。

全长HCV复制子RNA复制型细胞或全长HCV基因组RNA复制型细胞或用在这些细胞中产生的病毒颗粒感染的丙型肝炎病毒感染细胞，可用作测试系统，用于筛选促进或抑制例如丙型肝炎病毒复制、病毒颗粒重构和病毒颗粒释放的物质(抗丙型肝炎病毒的物质)。具体地说，例如，可通过确定测试物是否促进或抑制全长HCV复制子RNA或全长HCV基因组RNA的复制或者病毒颗粒的形成或释放，在测试物存在下培养这些细胞，并检测所获培养物中的全长HCV复制子RNA或全长HCV基因组RNA或病毒颗粒，筛选促进或抑制丙型肝炎病毒生长的物质。在此情况下，可通过测定由上述细胞提取的RNA制品中的全长HCV复制子RNA或全长HCV基因组RNA的量、比率或存在情况，对培养物中的全长HCV复制子RNA或全长HCV基因组RNA进行检测。可通过检测培养上清液中HCV蛋白的量、比率或存在情况，对培养物(主要是培养上清液)中的病毒颗粒进行检测。

而且，可通过检测该培养物中的病毒颗粒，研究由HCV感染患者的血清纯化的免疫球蛋白是否可预防本发明HCV颗粒的感染。在此测试中，可使用已用本发明的HCV病毒颗粒免疫的小鼠、大鼠、兔等的血清。可使用采用部分HCV蛋白、HCV基因等进行的免疫。可以相似的方式对其它感染预防性物质进行该测试。

产生的抗本发明HCV病毒颗粒的本发明抗体包括多克隆抗体和单克隆抗体。当优选多克隆抗体时，第一步用本发明的HCV颗粒免疫选定的哺乳动物(例如小鼠、兔、山羊、绵羊、马等)。由被免疫动物收集血清，并用已知方法加工。如果含针对HCV表位的多克隆抗体的血清包含针对其它抗原的抗体，则可通过免疫亲和层析纯化这些血清。生产多克隆抗血清的方法及其治疗方法是本领域已知的。可由已感染HCV的哺乳动物分离多克隆抗体。

本领域技术人员可容易地生产针对HCV表位的单克隆抗体。已知的常用方法是生产产生单克隆抗体的杂交瘤。例如，可使用在Current Protocols in Immunology(John Wiley & Sons，Inc.)中描述的方法。

可通过细胞融合或其它方法，例如用肿瘤基因DNA直接转化B淋巴细胞或用EB病毒转导，生产产生单克隆抗体的细胞系。

通过这些方法获得的单克隆抗体和多克隆抗体用于诊断、治疗和预防HCV。

使用本发明的HCV颗粒生产的抗体可与药物可接受的增溶剂、添加剂、稳定剂、缓冲剂等一起给予。可选择任何给予途径，但优选皮下、皮内和肌内给予，更优选静脉内给予。

在本发明的全长HCV复制子RNA复制型细胞或全长HCV基因组RNA复制型细胞中产生的HCV颗粒和HCV允许细胞可作为测试系统，用于筛选可刺激或抑制HCV与细胞结合的物质。具体地说，例如，可通过在测试物存在下培养本发明的全长HCV复制子RNA复制型细胞中产生的HCV颗粒和HCV允许细胞，检测所获培养物中的全长HCV复制子RNA或病毒颗粒，并确定测试物是否促进或抑制复制子RNA复制或病毒颗粒形成，筛选可促进或抑制丙型肝炎病毒生长的物质。

可按照上述技术或以下实施例对全长HCV复制子RNA或全长HCV基因组RNA进行这样的检测。上述测试系统可用于生产和评价丙型肝炎病毒感染的预防剂、治疗剂或诊断剂。

具体地说，上述本发明测试系统的应用实例包括以下几点：

(1)筛选抑制HCV生长和感染的物质

抑制HCV生长和感染的物质包括例如直接或间接影响HCV生长和感染的有机化合物、通过与HCV基因组或其互补链中的靶序列杂交而直接或间接影响HCV生长或HCV蛋白翻译的反义寡核苷酸等。

(2)评价在细胞培养物中具有抗病毒活性的各种物质

前述各种物质包括通过合理药物设计或高通量筛选获得的物质(例如纯化和分离的酶)。

(3)鉴别治疗HCV感染患者的新靶标

例如，本发明的全长HCV复制子RNA复制型细胞或全长HCV基因组RNA复制型细胞可用于鉴别可在HCV生长中起重要作用的宿主细胞蛋白。

(4)评价HCV对药物等的抗性获得能力，并鉴别与抗性相关的突变

(5)生产病毒蛋白作为抗原，用于开发、生产和评价丙型肝炎病毒感染的诊断剂和治疗剂

(6)生产病毒蛋白作为抗原，用于开发、生产和评价用于丙型肝炎病毒感染的疫苗和生产减毒HCV

(7)生产用于诊断和治疗丙型肝炎病毒感染的单克隆或多克隆抗体。

基于以下的实施例和附图将更具体地描述本发明。但本发明的技术范围不限于这些实施例。

实施例1：得自全长HCV基因组RNA的全长HCV复制子RNA的制备

(A)构建表达载体

构建质粒DNA，其中DNA(JFH-1克隆)插入到pUC19质粒中T7RNA启动子序列的下游，此DNA(JFH-1克隆)含已由暴发性肝衰竭患者中分离出的丙型肝炎病毒JFH-1毒株(基因型2a)的全长基因组cDNA。

具体地说，将通过扩增JFH-1毒株的病毒RNA所获得的RT-PCR片段克隆入pGEM-T EASY载体(Promega)中，以获得质粒pGEM1-258、pGEM44-486、pGEM317-849、pGEM617-1323、pGEM1141-2367、pGEM2285-3509、pGEM3471-4665、pGEM4547-5970、pGEM5883-7003、pGEM6950-8035、pGEM7984-8892、pGEM8680-9283、pGEM9231-9634和pGEM9594-9678(参见非特许文献6)。通过使用PCR法和限制酶，将包含在这些质粒中的病毒基因组RNA来源的cDNA连接在一起，以克隆全长病毒基因组cDNA。将T7R RNA启动子序列插入到全长病毒基因组cDNA的上游。在下文中，以此方式构建的质粒DNA称为pJFH1(图1的上部)。上述JFH-1克隆的制备已描述于特许文献1和非特许文献3。此外，JFH-1克隆的全长cDNA的核苷酸序列以检索号AB047639登记在国际DNA数据库(DDBJ/EMBL/GenBank)中。

接着，通过将EMCV-IRES(脑心肌炎病毒的内部核糖体进入位点)和新霉素抗性基因(neo；也称为新霉素磷酸转移酶基因)插入到pJFH1质粒DNA(图1的下部)的5′非翻译区和核心蛋白区之间，构建质粒DNA pFGREP-JFH1。该构建方法与先前的公开内容(非特许文献4)一致。此外，通过导入突变制备突变质粒克隆pJFH1/GND和pFGREP-JFH1/GND，其中所述突变将对应于RNA聚合酶(由pJFH1和pFGREP-JFH1中的NS5B区编码)活性中心的氨基酸基序GDD变为GND。因为由突变克隆pJFH1/GND和pFGREP-JFH1/GND编码的NS5B蛋白活性位点的氨基酸序列被改变，所以不能由突变克隆表达复制复制子RNA所需的活性NS5B蛋白。

此外，通过将荧光素酶基因插入到pFGREP-JFH1的第415-420的MluI位点和第2075-2082的PmeI位点之间，以用荧光素酶基因替代pFGREP-JFH1的新霉素抗性基因，制备pFGREP-JFH1/Luc，作为导入报告基因的表达载体。另外，通过将pFGREP-JFH1/Luc第10933位的G突变为A，制备突变体pFGREP-JFH1/Luc/GND，其中NS5bRNA聚合酶活性中心的GDD基序被变为GND。

通过将绿色荧光蛋白基因插入到pFGREP-JFH1的第415-420的MluI位点和第1142-1149的PmeI位点之间，制备pFGREP-JFH1/EGFP，其中pFGREP-JFH1的新霉素抗性基因被绿色荧光蛋白基因取代。另外，通过将pFGREP-JFH1/EGFP第10000位的G突变为A，制备突变体pFGREP-JFH1/EGFP/GND，其中NS5b RNA聚合酶活性中心的GDD基序被变为GND。

通过将分泌型胎盘碱性磷酸酶基因插入到pFGREP-JFH1的第415-420的MluI位点和第1982-1989的PmeI位点之间，以用分泌型胎盘碱性磷酸酶基因取代pFGREP-JFH1的新霉素抗性基因，制备pFGREP-JFH1/SEAP。另外，通过将pFGREP-JFH1/SEAP第10840位的G突变为A，制备突变体pFGREP-JFH1/SEAP/GND，其中NS5bRNA聚合酶活性中心的GDD基序被变为GND。

(B)全长HCV基因组RNA和全长HCV复制子RNA的制备

如上构建表达载体，用限制酶XbaI消化pJFH1、pJFH1/GND、pFGREP-JFH1和pFGREP-JFH1/GND，以制备模板DNA，用于合成全长HCV基因组RNA和全长HCV复制子RNA。随后，通过于30℃温育30分钟，用20U绿豆核酸酶的50μl反应溶液处理10-20μg各XbaI片段。绿豆核酸酶是这样一种酶：其催化涉及选择性消化双链DNA的单链部分的反应。通常，如果使用以上XbaI片段原样作为模板合成RNA，则合成在3′端具有4个额外碱基CUGA的复制子RNA，其为XbaI识别位点的一部分。因此，在该实施例中，通过用绿豆核酸酶处理XbaI片段，由XbaI片段去除4碱基CUGA。随后，对绿豆核酸酶处理后的含XbaI片段的溶液进行标准蛋白去除处理，以获得无4碱基CUGA的纯化XbaI片段，作为以下使用的模板DNA。

接着，使用T7 RNA聚合酶由该模板体外合成RNA。使用MEGAscript(Ambion Co.)进行RNA合成。按照生产商的说明书使20μl含0.5-1.0μg模板DNA的反应混合物反应。

在RNA合成后，将DNA酶(2U)加入到反应混合物中，于37℃反应15分钟，然后用酸-苯酚处理提取RNA，以去除模板DNA。以此方式由源自pJFH1、pJFH1/GND、pFGREP-JFH1和pFGREP-JFH1/GND的模板DNA合成的RNA分别称为rJFH1、rJFH1/GND、rFGREP-JFH1和rFGREP-JFH1/GND。这些RNA的核苷酸序列分别示于SEQ ID NO：12、13、14和15的rJFH-1、rFGREP-JFH1、rJFH1/GND和rFGREP-JFH1/GND。rJFH1是本发明全长HCV基因组RNA的实例，与JFH-1毒株的全长HCV基因组具有相同的序列结构。rFGREP-JFH1是本发明全长HCV复制子RNA的实例。

随后，通过分别使用如上制备的表达载体pFGREP-JFH1/Luc、pFGREP-JFH1/Luc/GND、pFGREP-JFH1/EGFP、pFGREP-JFH1/EGFP/GND、pFGREP-JFH1/S EAP和pFGREP-JFH1/SEAP/GND作为模板，生产为HCV复制子RNA的rFGR-JFH1/Luc(SEQ ID NO：21)、rFGR-JFH1/Luc/GND((SEQ ID NO：22)、rFGR-JFH1/EGFP(SEQID NO：23)、rFGR-JFH1/EGFP/GND(SEQ ID NO：24)、rFGR-JFH1/SEAP(SEQ ID NO：25)和rFGR-JFH1/SEAP/GND(SEQ ID NO：26)。

实施例2：全长HCV基因组RNA在细胞中的复制和病毒颗粒的产生

(C)全长HCV基因组RNA在细胞中的复制和病毒颗粒的产生

将如上合成的各种量的全长HCV基因组RNA(rJFH1或rJFH1/GND)与由Huh7细胞提取的总RNA混合，使RNA量达到10μg。随后，利用电穿孔法将混合的RNA导入到Huh7细胞中。将进行电穿孔处理的Huh7细胞接种入培养皿中。在温育12、24、48和72小时后，收集细胞，提取RNA，并通过RNA印迹法分析。按照Molecular Cloning，A laboratory Manual，第2版，J.Sambrook，E.F.Fritsch，T.Maniatis，Cold Spring Harbor Laboratory Press(1989)进行RNA印迹分析。具体地说，对由温育后细胞提取的RNA进行变性琼脂糖凝胶电泳，并在电泳后将RNA转移至带正电荷的尼龙膜上。使由pJFH1制备的32P标记DNA或RNA探针与前述转移至膜上的RNA杂交。漂洗膜，并使膜对胶片曝光，以检测对JFH-1克隆的全长HCV基因组RNA特异性的RNA条带。

如图2所示，当rJFH1/GND转染至细胞中时，在转染后4小时导入RNA的条带被证实为弱信号，但信号随着传代次数变得更弱，转染后24小时的条带信号几乎不可检测。相比之下，当转染rJFH1时，导入RNA条带的信号强度首先变弱，和rJFH1/GND转染后4-12小时的情况一样，但在转染后24小时证实了清晰的RNA信号。证实的信号是HCV基因组RNA特异性的。也就是说，一般认为某些导入的全长HCV基因组RNA复制并生长。对于其中为RNA复制酶的NS5B活性基序突变的rJFH1/GND，未观察到复制，这表明NS5B活性对全长HCV基因组RNA复制很重要。此外，对得自丙型肝炎病毒，例如H77毒株(非特许文献7)、J6毒株(非特许文献8)和本发明发明人由慢性肝炎患者分离的JCH1毒株(非特许文献6)的全长基因组RNA(先前已全部被分离)进行相同的实验，但证实这些毒株的全长HCV基因组RNA未复制。

(D)转染细胞培养物的培养基中HCV病毒颗粒的检测

将如上所述的电穿孔处理的Huh7细胞接种在培养皿中，培养12、24、48和72小时，然后检测培养上清液中的HCV核心蛋白。按照Ortho HCV抗原IRMA检验(非特许文献9)进行检测。如图3所示，在用rJFH1转染后48和72小时检测培养上清液中的核心蛋白。为检验此核心蛋白是否作为病毒颗粒被分泌，通过蔗糖密度梯度分级分离rJFH1转染后72小时的培养基。在离心管中，2ml 60％(重量/重量)蔗糖溶液(溶解在50mM Tris pH 7.5/0.1M NaCl/1mM EDTA中)、1ml 50％蔗糖溶液、1ml 40％蔗糖溶液、1ml 30％蔗糖溶液、1mlof 20％蔗糖溶液和1ml 10％蔗糖溶液分层，4ml样品培养上清液覆盖于其上。在Beckman转子SW41 Ti中以400,000RPM于4℃离心16小时。在离心后，由管底部收集各0.5ml的级分。测定各个级分中的密度、HCV核心蛋白浓度和全长HCV基因组RNA的量。按照Takeuchi T，Katsume A，Tanaka T，Abe A，Inoue K，Tsukiyama-KoharaK，Kawaguchi R，Tanaka S，Kohara M，“Real-Time detection system forquantification of Hepatitis C virus genome”，Gastroenterology 116：636-642(1999)，通过检测全长HCV基因组RNA的5′非翻译区的RNA，用定量RT-PCR法进行全长HCV基因组RNA的检测。具体地说，使用合成引物R6-130-S17：5′-CGGGAGAGCCATAGTGG-3′(SEQ IDNO：16)、R6-290-R19：5′-AGTACCACAAGGCCTTTCG-3′(SEQ ID NO：17)和TaqMan探针：R6-148-S21FT，5′-CTGCGGAACCGGTGAGTACAC-3′(SEQ ID NO：18)以及EZ rTthRNA PCR试剂盒，PCR扩增由细胞提取的RNA中包含的全长HCV基因组RNA，然后通过ABI Prism 7700序列检测系统检测。

如图4所示，在级分11中核心蛋白的峰与全长HCV基因组RNA的峰一致。该级分的密度约为1.18mg/ml，其表明比重比迄今为止所报道的核心蛋白和核酸缀合物的比重低。此外，当在用0.25％NP40处理培养上清液后进行相似的分级分离时，核心蛋白和全长HCV基因组RNA的峰迁移至约1.28mg/ml的比重。也就是说，一般认为NP40处理剥落了病毒颗粒的表面膜(其含脂质，于是具有较低比重)，产生仅由核酸和核心蛋白组成的核心颗粒，因此比重增加。以上结果表明，通过将rJFH1转染至Huh7细胞中，全长HCV基因组RNA在细胞中复制，结果形成病毒颗粒，并分泌入培养上清液中。

实施例3

(E)全长HCV复制子RNA复制型细胞的制备和细胞克隆的建立

通过如实施例2所述，将实施例1中制备的rFGREP-JFH1和rFGREP-JFH1/GND转染入Huh7细胞，制备全长HCV复制子RNA复制型细胞，然后尝试建立全长HCV复制子RNA复制型细胞克隆。

首先，在分别将rFGREP-JFH1和rFGREP-JFH1/GND转染入Huh7细胞中后，将细胞接种在培养皿中。在培养16-24小时后，加入不同浓度的G418。继续培养，同时每周更换两次培养基。在培养21天后，用结晶紫染色存活细胞。计数染色的集落，计算获得的集落数/用于转染的RNA重量。对于某些培养皿又继续培养，以克隆存活细胞的集落。分别由克隆细胞中提取RNA、基因组DNA和蛋白，然后检测全长HCV复制子RNA，研究新霉素抗性基因在基因组DNA中的整合以及HCV蛋白的表达。这些结果在下文详细给出。

(F)集落形成能力

以上转染的结果表明，对于在1.0mg/ml G418浓度下用rFGREP-JFH1转染的Huh7细胞(图5的左部)，集落形成能力/1μg用于转染的复制子RNA是368CFU(集落形成单位)/μg RNA。相反，对于用rFGREP-JFH1/GND转染的Huh7细胞(图5的右部)，未观测到集落形成。这表明，用rFGREP-JFH1复制子RNA转染的Huh7细胞的集落形成能力依赖于由rFGREP-JFH1表达的NS5B(RNA聚合酶)的活性。也就是说，一般认为在集落形成细胞中，借助于由rFGREP-JFH1表达的NS5B的作用，rFGREP-JFH1复制子RNA自主复制，引起新霉素抗性基因持续表达，因此保持了G418抗性，结果使细胞生长成为可能。

(G)在已建立细胞克隆中的全长HCV复制子RNA的检测

利用酸-苯酚提取法，由全长HCV复制子RNA复制型细胞克隆中提取总RNA，该细胞按照以上章节(E)通过将rFGREP-JFH1转染入Huh7细胞中而建立。随后，利用RNA印迹法检测此总RNA。在该方法中，使用pFGREP-JFH1特异性探针。以相似的方式由未转染的Huh7细胞提取的总RNA(在图6中显示为“Huh7”)，一个除由Huh7细胞提取的总RNA以外还含体外合成的10⁷个拷贝的复制子RNA的样品(在图6中显示为“10⁷”)和一个除由Huh7细胞提取的总RNA以外还含体外合成的10⁸个拷贝的复制子RNA的样品(在图6中显示为“108”)用作对照。在图6中，1-4表示细胞克隆号。

结果，用rFGREP-JFH1特异性探针检测与rFGREP-JFH1具有相似大小的RNA(图6)。由该结果证实转染的rFGREP-JFH1复制子RNA在细胞克隆中复制和生长。还表明细胞克隆中的复制子RNA的量有差异。如图6所示，例如，克隆2中的复制子RNA的量少于其它克隆。

(H)验证新霉素抗性基因有无整合入基因组DNA中

对于按照(E)所获得的细胞克隆1-8(如图7中的FGR-JFH1/2-1至FGR-JFH1/2-8所示)，使用新霉素抗性基因特异性引物(有义引物NEO-S3：5′-AACAAGATGGATTGCACGCA-3′(SEQ ID NO：19)，反义引物NEO-R：5′-CGTCAAGAAGGCGATAGAAG-3′(SEQ ID NO：20))，并使用由每个细胞克隆提取的宿主基因组DNA作为模板，进行PCR扩增，以便证实各个细胞克隆抗G418的抗性并非是由于新霉素抗性基因整合入宿主细胞基因组中。结果，如图7所示，未观测到显示扩增新霉素抗性基因的阳性克隆。

(H)的结果证实，全长HCV复制子RNA在通过本发明的全长HCV复制子RNA转染建立的细胞克隆中复制。

(I)HCV蛋白的检测

通过标准方法由通过rFGREP-JFH1转染建立的细胞克隆提取蛋白，然后通过SDS-PAGE和蛋白质印迹法分析。在此情况下检测的细胞克隆与以上章节(G)中使用的克隆相同。通过将制备的全长HCV基因组RNA瞬时转染入Huh7细胞中获得细胞提取物，将其用作阳性对照(在图8、9和10中显示为JFH-1)。通过转染HCV亚基因组RNA复制子(SGR-JFH1)获得的克隆的细胞提取物用作核心蛋白的阴性对照，并用作NS3和NS5a蛋白的阳性对照(在图8、9和10中显示为SGR-JFH1)。未转染的Huh7细胞的细胞提取物用作所有蛋白的阴性对照(在图8、9和10中显示为Huh7)。将由各个细胞克隆提取的蛋白样品印迹在PVDF膜(Immobilon-P，Millipore)上，然后使用抗核心蛋白特异性抗体和抗NS3特异性抗体(由Dr.Moradpour惠赠；Wolk B等，J.Virology，2000，74：2293-2304)检测由其中的全长HCV复制子RNA编码的核心蛋白和NS3蛋白。如图8和9所示，对于通过rFGREP-JFH1转染建立的细胞克隆1-4，检测各个蛋白中与阳性对照大小相同的蛋白。因为在未转染Huh7细胞中既没有检测到核心蛋白，也没有检测到NS3蛋白，所以证实在细胞克隆1-4中已转染的全长HCV复制子RNA自主复制，表达核心蛋白和NS3蛋白。

此外，对于每个细胞克隆，已如上所述检验NS3蛋白的表达，还使用丙型肝炎患者的血清作为抗体检验由全长HCV复制子RNA表达的NS5A蛋白(图10)。

由上述(H)和(I)的结果证实，在已通过全长HCV复制子RNA转染建立的细胞克隆中，全长HCV复制子RNA复制，病毒蛋白也表达。

(J)全长HCV复制子RNA复制型细胞中的病毒颗粒生产

按照以上章节(E)将rFGREP-JFH1转染入Huh7细胞中，建立全长HCV复制子RNA复制型细胞克隆2和3(FGR-JFH1/2-3)，然后回收其培养上清液。按照与上述(D)相似的方法检测培养上清液中的HCV病毒颗粒。结果示于图11。在图11中，阴影圆代表各个级分的比重(g/ml)。实心圆代表核心蛋白的量(fmol/L)。空心圆代表全长HCV复制子RNA的滴度(×0.1拷贝/mL)。

如图11所示，核心蛋白峰与比重约1.18-1.20mg/ml的级分中的全长HCV复制子RNA的峰一致。还在较轻级分中发现小峰。以上结果表明，全长HCV复制子RNA在用rFGREP-JFH11转染的Huh7细胞中复制，形成病毒颗粒并分泌入其培养上清液中。

实施例4

(K)采用培养上清液中的病毒颗粒的感染实验

通过将在(H)中使用的细胞克隆1-8(即FGR-JFH1/2-1、FGR-JFH1/2-2、FGR-JFH1/2-3、FGR-JFH1/2-4、FGR-JFH1/2-5、FGR-JGF1/2-6、FGR-JFH1/2-7、FGR-JFH1/2-8)的各个培养上清液加入Huh7细胞中，用在培养上清液中的病毒颗粒感染Huh7细胞。在第2天，以0.3mg/ml向感染的Huh7细胞的培养基中加入G418，再培养Huh7细胞21天。在结束培养后，固定细胞，并用结晶紫染色。对于分别用FGR-JFH1/2-3、FGR-JFH1/2-5和FGR-JFH1/2-6的培养上清液感染的细胞，观测到集落形成。另一方面，对于用作对照的亚基因组复制子细胞SGR-JFH1/4-1(描述于非特许文献6)的培养上清液感染的细胞，未观测到集落形成。图12显示了与加入的4ml或8mlFGR-JFH1/2-3或SGR-JFH1/4-1培养上清液培养21天后染色培养皿的照片。在其中接种与4ml或8ml FGR-JFH1/2-3培养上清液混合的细胞的培养皿中，分别发现3个和9个集落。但是，在其中接种与SGR-JFH1/4-1的培养上清液混合的细胞的培养皿中，未观测到集落。

随后，克隆分别使用FGR-JFH1/2-3和FGR-JFH1/2-5的培养上清液以丙型肝炎病毒感染形成的集落。由用FGR-JFH1/2-3的培养上清液感染的培养皿建立3个克隆FGR-JFH1/C2-3-11、FGR-JFH1/C2-3-12和FGR-JFH1/C2-3-13，并由用FGR-JFH1/C2-5的培养上清液感染的培养皿建立2个克隆FGR-JFH1/C2-5-11和FGR-JFH1/C2-5-12。

当用FGR-JFH1/C2-3-11、FGR-JFH1/C2-3-12、FGR-JFH1/C2-3-13、FGR-JFH1/C2-5-11和FGR-JFH1/C2-5-12各个细胞克隆的培养上清液感染Huh7细胞时，在分别用FGR-JFH1/C2-3-12和FGR-JFH1/C2-5-12的培养上清液感染的培养皿中观测到集落形成。由用FGR-JFH1/C2-3-12的培养上清液感染的细胞建立另两个克隆FGR-JFH1/C2-3-12-1和FGR-JFH1/C2-3-12-2。由用FGR-JFH1/C2-5-12的培养上清液感染的细胞建立另两个克隆FGR-JFH1/C2-5-12-1和FGR-JFH1/C2-5-12-2。

已用全长HCV复制子RNA复制型细胞的培养上清液感染的细胞建立细胞，由这些细胞克隆提取RNA、蛋白和基因组DNA。通过使用基因组DNA作为模板的PCR，检验新霉素抗性基因在这些细胞克隆的基因组DNA中的整合，结果全为阴性。而且，通过使用RNA作为模板的定量PCR可检测细胞中复制的全长HCV复制子RNA。再进一步，可检测培养上清液中的核心蛋白。这些结果表明，由本发明的全长HCV复制子RNA复制型细胞生产的含全长HCV复制子RNA的病毒颗粒可感染另一细胞。

工业实用性

按照本发明的方法，可在细胞培养系统中制备HCV病毒颗粒。使用本发明的复制子RNA，可在细胞培养系统中有效生产含全长HCV基因组RNA的RNA。而且，通过使用其中导入本发明的全长HCV复制子RNA或全长HCV基因组RNA的细胞，全长HCV复制子RNA或全长HCV基因组RNA可在细胞培养系统中复制，本发明的HCV病毒颗粒可在细胞培养系统中连续生产。其中导入本发明的全长HCV复制子RNA或全长HCV基因组RNA的细胞还可用作测试系统，用于筛选影响HCV复制过程、病毒颗粒形成过程和病毒颗粒胞外释放过程的各种物质。本发明的全长HCV复制子RNA和全长HCV基因组RNA以及病毒颗粒还用作外源基因的病毒载体。本发明的病毒颗粒或其部分可包含在疫苗中，作为抗丙型肝炎病毒的疫苗抗原。此外，其中本发明的病毒颗粒和其它细胞一起培养的系统可用作测试系统，用于筛选对病毒颗粒感染细胞有影响的各种物质。本发明的全长HCV复制子RNA或全长HCV基因组RNA用作模板，该模板能使HCV全长基因组序列简单再生。

本文提及的所有出版物、特许和特许申请都全文在此引入作为参考。

无内容序列表

SEQ ID NO：1代表来自JFH-1克隆的HCV基因组RNA的5′非翻译区序列。

SEQ ID NO：2代表来自JFH-1克隆的HCV基因组RNA的核心蛋白编码序列。

SEQ ID NO：3代表来自JFH-1克隆的HCV基因组RNA的E1蛋白编码序列。

SEQ ID NO：4代表来自JFH-1克隆的HCV基因组RNA的E2蛋白编码序列。

SEQ ID NO：5代表来自JFH-1克隆的HCV基因组RNA的NS2蛋白编码序列。

SEQ ID NO：6代表来自JFH-1克隆的HCV基因组RNA的NS3蛋白编码序列。

SEQ ID NO：7代表来自JFH-1克隆的HCV基因组RNA的NS4A蛋白编码序列。

SEQ ID NO：8代表来自JFH-1克隆的HCV基因组RNA的NS4B蛋白编码序列。

SEQ ID NO：9代表来自JFH-1克隆的HCV基因组RNA的NS5A蛋白编码序列。

SEQ ID NO：10代表来自JFH-1克隆的HCV基因组RNA的NS5B蛋白编码序列。

SEQ ID NO：11代表来自JFH-1克隆的HCV基因组RNA的3′非翻译区序列。

SEQ ID NO：12代表来自JFH-1克隆的全长HCV基因组RNA序列。

SEQ ID NO：13代表包含来自JFH-1克隆的全长HCV基因组RNA的复制子RNA序列。

SEQ ID NO：14代表来自JFH-1克隆的全长HCV基因组RNA序列，其中氨基酸基序GDD已突变为GND。

SEQ ID NO：15代表包含来自JFH-1克隆的全长HCV基因组RNA的复制子RNA序列，其中氨基酸基序GDD已突变为GND。

SEQ ID NO：16-20代表引物序列。

SEQ ID NO：21代表来源于表达载体pFGREP-JFH1/Luc的复制子RNA的序列。

SEQ ID NO：22代表来源于表达载体pFGREP-JFH1/Luc/GND的复制子RNA的序列。

SEQ ID NO：23代表来源于表达载体pFGREP-JFH1/EGFP的复制子RNA的序列。

SEQ ID NO：24代表来源于表达载体pFGREP-JFH1/EGFP/GND的复制子RNA的序列。

SEQ ID NO：25代表来源于表达载体pFGREP-JFH1/SEAP的复制子RNA的序列。

SEQ ID NO：26代表来源于表达载体pFGREP-JFH1/SEAP/GND的复制子RNA的序列。

                                序列表

<110>东京都医学研究机构

     日本东丽工业株式会社

<120>含人丙型肝炎病毒全长基因组的核酸构建物、核酸构建物转入其中的重组全长病毒基因组复制型细胞和生产人丙型肝炎病毒颗粒的方法

<130>PH-2372-PCT

<140>PCT/JP2005/003232

<141>2005-02-21

<150>JP 2004-045489

<151>2004-02-20

<160>26

<170>PatentIn Ver.2.1

<210>1

<211>340

<212>RNA

<213>丙型肝炎病毒

<220>

<223>来自JFH-1克隆的丙型肝炎病毒基因组RNA的5′非翻译区

<220>

<223>发明人：Wakita，Takaji

     发明人：Kato，Takanobu

     发明人：Date，Tomoko

     发明人：Miyamoto，Michiko

     发明人：Tanabe，Junichi

     发明人：Sone，Saburo

<400>1

accugccccu aauaggggcg acacuccgcc augaaucacu ccccugugag gaacuacugu       60

cuucacgcag aaagcgccua gccauggcgu uaguaugagu gucguacagc cuccaggccc       120

cccccucccg ggagagccau aguggucugc ggaaccggug aguacaccgg aauugccggg       180

aagacugggu ccuuucuugg auaaacccac ucuaugcccg gccauuuggg cgugcccccg       240

caagacugcu agccgaguag cguuggguug cgaaaggccu ugugguacug ccugauaggg       300

cgcuugcgag ugccccggga ggucucguag accgugcacc                             340

<210>2

<211>573

<212>RNA

<213>丙型肝炎病毒

<220>

<223>来自JFH-1克隆的丙型肝炎病毒基因组RNA的核心蛋白编码序列

<400>2

augagcacaa auccuaaacc ucaaagaaaa accaaaagaa acaccaaccg ucgcccagaa       60

gacguuaagu ucccgggcgg cggccagauc guuggcggag uauacuuguu gccgcgcagg       120

ggccccaggu ugggugugcg cacgacaagg aaaacuucgg agcgguccca gccacguggg       180

agacgccagc ccauccccaa agaucggcgc uccacuggca aggccugggg aaaaccaggu       240

cgccccuggc cccuauaugg gaaugaggga cucggcuggg caggauggcu ccuguccccc       300

cgaggcucuc gccccuccug gggccccacu gacccccggc auaggucgcg caacgugggu       360

aaagucaucg acacccuaac guguggcuuu gccgaccuca ugggguacau ccccgucgua       420

ggcgccccgc uuaguggcgc cgccagagcu gucgcgcacg gcgugagagu ccuggaggac       480

gggguuaauu augcaacagg gaaccuaccc gguuuccccu uuucuaucuu cuugcuggcc       540

cuguuguccu gcaucaccgu uccggucucu gcu                                    573

<210>3

<211>576

<212>RNA

<213>丙型肝炎病毒

<220>

<223>来自JFH-1克隆的丙型肝炎病毒基因组RNA的E1蛋白编码序列

<400>3

gcccagguga agaauaccag uagcagcuac auggugacca augacugcuc caaugacagc       60

aucacuuggc agcucgaggc ugcgguucuc cacguccccg ggugcguccc gugcgagaga       120

guggggaaua cgucacggug uugggugcca gucucgccaa acauggcugu gcggcagccc       180

ggugcccuca cgcagggucu gcggacgcac aucgauaugg uugugauguc cgccaccuuc       240

ugcucugcuc ucuacguggg ggaccucugu ggcgggguga ugcucgcggc ccagguguuc       300

aucgucucgc cgcaguacca cugguuugug caagaaugca auugcuccau cuacccuggc       360

accaucacug gacaccgcau ggcaugggac augaugauga acuggucgcc cacggccacc       420

augauccugg cguacgugau gcgcgucccc gaggucauca uagacaucgu uagcggggcu       480

cacuggggcg ucauguucgg cuuggccuac uucucuaugc agggagcgug ggcgaagguc       540

auugucaucc uucugcuggc cgcuggggug gacgcg                                 576

<210>4

<211>1290

<212>RNA

<213>丙型肝炎病毒

<220>

<223>来自JFH-1克隆的丙型肝炎病毒基因组RNA的E2蛋白编码序列

<400>4

ggcaccacca ccguuggagg cgcuguugca cguuccacca acgugauugc cggcguguuc       60

agccauggcc cucagcagaa cauucagcuc auuaacacca acggcaguug gcacaucaac       120

cguacugccu ugaauugcaa ugacuccuug aacaccggcu uucucgcggc cuuguucuac       180

accaaccgcu uuaacucguc agggugucca gggcgccugu ccgccugccg caacaucgag       240

gcuuuccgga uagggugggg cacccuacag uacgaggaua augucaccaa uccagaggau       300

augaggccgu acugcuggca cuacccccca aagccgugug gcguaguccc cgcgaggucu       360

guguguggcc caguguacug uuucaccccc agcccgguag uagugggcac gaccgacaga       420

cguggagugc ccaccuacac auggggagag aaugagacag augucuuccu acugaacagc       480

acccgaccgc cgcagggcuc augguucggc ugcacgugga ugaacuccac ugguuucacc       540

aagacuugug gcgcgccacc uugccgcacc agagcugacu ucaacgccag cacggacuug       600

uugugcccua cggauuguuu uaggaagcau ccugaugcca cuuauauuaa gugugguucu       660

gggcccuggc ucacaccaaa gugccugguc cacuacccuu acagacucug gcauuacccc       720

ugcacaguca auuuuaccau cuucaagaua agaauguaug uagggggggu ugagcacagg       780

cucacggccg caugcaacuu cacucguggg gaucgcugcg acuuggagga cagggacagg       840

agucagcugu cuccucuguu gcacucuacc acggaauggg ccauccugcc cugcaccuac       900

ucagacuuac ccgcuuuguc aacuggucuu cuccaccuuc accagaacau cguggacgua       960

caauacaugu auggccucuc accugcuauc acaaaauacg ucguucgaug ggagugggug       1020

guacucuuau uccugcucuu agcggacgcc agagucugcg ccugcuugug gaugcucauc       1080

uuguugggcc aggccgaagc agcauuggag aaguuggucg ucuugcacgc ugcgagugcg       1140

gcuaacugcc auggccuccu auauuuugcc aucuucuucg uggcagcuug gcacaucagg       1200

ggucgggugg uccccuugac caccuauugc cucacuggcc uauggcccuu cugccuacug       1260

cucauggcac ugccccggca ggcuuaugcc                                        1290

<210>5

<211>651

<212>RNA

<213>丙型肝炎病毒

<220>

<223>来自JFH-1克隆的丙型肝炎病毒基因组RNA的NS2蛋白编码序列

<400>5

uaugacgcac cugugcacgg acagauaggc guggguuugu ugauauugau cacccucuuc       60

acacucaccc cgggguauaa gacccuccuc ggccaguguc uguggugguu gugcuaucuc       120

cugacccugg gggaagccau gauucaggag uggguaccac ccaugcaggu gcgcggcggc       180

cgcgauggca ucgcgugggc cgucacuaua uucugcccgg gugugguguu ugacauuacc       240

aaauggcuuu uggcguugcu ugggccugcu uaccucuuaa gggccgcuuu gacacaugug       300

ccguacuucg ucagagcuca cgcucugaua aggguaugcg cuuuggugaa gcagcucgcg       360

ggggguaggu auguucaggu ggcgcuauug gcccuuggca gguggacugg caccuacauc       420

uaugaccacc ucacaccuau gucggacugg gccgcuagcg gccugcgcga cuuagcgguc       480

gccguggaac ccaucaucuu caguccgaug gagaagaagg ucaucgucug gggagcggag       540

acggcugcau guggggacau ucuacaugga cuucccgugu ccgcccgacu cggccaggag       600

auccuccucg gcccagcuga uggcuacacc uccaaggggu ggaagcuccu u                651

<210>6

<211>1893

<212>RNA

<213>丙型肝炎病毒

<220>

<223>来自JFH-1克隆的丙型肝炎病毒基因组RNA的NS3蛋白编码序列

<400>6

gcucccauca cugcuuaugc ccagcaaaca cgaggccucc ugggcgccau aguggugagu       60

augacggggc gugacaggac agaacaggcc ggggaagucc aaauccuguc cacagucucu       120

caguccuucc ucggaacaac caucucgggg guuuugugga cuguuuacca cggagcuggc       180

aacaagacuc uagccggcuu acgggguccg gucacgcaga uguacucgag ugcugagggg       240

gacuugguag gcuggcccag ccccccuggg accaagucuu uggagccgug caagugugga       300

gccgucgacc uauaucuggu cacgcggaac gcugauguca ucccggcucg gagacgcggg       360

gacaagcggg gagcauugcu cuccccgaga cccauuucga ccuugaaggg guccucgggg       420

gggccggugc ucugcccuag gggccacguc guugggcucu uccgagcagc ugugugcucu       480

cggggcgugg ccaaauccau cgauuucauc cccguugaga cacucgacgu uguuacaagg       540

ucucccacuu ucagugacaa cagcacgcca ccggcugugc cccagaccua ucaggucggg       600

uacuugcaug cuccaacugg caguggaaag agcaccaagg ucccugucgc guaugccgcc       660

cagggguaca aaguacuagu gcuuaacccc ucgguagcug ccacccuggg guuuggggcg       720

uaccuaucca aggcacaugg caucaauccc aacauuagga cuggagucag gaccgugaug       780

accggggagg ccaucacgua cuccacauau ggcaaauuuc ucgccgaugg gggcugcgcu       840

agcggcgccu augacaucau cauaugcgau gaaugccacg cuguggaugc uaccuccauu       900

cucggcaucg gaacgguccu ugaucaagca gagacagccg gggucagacu aacugugcug       960

gcuacggcca caccccccgg gucagugaca accccccauc ccgauauaga agagguaggc       1020

cucgggcggg agggugagau ccccuucuau gggagggcga uuccccuauc cugcaucaag       1080

ggagggagac accugauuuu cugccacuca aagaaaaagu gugacgagcu cgcggcggcc       1140

cuucggggca ugggcuugaa ugccguggca uacuauagag gguuggacgu cuccauaaua       1200

ccagcucagg gagauguggu ggucgucgcc accgacgccc ucaugacggg guacacugga       1260

gacuuugacu ccgugaucga cugcaaugua gcggucaccc aagcugucga cuucagccug       1320

gaccccaccu ucacuauaac cacacagacu gucccacaag acgcugucuc acgcagucag       1380

cgccgcgggc gcacagguag aggaagacag ggcacuuaua gguauguuuc cacuggugaa       1440

cgagccucag gaauguuuga caguguagug cuuugugagu gcuacgacgc aggggcugcg       1500

ugguacgauc ucacaccagc ggagaccacc gucaggcuua gagcguauuu caacacgccc       1560

ggccuacccg ugugucaaga ccaucuugaa uuuugggagg caguuuucac cggccucaca       1620

cacauagacg cccacuuccu cucccaaaca aagcaagcgg gggagaacuu cgcguaccua       1680

guagccuacc aagcuacggu gugcgccaga gccaaggccc cucccccguc cugggacgcc       1740

auguggaagu gccuggcccg acucaagccu acgcuugcgg gccccacacc ucuccuguac       1800

cguuugggcc cuauuaccaa ugaggucacc cucacacacc cugggacgaa guacaucgcc       1860

acaugcaugc aagcugaccu ugaggucaug acc                                    1893

<210>7

<211>162

<212>RNA

<213>丙型肝炎病毒

<220>

<223>来自JFH-1克隆的丙型肝炎病毒基因组RNA的NS4A蛋白编码序列

<400>7

agcacguggg uccuagcugg aggaguccug gcagccgucg ccgcauauug ccuggcgacu       60

ggaugcguuu ccaucaucgg ccgcuugcac gucaaccagc gagucgucgu ugcgccggau       120

aaggaggucc uguaugaggc uuuugaugag auggaggaau gc                          162

<210>8

<211>783

<212>RNA

<213>丙型肝炎病毒

<220>

<223>来自JFH-1克隆的丙型肝炎病毒基因组RNA的NS4B蛋白编码序列

<400>8

gccucuaggg cggcucucau cgaagagggg cagcggauag ccgagauguu gaaguccaag       60

auccaaggcu ugcugcagca ggccucuaag caggcccagg acauacaacc cgcuaugcag       120

gcuucauggc ccaaagugga acaauuuugg gccagacaca uguggaacuu cauuagcggc       180

auccaauacc ucgcaggauu gucaacacug ccagggaacc ccgcgguggc uuccaugaug       240

gcauucagug ccgcccucac caguccguug ucgaccagua ccaccauccu ucucaacauc       300

augggaggcu gguuagcguc ccagaucgca ccacccgcgg gggccaccgg cuuugucguc       360

aguggccugg ugggggcugc cgugggcagc auaggccugg guaaggugcu gguggacauc       420

cuggcaggau auggugcggg cauuucgggg gcccucgucg cauucaagau caugucuggc       480

gagaagcccu cuauggaaga ugucaucaau cuacugccug ggauccuguc uccgggagcc       540

cugguggugg gggucaucug cgcggccauu cugcgccgcc acgugggacc gggggagggc       600

gcgguccaau ggaugaacag gcuuauugcc uuugcuucca gaggaaacca cgucgccccu       660

acucacuacg ugacggaguc ggaugcgucg cagcguguga cccaacuacu uggcucucuu       720

acuauaacca gccuacucag aagacuccac aauuggauaa cugaggacug ccccauccca       780

ugc                                                                     783

<210>9

<211>1398

<212>RNA

<213>丙型肝炎病毒

<220>

<223>来自JFH-1克隆的丙型肝炎病毒基因组RNA的NS5A蛋白编码序列

<400>9

uccggauccu ggcuccgcga cgugugggac uggguuugca ccaucuugac agacuucaaa       60

aauuggcuga ccucuaaauu guuccccaag cugcccggcc uccccuucau cucuugucaa       120

aagggguaca agggugugug ggccggcacu ggcaucauga ccacgcgcug cccuugcggc       180

gccaacaucu cuggcaaugu ccgccugggc ucuaugagga ucacagggcc uaaaaccugc       240

augaacaccu ggcaggggac cuuuccuauc aauugcuaca cggagggcca gugcgcgccg       300

aaacccccca cgaacuacaa gaccgccauc uggagggugg cggccucgga guacgcggag       360

gugacgcagc augggucgua cuccuaugua acaggacuga ccacugacaa ucugaaaauu       420

ccuugccaac uaccuucucc agaguuuuuc uccugggugg acggugugca gauccauagg       480

uuugcaccca caccaaagcc guuuuuccgg gaugaggucu cguucugcgu ugggcuuaau       540

uccuaugcug ucggguccca gcuucccugu gaaccugagc ccgacgcaga cguauugagg       600

uccaugcuaa cagauccgcc ccacaucacg gcggagacug cggcgcggcg cuuggcacgg       660

ggaucaccuc caucugaggc gagcuccuca gugagccagc uaucagcacc gucgcugcgg       720

gccaccugca ccacccacag caacaccuau gacguggaca uggucgaugc caaccugcuc       780

auggagggcg guguggcuca gacagagccu gaguccaggg ugcccguucu ggacuuucuc       840

gagccaaugg ccgaggaaga gagcgaccuu gagcccucaa uaccaucgga gugcaugcuc       900

cccaggagcg gguuuccacg ggccuuaccg gcuugggcac ggccugacua caacccgccg       960

cucguggaau cguggaggag gccagauuac caaccgccca ccguugcugg uugugcucuc       1020

ccccccccca agaaggcccc gacgccuccc ccaaggagac gccggacagu gggucugagc       1080

gagagcacca uaucagaagc ccuccagcaa cuggccauca agaccuuugg ccagcccccc       1140

ucgagcggug augcaggcuc guccacgggg gcgggcgccg ccgaauccgg cgguccgacg       1200

uccccuggug agccggcccc cucagagaca gguuccgccu ccucuaugcc cccccucgag       1260

ggggagccug gagauccgga ccuggagucu gaucagguag agcuucaacc ucccccccag       1320

gggggggggg uagcucccgg uucgggcucg gggucuuggu cuacuugcuc cgaggaggac       1380

gauaccaccg ugugcugc                                                     1398

<210>10

<211>1773

<212>RNA

<213>丙型肝炎病毒

<220>

<223>来自JFH-1克隆的丙型肝炎病毒基因组RNA的NS5B蛋白编码序列

<400>10

uccaugucau acuccuggac cggggcucua auaacucccu guagccccga agaggaaaag       60

uugccaauca acccuuugag uaacucgcug uugcgauacc auaacaaggu guacuguaca       120

acaucaaaga gcgccucaca gagggcuaaa aagguaacuu uugacaggac gcaagugcuc       180

gacgcccauu augacucagu cuuaaaggac aucaagcuag cggcuuccaa ggucagcgca       240

aggcuccuca ccuuggagga ggcgugccag uugacuccac cccauucugc aagauccaag       300

uauggauucg gggccaagga gguccgcagc uuguccggga gggccguuaa ccacaucaag       360

uccgugugga aggaccuccu ggaagaccca caaacaccaa uucccacaac caucauggcc       420

aaaaaugagg uguucugcgu ggaccccgcc aaggggggua agaaaccagc ucgccucauc       480

guuuacccug accucggcgu ccgggucugc gagaaaaugg cccucuauga cauuacacaa       540

aagcuuccuc aggcgguaau gggagcuucc uauggcuucc aguacucccc ugcccaacgg       600

guggaguauc ucuugaaagc augggcggaa aagaaggacc ccauggguuu uucguaugau       660

acccgaugcu ucgacucaac cgucacugag agagacauca ggaccgagga guccauauac       720

caggccugcu cccugcccga ggaggcccgc acugccauac acucgcugac ugagagacuu       780

uacguaggag ggcccauguu caacagcaag ggucaaaccu gcgguuacag acguugccgc       840

gccagcgggg ugcuaaccac uagcaugggu aacaccauca caugcuaugu gaaagcccua       900

gcggccugca aggcugcggg gauaguugcg cccacaaugc ugguaugcgg cgaugaccua       960

guagucaucu cagaaagcca ggggacugag gaggacgagc ggaaccugag agccuucacg       1020

gaggccauga ccagguacuc ugccccuccu ggugaucccc ccagaccgga auaugaccug       1080

gagcuaauaa cauccuguuc cucaaaugug ucuguggcgu ugggcccgcg gggccgccgc       1140

agauacuacc ugaccagaga cccaaccacu ccacucgccc gggcugccug ggaaacaguu       1200

agacacuccc cuaucaauuc auggcuggga aacaucaucc aguaugcucc aaccauaugg       1260

guucgcaugg uccuaaugac acacuucuuc uccauucuca ugguccaaga cacccuggac       1320

cagaaccuca acuuugagau guauggauca guauacuccg ugaauccuuu ggaccuucca       1380

gccauaauug agagguuaca cgggcuugac gccuuuucua ugcacacaua cucucaccac       1440

gaacugacgc ggguggcuuc agcccucaga aaacuugggg cgccaccccu cagggugugg       1500

aagagucggg cucgcgcagu cagggcgucc cucaucuccc guggagggaa agcggccguu       1560

ugcggccgau aucucuucaa uugggcggug aagaccaagc ucaaacucac uccauugccg       1620

gaggcgcgcc uacuggacuu auccaguugg uucaccgucg gcgccggcgg gggcgacauu       1680

uuucacagcg ugucgcgcgc ccgaccccgc ucauuacucu ucggccuacu ccuacuuuuc       1740

guagggguag gccucuuccu acuccccgcu cgg                                    1773

<210>11

<211>239

<212>RNA

<213>丙型肝炎病毒

<220>

<223>来自JFH-1克隆的丙型肝炎病毒基因组RNA的3′非翻译区

<400>11

uagagcggca cacacuaggu acacuccaua gcuaacuguu ccuuuuuuuu uuuuuuuuuu       60

uuuuuuuuuu uuuuuuuuuu uuuuucuuuu uuuuuuuuuu cccucuuucu ucccuucuca       120

ucuuauucua cuuucuuucu ugguggcucc aucuuagccc uagucacggc uagcugugaa       180

agguccguga gccgcaugac ugcagagagu gccguaacug gucucucugc agaucaugu        239

<210>12

<211>9707

<212>RNA

<213>丙型肝炎病毒

<220>

<223>来自JFH-1克隆的全长丙型肝炎病毒基因组RNA

<400>12

gaauucuaau acgacucacu auagaccugc cccuaauagg ggcgacacuc cgccaugaau       60

cacuccccug ugaggaacua cugucuucac gcagaaagcg ccuagccaug gcguuaguau       120

gagugucgua cagccuccag gccccccccu cccgggagag ccauaguggu cugcggaacc       180

ggugaguaca ccggaauugc cgggaagacu ggguccuuuc uuggauaaac ccacucuaug       240

cccggccauu ugggcgugcc cccgcaagac ugcuagccga guagcguugg guugcgaaag       300

gccuuguggu acugccugau agggcgcuug cgagugcccc gggaggucuc guagaccgug       360

caccaugagc acaaauccua aaccucaaag aaaaaccaaa agaaacacca accgucgccc       420

agaagacguu aaguucccgg gcggcggcca gaucguuggc ggaguauacu uguugccgcg       480

caggggcccc agguugggug ugcgcacgac aaggaaaacu ucggagcggu cccagccacg       540

ugggagacgc cagcccaucc ccaaagaucg gcgcuccacu ggcaaggccu ggggaaaacc       600

aggucgcccc uggccccuau augggaauga gggacucggc ugggcaggau ggcuccuguc       660

cccccgaggc ucucgccccu ccuggggccc cacugacccc cggcauaggu cgcgcaacgu       720

ggguaaaguc aucgacaccc uaacgugugg cuuugccgac cucauggggu acauccccgu       780

cguaggcgcc ccgcuuagug gcgccgccag agcugucgcg cacggcguga gaguccugga       840

ggacgggguu aauuaugcaa cagggaaccu acccgguuuc cccuuuucua ucuucuugcu       900

ggcccuguug uccugcauca ccguuccggu cucugcugcc caggugaaga auaccaguag       960

cagcuacaug gugaccaaug acugcuccaa ugacagcauc acuuggcagc ucgaggcugc       1020

gguucuccac guccccgggu gcgucccgug cgagagagug gggaauacgu cacgguguug       1080

ggugccaguc ucgccaaaca uggcugugcg gcagcccggu gcccucacgc agggucugcg       1140

gacgcacauc gauaugguug ugauguccgc caccuucugc ucugcucucu acguggggga       1200

ccucuguggc ggggugaugc ucgcggccca gguguucauc gucucgccgc aguaccacug       1260

guuugugcaa gaaugcaauu gcuccaucua cccuggcacc aucacuggac accgcauggc       1320

augggacaug augaugaacu ggucgcccac ggccaccaug auccuggcgu acgugaugcg       1380

cguccccgag gucaucauag acaucguuag cggggcucac uggggcguca uguucggcuu       1440

ggccuacuuc ucuaugcagg gagcgugggc gaaggucauu gucauccuuc ugcuggccgc       1500

ugggguggac gcgggcacca ccaccguugg aggcgcuguu gcacguucca ccaacgugau       1560

ugccggcgug uucagccaug gcccucagca gaacauucag cucauuaaca ccaacggcag       1620

uuggcacauc aaccguacug ccuugaauug caaugacucc uugaacaccg gcuuucucgc       1680

ggccuuguuc uacaccaacc gcuuuaacuc gucagggugu ccagggcgcc uguccgccug       1740

ccgcaacauc gaggcuuucc ggauagggug gggcacccua caguacgagg auaaugucac       1800

caauccagag gauaugaggc cguacugcug gcacuacccc ccaaagccgu guggcguagu       1860

ccccgcgagg ucugugugug gcccagugua cuguuucacc cccagcccgg uaguaguggg       1920

cacgaccgac agacguggag ugcccaccua cacaugggga gagaaugaga cagaugucuu       1980

ccuacugaac agcacccgac cgccgcaggg cucaugguuc ggcugcacgu ggaugaacuc       2040

cacugguuuc accaagacuu guggcgcgcc accuugccgc accagagcug acuucaacgc       2100

cagcacggac uuguugugcc cuacggauug uuuuaggaag cauccugaug ccacuuauau       2160

uaaguguggu ucugggcccu ggcucacacc aaagugccug guccacuacc cuuacagacu       2220

cuggcauuac cccugcacag ucaauuuuac caucuucaag auaagaaugu auguaggggg       2280

gguugagcac aggcucacgg ccgcaugcaa cuucacucgu ggggaucgcu gcgacuugga       2340

ggacagggac aggagucagc ugucuccucu guugcacucu accacggaau gggccauccu       2400

gcccugcacc uacucagacu uacccgcuuu gucaacuggu cuucuccacc uucaccagaa       2460

caucguggac guacaauaca uguauggccu cucaccugcu aucacaaaau acgucguucg       2520

augggagugg gugguacucu uauuccugcu cuuagcggac gccagagucu gcgccugcuu       2580

guggaugcuc aucuuguugg gccaggccga agcagcauug gagaaguugg ucgucuugca       2640

cgcugcgagu gcggcuaacu gccauggccu ccuauauuuu gccaucuucu ucguggcagc       2700

uuggcacauc aggggucggg ugguccccuu gaccaccuau ugccucacug gccuauggcc       2760

cuucugccua cugcucaugg cacugccccg gcaggcuuau gccuaugacg caccugugca       2820

cggacagaua ggcguggguu uguugauauu gaucacccuc uucacacuca ccccggggua       2880

uaagacccuc cucggccagu gucuguggug guugugcuau cuccugaccc ugggggaagc       2940

caugauucag gaguggguac cacccaugca ggugcgcggc ggccgcgaug gcaucgcgug       3000

ggccgucacu auauucugcc cggguguggu guuugacauu accaaauggc uuuuggcguu       3060

gcuugggccu gcuuaccucu uaagggccgc uuugacacau gugccguacu ucgucagagc       3120

ucacgcucug auaaggguau gcgcuuuggu gaagcagcuc gcggggggua gguauguuca       3180

gguggcgcua uuggcccuug gcagguggac uggcaccuac aucuaugacc accucacacc       3240

uaugucggac ugggccgcua gcggccugcg cgacuuagcg gucgccgugg aacccaucau       3300

cuucaguccg auggagaaga aggucaucgu cuggggagcg gagacggcug caugugggga       3360

cauucuacau ggacuucccg uguccgcccg acucggccag gagauccucc ucggcccagc       3420

ugauggcuac accuccaagg gguggaagcu ccuugcuccc aucacugcuu augcccagca       3480

aacacgaggc cuccugggcg ccauaguggu gaguaugacg gggcgugaca ggacagaaca       3540

ggccggggaa guccaaaucc uguccacagu cucucagucc uuccucggaa caaccaucuc       3600

ggggguuuug uggacuguuu accacggagc uggcaacaag acucuagccg gcuuacgggg       3660

uccggucacg cagauguacu cgagugcuga gggggacuug guaggcuggc ccagcccccc       3720

ugggaccaag ucuuuggagc cgugcaagug uggagccguc gaccuauauc uggucacgcg       3780

gaacgcugau gucaucccgg cucggagacg cggggacaag cggggagcau ugcucucccc       3840

gagacccauu ucgaccuuga agggguccuc gggggggccg gugcucugcc cuaggggcca       3900

cgucguuggg cucuuccgag cagcugugug cucucggggc guggccaaau ccaucgauuu       3960

cauccccguu gagacacucg acguuguuac aaggucuccc acuuucagug acaacagcac       4020

gccaccggcu gugccccaga ccuaucaggu cggguacuug caugcuccaa cuggcagugg       4080

aaagagcacc aaggucccug ucgcguaugc cgcccagggg uacaaaguac uagugcuuaa       4140

ccccucggua gcugccaccc ugggguuugg ggcguaccua uccaaggcac auggcaucaa       4200

ucccaacauu aggacuggag ucaggaccgu gaugaccggg gaggccauca cguacuccac       4260

auauggcaaa uuucucgccg augggggcug cgcuagcggc gccuaugaca ucaucauaug       4320

cgaugaaugc cacgcugugg augcuaccuc cauucucggc aucggaacgg uccuugauca       4380

agcagagaca gccgggguca gacuaacugu gcuggcuacg gccacacccc ccgggucagu       4440

gacaaccccc caucccgaua uagaagaggu aggccucggg cgggagggug agauccccuu       4500

cuaugggagg gcgauucccc uauccugcau caagggaggg agacaccuga uuuucugcca       4560

cucaaagaaa aagugugacg agcucgcggc ggcccuucgg ggcaugggcu ugaaugccgu       4620

ggcauacuau agaggguugg acgucuccau aauaccagcu cagggagaug ugguggucgu       4680

cgccaccgac gcccucauga cgggguacac uggagacuuu gacuccguga ucgacugcaa       4740

uguagcgguc acccaagcug ucgacuucag ccuggacccc accuucacua uaaccacaca       4800

gacuguccca caagacgcug ucucacgcag ucagcgccgc gggcgcacag guagaggaag       4860

acagggcacu uauagguaug uuuccacugg ugaacgagcc ucaggaaugu uugacagugu       4920

agugcuuugu gagugcuacg acgcaggggc ugcgugguac gaucucacac cagcggagac       4980

caccgucagg cuuagagcgu auuucaacac gcccggccua cccguguguc aagaccaucu       5040

ugaauuuugg gaggcaguuu ucaccggccu cacacacaua gacgcccacu uccucuccca       5100

aacaaagcaa gcgggggaga acuucgcgua ccuaguagcc uaccaagcua cggugugcgc       5160

cagagccaag gccccucccc cguccuggga cgccaugugg aagugccugg cccgacucaa       5220

gccuacgcuu gcgggcccca caccucuccu guaccguuug ggcccuauua ccaaugaggu       5280

cacccucaca cacccuggga cgaaguacau cgccacaugc augcaagcug accuugaggu       5340

caugaccagc acgugggucc uagcuggagg aguccuggca gccgucgccg cauauugccu       5400

ggcgacugga ugcguuucca ucaucggccg cuugcacguc aaccagcgag ucgucguugc       5460

gccggauaag gagguccugu augaggcuuu ugaugagaug gaggaaugcg ccucuagggc       5520

ggcucucauc gaagaggggc agcggauagc cgagauguug aaguccaaga uccaaggcuu       5580

gcugcagcag gccucuaagc aggcccagga cauacaaccc gcuaugcagg cuucauggcc       5640

caaaguggaa caauuuuggg ccagacacau guggaacuuc auuagcggca uccaauaccu       5700

cgcaggauug ucaacacugc cagggaaccc cgcgguggcu uccaugaugg cauucagugc       5760

cgcccucacc aguccguugu cgaccaguac caccauccuu cucaacauca ugggaggcug       5820

guuagcgucc cagaucgcac cacccgcggg ggccaccggc uuugucguca guggccuggu       5880

gggggcugcc gugggcagca uaggccuggg uaaggugcug guggacaucc uggcaggaua       5940

uggugcgggc auuucggggg cccucgucgc auucaagauc augucuggcg agaagcccuc       6000

uauggaagau gucaucaauc uacugccugg gauccugucu ccgggagccc uggugguggg       6060

ggucaucugc gcggccauuc ugcgccgcca cgugggaccg ggggagggcg cgguccaaug       6120

gaugaacagg cuuauugccu uugcuuccag aggaaaccac gucgccccua cucacuacgu       6180

gacggagucg gaugcgucgc agcgugugac ccaacuacuu ggcucucuua cuauaaccag       6240

ccuacucaga agacuccaca auuggauaac ugaggacugc cccaucccau gcuccggauc       6300

cuggcuccgc gacguguggg acuggguuug caccaucuug acagacuuca aaaauuggcu       6360

gaccucuaaa uuguucccca agcugcccgg ccuccccuuc aucucuuguc aaaaggggua       6420

caagggugug ugggccggca cuggcaucau gaccacgcgc ugcccuugcg gcgccaacau       6480

cucuggcaau guccgccugg gcucuaugag gaucacaggg ccuaaaaccu gcaugaacac       6540

cuggcagggg accuuuccua ucaauugcua cacggagggc cagugcgcgc cgaaaccccc       6600

cacgaacuac aagaccgcca ucuggagggu ggcggccucg gaguacgcgg aggugacgca       6660

gcaugggucg uacuccuaug uaacaggacu gaccacugac aaucugaaaa uuccuugcca       6720

acuaccuucu ccagaguuuu ucuccugggu ggacggugug cagauccaua gguuugcacc       6780

cacaccaaag ccguuuuucc gggaugaggu cucguucugc guugggcuua auuccuaugc       6840

ugucgggucc cagcuucccu gugaaccuga gcccgacgca gacguauuga gguccaugcu       6900

aacagauccg ccccacauca cggcggagac ugcggcgcgg cgcuuggcac ggggaucacc       6960

uccaucugag gcgagcuccu cagugagcca gcuaucagca ccgucgcugc gggccaccug       7020

caccacccac agcaacaccu augacgugga cauggucgau gccaaccugc ucauggaggg       7080

cgguguggcu cagacagagc cugaguccag ggugcccguu cuggacuuuc ucgagccaau       7140

ggccgaggaa gagagcgacc uugagcccuc aauaccaucg gagugcaugc uccccaggag       7200

cggguuucca cgggccuuac cggcuugggc acggccugac uacaacccgc cgcucgugga       7260

aucguggagg aggccagauu accaaccgcc caccguugcu gguugugcuc uccccccccc       7320

caagaaggcc ccgacgccuc ccccaaggag acgccggaca gugggucuga gcgagagcac       7380

cauaucagaa gcccuccagc aacuggccau caagaccuuu ggccagcccc ccucgagcgg       7440

ugaugcaggc ucguccacgg gggcgggcgc cgccgaaucc ggcgguccga cguccccugg       7500

ugagccggcc cccucagaga cagguuccgc cuccucuaug cccccccucg agggggagcc       7560

uggagauccg gaccuggagu cugaucaggu agagcuucaa ccuccccccc aggggggggg       7620

gguagcuccc gguucgggcu cggggucuug gucuacuugc uccgaggagg acgauaccac       7680

cgugugcugc uccaugucau acuccuggac cggggcucua auaacucccu guagccccga       7740

agaggaaaag uugccaauca acccuuugag uaacucgcug uugcgauacc auaacaaggu       7800

guacuguaca acaucaaaga gcgccucaca gagggcuaaa aagguaacuu uugacaggac       7860

gcaagugcuc gacgcccauu augacucagu cuuaaaggac aucaagcuag cggcuuccaa       7920

ggucagcgca aggcuccuca ccuuggagga ggcgugccag uugacuccac cccauucugc       7980

aagauccaag uauggauucg gggccaagga gguccgcagc uuguccggga gggccguuaa       8040

ccacaucaag uccgugugga aggaccuccu ggaagaccca caaacaccaa uucccacaac       8100

caucauggcc aaaaaugagg uguucugcgu ggaccccgcc aaggggggua agaaaccagc       8160

ucgccucauc guuuacccug accucggcgu ccgggucugc gagaaaaugg cccucuauga       8220

cauuacacaa aagcuuccuc aggcgguaau gggagcuucc uauggcuucc aguacucccc       8280

ugcccaacgg guggaguauc ucuugaaagc augggcggaa aagaaggacc ccauggguuu       8340

uucguaugau acccgaugcu ucgacucaac cgucacugag agagacauca ggaccgagga       8400

guccauauac caggccugcu cccugcccga ggaggcccgc acugccauac acucgcugac       8460

ugagagacuu uacguaggag ggcccauguu caacagcaag ggucaaaccu gcgguuacag       8520

acguugccgc gccagcgggg ugcuaaccac uagcaugggu aacaccauca caugcuaugu       8580

gaaagcccua gcggccugca aggcugcggg gauaguugcg cccacaaugc ugguaugcgg       8640

cgaugaccua guagucaucu cagaaagcca ggggacugag gaggacgagc ggaaccugag       8700

agccuucacg gaggccauga ccagguacuc ugccccuccu ggugaucccc ccagaccgga       8760

auaugaccug gagcuaauaa cauccuguuc cucaaaugug ucuguggcgu ugggcccgcg       8820

gggccgccgc agauacuacc ugaccagaga cccaaccacu ccacucgccc gggcugccug       8880

ggaaacaguu agacacuccc cuaucaauuc auggcuggga aacaucaucc aguaugcucc       8940

aaccauaugg guucgcaugg uccuaaugac acacuucuuc uccauucuca ugguccaaga       9000

cacccuggac cagaaccuca acuuugagau guauggauca guauacuccg ugaauccuuu       9060

ggaccuucca gccauaauug agagguuaca cgggcuugac gccuuuucua ugcacacaua       9120

cucucaccac gaacugacgc ggguggcuuc agcccucaga aaacuugggg cgccaccccu       9180

cagggugugg aagagucggg cucgcgcagu cagggcgucc cucaucuccc guggagggaa       9240

agcggccguu ugcggccgau aucucuucaa uugggcggug aagaccaagc ucaaacucac       9300

uccauugccg gaggcgcgcc uacuggacuu auccaguugg uucaccgucg gcgccggcgg       9360

gggcgacauu uuucacagcg ugucgcgcgc ccgaccccgc ucauuacucu ucggccuacu       9420

ccuacuuuuc guagggguag gccucuuccu acuccccgcu cgguagagcg gcacacacua       9480

gguacacucc auagcuaacu guuccuuuuu uuuuuuuuuu uuuuuuuuuu uuuuuuuuuu       9540

uuuuuuuucu uuuuuuuuuu uuucccucuu ucuucccuuc ucaucuuauu cuacuuucuu       9600

ucuugguggc uccaucuuag cccuagucac ggcuagcugu gaaagguccg ugagccgcau       9660

gacugcagag agugccguaa cuggucucuc ugcagaucau gucuaga                     9707

<210>13

<211>11111

<212>RNA

<213>人工序列

<220>

<223>人工序列描述：含来自JFH-1克隆的全长丙型肝炎病毒基因组RNA的复制子RNA

<400>13

accugccccu aauaggggcg acacuccgcc augaaucacu ccccugugag gaacuacugu       60

cuucacgcag aaagcgccua gccauggcgu uaguaugagu gucguacagc cuccaggccc       120

cccccucccg ggagagccau aguggucugc ggaaccggug aguacaccgg aauugccggg       180

aagacugggu ccuuucuugg auaaacccac ucuaugcccg gccauuuggg cgugcccccg       240

caagacugcu agccgaguag cguuggguug cgaaaggccu ugugguacug ccugauaggg       300

cgcuugcgag ugccccggga ggucucguag accgugcacc augagcacaa auccuaaacc       360

ucaaagaaaa accaaaagaa acaccaaccg ucgcccaaug auugaacaag auggauugca       420

cgcagguucu ccggccgcuu ggguggagag gcuauucggc uaugacuggg cacaacagac       480

aaucggcugc ucugaugccg ccguguuccg gcugucagcg caggggcgcc cgguucuuuu       540

ugucaagacc gaccuguccg gugcccugaa ugaacugcag gacgaggcag cgcggcuauc       600

guggcuggcc acgacgggcg uuccuugcgc agcugugcuc gacguuguca cugaagcggg       660

aagggacugg cugcuauugg gcgaagugcc ggggcaggau cuccugucau cucaccuugc       720

uccugccgag aaaguaucca ucauggcuga ugcaaugcgg cggcugcaua cgcuugaucc       780

ggcuaccugc ccauucgacc accaagcgaa acaucgcauc gagcgagcac guacucggau       840

ggaagccggu cuugucgauc aggaugaucu ggacgaagag caucaggggc ucgcgccagc       900

cgaacuguuc gccaggcuca aggcgcgcau gcccgacggc gaggaucucg ucgugaccca       960

uggcgaugcc ugcuugccga auaucauggu ggaaaauggc cgcuuuucug gauucaucga       1020

cuguggccgg cugggugugg cggaccgcua ucaggacaua gcguuggcua cccgugauau       1080

ugcugaagag cuuggcggcg aaugggcuga ccgcuuccuc gugcuuuacg guaucgccgc       1140

ucccgauucg cagcgcaucg ccuucuaucg ccuucuugac gaguucuucu gaguuuaaac       1200

ccucucccuc cccccccccu aacguuacug gccgaagccg cuuggaauaa ggccggugug       1260

cguuugucua uauguuauuu uccaccauau ugccgucuuu uggcaaugug agggcccgga       1320

aaccuggccc ugucuucuug acgagcauuc cuaggggucu uuccccucuc gccaaaggaa       1380

ugcaaggucu guugaauguc gugaaggaag caguuccucu ggaagcuucu ugaagacaaa       1440

caacgucugu agcgacccuu ugcaggcagc ggaacccccc accuggcgac aggugccucu       1500

gcggccaaaa gccacgugua uaagauacac cugcaaaggc ggcacaaccc cagugccacg       1560

uugugaguug gauaguugug gaaagaguca aauggcucuc cucaagcgua uucaacaagg       1620

ggcugaagga ugcccagaag guaccccauu guaugggauc ugaucugggg ccucggugca       1680

caugcuuuac auguguuuag ucgagguuaa aaaaacgucu aggccccccg aaccacgggg       1740

acgugguuuu ccuuugaaaa acacgaugau accaugagca caaauccuaa accucaaaga       1800

aaaaccaaaa gaaacaccaa ccgucgccca gaagacguua aguucccggg cggcggccag       1860

aucguuggcg gaguauacuu guugccgcgc aggggcccca gguugggugu gcgcacgaca       1920

aggaaaacuu cggagcgguc ccagccacgu gggagacgcc agcccauccc caaagaucgg       1980

cgcuccacug gcaaggccug gggaaaacca ggucgccccu ggccccuaua ugggaaugag       2040

ggacucggcu gggcaggaug gcuccugucc ccccgaggcu cucgccccuc cuggggcccc       2100

acugaccccc ggcauagguc gcgcaacgug gguaaaguca ucgacacccu aacguguggc       2160

uuugccgacc ucauggggua cauccccguc guaggcgccc cgcuuagugg cgccgccaga       2220

gcugucgcgc acggcgugag aguccuggag gacgggguua auuaugcaac agggaaccua       2280

cccgguuucc ccuuuucuau cuucuugcug gcccuguugu ccugcaucac cguuccgguc       2340

ucugcugccc aggugaagaa uaccaguagc agcuacaugg ugaccaauga cugcuccaau       2400

gacagcauca cuuggcagcu cgaggcugcg guucuccacg uccccgggug cgucccgugc       2460

gagagagugg ggaauacguc acgguguugg gugccagucu cgccaaacau ggcugugcgg       2520

cagcccggug cccucacgca gggucugcgg acgcacaucg auaugguugu gauguccgcc       2580

accuucugcu cugcucucua cgugggggac cucuguggcg gggugaugcu cgcggcccag       2640

guguucaucg ucucgccgca guaccacugg uuugugcaag aaugcaauug cuccaucuac       2700

ccuggcacca ucacuggaca ccgcauggca ugggacauga ugaugaacug gucgcccacg       2760

gccaccauga uccuggcgua cgugaugcgc guccccgagg ucaucauaga caucguuagc       2820

ggggcucacu ggggcgucau guucggcuug gccuacuucu cuaugcaggg agcgugggcg       2880

aaggucauug ucauccuucu gcuggccgcu gggguggacg cgggcaccac caccguugga       2940

ggcgcuguug cacguuccac caacgugauu gccggcgugu ucagccaugg cccucagcag       3000

aacauucagc ucauuaacac caacggcagu uggcacauca accguacugc cuugaauugc       3060

aaugacuccu ugaacaccgg cuuucucgcg gccuuguucu acaccaaccg cuuuaacucg       3120

ucaggguguc cagggcgccu guccgccugc cgcaacaucg aggcuuuccg gauagggugg       3180

ggcacccuac aguacgagga uaaugucacc aauccagagg auaugaggcc guacugcugg       3240

cacuaccccc caaagccgug uggcguaguc cccgcgaggu cugugugugg cccaguguac       3300

uguuucaccc ccagcccggu aguagugggc acgaccgaca gacguggagu gcccaccuac       3360

acauggggag agaaugagac agaugucuuc cuacugaaca gcacccgacc gccgcagggc       3420

ucaugguucg gcugcacgug gaugaacucc acugguuuca ccaagacuug uggcgcgcca       3480

ccuugccgca ccagagcuga cuucaacgcc agcacggacu uguugugccc uacggauugu       3540

uuuaggaagc auccugaugc cacuuauauu aagugugguu cugggcccug gcucacacca       3600

aagugccugg uccacuaccc uuacagacuc uggcauuacc ccugcacagu caauuuuacc       3660

aucuucaaga uaagaaugua uguagggggg guugagcaca ggcucacggc cgcaugcaac       3720

uucacucgug gggaucgcug cgacuuggag gacagggaca ggagucagcu gucuccucug       3780

uugcacucua ccacggaaug ggccauccug cccugcaccu acucagacuu acccgcuuug       3840

ucaacugguc uucuccaccu ucaccagaac aucguggacg uacaauacau guauggccuc       3900

ucaccugcua ucacaaaaua cgucguucga ugggaguggg ugguacucuu auuccugcuc       3960

uuagcggacg ccagagucug cgccugcuug uggaugcuca ucuuguuggg ccaggccgaa       4020

gcagcauugg agaaguuggu cgucuugcac gcugcgagug cggcuaacug ccauggccuc       4080

cuauauuuug ccaucuucuu cguggcagcu uggcacauca ggggucgggu gguccccuug       4140

accaccuauu gccucacugg ccuauggccc uucugccuac ugcucauggc acugccccgg       4200

caggcuuaug ccuaugacgc accugugcac ggacagauag gcguggguuu guugauauug       4260

aucacccucu ucacacucac cccgggguau aagacccucc ucggccagug ucuguggugg       4320

uugugcuauc uccugacccu gggggaagcc augauucagg aguggguacc acccaugcag       4380

gugcgcggcg gccgcgaugg caucgcgugg gccgucacua uauucugccc ggguguggug       4440

uuugacauuaccaaauggcu  uuuggcguug cuugggccug cuuaccucuu aagggccgcu       4500

uugacacaug ugccguacuu cgucagagcu cacgcucuga uaaggguaug cgcuuuggug       4560

aagcagcucg cgggggguag guauguucag guggcgcuau uggcccuugg cagguggacu       4620

ggcaccuaca ucuaugacca ccucacaccu augucggacu gggccgcuag cggccugcgc       4680

gacuuagcgg ucgccgugga acccaucauc uucaguccga uggagaagaa ggucaucguc       4740

uggggagcgg agacggcugc auguggggac auucuacaug gacuucccgu guccgcccga       4800

cucggccagg agauccuccu cggcccagcu gauggcuaca ccuccaaggg guggaagcuc       4860

cuugcuccca ucacugcuua ugcccagcaa acacgaggcc uccugggcgc cauaguggug       4920

aguaugacgg ggcgugacag gacagaacag gccggggaag uccaaauccu guccacaguc       4980

ucucaguccu uccucggaac aaccaucucg gggguuuugu ggacuguuua ccacggagcu       5040

ggcaacaaga cucuagccgg cuuacggggu ccggucacgc agauguacuc gagugcugag       5100

ggggacuugg uaggcuggcc cagccccccu gggaccaagu cuuuggagcc gugcaagugu       5160

ggagccgucg accuauaucu ggucacgcgg aacgcugaug ucaucccggc ucggagacgc       5220

ggggacaagc ggggagcauu gcucuccccg agacccauuu cgaccuugaa gggguccucg       5280

ggggggccgg ugcucugccc uaggggccac gucguugggc ucuuccgagc agcugugugc       5340

ucucggggcg uggccaaauc caucgauuuc auccccguug agacacucga cguuguuaca       5400

aggucuccca cuuucaguga caacagcacg ccaccggcug ugccccagac cuaucagguc       5460

ggguacuugc augcuccaac uggcagugga aagagcacca aggucccugu cgcguaugcc       5520

gcccaggggu acaaaguacu agugcuuaac cccucgguag cugccacccu gggguuuggg       5580

gcguaccuau ccaaggcaca uggcaucaau cccaacauua ggacuggagu caggaccgug       5640

augaccgggg aggccaucac guacuccaca uauggcaaau uucucgccga ugggggcugc       5700

gcuagcggcg ccuaugacau caucauaugc gaugaaugcc acgcugugga ugcuaccucc       5760

auucucggca ucggaacggu ccuugaucaa gcagagacag ccggggucag acuaacugug       5820

cuggcuacgg ccacaccccc cgggucagug acaacccccc aucccgauau agaagaggua       5880

ggccucgggc gggaggguga gauccccuuc uaugggaggg cgauuccccu auccugcauc       5940

aagggaggga gacaccugau uuucugccac ucaaagaaaa agugugacga gcucgcggcg       6000

gcccuucggg gcaugggcuu gaaugccgug gcauacuaua gaggguugga cgucuccaua       6060

auaccagcuc agggagaugu gguggucguc gccaccgacg cccucaugac gggguacacu       6120

ggagacuuug acuccgugau cgacugcaau guagcgguca cccaagcugu cgacuucagc       6180

cuggacccca ccuucacuau aaccacacag acugucccac aagacgcugu cucacgcagu       6240

cagcgccgcg ggcgcacagg uagaggaaga cagggcacuu auagguaugu uuccacuggu       6300

gaacgagccu caggaauguu ugacagugua gugcuuugug agugcuacga cgcaggggcu       6360

gcgugguacg aucucacacc agcggagacc accgucaggc uuagagcgua uuucaacacg       6420

cccggccuac ccguguguca agaccaucuu gaauuuuggg aggcaguuuu caccggccuc       6480

acacacauag acgcccacuu ccucucccaa acaaagcaag cgggggagaa cuucgcguac       6540

cuaguagccu accaagcuac ggugugcgcc agagccaagg ccccuccccc guccugggac       6600

gccaugugga agugccuggc ccgacucaag ccuacgcuug cgggccccac accucuccug       6660

uaccguuugg gcccuauuac caaugagguc acccucacac acccugggac gaaguacauc       6720

gccacaugca ugcaagcuga ccuugagguc augaccagca cguggguccu agcuggagga       6780

guccuggcag ccgucgccgc auauugccug gcgacuggau gcguuuccau caucggccgc       6840

uugcacguca accagcgagu cgucguugcg ccggauaagg agguccugua ugaggcuuuu       6900

gaugagaugg aggaaugcgc cucuagggcg gcucucaucg aagaggggca gcggauagcc       6960

gagauguuga aguccaagau ccaaggcuug cugcagcagg ccucuaagca ggcccaggac       7020

auacaacccg cuaugcaggc uucauggccc aaaguggaac aauuuugggc cagacacaug       7080

uggaacuuca uuagcggcau ccaauaccuc gcaggauugu caacacugcc agggaacccc       7140

gcgguggcuu ccaugauggc auucagugcc gcccucacca guccguuguc gaccaguacc       7200

accauccuuc ucaacaucau gggaggcugg uuagcguccc agaucgcacc acccgcgggg       7260

gccaccggcu uugucgucag uggccuggug ggggcugccg ugggcagcau aggccugggu       7320

aaggugcugg uggacauccu ggcaggauau ggugcgggca uuucgggggc ccucgucgca       7380

uucaagauca ugucuggcga gaagcccucu auggaagaug ucaucaaucu acugccuggg       7440

auccugucuc cgggagcccu gguggugggg gucaucugcg cggccauucu gcgccgccac       7500

gugggaccgg gggagggcgc gguccaaugg augaacaggc uuauugccuu ugcuuccaga       7560

ggaaaccacg ucgccccuac ucacuacgug acggagucgg augcgucgca gcgugugacc       7620

caacuacuug gcucucuuac uauaaccagc cuacucagaa gacuccacaa uuggauaacu       7680

gaggacugcc ccaucccaug cuccggaucc uggcuccgcg acguguggga cuggguuugc       7740

accaucuuga cagacuucaa aaauuggcug accucuaaau uguuccccaa gcugcccggc       7800

cuccccuuca ucucuuguca aaagggguac aagggugugu gggccggcac uggcaucaug       7860

accacgcgcu gcccuugcgg cgccaacauc ucuggcaaug uccgccuggg cucuaugagg       7920

aucacagggc cuaaaaccug caugaacacc uggcagggga ccuuuccuau caauugcuac       7980

acggagggcc agugcgcgcc gaaacccccc acgaacuaca agaccgccau cuggagggug       8040

gcggccucgg aguacgcgga ggugacgcag caugggucgu acuccuaugu aacaggacug       8100

accacugaca aucugaaaau uccuugccaa cuaccuucuc cagaguuuuu cuccugggug       8160

gacggugugc agauccauag guuugcaccc acaccaaagc cguuuuuccg ggaugagguc       8220

ucguucugcg uugggcuuaa uuccuaugcu gucggguccc agcuucccug ugaaccugag       8280

cccgacgcag acguauugag guccaugcua acagauccgc cccacaucac ggcggagacu       8340

gcggcgcggc gcuuggcacg gggaucaccu ccaucugagg cgagcuccuc agugagccag       8400

cuaucagcac cgucgcugcg ggccaccugc accacccaca gcaacaccua ugacguggac       8460

auggucgaug ccaaccugcu cauggagggc gguguggcuc agacagagcc ugaguccagg       8520

gugcccguuc uggacuuucu cgagccaaug gccgaggaag agagcgaccu ugagcccuca       8580

auaccaucgg agugcaugcu ccccaggagc ggguuuccac gggccuuacc ggcuugggca       8640

cggccugacu acaacccgcc gcucguggaa ucguggagga ggccagauua ccaaccgccc       8700

accguugcug guugugcucu cccccccccc aagaaggccc cgacgccucc cccaaggaga       8760

cgccggacag ugggucugag cgagagcacc auaucagaag cccuccagca acuggccauc       8820

aagaccuuug gccagccccc cucgagcggu gaugcaggcu cguccacggg ggcgggcgcc       8880

gccgaauccg gcgguccgac guccccuggu gagccggccc ccucagagac agguuccgcc       8940

uccucuaugc ccccccucga gggggagccu ggagauccgg accuggaguc ugaucaggua       9000

gagcuucaac cuccccccca gggggggggg guagcucccg guucgggcuc ggggucuugg       9060

ucuacuugcu ccgaggagga cgauaccacc gugugcugcu ccaugucaua cuccuggacc       9120

ggggcucuaa uaacucccug uagccccgaa gaggaaaagu ugccaaucaa cccuuugagu       9180

aacucgcugu ugcgauacca uaacaaggug uacuguacaa caucaaagag cgccucacag       9240

agggcuaaaa agguaacuuu ugacaggacg caagugcucg acgcccauua ugacucaguc       9300

uuaaaggaca ucaagcuagc ggcuuccaag gucagcgcaa ggcuccucac cuuggaggag       9360

gcgugccagu ugacuccacc ccauucugca agauccaagu auggauucgg ggccaaggag       9420

guccgcagcu uguccgggag ggccguuaac cacaucaagu ccguguggaa ggaccuccug       9480

gaagacccac aaacaccaau ucccacaacc aucauggcca aaaaugaggu guucugcgug       9540

gaccccgcca agggggguaa gaaaccagcu cgccucaucg uuuacccuga ccucggcguc       9600

cgggucugcg agaaaauggc ccucuaugac auuacacaaa agcuuccuca ggcgguaaug       9660

ggagcuuccu auggcuucca guacuccccu gcccaacggg uggaguaucu cuugaaagca       9720

ugggcggaaa agaaggaccc cauggguuuu ucguaugaua cccgaugcuu cgacucaacc       9780

gucacugaga gagacaucag gaccgaggag uccauauacc aggccugcuc ccugcccgag       9840

gaggcccgca cugccauaca cucgcugacu gagagacuuu acguaggagg gcccauguuc       9900

aacagcaagg gucaaaccug cgguuacaga cguugccgcg ccagcggggu gcuaaccacu       9960

agcaugggua acaccaucac augcuaugug aaagcccuag cggccugcaa ggcugcgggg       10020

auaguugcgc ccacaaugcu gguaugcggc gaugaccuag uagucaucuc agaaagccag       10080

gggacugagg aggacgagcg gaaccugaga gccuucacgg aggccaugac cagguacucu       10140

gccccuccug gugauccccc cagaccggaa uaugaccugg agcuaauaac auccuguucc       10200

ucaaaugugu cuguggcguu gggcccgcgg ggccgccgca gauacuaccu gaccagagac       10260

ccaaccacuc cacucgcccg ggcugccugg gaaacaguua gacacucccc uaucaauuca       10320

uggcugggaa acaucaucca guaugcucca accauauggg uucgcauggu ccuaaugaca       10380

cacuucuucu ccauucucau gguccaagac acccuggacc agaaccucaa cuuugagaug       10440

uauggaucag uauacuccgu gaauccuuug gaccuuccag ccauaauuga gagguuacac       10500

gggcuugacg ccuuuucuau gcacacauac ucucaccacg aacugacgcg gguggcuuca       10560

gcccucagaa aacuuggggc gccaccccuc agggugugga agagucgggc ucgcgcaguc       10620

agggcguccc ucaucucccg uggagggaaa gcggccguuu gcggccgaua ucucuucaau       10680

ugggcgguga agaccaagcu caaacucacu ccauugccgg aggcgcgccu acuggacuua       10740

uccaguuggu ucaccgucgg cgccggcggg ggcgacauuu uucacagcgu gucgcgcgcc       10800

cgaccccgcu cauuacucuu cggccuacuc cuacuuuucg uagggguagg ccucuuccua       10860

cuccccgcuc gguagagcgg cacacacuag guacacucca uagcuaacug uuccuuuuuu       10920

uuuuuuuuuu uuuuuuuuuu uuuuuuuuuu uuuuuuucuu uuuuuuuuuu uucccucuuu       10980

cuucccuucu caucuuauuc uacuuucuuu cuugguggcu ccaucuuagc ccuagucacg       11040

gcuagcugug aaagguccgu gagccgcaug acugcagaga gugccguaac uggucucucu       11100

gcagaucaug u                                                            11111

<210>14

<211>11111

<212>RNA

<213>人工序列

<220>

<223>人工序列的描述：来自JFH-1克隆的全长丙型肝炎病毒基因组RNA，其中氨基酸基序GDD已突变为GND

<400>14

accugccccu aauaggggcg acacuccgcc augaaucacu ccccugugag gaacuacugu       60

cuucacgcag aaagcgccua gccauggcgu uaguaugagu gucguacagc cuccaggccc       120

cccccucccg ggagagccau aguggucugc ggaaccggug aguacaccgg aauugccggg       180

aagacugggu ccuuucuugg auaaacccac ucuaugcccg gccauuuggg cgugcccccg       240

caagacugcu agccgaguag cguuggguug cgaaaggccu ugugguacug ccugauaggg       300

cgcuugcgag ugccccggga ggucucguag accgugcacc augagcacaa auccuaaacc       360

ucaaagaaaa accaaaagaa acaccaaccg ucgcccaaug auugaacaag auggauugca       420

cgcagguucu ccggccgcuu ggguggagag gcuauucggc uaugacuggg cacaacagac       480

aaucggcugc ucugaugccg ccguguuccg gcugucagcg caggggcgcc cgguucuuuu       540

ugucaagacc gaccuguccg gugcccugaa ugaacugcag gacgaggcag cgcggcuauc       600

guggcuggcc acgacgggcg uuccuugcgc agcugugcuc gacguuguca cugaagcggg       660

aagggacugg cugcuauugg gcgaagugcc ggggcaggau cuccugucau cucaccuugc       720

uccugccgag aaaguaucca ucauggcuga ugcaaugcgg cggcugcaua cgcuugaucc       780

ggcuaccugc ccauucgacc accaagcgaa acaucgcauc gagcgagcac guacucggau       840

ggaagccggu cuugucgauc aggaugaucu ggacgaagag caucaggggc ucgcgccagc       900

cgaacuguuc gccaggcuca aggcgcgcau gcccgacggc gaggaucucg ucgugaccca       960

uggcgaugcc ugcuugccga auaucauggu ggaaaauggc cgcuuuucug gauucaucga       1020

cuguggccgg cugggugugg cggaccgcua ucaggacaua gcguuggcua cccgugauau       1080

ugcugaagag cuuggcggcg aaugggcuga ccgcuuccuc gugcuuuacg guaucgccgc       1140

ucccgauucg cagcgcaucg ccuucuaucg ccuucuugac gaguucuucu gaguuuaaac       1200

ccucucccuc cccccccccu aacguuacug gccgaagccg cuuggaauaa ggccggugug       1260

cguuugucua uauguuauuu uccaccauau ugccgucuuu uggcaaugug agggcccgga       1320

aaccuggccc ugucuucuug acgagcauuc cuaggggucu uuccccucuc gccaaaggaa       1380

ugcaaggucu guugaauguc gugaaggaag caguuccucu ggaagcuucu ugaagacaaa       1440

caacgucugu agcgacccuu ugcaggcagc ggaacccccc accuggcgac aggugccucu       1500

gcggccaaaa gccacgugua uaagauacac cugcaaaggc ggcacaaccc cagugccacg       1560

uugugaguug gauaguugug gaaagaguca aauggcucuc cucaagcgua uucaacaagg       1620

ggcugaagga ugcccagaag guaccccauu guaugggauc ugaucugggg ccucggugca       1680

caugcuuuac auguguuuag ucgagguuaa aaaaacgucu aggccccccg aaccacgggg       1740

acgugguuuu ccuuugaaaa acacgaugau accaugagca caaauccuaa accucaaaga       1800

aaaaccaaaa gaaacaccaa ccgucgccca gaagacguua aguucccggg cggcggccag       1860

aucguuggcg gaguauacuu guugccgcgc aggggcccca gguugggugu gcgcacgaca       1920

aggaaaacuu cggagcgguc ccagccacgu gggagacgcc agcccauccc caaagaucgg       1980

cgcuccacug gcaaggccug gggaaaacca ggucgccccu ggccccuaua ugggaaugag       2040

ggacucggcu gggcaggaug gcuccugucc ccccgaggcu cucgccccuc cuggggcccc       2100

acugaccccc ggcauagguc gcgcaacgug gguaaaguca ucgacacccu aacguguggc       2160

uuugccgacc ucauggggua cauccccguc guaggcgccc cgcuuagugg cgccgccaga       2220

gcugucgcgc acggcgugag aguccuggag gacgggguua auuaugcaac agggaaccua       2280

cccgguuucc ccuuuucuau cuucuugcug gcccuguugu ccugcaucac cguuccgguc       2340

ucugcugccc aggugaagaa uaccaguagc agcuacaugg ugaccaauga cugcuccaau       2400

gacagcauca cuuggcagcu cgaggcugcg guucuccacg uccccgggug cgucccgugc       2460

gagagagugg ggaauacguc acgguguugg gugccagucu cgccaaacau ggcugugcgg       2520

cagcccggug cccucacgca gggucugcgg acgcacaucg auaugguugu gauguccgcc       2580

accuucugcu cugcucucua cgugggggac cucuguggcg gggugaugcu cgcggcccag       2640

guguucaucg ucucgccgca guaccacugg uuugugcaag aaugcaauug cuccaucuac       2700

ccuggcacca ucacuggaca ccgcauggca ugggacauga ugaugaacug gucgcccacg       2760

gccaccauga uccuggcgua cgugaugcgc guccccgagg ucaucauaga caucguuagc       2820

ggggcucacu ggggcgucau guucggcuug gccuacuucu cuaugcaggg agcgugggcg       2880

aaggucauug ucauccuucu gcuggccgcu gggguggacg cgggcaccac caccguugga       2940

ggcgcuguug cacguuccac caacgugauu gccggcgugu ucagccaugg cccucagcag       3000

aacauucagc ucauuaacac caacggcagu uggcacauca accguacugc cuugaauugc       3060

aaugacuccu ugaacaccgg cuuucucgcg gccuuguucu acaccaaccg cuuuaacucg       3120

ucaggguguc cagggcgccu guccgccugc cgcaacaucg aggcuuuccg gauagggugg       3180

ggcacccuac aguacgagga uaaugucacc aauccagagg auaugaggcc guacugcugg       3240

cacuaccccc caaagccgug uggcguaguc cccgcgaggu cugugugugg cccaguguac       3300

uguuucaccc ccagcccggu aguagugggc acgaccgaca gacguggagu gcccaccuac       3360

acauggggag agaaugagac agaugucuuc cuacugaaca gcacccgacc gccgcagggc       3420

ucaugguucg gcugcacgug gaugaacucc acugguuuca ccaagacuug uggcgcgcca       3480

ccuugccgca ccagagcuga cuucaacgcc agcacggacu uguugugccc uacggauugu       3540

uuuaggaagc auccugaugc cacuuauauu aagugugguu cugggcccug gcucacacca       3600

aagugccugg uccacuaccc uuacagacuc uggcauuacc ccugcacagu caauuuuacc       3660

aucuucaaga uaagaaugua uguagggggg guugagcaca ggcucacggc cgcaugcaac       3720

uucacucgug gggaucgcug cgacuuggag gacagggaca ggagucagcu gucuccucug       3780

uugcacucua ccacggaaug ggccauccug cccugcaccu acucagacuu acccgcuuug       3840

ucaacugguc uucuccaccu ucaccagaac aucguggacg uacaauacau guauggccuc       3900

ucaccugcua ucacaaaaua cgucguucga ugggaguggg ugguacucuu auuccugcuc       3960

uuagcggacg ccagagucug cgccugcuug uggaugcuca ucuuguuggg ccaggccgaa       4020

gcagcauugg agaaguuggu cgucuugcac gcugcgagug cggcuaacug ccauggccuc       4080

cuauauuuug ccaucuucuu cguggcagcu uggcacauca ggggucgggu gguccccuug       4140

accaccuauu gccucacugg ccuauggccc uucugccuac ugcucauggc acugccccgg       4200

caggcuuaug ccuaugacgc accugugcac ggacagauag gcguggguuu guugauauug       4260

aucacccucu ucacacucac cccgggguau aagacccucc ucggccagug ucuguggugg       4320

uugugcuauc uccugacccu gggggaagcc augauucagg aguggguacc acccaugcag       4380

gugcgcggcg gccgcgaugg caucgcgugg gccgucacua uauucugccc ggguguggug       4440

uuugacauua ccaaauggcu uuuggcguug cuugggccug cuuaccucuu aagggccgcu       4500

uugacacaug ugccguacuu cgucagagcu cacgcucuga uaaggguaug cgcuuuggug       4560

aagcagcucg cgggggguag guauguucag guggcgcuau uggcccuugg cagguggacu       4620

ggcaccuaca ucuaugacca ccucacaccu augucggacu gggccgcuag cggccugcgc       4680

gacuuagcgg ucgccgugga acccaucauc uucaguccga uggagaagaa ggucaucguc       4740

uggggagcgg agacggcugc auguggggac auucuacaug gacuucccgu guccgcccga       4800

cucggccagg agauccuccu cggcccagcu gauggcuaca ccuccaaggg guggaagcuc       4860

cuugcuccca ucacugcuua ugcccagcaa acacgaggcc uccugggcgc cauaguggug       4920

aguaugacgg ggcgugacag gacagaacag gccggggaag uccaaauccu guccacaguc       4980

ucucaguccu uccucggaac aaccaucucg gggguuuugu ggacuguuua ccacggagcu       5040

ggcaacaaga cucuagccgg cuuacggggu ccggucacgc agauguacuc gagugcugag       5100

ggggacuugg uaggcuggcc cagccccccu gggaccaagu cuuuggagcc gugcaagugu       5160

ggagccgucg accuauaucu ggucacgcgg aacgcugaug ucaucccggc ucggagacgc       5220

ggggacaagc ggggagcauu gcucuccccg agacccauuu cgaccuugaa gggguccucg       5280

ggggggccgg ugcucugccc uaggggccac gucguugggc ucuuccgagc agcugugugc       5340

ucucggggcg uggccaaauc caucgauuuc auccccguug agacacucga cguuguuaca       5400

aggucuccca cuuucaguga caacagcacg ccaccggcug ugccccagac cuaucagguc       5460

ggguacuugc augcuccaac uggcagugga aagagcacca aggucccugu cgcguaugcc       5520

gcccaggggu acaaaguacu agugcuuaac cccucgguag cugccacccu gggguuuggg       5580

gcguaccuau ccaaggcaca uggcaucaau cccaacauua ggacuggagu caggaccgug       5640

augaccgggg aggccaucac guacuccaca uauggcaaau uucucgccga ugggggcugc       5700

gcuagcggcg ccuaugacau caucauaugc gaugaaugcc acgcugugga ugcuaccucc       5760

auucucggca ucggaacggu ccuugaucaa gcagagacag ccggggucag acuaacugug       5820

cuggcuacgg ccacaccccc cgggucagug acaacccccc aucccgauau agaagaggua       5880

ggccucgggc gggaggguga gauccccuuc uaugggaggg cgauuccccu auccugcauc       5940

aagggaggga gacaccugau uuucugccac ucaaagaaaa agugugacga gcucgcggcg       6000

gcccuucggg gcaugggcuu gaaugccgug gcauacuaua gaggguugga cgucuccaua       6060

auaccagcuc agggagaugu gguggucguc gccaccgacg cccucaugac gggguacacu       6120

ggagacuuug acuccgugau cgacugcaau guagcgguca cccaagcugu cgacuucagc       6180

cuggacccca ccuucacuau aaccacacag acugucccac aagacgcugu cucacgcagu       6240

cagcgccgcg ggcgcacagg uagaggaaga cagggcacuu auagguaugu uuccacuggu       6300

gaacgagccu caggaauguu ugacagugua gugcuuugug agugcuacga cgcaggggcu       6360

gcgugguacg aucucacacc agcggagacc accgucaggc uuagagcgua uuucaacacg       6420

cccggccuac ccguguguca agaccaucuu gaauuuuggg aggcaguuuu caccggccuc       6480

acacacauag acgcccacuu ccucucccaa acaaagcaag cgggggagaa cuucgcguac       6540

cuaguagccu accaagcuac ggugugcgcc agagccaagg ccccuccccc guccugggac       6600

gccaugugga agugccuggc ccgacucaag ccuacgcuug cgggccccac accucuccug       6660

uaccguuugg gcccuauuac caaugagguc acccucacac acccugggac gaaguacauc       6720

gccacaugca ugcaagcuga ccuugagguc augaccagca cguggguccu agcuggagga       6780

guccuggcag ccgucgccgc auauugccug gcgacuggau gcguuuccau caucggccgc       6840

uugcacguca accagcgagu cgucguugcg ccggauaagg agguccugua ugaggcuuuu       6900

gaugagaugg aggaaugcgc cucuagggcg gcucucaucg aagaggggca gcggauagcc       6960

gagauguuga aguccaagau ccaaggcuug cugcagcagg ccucuaagca ggcccaggac       7020

auacaacccg cuaugcaggc uucauggccc aaaguggaac aauuuugggc cagacacaug       7080

uggaacuuca uuagcggcau ccaauaccuc gcaggauugu caacacugcc agggaacccc       7140

gcgguggcuu ccaugauggc auucagugcc gcccucacca guccguuguc gaccaguacc       7200

accauccuuc ucaacaucau gggaggcugg uuagcguccc agaucgcacc acccgcgggg       7260

gccaccggcu uugucgucag uggccuggug ggggcugccg ugggcagcau aggccugggu       7320

aaggugcugg uggacauccu ggcaggauau ggugcgggca uuucgggggc ccucgucgca       7380

uucaagauca ugucuggcga gaagcccucu auggaagaug ucaucaaucu acugccuggg       7440

auccugucuc cgggagcccu gguggugggg gucaucugcg cggccauucu gcgccgccac       7500

gugggaccgg gggagggcgc gguccaaugg augaacaggc uuauugccuu ugcuuccaga       7560

ggaaaccacg ucgccccuac ucacuacgug acggagucgg augcgucgca gcgugugacc       7620

caacuacuug gcucucuuac uauaaccagc cuacucagaa gacuccacaa uuggauaacu       7680

gaggacugcc ccaucccaug cuccggaucc uggcuccgcg acguguggga cuggguuugc       7740

accaucuuga cagacuucaa aaauuggcug accucuaaau uguuccccaa gcugcccggc       7800

cuccccuuca ucucuuguca aaagggguac aagggugugu gggccggcac uggcaucaug       7860

accacgcgcu gcccuugcgg cgccaacauc ucuggcaaug uccgccuggg cucuaugagg       7920

aucacagggc cuaaaaccug caugaacacc uggcagggga ccuuuccuau caauugcuac       7980

acggagggcc agugcgcgcc gaaacccccc acgaacuaca agaccgccau cuggagggug       8040

gcggccucgg aguacgcgga ggugacgcag caugggucgu acuccuaugu aacaggacug       8100

accacugaca aucugaaaau uccuugccaa cuaccuucuc cagaguuuuu cuccugggug       8160

gacggugugc agauccauag guuugcaccc acaccaaagc cguuuuuccg ggaugagguc       8220

ucguucugcg uugggcuuaa uuccuaugcu gucggguccc agcuucccug ugaaccugag       8280

cccgacgcag acguauugag guccaugcua acagauccgc cccacaucac ggcggagacu       8340

gcggcgcggc gcuuggcacg gggaucaccu ccaucugagg cgagcuccuc agugagccag       8400

cuaucagcac cgucgcugcg ggccaccugc accacccaca gcaacaccua ugacguggac       8460

auggucgaug ccaaccugcu cauggagggc gguguggcuc agacagagcc ugaguccagg       8520

gugcccguuc uggacuuucu cgagccaaug gccgaggaag agagcgaccu ugagcccuca       8580

auaccaucgg agugcaugcu ccccaggagc ggguuuccac gggccuuacc ggcuugggca       8640

cggccugacu acaacccgcc gcucguggaa ucguggagga ggccagauua ccaaccgccc       8700

accguugcug guugugcucu cccccccccc aagaaggccc cgacgccucc cccaaggaga       8760

cgccggacag ugggucugag cgagagcacc auaucagaag cccuccagca acuggccauc       8820

aagaccuuug gccagccccc cucgagcggu gaugcaggcu cguccacggg ggcgggcgcc       8880

gccgaauccg gcgguccgac guccccuggu gagccggccc ccucagagac agguuccgcc       8940

uccucuaugc ccccccucga gggggagccu ggagauccgg accuggaguc ugaucaggua       9000

gagcuucaac cuccccccca gggggggggg guagcucccg guucgggcuc ggggucuugg       9060

ucuacuugcu ccgaggagga cgauaccacc gugugcugcu ccaugucaua cuccuggacc       9120

ggggcucuaa uaacucccug uagccccgaa gaggaaaagu ugccaaucaa cccuuugagu       9180

aacucgcugu ugcgauacca uaacaaggug uacuguacaa caucaaagag cgccucacag       9240

agggcuaaaa agguaacuuu ugacaggacg caagugcucg acgcccauua ugacucaguc       9300

uuaaaggaca ucaagcuagc ggcuuccaag gucagcgcaa ggcuccucac cuuggaggag       9360

gcgugccagu ugacuccacc ccauucugca agauccaagu auggauucgg ggccaaggag       9420

guccgcagcu uguccgggag ggccguuaac cacaucaagu ccguguggaa ggaccuccug       9480

gaagacccac aaacaccaau ucccacaacc aucauggcca aaaaugaggu guucugcgug       9540

gaccccgcca agggggguaa gaaaccagcu cgccucaucg uuuacccuga ccucggcguc       9600

cgggucugcg agaaaauggc ccucuaugac auuacacaaa agcuuccuca ggcgguaaug       9660

ggagcuuccu auggcuucca guacuccccu gcccaacggg uggaguaucu cuugaaagca       9720

ugggcggaaa agaaggaccc cauggguuuu ucguaugaua cccgaugcuu cgacucaacc       9780

gucacugaga gagacaucag gaccgaggag uccauauacc aggccugcuc ccugcccgag       9840

gaggcccgca cugccauaca cucgcugacu gagagacuuu acguaggagg gcccauguuc       9900

aacagcaagg gucaaaccug cgguuacaga cguugccgcg ccagcggggu gcuaaccacu       9960

agcaugggua acaccaucac augcuaugug aaagcccuag cggccugcaa ggcugcgggg       10020

auaguugcgc ccacaaugcu gguaugcggc aaugaccuag uagucaucuc agaaagccag       10080

gggacugagg aggacgagcg gaaccugaga gccuucacgg aggccaugac cagguacucu       10140

gccccuccug gugauccccc cagaccggaa uaugaccugg agcuaauaac auccuguucc       10200

ucaaaugugu cuguggcguu gggcccgcgg ggccgccgca gauacuaccu gaccagagac       10260

ccaaccacuc cacucgcccg ggcugccugg gaaacaguua gacacucccc uaucaauuca       10320

uggcugggaa acaucaucca guaugcucca accauauggg uucgcauggu ccuaaugaca       10380

cacuucuucu ccauucucau gguccaagac acccuggacc agaaccucaa cuuugagaug       10440

uauggaucag uauacuccgu gaauccuuug gaccuuccag ccauaauuga gagguuacac       10500

gggcuugacg ccuuuucuau gcacacauac ucucaccacg aacugacgcg gguggcuuca       10560

gcccucagaa aacuuggggc gccaccccuc agggugugga agagucgggc ucgcgcaguc       10620

agggcguccc ucaucucccg uggagggaaa gcggccguuu gcggccgaua ucucuucaau       10680

ugggcgguga agaccaagcu caaacucacu ccauugccgg aggcgcgccu acuggacuua       10740

uccaguuggu ucaccgucgg cgccggcggg ggcgacauuu uucacagcgu gucgcgcgcc       10800

cgaccccgcu cauuacucuu cggccuacuc cuacuuuucg uagggguagg ccucuuccua       10860

cuccccgcuc gguagagcgg cacacacuag guacacucca uagcuaacug uuccuuuuuu       10920

uuuuuuuuuu uuuuuuuuuu uuuuuuuuuu uuuuuuucuu uuuuuuuuuu uucccucuuu       10980

cuucccuucu caucuuauuc uacuuucuuu cuugguggcu ccaucuuagc ccuagucacg       11040

gcuagcugug aaagguccgu gagccgcaug acugcagaga gugccguaac uggucucucu       11100

gcagaucaug u                                                            11111

<210>15

<211>9707

<212>RNA

<213>人工序列

<220>

<223>人工序列的描述：含来自JFH-1克隆的全长丙型肝炎病毒基因组RNA的复制子RNA，其中氨基酸基序GDD已突变为GND

<400>15

gaauucuaau acgacucacu auagaccugc cccuaauagg ggcgacacuc cgccaugaau       60

cacuccccug ugaggaacua cugucuucac gcagaaagcg ccuagccaug gcguuaguau       120

gagugucgua cagccuccag gccccccccu cccgggagag ccauaguggu cugcggaacc       180

ggugaguaca ccggaauugc cgggaagacu ggguccuuuc uuggauaaac ccacucuaug       240

cccggccauu ugggcgugcc cccgcaagac ugcuagccga guagcguugg guugcgaaag       300

gccuuguggu acugccugau agggcgcuug cgagugcccc gggaggucuc guagaccgug       360

caccaugagc acaaauccu aaaccucaaag aaaaaccaaa agaaacacca accgucgccc       420

agaagacguu aaguucccgg gcggcggcca gaucguuggc ggaguauacu uguugccgcg       480

caggggcccc agguugggug ugcgcacgac aaggaaaacu ucggagcggu cccagccacg       540

ugggagacgc cagcccaucc ccaaagaucg gcgcuccacu ggcaaggccu ggggaaaacc       600

aggucgcccc uggccccuau augggaauga gggacucggc ugggcaggau ggcuccuguc       660

cccccgaggc ucucgccccu ccuggggccc cacugacccc cggcauaggu cgcgcaacgu       720

ggguaaaguc aucgacaccc uaacgugugg cuuugccgac cucauggggu acauccccgu       780

cguaggcgcc ccgcuuagug gcgccgccag agcugucgcg cacggcguga gaguccugga       840

ggacgggguu aauuaugcaa cagggaaccu acccgguuuc cccuuuucua ucuucuugcu       900

ggcccuguug uccugcauca ccguuccggu cucugcugcc caggugaaga auaccaguag       960

cagcuacaug gugaccaaug acugcuccaa ugacagcauc acuuggcagc ucgaggcugc       1020

gguucuccac guccccgggu gcgucccgug cgagagagug gggaauacgu cacgguguug       1080

ggugccaguc ucgccaaaca uggcugugcg gcagcccggu gcccucacgc agggucugcg       1140

gacgcacauc gauaugguug ugauguccgc caccuucugc ucugcucucu acguggggga       1200

ccucuguggc ggggugaugc ucgcggccca gguguucauc gucucgccgc aguaccacug       1260

guuugugcaa gaaugcaauu gcuccaucua cccuggcacc aucacuggac accgcauggc       1320

augggacaug augaugaacu ggucgcccac ggccaccaug auccuggcgu acgugaugcg       1380

cguccccgag gucaucauag acaucguuag cggggcucac uggggcguca uguucggcuu       1440

ggccuacuuc ucuaugcagg gagcgugggc gaaggucauu gucauccuuc ugcuggccgc       1500

ugggguggac gcgggcacca ccaccguugg aggcgcuguu gcacguucca ccaacgugau       1560

ugccggcgug uucagccaug gcccucagca gaacauucag cucauuaaca ccaacggcag       1620

uuggcacauc aaccguacug ccuugaauug caaugacucc uugaacaccg gcuuucucgc       1680

ggccuuguuc uacaccaacc gcuuuaacuc gucagggugu ccagggcgcc uguccgccug       1740

ccgcaacauc gaggcuuucc ggauagggug gggcacccua caguacgagg auaaugucac       1800

caauccagag gauaugaggc cguacugcug gcacuacccc ccaaagccgu guggcguagu       1860

ccccgcgagg ucugugugug gcccagugua cuguuucacc cccagcccgg uaguaguggg       1920

cacgaccgac agacguggag ugcccaccua cacaugggga gagaaugaga cagaugucuu       1980

ccuacugaac agcacccgac cgccgcaggg cucaugguuc ggcugcacgu ggaugaacuc       2040

cacugguuuc accaagacuu guggcgcgcc accuugccgc accagagcug acuucaacgc       2100

cagcacggac uuguugugcc cuacggauug uuuuaggaag cauccugaug ccacuuauau       2160

uaaguguggu ucugggcccu ggcucacacc aaagugccug guccacuacc cuuacagacu       2220

cuggcauuac cccugcacag ucaauuuuac caucuucaag auaagaaugu auguaggggg       2280

gguugagcac aggcucacgg ccgcaugcaa cuucacucgu ggggaucgcu gcgacuugga       2340

ggacagggac aggagucagc ugucuccucu guugcacucu accacggaau gggccauccu       2400

gcccugcacc uacucagacu uacccgcuuu gucaacuggu cuucuccacc uucaccagaa       2460

caucguggac guacaauaca uguauggccu cucaccugcu aucacaaaau acgucguucg       2520

augggagugg gugguacucu uauuccugcu cuuagcggac gccagagucu gcgccugcuu       2580

guggaugcuc aucuuguugg gccaggccga agcagcauug gagaaguugg ucgucuugca       2640

cgcugcgagu gcggcuaacu gccauggccu ccuauauuuu gccaucuucu ucguggcagc       2700

uuggcacauc aggggucggg ugguccccuu gaccaccuau ugccucacug gccuauggcc       2760

cuucugccua cugcucaugg cacugccccg gcaggcuuau gccuaugacg caccugugca       2820

cggacagaua ggcguggguu uguugauauu gaucacccuc uucacacuca ccccggggua       2880

uaagacccuc cucggccagu gucuguggug guugugcuau cuccugaccc ugggggaagc       2940

caugauucag gaguggguac cacccaugca ggugcgcggc ggccgcgaug gcaucgcgug       3000

ggccgucacu auauucugcc cggguguggu guuugacauu accaaauggc uuuuggcguu       3060

gcuugggccu gcuuaccucu uaagggccgc uuugacacau gugccguacu ucgucagagc       3120

ucacgcucug auaaggguau gcgcuuuggu gaagcagcuc gcggggggua gguauguuca       3180

gguggcgcua uuggcccuug gcagguggac uggcaccuac aucuaugacc accucacacc       3240

uaugucggac ugggccgcua gcggccugcg cgacuuagcg gucgccgugg aacccaucau       3300

cuucaguccg auggagaaga aggucaucgu cuggggagcg gagacggcug caugugggga       3360

cauucuacau ggacuucccg uguccgcccg acucggccag gagauccucc ucggcccagc       3420

ugauggcuac accuccaagg gguggaagcu ccuugcuccc aucacugcuu augcccagca       3480

aacacgaggc cuccugggcg ccauaguggu gaguaugacg gggcgugaca ggacagaaca       3540

ggccggggaa guccaaaucc uguccacagu cucucagucc uuccucggaa caaccaucuc       3600

ggggguuuug uggacuguuu accacggagc uggcaacaag acucuagccg gcuuacgggg       3660

uccggucacg cagauguacu cgagugcuga gggggacuug guaggcuggc ccagcccccc       3720

ugggaccaag ucuuuggagc cgugcaagug uggagccguc gaccuauauc uggucacgcg       3780

gaacgcugau gucaucccgg cucggagacg cggggacaag cggggagcau ugcucucccc       3840

gagacccauu ucgaccuuga agggguccuc gggggggccg gugcucugcc cuaggggcca       3900

cgucguuggg cucuuccgag cagcugugug cucucggggc guggccaaau ccaucgauuu       3960

cauccccguu gagacacucg acguuguuac aaggucuccc acuuucagug acaacagcac       4020

gccaccggcu gugccccaga ccuaucaggu cggguacuug caugcuccaa cuggcagugg       4080

aaagagcacc aaggucccug ucgcguaugc cgcccagggg uacaaaguac uagugcuuaa       4140

ccccucggua gcugccaccc ugggguuugg ggcguaccua uccaaggcac auggcaucaa       4200

ucccaacauu aggacuggag ucaggaccgu gaugaccggg gaggccauca cguacuccac       4260

auauggcaaa uuucucgccg augggggcug cgcuagcggc gccuaugaca ucaucauaug       4320

cgaugaaugc cacgcugugg augcuaccuc cauucucggc aucggaacgg uccuugauca       4380

agcagagaca gccgggguca gacuaacugu gcuggcuacg gccacacccc ccgggucagu       4440

gacaaccccc caucccgaua uagaagaggu aggccucggg cgggagggug agauccccuu       4500

cuaugggagg gcgauucccc uauccugcau caagggaggg agacaccuga uuuucugcca       4560

cucaaagaaa aagugugacg agcucgcggc ggcccuucgg ggcaugggcu ugaaugccgu       4620

ggcauacuau agaggguugg acgucuccau aauaccagcu cagggagaug ugguggucgu       4680

cgccaccgac gcccucauga cgggguacac uggagacuuu gacuccguga ucgacugcaa       4740

uguagcgguc acccaagcug ucgacuucag ccuggacccc accuucacua uaaccacaca       4800

gacuguccca caagacgcug ucucacgcag ucagcgccgc gggcgcacag guagaggaag       4860

acagggcacu uauagguaug uuuccacugg ugaacgagcc ucaggaaugu uugacagugu       4920

agugcuuugu gagugcuacg acgcaggggc ugcgugguac gaucucacac cagcggagac       4980

caccgucagg cuuagagcgu auuucaacac gcccggccua cccguguguc aagaccaucu       5040

ugaauuuugg gaggcaguuu ucaccggccu cacacacaua gacgcccacu uccucuccca       5100

aacaaagcaa gcgggggaga acuucgcgua ccuaguagcc uaccaagcua cggugugcgc       5160

cagagccaag gccccucccc cguccuggga cgccaugugg aagugccugg cccgacucaa       5220

gccuacgcuu gcgggcccca caccucuccu guaccguuug ggcccuauua ccaaugaggu       5280

cacccucaca cacccuggga cgaaguacau cgccacaugc augcaagcug accuugaggu       5340

caugaccagc acgugggucc uagcuggagg aguccuggca gccgucgccg cauauugccu       5400

ggcgacugga ugcguuucca ucaucggccg cuugcacguc aaccagcgag ucgucguugc       5460

gccggauaag gagguccugu augaggcuuu ugaugagaug gaggaaugcg ccucuagggc       5520

ggcucucauc gaagaggggc agcggauagc cgagauguug aaguccaaga uccaaggcuu       5580

gcugcagcag gccucuaagc aggcccagga cauacaaccc gcuaugcagg cuucauggcc       5640

caaaguggaa caauuuuggg ccagacacau guggaacuuc auuagcggca uccaauaccu       5700

cgcaggauug ucaacacugc cagggaaccc cgcgguggcu uccaugaugg cauucagugc       5760

cgcccucacc aguccguugu cgaccaguac caccauccuu cucaacauca ugggaggcug       5820

guuagcgucc cagaucgcac cacccgcggg ggccaccggc uuugucguca guggccuggu       5880

gggggcugcc gugggcagca uaggccuggg uaaggugcug guggacaucc uggcaggaua       5940

uggugcgggc auuucggggg cccucgucgc auucaagauc augucuggcg agaagcccuc       6000

uauggaagau gucaucaauc uacugccugg gauccugucu ccgggagccc uggugguggg       6060

ggucaucugc gcggccauuc ugcgccgcca cgugggaccg ggggagggcg cgguccaaug       6120

gaugaacagg cuuauugccu uugcuuccag aggaaaccac gucgccccua cucacuacgu       6180

gacggagucg gaugcgucgc agcgugugac ccaacuacuu ggcucucuua cuauaaccag       6240

ccuacucaga agacuccaca auuggauaac ugaggacugc cccaucccau gcuccggauc       6300

cuggcuccgc gacguguggg acuggguuug caccaucuug acagacuuca aaaauuggcu       6360

gaccucuaaa uuguucccca agcugcccgg ccuccccuuc aucucuuguc aaaaggggua       6420

caagggugug ugggccggca cuggcaucau gaccacgcgc ugcccuugcg gcgccaacau       6480

cucuggcaau guccgccugg gcucuaugag gaucacaggg ccuaaaaccu gcaugaacac       6540

cuggcagggg accuuuccua ucaauugcua cacggagggc cagugcgcgc cgaaaccccc       6600

cacgaacuac aagaccgcca ucuggagggu ggcggccucg gaguacgcgg aggugacgca       6660

gcaugggucg uacuccuaug uaacaggacu gaccacugac aaucugaaaa uuccuugcca       6720

acuaccuucu ccagaguuuu ucuccugggu ggacggugug cagauccaua gguuugcacc       6780

cacaccaaag ccguuuuucc gggaugaggu cucguucugc guugggcuua auuccuaugc       6840

ugucgggucc cagcuucccu gugaaccuga gcccgacgca gacguauuga gguccaugcu       6900

aacagauccg ccccacauca cggcggagac ugcggcgcgg cgcuuggcac ggggaucacc       6960

uccaucugag gcgagcuccu cagugagcca gcuaucagca ccgucgcugc gggccaccug       7020

caccacccac agcaacaccu augacgugga cauggucgau gccaaccugc ucauggaggg       7080

cgguguggcu cagacagagc cugaguccag ggugcccguu cuggacuuuc ucgagccaau       7140

ggccgaggaa gagagcgacc uugagcccuc aauaccaucg gagugcaugc uccccaggag       7200

cggguuucca cgggccuuac cggcuugggc acggccugac uacaacccgc cgcucgugga       7260

aucguggagg aggccagauu accaaccgcc caccguugcu gguugugcuc uccccccccc       7320

caagaaggcc ccgacgccuc ccccaaggag acgccggaca gugggucuga gcgagagcac       7380

cauaucagaa gcccuccagc aacuggccau caagaccuuu ggccagcccc ccucgagcgg       7440

ugaugcaggc ucguccacgg gggcgggcgc cgccgaaucc ggcgguccga cguccccugg       7500

ugagccggcc cccucagaga cagguuccgc cuccucuaug cccccccucg agggggagcc       7560

uggagauccg gaccuggagu cugaucaggu agagcuucaa ccuccccccc aggggggggg       7620

gguagcuccc gguucgggcu cggggucuug gucuacuugc uccgaggagg acgauaccac       7680

cgugugcugc uccaugucau acuccuggac cggggcucua auaacucccu guagccccga       7740

agaggaaaag uugccaauca acccuuugag uaacucgcug uugcgauacc auaacaaggu       7800

guacuguaca acaucaaaga gcgccucaca gagggcuaaa aagguaacuu uugacaggac       7860

gcaagugcuc gacgcccauu augacucagu cuuaaaggac aucaagcuag cggcuuccaa       7920

ggucagcgca aggcuccuca ccuuggagga ggcgugccag uugacuccac cccauucugc       7980

aagauccaag uauggauucg gggccaagga gguccgcagc uuguccggga gggccguuaa       8040

ccacaucaag uccgugugga aggaccuccu ggaagaccca caaacaccaa uucccacaac       8100

caucauggcc aaaaaugagg uguucugcgu ggaccccgcc aaggggggua agaaaccagc       8160

ucgccucauc guuuacccug accucggcgu ccgggucugc gagaaaaugg cccucuauga       8220

cauuacacaa aagcuuccuc aggcgguaau gggagcuucc uauggcuucc aguacucccc       8280

ugcccaacgg guggaguauc ucuugaaagc augggcggaa aagaaggacc ccauggguuu       8340

uucguaugau acccgaugcu ucgacucaac cgucacugag agagacauca ggaccgagga       8400

guccauauac caggccugcu cccugcccga ggaggcccgc acugccauac acucgcugac       8460

ugagagacuu uacguaggag ggcccauguu caacagcaag ggucaaaccu gcgguuacag       8520

acguugccgc gccagcgggg ugcuaaccac uagcaugggu aacaccauca caugcuaugu       8580

gaaagcccua gcggccugca aggcugcggg gauaguugcg cccacaaugc ugguaugcgg       8640

caaugaccua guagucaucu cagaaagcca ggggacugag gaggacgagc ggaaccugag       8700

agccuucacg gaggccauga ccagguacuc ugccccuccu ggugaucccc ccagaccgga       8760

auaugaccug gagcuaauaa cauccuguuc cucaaaugug ucuguggcgu ugggcccgcg       8820

gggccgccgc agauacuacc ugaccagaga cccaaccacu ccacucgccc gggcugccug       8880

ggaaacaguu agacacuccc cuaucaauuc auggcuggga aacaucaucc aguaugcucc       8940

aaccauaugg guucgcaugg uccuaaugac acacuucuuc uccauucuca ugguccaaga       9000

cacccuggac cagaaccuca acuuugagau guauggauca guauacuccg ugaauccuuu       9060

ggaccuucca gccauaauug agagguuaca cgggcuugac gccuuuucua ugcacacaua       9120

cucucaccac gaacugacgc ggguggcuuc agcccucaga aaacuugggg cgccaccccu       9180

cagggugugg aagagucggg cucgcgcagu cagggcgucc cucaucuccc guggagggaa       9240

agcggccguu ugcggccgau aucucuucaa uugggcggug aagaccaagc ucaaacucac       9300

uccauugccg gaggcgcgcc uacuggacuu auccaguugg uucaccgucg gcgccggcgg       9360

gggcgacauu uuucacagcg ugucgcgcgc ccgaccccgc ucauuacucu ucggccuacu       9420

ccuacuuuuc guagggguag gccucuuccu acuccccgcu cgguagagcg gcacacacua       9480

gguacacucc auagcuaacu guuccuuuuu uuuuuuuuuu uuuuuuuuuu uuuuuuuuuu       9540

uuuuuuuucu uuuuuuuuuu uuucccucuu ucuucccuuc ucaucuuauu cuacuuucuu       9600

ucuugguggc uccaucuuag cccuagucac ggcuagcugu gaaagguccg ugagccgcau       9660

gacugcagag agugccguaa cuggucucuc ugcagaucau gucuaga                     9707

<210>16

<211>17

<212>DNA

<213>人工序列

<220>

<223>人工序列的描述：引物

<400>16

cgggagagcc atagtgg                                                      17

<210>17

<211>19

<212>DNA

<213>人工序列

<220>

<223>人工序列的描述：引物

<400>17

agtaccacaa ggcctttcg                                                    19

<210>18

<211>21

<212>DNA

<213>人工序列

<220>

<223>人工序列的描述：引物

<400>18

ctgcggaacc ggtgagtaca c                                                 21

<210>19

<211>20

<212>DNA

<213>人工序列

<220>

<223>人工序列的描述：引物

<400>19

aacaagatgg attgcacgca                                                   20

<210>20

<211>20

<212>DNA

<213>人工序列

<220>

<223>人工序列的描述：引物

<400>20

cgtcaagaag gcgatagaag                                                   20

<210>21

<211>11969

<212>RNA

<213>人工序列

<220>

<223>人工序列的描述：来源于表达载体rFGR-JFH1/Luc的复制子RNA

<400>21

accugccccu aauaggggcg acacuccgcc augaaucacu ccccugugag gaacuacugu       60

cuucacgcag aaagcgccua gccauggcgu uaguaugagu gucguacagc cuccaggccc       120

cccccucccg ggagagccau aguggucugc ggaaccggug aguacaccgg aauugccggg       180

aagacugggu ccuuucuugg auaaacccac ucuaugcccg gccauuuggg cgugcccccg       240

caagacugcu agccgaguag cguuggguug cgaaaggccu ugugguacug ccugauaggg       300

cgcuugcgag ugccccggga ggucucguag accgugcacc augagcacaa auccuaaacc       360

ucaaagaaaa accaaaagaa acaccaaccg acgcguaaug gaagacgcca aaaacauaaa       420

gaaaggcccg gcgccauucu auccucugga ggauggaacc gcuggagagc aacugcauaa       480

ggcuaugaag agauacgccc ugguuccugg aacaauugcu uuuacagaug cacauaucga       540

ggugaacauc acguacgcgg aauacuucga aauguccguu cgguuggcag aagcuaugaa       600

acgauauggg cugaauacaa aucacagaau cgucguaugc agugaaaacu cucuucaauu       660

cuuuaugccg guguugggcg cguuauuuau cggaguugca guugcgcccg cgaacgacau       720

uuauaaugaa cgugaauugc ucaacaguau gaacauuucg cagccuaccg uaguguuugu       780

uuccaaaaag ggguugcaaa aaauuuugaa cgugcaaaaa aaauuaccaa uaauccagaa       840

aauuauuauc auggauucua aaacggauua ccagggauuu cagucgaugu acacguucgu       900

cacaucucau cuaccucccg guuuuaauga auacgauuuu guaccagagu ccuuugaucg       960

ugacaaaaca auugcacuga uaaugaacuc cucuggaucu acuggguuac cuaagggugu       1020

ggcccuuccg cauagaacug ccugcgucag auucucgcau gccagagauc cuauuuuugg       1080

caaucaaauc auuccggaua cugcgauuuu aaguguuguu ccauuccauc acgguuuugg       1140

aauguuuacu acacucggau auuugauaug uggauuucga gucgucuuaa uguauagauu       1200

ugaagaagag cuguuuuuac gaucccuuca ggauuacaaa auucaaagug cguugcuagu       1260

accaacccua uuuucauucu ucgccaaaag cacucugauu gacaaauacg auuuaucuaa       1320

uuuacacgaa auugcuucug ggggcgcacc ucuuucgaaa gaagucgggg aagcgguugc       1380

aaaacgcuuc caucuuccag ggauacgaca aggauauggg cucacugaga cuacaucagc       1440

uauucugauu acacccgagg gggaugauaa accgggcgcg gucgguaaag uuguuccauu       1500

uuuugaagcg aagguugugg aucuggauac cgggaaaacg cugggcguua aucagagagg       1560

cgaauuaugu gucagaggac cuaugauuau guccgguuau guaaacaauc cggaagcgac       1620

caacgccuug auugacaagg auggauggcu acauucugga gacauagcuu acugggacga       1680

agacgaacac uucuucauag uugaccgcuu gaagucuuua auuaaauaca aaggauauca       1740

gguggccccc gcugaauugg aaucgauauu guuacaacac cccaacaucu ucgacgcggg       1800

cguggcaggu cuucccgacg augacgccgg ugaacuuccc gccgccguug uuguuuugga       1860

gcacggaaag acgaugacgg aaaaagagau cguggauuac gucgccaguc aaguaacaac       1920

cgcgaaaaag uugcgcggag gaguuguguu uguggacgaa guaccgaaag gucuuaccgg       1980

aaaacucgac gcaagaaaaa ucagagagau ccucauaaag gccaagaagg gcggaaaguc       2040

caaauuguaa guuuaaaccc ucucccuccc cccccccuaa cguuacuggc cgaagccgcu       2100

uggaauaagg ccggugugcg uuugucuaua uguuauuuuc caccauauug ccgucuuuug       2160

gcaaugugag ggcccggaaa ccuggcccug ucuucuugac gagcauuccu aggggucuuu       2220

ccccucucgc caaaggaaug caaggucugu ugaaugucgu gaaggaagca guuccucugg       2280

aagcuucuug aagacaaaca acgucuguag cgacccuuug caggcagcgg aaccccccac       2340

cuggcgacag gugccucugc ggccaaaagc cacguguaua agauacaccu gcaaaggcgg       2400

cacaacccca gugccacguu gugaguugga uaguugugga aagagucaaa uggcucuccu       2460

caagcguauu caacaagggg cugaaggaug cccagaaggu accccauugu augggaucug       2520

aucuggggcc ucggugcaca ugcuuuacau guguuuaguc gagguuaaaa aaacgucuag       2580

gccccccgaa ccacggggac gugguuuucc uuugaaaaac acgaugauac caugagcaca       2640

aauccuaaac cucaaagaaa aaccaaaaga aacaccaacc gucgcccaga agacguuaag       2700

uucccgggcg gcggccagau cguuggcgga guauacuugu ugccgcgcag gggccccagg       2760

uugggugugc gcacgacaag gaaaacuucg gagcgguccc agccacgugg gagacgccag       2820

cccaucccca aagaucggcg cuccacuggc aaggccuggg gaaaaccagg ucgccccugg       2880

ccccuauaug ggaaugaggg acucggcugg gcaggauggc uccugucccc ccgaggcucu       2940

cgccccuccu ggggccccac ugacccccgg cauaggucgc gcaacguggg uaaagucauc       3000

gacacccuaa cguguggcuu ugccgaccuc augggguaca uccccgucgu aggcgccccg       3060

cuuaguggcg ccgccagagc ugucgcgcac ggcgugagag uccuggagga cgggguuaau       3120

uaugcaacag ggaaccuacc cgguuucccc uuuucuaucu ucuugcuggc ccuguugucc       3180

ugcaucaccg uuccggucuc ugcugcccag gugaagaaua ccaguagcag cuacauggug       3240

accaaugacu gcuccaauga cagcaucacu uggcagcucg aggcugcggu ucuccacguc       3300

cccgggugcg ucccgugcga gagagugggg aauacgucac gguguugggu gccagucucg       3360

ccaaacaugg cugugcggca gcccggugcc cucacgcagg gucugcggac gcacaucgau       3420

augguuguga uguccgccac cuucugcucu gcucucuacg ugggggaccu cuguggcggg       3480

gugaugcucg cggcccaggu guucaucguc ucgccgcagu accacugguu ugugcaagaa       3540

ugcaauugcu ccaucuaccc uggcaccauc acuggacacc gcauggcaug ggacaugaug       3600

augaacuggu cgcccacggc caccaugauc cuggcguacg ugaugcgcgu ccccgagguc       3660

aucauagaca ucguuagcgg ggcucacugg ggcgucaugu ucggcuuggc cuacuucucu       3720

augcagggag cgugggcgaa ggucauuguc auccuucugc uggccgcugg gguggacgcg       3780

ggcaccacca ccguuggagg cgcuguugca cguuccacca acgugauugc cggcguguuc       3840

agccauggcc cucagcagaa cauucagcuc auuaacacca acggcaguug gcacaucaac       3900

cguacugccu ugaauugcaa ugacuccuug aacaccggcu uucucgcggc cuuguucuac       3960

accaaccgcu uuaacucguc agggugucca gggcgccugu ccgccugccg caacaucgag       4020

gcuuuccgga uagggugggg cacccuacag uacgaggaua augucaccaa uccagaggau       4080

augaggccgu acugcuggca cuacccccca aagccgugug gcguaguccc cgcgaggucu       4140

guguguggcc caguguacug uuucaccccc agcccgguag uagugggcac gaccgacaga       4200

cguggagugc ccaccuacac auggggagag aaugagacag augucuuccu acugaacagc       4260

acccgaccgc cgcagggcuc augguucggc ugcacgugga ugaacuccac ugguuucacc       4320

aagacuugug gcgcgccacc uugccgcacc agagcugacu ucaacgccag cacggacuug       4380

uugugcccua cggauuguuu uaggaagcau ccugaugcca cuuauauuaa gugugguucu       4440

gggcccuggc ucacaccaaa gugccugguc cacuacccuu acagacucug gcauuacccc       4500

ugcacaguca auuuuaccau cuucaagaua agaauguaug uagggggggu ugagcacagg       4560

cucacggccg caugcaacuu cacucguggg gaucgcugcg acuuggagga cagggacagg       4620

agucagcugu cuccucuguu gcacucuacc acggaauggg ccauccugcc cugcaccuac       4680

ucagacuuac ccgcuuuguc aacuggucuu cuccaccuuc accagaacau cguggacgua       4740

caauacaugu auggccucuc accugcuauc acaaaauacg ucguucgaug ggagugggug       4800

guacucuuau uccugcucuu agcggacgcc agagucugcg ccugcuugug gaugcucauc       4860

uuguugggcc aggccgaagc agcauuggag aaguuggucg ucuugcacgc ugcgagugcg       4920

gcuaacugcc auggccuccu auauuuugcc aucuucuucg uggcagcuug gcacaucagg       4980

ggucgggugg uccccuugac caccuauugc cucacuggcc uauggcccuu cugccuacug       5040

cucauggcac ugccccggca ggcuuaugcc uaugacgcac cugugcacgg acagauaggc       5100

guggguuugu ugauauugau cacccucuuc acacucaccc cgggguauaa gacccuccuc       5160

ggccaguguc uguggugguu gugcuaucuc cugacccugg gggaagccau gauucaggag       5220

uggguaccac ccaugcaggu gcgcggcggc cgcgauggca ucgcgugggc cgucacuaua       5280

uucugcccgg gugugguguu ugacauuacc aaauggcuuu uggcguugcu ugggccugcu       5340

uaccucuuaa gggccgcuuu gacacaugug ccguacuucg ucagagcuca cgcucugaua       5400

aggguaugcg cuuuggugaa gcagcucgcg ggggguaggu auguucaggu ggcgcuauug       5460

gcccuuggca gguggacugg caccuacauc uaugaccacc ucacaccuau gucggacugg       5520

gccgcuagcg gccugcgcga cuuagcgguc gccguggaac ccaucaucuu caguccgaug       5580

gagaagaagg ucaucgucug gggagcggag acggcugcau guggggacau ucuacaugga       5640

cuucccgugu ccgcccgacu cggccaggag auccuccucg gcccagcuga uggcuacacc       5700

uccaaggggu ggaagcuccu ugcucccauc acugcuuaug cccagcaaac acgaggccuc       5760

cugggcgcca uaguggugag uaugacgggg cgugacagga cagaacaggc cggggaaguc       5820

caaauccugu ccacagucuc ucaguccuuc cucggaacaa ccaucucggg gguuuugugg       5880

acuguuuacc acggagcugg caacaagacu cuagccggcu uacggggucc ggucacgcag       5940

auguacucga gugcugaggg ggacuuggua ggcuggccca gccccccugg gaccaagucu       6000

uuggagccgu gcaagugugg agccgucgac cuauaucugg ucacgcggaa cgcugauguc       6060

aucccggcuc ggagacgcgg ggacaagcgg ggagcauugc ucuccccgag acccauuucg       6120

accuugaagg gguccucggg ggggccggug cucugcccua ggggccacgu cguugggcuc       6180

uuccgagcag cugugugcuc ucggggcgug gccaaaucca ucgauuucau ccccguugag       6240

acacucgacg uuguuacaag gucucccacu uucagugaca acagcacgcc accggcugug       6300

ccccagaccu aucaggucgg guacuugcau gcuccaacug gcaguggaaa gagcaccaag       6360

gucccugucg cguaugccgc ccagggguac aaaguacuag ugcuuaaccc cucgguagcu       6420

gccacccugg gguuuggggc guaccuaucc aaggcacaug gcaucaaucc caacauuagg       6480

acuggaguca ggaccgugau gaccggggag gccaucacgu acuccacaua uggcaaauuu       6540

cucgccgaug ggggcugcgc uagcggcgcc uaugacauca ucauaugcga ugaaugccac       6600

gcuguggaug cuaccuccau ucucggcauc ggaacggucc uugaucaagc agagacagcc       6660

ggggucagac uaacugugcu ggcuacggcc acaccccccg ggucagugac aaccccccau       6720

cccgauauag aagagguagg ccucgggcgg gagggugaga uccccuucua ugggagggcg       6780

auuccccuau ccugcaucaa gggagggaga caccugauuu ucugccacuc aaagaaaaag       6840

ugugacgagc ucgcggcggc ccuucggggc augggcuuga augccguggc auacuauaga       6900

ggguuggacg ucuccauaau accagcucag ggagaugugg uggucgucgc caccgacgcc       6960

cucaugacgg gguacacugg agacuuugac uccgugaucg acugcaaugu agcggucacc       7020

caagcugucg acuucagccu ggaccccacc uucacuauaa ccacacagac ugucccacaa       7080

gacgcugucu cacgcaguca gcgccgcggg cgcacaggua gaggaagaca gggcacuuau       7140

agguauguuu ccacugguga acgagccuca ggaauguuug acaguguagu gcuuugugag       7200

ugcuacgacg caggggcugc gugguacgau cucacaccag cggagaccac cgucaggcuu       7260

agagcguauu ucaacacgcc cggccuaccc gugugucaag accaucuuga auuuugggag       7320

gcaguuuuca ccggccucac acacauagac gcccacuucc ucucccaaac aaagcaagcg       7380

ggggagaacu ucgcguaccu aguagccuac caagcuacgg ugugcgccag agccaaggcc       7440

ccucccccgu ccugggacgc cauguggaag ugccuggccc gacucaagcc uacgcuugcg       7500

ggccccacac cucuccugua ccguuugggc ccuauuacca augaggucac ccucacacac       7560

ccugggacga aguacaucgc cacaugcaug caagcugacc uugaggucau gaccagcacg       7620

uggguccuag cuggaggagu ccuggcagcc gucgccgcau auugccuggc gacuggaugc       7680

guuuccauca ucggccgcuu gcacgucaac cagcgagucg ucguugcgcc ggauaaggag       7740

guccuguaug aggcuuuuga ugagauggag gaaugcgccu cuagggcggc ucucaucgaa       7800

gaggggcagc ggauagccga gauguugaag uccaagaucc aaggcuugcu gcagcaggcc       7860

ucuaagcagg cccaggacau acaacccgcu augcaggcuu cauggcccaa aguggaacaa       7920

uuuugggcca gacacaugug gaacuucauu agcggcaucc aauaccucgc aggauuguca       7980

acacugccag ggaaccccgc gguggcuucc augauggcau ucagugccgc ccucaccagu       8040

ccguugucga ccaguaccac cauccuucuc aacaucaugg gaggcugguu agcgucccag       8100

aucgcaccac ccgcgggggc caccggcuuu gucgucagug gccugguggg ggcugccgug       8160

ggcagcauag gccuggguaa ggugcuggug gacauccugg caggauaugg ugcgggcauu       8220

ucgggggccc ucgucgcauu caagaucaug ucuggcgaga agcccucuau ggaagauguc       8280

aucaaucuac ugccugggau ccugucuccg ggagcccugg uggugggggu caucugcgcg       8340

gccauucugc gccgccacgu gggaccgggg gagggcgcgg uccaauggau gaacaggcuu       8400

auugccuuug cuuccagagg aaaccacguc gccccuacuc acuacgugac ggagucggau       8460

gcgucgcagc gugugaccca acuacuuggc ucucuuacua uaaccagccu acucagaaga       8520

cuccacaauu ggauaacuga ggacugcccc aucccaugcu ccggauccug gcuccgcgac       8580

gugugggacu ggguuugcac caucuugaca gacuucaaaa auuggcugac cucuaaauug       8640

uuccccaagc ugcccggccu ccccuucauc ucuugucaaa agggguacaa gggugugugg       8700

gccggcacug gcaucaugac cacgcgcugc ccuugcggcg ccaacaucuc uggcaauguc       8760

cgccugggcu cuaugaggau cacagggccu aaaaccugca ugaacaccug gcaggggacc       8820

uuuccuauca auugcuacac ggagggccag ugcgcgccga aaccccccac gaacuacaag       8880

accgccaucu ggaggguggc ggccucggag uacgcggagg ugacgcagca ugggucguac       8940

uccuauguaa caggacugac cacugacaau cugaaaauuc cuugccaacu accuucucca       9000

gaguuuuucu ccugggugga cggugugcag auccauaggu uugcacccac accaaagccg       9060

uuuuuccggg augaggucuc guucugcguu gggcuuaauu ccuaugcugu cgggucccag       9120

cuucccugug aaccugagcc cgacgcagac guauugaggu ccaugcuaac agauccgccc       9180

cacaucacgg cggagacugc ggcgcggcgc uuggcacggg gaucaccucc aucugaggcg       9240

agcuccucag ugagccagcu aucagcaccg ucgcugcggg ccaccugcac cacccacagc       9300

aacaccuaug acguggacau ggucgaugcc aaccugcuca uggagggcgg uguggcucag       9360

acagagccug aguccagggu gcccguucug gacuuucucg agccaauggc cgaggaagag       9420

agcgaccuug agcccucaau accaucggag ugcaugcucc ccaggagcgg guuuccacgg       9480

gccuuaccgg cuugggcacg gccugacuac aacccgccgc ucguggaauc guggaggagg       9540

ccagauuacc aaccgcccac cguugcuggu ugugcucucc ccccccccaa gaaggccccg       9600

acgccucccc caaggagacg ccggacagug ggucugagcg agagcaccau aucagaagcc       9660

cuccagcaac uggccaucaa gaccuuuggc cagccccccu cgagcgguga ugcaggcucg       9720

uccacggggg cgggcgccgc cgaauccggc gguccgacgu ccccugguga gccggccccc       9780

ucagagacag guuccgccuc cucuaugccc ccccucgagg gggagccugg agauccggac       9840

cuggagucug aucagguaga gcuucaaccu cccccccagg gggggggggu agcucccggu       9900

ucgggcucgg ggucuugguc uacuugcucc gaggaggacg auaccaccgu gugcugcucc       9960

augucauacu ccuggaccgg ggcucuaaua acucccugua gccccgaaga ggaaaaguug       10020

ccaaucaacc cuuugaguaa cucgcuguug cgauaccaua acaaggugua cuguacaaca       10080

ucaaagagcg ccucacagag ggcuaaaaag guaacuuuug acaggacgca agugcucgac       10140

gcccauuaug acucagucuu aaaggacauc aagcuagcgg cuuccaaggu cagcgcaagg       10200

cuccucaccu uggaggaggc gugccaguug acuccacccc auucugcaag auccaaguau       10260

ggauucgggg ccaaggaggu ccgcagcuug uccgggaggg ccguuaacca caucaagucc       10320

guguggaagg accuccugga agacccacaa acaccaauuc ccacaaccau cauggccaaa       10380

aaugaggugu ucugcgugga ccccgccaag ggggguaaga aaccagcucg ccucaucguu       10440

uacccugacc ucggcguccg ggucugcgag aaaauggccc ucuaugacau uacacaaaag       10500

cuuccucagg cgguaauggg agcuuccuau ggcuuccagu acuccccugc ccaacgggug       10560

gaguaucucu ugaaagcaug ggcggaaaag aaggacccca uggguuuuuc guaugauacc       10620

cgaugcuucg acucaaccgu cacugagaga gacaucagga ccgaggaguc cauauaccag       10680

gccugcuccc ugcccgagga ggcccgcacu gccauacacu cgcugacuga gagacuuuac       10740

guaggagggc ccauguucaa cagcaagggu caaaccugcg guuacagacg uugccgcgcc       10800

agcggggugc uaaccacuag cauggguaac accaucacau gcuaugugaa agcccuagcg       10860

gccugcaagg cugcggggau aguugcgccc acaaugcugg uaugcggcga ugaccuagua       10920

gucaucucag aaagccaggg gacugaggag gacgagcgga accugagagc cuucacggag       10980

gccaugacca gguacucugc cccuccuggu gaucccccca gaccggaaua ugaccuggag       11040

cuaauaacau ccuguuccuc aaaugugucu guggcguugg gcccgcgggg ccgccgcaga       11100

uacuaccuga ccagagaccc aaccacucca cucgcccggg cugccuggga aacaguuaga       11160

cacuccccua ucaauucaug gcugggaaac aucauccagu augcuccaac cauauggguu       11220

cgcauggucc uaaugacaca cuucuucucc auucucaugg uccaagacac ccuggaccag       11280

aaccucaacu uugagaugua uggaucagua uacuccguga auccuuugga ccuuccagcc       11340

auaauugaga gguuacacgg gcuugacgcc uuuucuaugc acacauacuc ucaccacgaa       11400

cugacgcggg uggcuucagc ccucagaaaa cuuggggcgc caccccucag gguguggaag       11460

agucgggcuc gcgcagucag ggcgucccuc aucucccgug gagggaaagc ggccguuugc       11520

ggccgauauc ucuucaauug ggcggugaag accaagcuca aacucacucc auugccggag       11580

gcgcgccuac uggacuuauc caguugguuc accgucggcg ccggcggggg cgacauuuuu       11640

cacagcgugu cgcgcgcccg accccgcuca uuacucuucg gccuacuccu acuuuucgua       11700

gggguaggcc ucuuccuacu ccccgcucgg uagagcggca cacacuaggu acacuccaua       11760

gcuaacuguu ccuuuuuuuu uuuuuuuuuu uuuuuuuuuu uuuuuuuuuu uuuuucuuuu       11820

uuuuuuuuuu cccucuuucu ucccuucuca ucuuauucua cuuucuuucu ugguggcucc       11880

aucuuagccc uagucacggc uagcugugaa agguccguga gccgcaugac ugcagagagu       11940

gccguaacug gucucucugc agaucaugu                                         11969

<210>22

<211>11969

<212>RNA

<213>人工序列

<220>

<223>人工序列的描述：来源于表达载体rFGR-JFH1/Luc/GND的复制子RNA

<400>22

accugccccu aauaggggcg acacuccgcc augaaucacu ccccugugag gaacuacugu       60

cuucacgcag aaagcgccua gccauggcgu uaguaugagu gucguacagc cuccaggccc       120

cccccucccg ggagagccau aguggucugc ggaaccggug aguacaccgg aauugccggg       180

aagacugggu ccuuucuugg auaaacccac ucuaugcccg gccauuuggg cgugcccccg       240

caagacugcu agccgaguag cguuggguug cgaaaggccu ugugguacug ccugauaggg       300

cgcuugcgag ugccccggga ggucucguag accgugcacc augagcacaa auccuaaacc       360

ucaaagaaaa accaaaagaa acaccaaccg acgcguaaug gaagacgcca aaaacauaaa       420

gaaaggcccg gcgccauucu auccucugga ggauggaacc gcuggagagc aacugcauaa       480

ggcuaugaag agauacgccc ugguuccugg aacaauugcu uuuacagaug cacauaucga       540

ggugaacauc acguacgcgg aauacuucga aauguccguu cgguuggcag aagcuaugaa       600

acgauauggg cugaauacaa aucacagaau cgucguaugc agugaaaacu cucuucaauu       660

cuuuaugccg guguugggcg cguuauuuau cggaguugca guugcgcccg cgaacgacau       720

uuauaaugaa cgugaauugc ucaacaguau gaacauuucg cagccuaccg uaguguuugu       780

uuccaaaaag ggguugcaaa aaauuuugaa cgugcaaaaa aaauuaccaa uaauccagaa       840

aauuauuauc auggauucua aaacggauua ccagggauuu cagucgaugu acacguucgu       900

cacaucucau cuaccucccg guuuuaauga auacgauuuu guaccagagu ccuuugaucg       960

ugacaaaaca auugcacuga uaaugaacuc cucuggaucu acuggguuac cuaagggugu       1020

ggcccuuccg cauagaacug ccugcgucag auucucgcau gccagagauc cuauuuuugg       1080

caaucaaauc auuccggaua cugcgauuuu aaguguuguu ccauuccauc acgguuuugg       1140

aauguuuacu acacucggau auuugauaug uggauuucga gucgucuuaa uguauagauu       1200

ugaagaagag cuguuuuuac gaucccuuca ggauuacaaa auucaaagug cguugcuagu       1260

accaacccua uuuucauucu ucgccaaaag cacucugauu gacaaauacg auuuaucuaa       1320

uuuacacgaa auugcuucug ggggcgcacc ucuuucgaaa gaagucgggg aagcgguugc       1380

aaaacgcuuc caucuuccag ggauacgaca aggauauggg cucacugaga cuacaucagc       1440

uauucugauu acacccgagg gggaugauaa accgggcgcg gucgguaaag uuguuccauu       1500

uuuugaagcg aagguugugg aucuggauac cgggaaaacg cugggcguua aucagagagg       1560

cgaauuaugu gucagaggac cuaugauuau guccgguuau guaaacaauc cggaagcgac       1620

caacgccuug auugacaagg auggauggcu acauucugga gacauagcuu acugggacga       1680

agacgaacac uucuucauag uugaccgcuu gaagucuuua auuaaauaca aaggauauca       1740

gguggccccc gcugaauugg aaucgauauu guuacaacac cccaacaucu ucgacgcggg       1800

cguggcaggu cuucccgacg augacgccgg ugaacuuccc gccgccguug uuguuuugga       1860

gcacggaaag acgaugacgg aaaaagagau cguggauuac gucgccaguc aaguaacaac       1920

cgcgaaaaag uugcgcggag gaguuguguu uguggacgaa guaccgaaag gucuuaccgg       1980

aaaacucgac gcaagaaaaa ucagagagau ccucauaaag gccaagaagg gcggaaaguc       2040

caaauuguaa guuuaaaccc ucucccuccc cccccccuaa cguuacuggc cgaagccgcu       2100

uggaauaagg ccggugugcg uuugucuaua uguuauuuuc caccauauug ccgucuuuug       2160

gcaaugugag ggcccggaaa ccuggcccug ucuucuugac gagcauuccu aggggucuuu       2220

ccccucucgc caaaggaaug caaggucugu ugaaugucgu gaaggaagca guuccucugg       2280

aagcuucuug aagacaaaca acgucuguag cgacccuuug caggcagcgg aaccccccac       2340

cuggcgacag gugccucugc ggccaaaagc cacguguaua agauacaccu gcaaaggcgg       2400

cacaacccca gugccacguu gugaguugga uaguugugga aagagucaaa uggcucuccu       2460

caagcguauu caacaagggg cugaaggaug cccagaaggu accccauugu augggaucug       2520

aucuggggcc ucggugcaca ugcuuuacau guguuuaguc gagguuaaaa aaacgucuag       2580

gccccccgaa ccacggggac gugguuuucc uuugaaaaac acgaugauac caugagcaca       2640

aauccuaaac cucaaagaaa aaccaaaaga aacaccaacc gucgcccaga agacguuaag       2700

uucccgggcg gcggccagau cguuggcgga guauacuugu ugccgcgcag gggccccagg       2760

uugggugugc gcacgacaag gaaaacuucg gagcgguccc agccacgugg gagacgccag       2820

cccaucccca aagaucggcg cuccacuggc aaggccuggg gaaaaccagg ucgccccugg       2880

ccccuauaug ggaaugaggg acucggcugg gcaggauggc uccugucccc ccgaggcucu       2940

cgccccuccu ggggccccac ugacccccgg cauaggucgc gcaacguggg uaaagucauc       3000

gacacccuaa cguguggcuu ugccgaccuc augggguaca uccccgucgu aggcgccccg       3060

cuuaguggcg ccgccagagc ugucgcgcac ggcgugagag uccuggagga cgggguuaau       3120

uaugcaacag ggaaccuacc cgguuucccc uuuucuaucu ucuugcuggc ccuguugucc       3180

ugcaucaccg uuccggucuc ugcugcccag gugaagaaua ccaguagcag cuacauggug       3240

accaaugacu gcuccaauga cagcaucacu uggcagcucg aggcugcggu ucuccacguc       3300

cccgggugcg ucccgugcga gagagugggg aauacgucac gguguugggu gccagucucg       3360

ccaaacaugg cugugcggca gcccggugcc cucacgcagg gucugcggac gcacaucgau       3420

augguuguga uguccgccac cuucugcucu gcucucuacg ugggggaccu cuguggcggg       3480

gugaugcucg cggcccaggu guucaucguc ucgccgcagu accacugguu ugugcaagaa       3540

ugcaauugcu ccaucuaccc uggcaccauc acuggacacc gcauggcaug ggacaugaug       3600

augaacuggu cgcccacggc caccaugauc cuggcguacg ugaugcgcgu ccccgagguc       3660

aucauagaca ucguuagcgg ggcucacugg ggcgucaugu ucggcuuggc cuacuucucu       3720

augcagggag cgugggcgaa ggucauuguc auccuucugc uggccgcugg gguggacgcg       3780

ggcaccacca ccguuggagg cgcuguugca cguuccacca acgugauugc cggcguguuc       3840

agccauggcc cucagcagaa cauucagcuc auuaacacca acggcaguug gcacaucaac       3900

cguacugccu ugaauugcaa ugacuccuug aacaccggcu uucucgcggc cuuguucuac       3960

accaaccgcu uuaacucguc agggugucca gggcgccugu ccgccugccg caacaucgag       4020

gcuuuccgga uagggugggg cacccuacag uacgaggaua augucaccaa uccagaggau       4080

augaggccgu acugcuggca cuacccccca aagccgugug gcguaguccc cgcgaggucu       4140

guguguggcc caguguacug uuucaccccc agcccgguag uagugggcac gaccgacaga       4200

cguggagugc ccaccuacac auggggagag aaugagacag augucuuccu acugaacagc       4260

acccgaccgc cgcagggcuc augguucggc ugcacgugga ugaacuccac ugguuucacc       4320

aagacuugug gcgcgccacc uugccgcacc agagcugacu ucaacgccag cacggacuug       4380

uugugcccua cggauuguuu uaggaagcau ccugaugcca cuuauauuaa gugugguucu       4440

gggcccuggc ucacaccaaa gugccugguc cacuacccuu acagacucug gcauuacccc       4500

ugcacaguca auuuuaccau cuucaagaua agaauguaug uagggggggu ugagcacagg       4560

cucacggccg caugcaacuu cacucguggg gaucgcugcg acuuggagga cagggacagg       4620

agucagcugu cuccucuguu gcacucuacc acggaauggg ccauccugcc cugcaccuac       4680

ucagacuuac ccgcuuuguc aacuggucuu cuccaccuuc accagaacau cguggacgua       4740

caauacaugu auggccucuc accugcuauc acaaaauacg ucguucgaug ggagugggug       4800

guacucuuau uccugcucuu agcggacgcc agagucugcg ccugcuugug gaugcucauc       4860

uuguugggcc aggccgaagc agcauuggag aaguuggucg ucuugcacgc ugcgagugcg       4920

gcuaacugcc auggccuccu auauuuugcc aucuucuucg uggcagcuug gcacaucagg       4980

ggucgggugg uccccuugac caccuauugc cucacuggcc uauggcccuu cugccuacug       5040

cucauggcac ugccccggca ggcuuaugcc uaugacgcac cugugcacgg acagauaggc       5100

guggguuugu ugauauugau cacccucuuc acacucaccc cgggguauaa gacccuccuc       5160

ggccaguguc uguggugguu gugcuaucuc cugacccugg gggaagccau gauucaggag       5220

uggguaccac ccaugcaggu gcgcggcggc cgcgauggca ucgcgugggc cgucacuaua       5280

uucugcccgg gugugguguu ugacauuacc aaauggcuuu uggcguugcu ugggccugcu       5340

uaccucuuaa gggccgcuuu gacacaugug ccguacuucg ucagagcuca cgcucugaua       5400

aggguaugcg cuuuggugaa gcagcucgcg ggggguaggu auguucaggu ggcgcuauug       5460

gcccuuggca gguggacugg caccuacauc uaugaccacc ucacaccuau gucggacugg       5520

gccgcuagcg gccugcgcga cuuagcgguc gccguggaac ccaucaucuu caguccgaug       5580

gagaagaagg ucaucgucug gggagcggag acggcugcau guggggacau ucuacaugga       5640

cuucccgugu ccgcccgacu cggccaggag auccuccucg gcccagcuga uggcuacacc       5700

uccaaggggu ggaagcuccu ugcucccauc acugcuuaug cccagcaaac acgaggccuc       5760

cugggcgcca uaguggugag uaugacgggg cgugacagga cagaacaggc cggggaaguc       5820

caaauccugu ccacagucuc ucaguccuuc cucggaacaa ccaucucggg gguuuugugg       5880

acuguuuacc acggagcugg caacaagacu cuagccggcu uacggggucc ggucacgcag       5940

auguacucga gugcugaggg ggacuuggua ggcuggccca gccccccugg gaccaagucu       6000

uuggagccgu gcaagugugg agccgucgac cuauaucugg ucacgcggaa cgcugauguc       6060

aucccggcuc ggagacgcgg ggacaagcgg ggagcauugc ucuccccgag acccauuucg       6120

accuugaagg gguccucggg ggggccggug cucugcccua ggggccacgu cguugggcuc       6180

uuccgagcag cugugugcuc ucggggcgug gccaaaucca ucgauuucau ccccguugag       6240

acacucgacg uuguuacaag gucucccacu uucagugaca acagcacgcc accggcugug       6300

ccccagaccu aucaggucgg guacuugcau gcuccaacug gcaguggaaa gagcaccaag       6360

gucccugucg cguaugccgc ccagggguac aaaguacuag ugcuuaaccc cucgguagcu       6420

gccacccugg gguuuggggc guaccuaucc aaggcacaug gcaucaaucc caacauuagg       6480

acuggaguca ggaccgugau gaccggggag gccaucacgu acuccacaua uggcaaauuu       6540

cucgccgaug ggggcugcgc uagcggcgcc uaugacauca ucauaugcga ugaaugccac       6600

gcuguggaug cuaccuccau ucucggcauc ggaacggucc uugaucaagc agagacagcc       6660

ggggucagac uaacugugcu ggcuacggcc acaccccccg ggucagugac aaccccccau       6720

cccgauauag aagagguagg ccucgggcgg gagggugaga uccccuucua ugggagggcg       6780

auuccccuau ccugcaucaa gggagggaga caccugauuu ucugccacuc aaagaaaaag       6840

ugugacgagc ucgcggcggc ccuucggggc augggcuuga augccguggc auacuauaga       6900

ggguuggacg ucuccauaau accagcucag ggagaugugg uggucgucgc caccgacgcc       6960

cucaugacgg gguacacugg agacuuugac uccgugaucg acugcaaugu agcggucacc       7020

caagcugucg acuucagccu ggaccccacc uucacuauaa ccacacagac ugucccacaa       7080

gacgcugucu cacgcaguca gcgccgcggg cgcacaggua gaggaagaca gggcacuuau       7140

agguauguuu ccacugguga acgagccuca ggaauguuug acaguguagu gcuuugugag       7200

ugcuacgacg caggggcugc gugguacgau cucacaccag cggagaccac cgucaggcuu       7260

agagcguauu ucaacacgcc cggccuaccc gugugucaag accaucuuga auuuugggag       7320

gcaguuuuca ccggccucac acacauagac gcccacuucc ucucccaaac aaagcaagcg       7380

ggggagaacu ucgcguaccu aguagccuac caagcuacgg ugugcgccag agccaaggcc       7440

ccucccccgu ccugggacgc cauguggaag ugccuggccc gacucaagcc uacgcuugcg       7500

ggccccacac cucuccugua ccguuugggc ccuauuacca augaggucac ccucacacac       7560

ccugggacga aguacaucgc cacaugcaug caagcugacc uugaggucau gaccagcacg       7620

uggguccuag cuggaggagu ccuggcagcc gucgccgcau auugccuggc gacuggaugc       7680

guuuccauca ucggccgcuu gcacgucaac cagcgagucg ucguugcgcc ggauaaggag       7740

guccuguaug aggcuuuuga ugagauggag gaaugcgccu cuagggcggc ucucaucgaa       7800

gaggggcagc ggauagccga gauguugaag uccaagaucc aaggcuugcu gcagcaggcc       7860

ucuaagcagg cccaggacau acaacccgcu augcaggcuu cauggcccaa aguggaacaa       7920

uuuugggcca gacacaugug gaacuucauu agcggcaucc aauaccucgc aggauuguca       7980

acacugccag ggaaccccgc gguggcuucc augauggcau ucagugccgc ccucaccagu       8040

ccguugucga ccaguaccac cauccuucuc aacaucaugg gaggcugguu agcgucccag       8100

aucgcaccac ccgcgggggc caccggcuuu gucgucagug gccugguggg ggcugccgug       8160

ggcagcauag gccuggguaa ggugcuggug gacauccugg caggauaugg ugcgggcauu       8220

ucgggggccc ucgucgcauu caagaucaug ucuggcgaga agcccucuau ggaagauguc       8280

aucaaucuac ugccugggau ccugucuccg ggagcccugg uggugggggu caucugcgcg       8340

gccauucugc gccgccacgu gggaccgggg gagggcgcgg uccaauggau gaacaggcuu       8400

auugccuuug cuuccagagg aaaccacguc gccccuacuc acuacgugac ggagucggau       8460

gcgucgcagc gugugaccca acuacuuggc ucucuuacua uaaccagccu acucagaaga       8520

cuccacaauu ggauaacuga ggacugcccc aucccaugcu ccggauccug gcuccgcgac       8580

gugugggacu ggguuugcac caucuugaca gacuucaaaa auuggcugac cucuaaauug       8640

uuccccaagc ugcccggccu ccccuucauc ucuugucaaa agggguacaa gggugugugg       8700

gccggcacug gcaucaugac cacgcgcugc ccuugcggcg ccaacaucuc uggcaauguc       8760

cgccugggcu cuaugaggau cacagggccu aaaaccugca ugaacaccug gcaggggacc       8820

uuuccuauca auugcuacac ggagggccag ugcgcgccga aaccccccac gaacuacaag       8880

accgccaucu ggaggguggc ggccucggag uacgcggagg ugacgcagca ugggucguac       8940

uccuauguaa caggacugac cacugacaau cugaaaauuc cuugccaacu accuucucca       9000

gaguuuuucu ccugggugga cggugugcag auccauaggu uugcacccac accaaagccg       9060

uuuuuccggg augaggucuc guucugcguu gggcuuaauu ccuaugcugu cgggucccag       9120

cuucccugug aaccugagcc cgacgcagac guauugaggu ccaugcuaac agauccgccc       9180

cacaucacgg cggagacugc ggcgcggcgc uuggcacggg gaucaccucc aucugaggcg       9240

agcuccucag ugagccagcu aucagcaccg ucgcugcggg ccaccugcac cacccacagc       9300

aacaccuaug acguggacau ggucgaugcc aaccugcuca uggagggcgg uguggcucag       9360

acagagccug aguccagggu gcccguucug gacuuucucg agccaauggc cgaggaagag       9420

agcgaccuug agcccucaau accaucggag ugcaugcucc ccaggagcgg guuuccacgg       9480

gccuuaccgg cuugggcacg gccugacuac aacccgccgc ucguggaauc guggaggagg       9540

ccagauuacc aaccgcccac cguugcuggu ugugcucucc ccccccccaa gaaggccccg       9600

acgccucccc caaggagacg ccggacagug ggucugagcg agagcaccau aucagaagcc       9660

cuccagcaac uggccaucaa gaccuuuggc cagccccccu cgagcgguga ugcaggcucg       9720

uccacggggg cgggcgccgc cgaauccggc gguccgacgu ccccugguga gccggccccc       9780

ucagagacag guuccgccuc cucuaugccc ccccucgagg gggagccugg agauccggac       9840

cuggagucug aucagguaga gcuucaaccu cccccccagg gggggggggu agcucccggu       9900

ucgggcucgg ggucuugguc uacuugcucc gaggaggacg auaccaccgu gugcugcucc       9960

augucauacu ccuggaccgg ggcucuaaua acucccugua gccccgaaga ggaaaaguug       10020

ccaaucaacc cuuugaguaa cucgcuguug cgauaccaua acaaggugua cuguacaaca       10080

ucaaagagcg ccucacagag ggcuaaaaag guaacuuuug acaggacgca agugcucgac       10140

gcccauuaug acucagucuu aaaggacauc aagcuagcgg cuuccaaggu cagcgcaagg       10200

cuccucaccu uggaggaggc gugccaguug acuccacccc auucugcaag auccaaguau       10260

ggauucgggg ccaaggaggu ccgcagcuug uccgggaggg ccguuaacca caucaagucc       10320

guguggaagg accuccugga agacccacaa acaccaauuc ccacaaccau cauggccaaa       10380

aaugaggugu ucugcgugga ccccgccaag ggggguaaga aaccagcucg ccucaucguu       10440

uacccugacc ucggcguccg ggucugcgag aaaauggccc ucuaugacau uacacaaaag       10500

cuuccucagg cgguaauggg agcuuccuau ggcuuccagu acuccccugc ccaacgggug       10560

gaguaucucu ugaaagcaug ggcggaaaag aaggacccca uggguuuuuc guaugauacc       10620

cgaugcuucg acucaaccgu cacugagaga gacaucagga ccgaggaguc cauauaccag       10680

gccugcuccc ugcccgagga ggcccgcacu gccauacacu cgcugacuga gagacuuuac       10740

guaggagggc ccauguucaa cagcaagggu caaaccugcg guuacagacg uugccgcgcc       10800

agcggggugc uaaccacuag cauggguaac accaucacau gcuaugugaa agcccuagcg       10860

gccugcaagg cugcggggau aguugcgccc acaaugcugg uaugcggcaa ugaccuagua       10920

gucaucucag aaagccaggg gacugaggag gacgagcgga accugagagc cuucacggag       10980

gccaugacca gguacucugc cccuccuggu gaucccccca gaccggaaua ugaccuggag       11040

cuaauaacau ccuguuccuc aaaugugucu guggcguugg gcccgcgggg ccgccgcaga       11100

uacuaccuga ccagagaccc aaccacucca cucgcccggg cugccuggga aacaguuaga       11160

cacuccccua ucaauucaug gcugggaaac aucauccagu augcuccaac cauauggguu       11220

cgcauggucc uaaugacaca cuucuucucc auucucaugg uccaagacac ccuggaccag       11280

aaccucaacu uugagaugua uggaucagua uacuccguga auccuuugga ccuuccagcc       11340

auaauugaga gguuacacgg gcuugacgcc uuuucuaugc acacauacuc ucaccacgaa       11400

cugacgcggg uggcuucagc ccucagaaaa cuuggggcgc caccccucag gguguggaag       11460

agucgggcuc gcgcagucag ggcgucccuc aucucccgug gagggaaagc ggccguuugc       11520

ggccgauauc ucuucaauug ggcggugaag accaagcuca aacucacucc auugccggag       11580

gcgcgccuac uggacuuauc caguugguuc accgucggcg ccggcggggg cgacauuuuu       11640

cacagcgugu cgcgcgcccg accccgcuca uuacucuucg gccuacuccu acuuuucgua       11700

gggguaggcc ucuuccuacu ccccgcucgg uagagcggca cacacuaggu acacuccaua       11760

gcuaacuguu ccuuuuuuuu uuuuuuuuuu uuuuuuuuuu uuuuuuuuuu uuuuucuuuu       11820

uuuuuuuuuu cccucuuucu ucccuucuca ucuuauucua cuuucuuucu ugguggcucc       11880

aucuuagccc uagucacggc uagcugugaa agguccguga gccgcaugac ugcagagagu       11940

gccguaacug gucucucugc agaucaugu                                         11969

<210>23

<211>11036

<212>RNA

<213>人工序列

<220>

<223>人工序列的描述：来源于表达载体rFGR-JFH1/EGFP的复制子RNA

<400>23

accugccccu aauaggggcg acacuccgcc augaaucacu ccccugugag gaacuacugu       60

cuucacgcag aaagcgccua gccauggcgu uaguaugagu gucguacagc cuccaggccc       120

cccccucccg ggagagccau aguggucugc ggaaccggug aguacaccgg aauugccggg       180

aagacugggu ccuuucuugg auaaacccac ucuaugcccg gccauuuggg cgugcccccg       240

caagacugcu agccgaguag cguuggguug cgaaaggccu ugugguacug ccugauaggg       300

cgcuugcgag ugccccggga ggucucguag accgugcacc augagcacaa auccuaaacc       360

ucaaagaaaa accaaaagaa acaccaaccg acgcguaaug gugagcaagg gcgaggagcu       420

guucaccggg guggugccca uccuggucga gcuggacggc gacguaaacg gccacaaguu       480

cagcgugucc ggcgagggcg agggcgaugc caccuacggc aagcugaccc ugaaguucau       540

cugcaccacc ggcaagcugc ccgugcccug gcccacccuc gugaccaccc ugaccuacgg       600

cgugcagugc uucagccgcu accccgacca caugaagcag cacgacuucu ucaaguccgc       660

caugcccgaa ggcuacgucc aggagcgcac caucuucuuc aaggacgacg gcaacuacaa       720

gacccgcgcc gaggugaagu ucgagggcga cacccuggug aaccgcaucg agcugaaggg       780

caucgacuuc aaggaggacg gcaacauccu ggggcacaag cuggaguaca acuacaacag       840

ccacaacguc uauaucaugg ccgacaagca gaagaacggc aucaagguga acuucaagau       900

ccgccacaac aucgaggacg gcagcgugca gcucgccgac cacuaccagc agaacacccc       960

caucggcgac ggccccgugc ugcugcccga caaccacuac cugagcaccc aguccgcccu       1020

gagcaaagac cccaacgaga agcgcgauca caugguccug cuggaguucg ugaccgccgc       1080

cgggaucacu cucggcaugg acgagcugua caaguaaguu uaaacccucu cccucccccc       1140

ccccuaacgu uacuggccga agccgcuugg aauaaggccg gugugcguuu gucuauaugu       1200

uauuuuccac cauauugccg ucuuuuggca augugagggc ccggaaaccu ggcccugucu       1260

ucuugacgag cauuccuagg ggucuuuccc cucucgccaa aggaaugcaa ggucuguuga       1320

augucgugaa ggaagcaguu ccucuggaag cuucuugaag acaaacaacg ucuguagcga       1380

cccuuugcag gcagcggaac cccccaccug gcgacaggug ccucugcggc caaaagccac       1440

guguauaaga uacaccugca aaggcggcac aaccccagug ccacguugug aguuggauag       1500

uuguggaaag agucaaaugg cucuccucaa gcguauucaa caaggggcug aaggaugccc       1560

agaagguacc ccauuguaug ggaucugauc uggggccucg gugcacaugc uuuacaugug       1620

uuuagucgag guuaaaaaaa cgucuaggcc ccccgaacca cggggacgug guuuuccuuu       1680

gaaaaacacg augauaccau gagcacaaau ccuaaaccuc aaagaaaaac caaaagaaac       1740

accaaccguc gcccagaaga cguuaaguuc ccgggcggcg gccagaucgu uggcggagua       1800

uacuuguugc cgcgcagggg ccccagguug ggugugcgca cgacaaggaa aacuucggag       1860

cggucccagc cacgugggag acgccagccc auccccaaag aucggcgcuc cacuggcaag       1920

gccuggggaa aaccaggucg ccccuggccc cuauauggga augagggacu cggcugggca       1980

ggauggcucc uguccccccg aggcucucgc cccuccuggg gccccacuga cccccggcau       2040

aggucgcgca acguggguaa agucaucgac acccuaacgu guggcuuugc cgaccucaug       2100

ggguacaucc ccgucguagg cgccccgcuu aguggcgccg ccagagcugu cgcgcacggc       2160

gugagagucc uggaggacgg gguuaauuau gcaacaggga accuacccgg uuuccccuuu       2220

ucuaucuucu ugcuggcccu guuguccugc aucaccguuc cggucucugc ugcccaggug       2280

aagaauacca guagcagcua cauggugacc aaugacugcu ccaaugacag caucacuugg       2340

cagcucgagg cugcgguucu ccacgucccc gggugcgucc cgugcgagag aguggggaau       2400

acgucacggu guugggugcc agucucgcca aacauggcug ugcggcagcc cggugcccuc       2460

acgcaggguc ugcggacgca caucgauaug guugugaugu ccgccaccuu cugcucugcu       2520

cucuacgugg gggaccucug uggcggggug augcucgcgg cccagguguu caucgucucg       2580

ccgcaguacc acugguuugu gcaagaaugc aauugcucca ucuacccugg caccaucacu       2640

ggacaccgca uggcauggga caugaugaug aacuggucgc ccacggccac caugauccug       2700

gcguacguga ugcgcguccc cgaggucauc auagacaucg uuagcggggc ucacuggggc       2760

gucauguucg gcuuggccua cuucucuaug cagggagcgu gggcgaaggu cauugucauc       2820

cuucugcugg ccgcuggggu ggacgcgggc accaccaccg uuggaggcgc uguugcacgu       2880

uccaccaacg ugauugccgg cguguucagc cauggcccuc agcagaacau ucagcucauu       2940

aacaccaacg gcaguuggca caucaaccgu acugccuuga auugcaauga cuccuugaac       3000

accggcuuuc ucgcggccuu guucuacacc aaccgcuuua acucgucagg guguccaggg       3060

cgccuguccg ccugccgcaa caucgaggcu uuccggauag gguggggcac ccuacaguac       3120

gaggauaaug ucaccaaucc agaggauaug aggccguacu gcuggcacua ccccccaaag       3180

ccguguggcg uaguccccgc gaggucugug uguggcccag uguacuguuu cacccccagc       3240

ccgguaguag ugggcacgac cgacagacgu ggagugccca ccuacacaug gggagagaau       3300

gagacagaug ucuuccuacu gaacagcacc cgaccgccgc agggcucaug guucggcugc       3360

acguggauga acuccacugg uuucaccaag acuuguggcg cgccaccuug ccgcaccaga       3420

gcugacuuca acgccagcac ggacuuguug ugcccuacgg auuguuuuag gaagcauccu       3480

gaugccacuu auauuaagug ugguucuggg cccuggcuca caccaaagug ccugguccac       3540

uacccuuaca gacucuggca uuaccccugc acagucaauu uuaccaucuu caagauaaga       3600

auguauguag gggggguuga gcacaggcuc acggccgcau gcaacuucac ucguggggau       3660

cgcugcgacu uggaggacag ggacaggagu cagcugucuc cucuguugca cucuaccacg       3720

gaaugggcca uccugcccug caccuacuca gacuuacccg cuuugucaac uggucuucuc       3780

caccuucacc agaacaucgu ggacguacaa uacauguaug gccucucacc ugcuaucaca       3840

aaauacgucg uucgauggga guggguggua cucuuauucc ugcucuuagc ggacgccaga       3900

gucugcgccu gcuuguggau gcucaucuug uugggccagg ccgaagcagc auuggagaag       3960

uuggucgucu ugcacgcugc gagugcggcu aacugccaug gccuccuaua uuuugccauc       4020

uucuucgugg cagcuuggca caucaggggu cggguggucc ccuugaccac cuauugccuc       4080

acuggccuau ggcccuucug ccuacugcuc auggcacugc cccggcaggc uuaugccuau       4140

gacgcaccug ugcacggaca gauaggcgug gguuuguuga uauugaucac ccucuucaca       4200

cucaccccgg gguauaagac ccuccucggc cagugucugu ggugguugug cuaucuccug       4260

acccuggggg aagccaugau ucaggagugg guaccaccca ugcaggugcg cggcggccgc       4320

gauggcaucg cgugggccgu cacuauauuc ugcccgggug ugguguuuga cauuaccaaa       4380

uggcuuuugg cguugcuugg gccugcuuac cucuuaaggg ccgcuuugac acaugugccg       4440

uacuucguca gagcucacgc ucugauaagg guaugcgcuu uggugaagca gcucgcgggg       4500

gguagguaug uucagguggc gcuauuggcc cuuggcaggu ggacuggcac cuacaucuau       4560

gaccaccuca caccuauguc ggacugggcc gcuagcggcc ugcgcgacuu agcggucgcc       4620

guggaaccca ucaucuucag uccgauggag aagaagguca ucgucugggg agcggagacg       4680

gcugcaugug gggacauucu acauggacuu cccguguccg cccgacucgg ccaggagauc       4740

cuccucggcc cagcugaugg cuacaccucc aaggggugga agcuccuugc ucccaucacu       4800

gcuuaugccc agcaaacacg aggccuccug ggcgccauag uggugaguau gacggggcgu       4860

gacaggacag aacaggccgg ggaaguccaa auccugucca cagucucuca guccuuccuc       4920

ggaacaacca ucucgggggu uuuguggacu guuuaccacg gagcuggcaa caagacucua       4980

gccggcuuac gggguccggu cacgcagaug uacucgagug cugaggggga cuugguaggc       5040

uggcccagcc ccccugggac caagucuuug gagccgugca aguguggagc cgucgaccua       5100

uaucugguca cgcggaacgc ugaugucauc ccggcucgga gacgcgggga caagcgggga       5160

gcauugcucu ccccgagacc cauuucgacc uugaaggggu ccucgggggg gccggugcuc       5220

ugcccuaggg gccacgucgu ugggcucuuc cgagcagcug ugugcucucg gggcguggcc       5280

aaauccaucg auuucauccc cguugagaca cucgacguug uuacaagguc ucccacuuuc       5340

agugacaaca gcacgccacc ggcugugccc cagaccuauc aggucgggua cuugcaugcu       5400

ccaacuggca guggaaagag caccaagguc ccugucgcgu augccgccca gggguacaaa       5460

guacuagugc uuaaccccuc gguagcugcc acccuggggu uuggggcgua ccuauccaag       5520

gcacauggca ucaaucccaa cauuaggacu ggagucagga ccgugaugac cggggaggcc       5580

aucacguacu ccacauaugg caaauuucuc gccgaugggg gcugcgcuag cggcgccuau       5640

gacaucauca uaugcgauga augccacgcu guggaugcua ccuccauucu cggcaucgga       5700

acgguccuug aucaagcaga gacagccggg gucagacuaa cugugcuggc uacggccaca       5760

ccccccgggu cagugacaac cccccauccc gauauagaag agguaggccu cgggcgggag       5820

ggugagaucc ccuucuaugg gagggcgauu ccccuauccu gcaucaaggg agggagacac       5880

cugauuuucu gccacucaaa gaaaaagugu gacgagcucg cggcggcccu ucggggcaug       5940

ggcuugaaug ccguggcaua cuauagaggg uuggacgucu ccauaauacc agcucaggga       6000

gauguggugg ucgucgccac cgacgcccuc augacggggu acacuggaga cuuugacucc       6060

gugaucgacu gcaauguagc ggucacccaa gcugucgacu ucagccugga ccccaccuuc       6120

acuauaacca cacagacugu cccacaagac gcugucucac gcagucagcg ccgcgggcgc       6180

acagguagag gaagacaggg cacuuauagg uauguuucca cuggugaacg agccucagga       6240

auguuugaca guguagugcu uugugagugc uacgacgcag gggcugcgug guacgaucuc       6300

acaccagcgg agaccaccgu caggcuuaga gcguauuuca acacgcccgg ccuacccgug       6360

ugucaagacc aucuugaauu uugggaggca guuuucaccg gccucacaca cauagacgcc       6420

cacuuccucu cccaaacaaa gcaagcgggg gagaacuucg cguaccuagu agccuaccaa       6480

gcuacggugu gcgccagagc caaggccccu cccccguccu gggacgccau guggaagugc       6540

cuggcccgac ucaagccuac gcuugcgggc cccacaccuc uccuguaccg uuugggcccu       6600

auuaccaaug aggucacccu cacacacccu gggacgaagu acaucgccac augcaugcaa       6660

gcugaccuug aggucaugac cagcacgugg guccuagcug gaggaguccu ggcagccguc       6720

gccgcauauu gccuggcgac uggaugcguu uccaucaucg gccgcuugca cgucaaccag       6780

cgagucgucg uugcgccgga uaaggagguc cuguaugagg cuuuugauga gauggaggaa       6840

ugcgccucua gggcggcucu caucgaagag gggcagcgga uagccgagau guugaagucc       6900

aagauccaag gcuugcugca gcaggccucu aagcaggccc aggacauaca acccgcuaug       6960

caggcuucau ggcccaaagu ggaacaauuu ugggccagac acauguggaa cuucauuagc       7020

ggcauccaau accucgcagg auugucaaca cugccaggga accccgcggu ggcuuccaug       7080

auggcauuca gugccgcccu caccaguccg uugucgacca guaccaccau ccuucucaac       7140

aucaugggag gcugguuagc gucccagauc gcaccacccg cgggggccac cggcuuuguc       7200

gucaguggcc uggugggggc ugccgugggc agcauaggcc uggguaaggu gcugguggac       7260

auccuggcag gauauggugc gggcauuucg ggggcccucg ucgcauucaa gaucaugucu       7320

ggcgagaagc ccucuaugga agaugucauc aaucuacugc cugggauccu gucuccggga       7380

gcccuggugg ugggggucau cugcgcggcc auucugcgcc gccacguggg accgggggag       7440

ggcgcggucc aauggaugaa caggcuuauu gccuuugcuu ccagaggaaa ccacgucgcc       7500

ccuacucacu acgugacgga gucggaugcg ucgcagcgug ugacccaacu acuuggcucu       7560

cuuacuauaa ccagccuacu cagaagacuc cacaauugga uaacugagga cugccccauc       7620

ccaugcuccg gauccuggcu ccgcgacgug ugggacuggg uuugcaccau cuugacagac       7680

uucaaaaauu ggcugaccuc uaaauuguuc cccaagcugc ccggccuccc cuucaucucu       7740

ugucaaaagg gguacaaggg ugugugggcc ggcacuggca ucaugaccac gcgcugcccu       7800

ugcggcgcca acaucucugg caauguccgc cugggcucua ugaggaucac agggccuaaa       7860

accugcauga acaccuggca ggggaccuuu ccuaucaauu gcuacacgga gggccagugc       7920

gcgccgaaac cccccacgaa cuacaagacc gccaucugga ggguggcggc cucggaguac       7980

gcggagguga cgcagcaugg gucguacucc uauguaacag gacugaccac ugacaaucug       8040

aaaauuccuu gccaacuacc uucuccagag uuuuucuccu ggguggacgg ugugcagauc       8100

cauagguuug cacccacacc aaagccguuu uuccgggaug aggucucguu cugcguuggg       8160

cuuaauuccu augcugucgg gucccagcuu cccugugaac cugagcccga cgcagacgua       8220

uugaggucca ugcuaacaga uccgccccac aucacggcgg agacugcggc gcggcgcuug       8280

gcacggggau caccuccauc ugaggcgagc uccucaguga gccagcuauc agcaccgucg       8340

cugcgggcca ccugcaccac ccacagcaac accuaugacg uggacauggu cgaugccaac       8400

cugcucaugg agggcggugu ggcucagaca gagccugagu ccagggugcc cguucuggac       8460

uuucucgagc caauggccga ggaagagagc gaccuugagc ccucaauacc aucggagugc       8520

augcucccca ggagcggguu uccacgggcc uuaccggcuu gggcacggcc ugacuacaac       8580

ccgccgcucg uggaaucgug gaggaggcca gauuaccaac cgcccaccgu ugcugguugu       8640

gcucuccccc cccccaagaa ggccccgacg ccucccccaa ggagacgccg gacagugggu       8700

cugagcgaga gcaccauauc agaagcccuc cagcaacugg ccaucaagac cuuuggccag       8760

ccccccucga gcggugaugc aggcucgucc acgggggcgg gcgccgccga auccggcggu       8820

ccgacguccc cuggugagcc ggcccccuca gagacagguu ccgccuccuc uaugcccccc       8880

cucgaggggg agccuggaga uccggaccug gagucugauc agguagagcu ucaaccuccc       8940

ccccaggggg gggggguagc ucccgguucg ggcucggggu cuuggucuac uugcuccgag       9000

gaggacgaua ccaccgugug cugcuccaug ucauacuccu ggaccggggc ucuaauaacu       9060

cccuguagcc ccgaagagga aaaguugcca aucaacccuu ugaguaacuc gcuguugcga       9120

uaccauaaca agguguacug uacaacauca aagagcgccu cacagagggc uaaaaaggua       9180

acuuuugaca ggacgcaagu gcucgacgcc cauuaugacu cagucuuaaa ggacaucaag       9240

cuagcggcuu ccaaggucag cgcaaggcuc cucaccuugg aggaggcgug ccaguugacu       9300

ccaccccauu cugcaagauc caaguaugga uucggggcca aggagguccg cagcuugucc       9360

gggagggccg uuaaccacau caaguccgug uggaaggacc uccuggaaga cccacaaaca       9420

ccaauuccca caaccaucau ggccaaaaau gagguguucu gcguggaccc cgccaagggg       9480

gguaagaaac cagcucgccu caucguuuac ccugaccucg gcguccgggu cugcgagaaa       9540

auggcccucu augacauuac acaaaagcuu ccucaggcgg uaaugggagc uuccuauggc       9600

uuccaguacu ccccugccca acggguggag uaucucuuga aagcaugggc ggaaaagaag       9660

gaccccaugg guuuuucgua ugauacccga ugcuucgacu caaccgucac ugagagagac       9720

aucaggaccg aggaguccau auaccaggcc ugcucccugc ccgaggaggc ccgcacugcc       9780

auacacucgc ugacugagag acuuuacgua ggagggccca uguucaacag caagggucaa       9840

accugcgguu acagacguug ccgcgccagc ggggugcuaa ccacuagcau ggguaacacc       9900

aucacaugcu augugaaagc ccuagcggcc ugcaaggcug cggggauagu ugcgcccaca       9960

augcugguau gcggcgauga ccuaguaguc aucucagaaa gccaggggac ugaggaggac       10020

gagcggaacc ugagagccuu cacggaggcc augaccaggu acucugcccc uccuggugau       10080

ccccccagac cggaauauga ccuggagcua auaacauccu guuccucaaa ugugucugug       10140

gcguugggcc cgcggggccg ccgcagauac uaccugacca gagacccaac cacuccacuc       10200

gcccgggcug ccugggaaac aguuagacac uccccuauca auucauggcu gggaaacauc       10260

auccaguaug cuccaaccau auggguucgc augguccuaa ugacacacuu cuucuccauu       10320

cucauggucc aagacacccu ggaccagaac cucaacuuug agauguaugg aucaguauac       10380

uccgugaauc cuuuggaccu uccagccaua auugagaggu uacacgggcu ugacgccuuu       10440

ucuaugcaca cauacucuca ccacgaacug acgcgggugg cuucagcccu cagaaaacuu       10500

ggggcgccac cccucagggu guggaagagu cgggcucgcg cagucagggc gucccucauc       10560

ucccguggag ggaaagcggc cguuugcggc cgauaucucu ucaauugggc ggugaagacc       10620

aagcucaaac ucacuccauu gccggaggcg cgccuacugg acuuauccag uugguucacc       10680

gucggcgccg gcgggggcga cauuuuucac agcgugucgc gcgcccgacc ccgcucauua       10740

cucuucggcc uacuccuacu uuucguaggg guaggccucu uccuacuccc cgcucgguag       10800

agcggcacac acuagguaca cuccauagcu aacuguuccu uuuuuuuuuu uuuuuuuuuu       10860

uuuuuuuuuu uuuuuuuuuu uucuuuuuuu uuuuuuuccc ucuuucuucc cuucucaucu       10920

uauucuacuu ucuuucuugg uggcuccauc uuagcccuag ucacggcuag cugugaaagg       10980

uccgugagcc gcaugacugc agagagugcc guaacugguc ucucugcaga ucaugu           11036

<210>24

<211>11036

<212>RNA

<213>人工序列

<220>

<223>人工序列的描述：来源于表达载体rFGR-JFH1/EGFP/GND的复制子RNA

<400>24

accugccccu aauaggggcg acacuccgcc augaaucacu ccccugugag gaacuacugu       60

cuucacgcag aaagcgccua gccauggcgu uaguaugagu gucguacagc cuccaggccc       120

cccccucccg ggagagccau aguggucugc ggaaccggug aguacaccgg aauugccggg       180

aagacugggu ccuuucuugg auaaacccac ucuaugcccg gccauuuggg cgugcccccg       240

caagacugcu agccgaguag cguuggguug cgaaaggccu ugugguacug ccugauaggg       300

cgcuugcgag ugccccggga ggucucguag accgugcacc augagcacaa auccuaaacc       360

ucaaagaaaa accaaaagaa acaccaaccg acgcguaaug gugagcaagg gcgaggagcu       420

guucaccggg guggugccca uccuggucga gcuggacggc gacguaaacg gccacaaguu       480

cagcgugucc ggcgagggcg agggcgaugc caccuacggc aagcugaccc ugaaguucau       540

cugcaccacc ggcaagcugc ccgugcccug gcccacccuc gugaccaccc ugaccuacgg       600

cgugcagugc uucagccgcu accccgacca caugaagcag cacgacuucu ucaaguccgc       660

caugcccgaa ggcuacgucc aggagcgcac caucuucuuc aaggacgacg gcaacuacaa       720

gacccgcgcc gaggugaagu ucgagggcga cacccuggug aaccgcaucg agcugaaggg       780

caucgacuuc aaggaggacg gcaacauccu ggggcacaag cuggaguaca acuacaacag       840

ccacaacguc uauaucaugg ccgacaagca gaagaacggc aucaagguga acuucaagau       900

ccgccacaac aucgaggacg gcagcgugca gcucgccgac cacuaccagc agaacacccc       960

caucggcgac ggccccgugc ugcugcccga caaccacuac cugagcaccc aguccgcccu       1020

gagcaaagac cccaacgaga agcgcgauca caugguccug cuggaguucg ugaccgccgc       1080

cgggaucacu cucggcaugg acgagcugua caaguaaguu uaaacccucu cccucccccc       1140

ccccuaacgu uacuggccga agccgcuugg aauaaggccg gugugcguuu gucuauaugu       1200

uauuuuccac cauauugccg ucuuuuggca augugagggc ccggaaaccu ggcccugucu       1260

ucuugacgag cauuccuagg ggucuuuccc cucucgccaa aggaaugcaa ggucuguuga       1320

augucgugaa ggaagcaguu ccucuggaag cuucuugaag acaaacaacg ucuguagcga       1380

cccuuugcag gcagcggaac cccccaccug gcgacaggug ccucugcggc caaaagccac       1440

guguauaaga uacaccugca aaggcggcac aaccccagug ccacguugug aguuggauag       1500

uuguggaaag agucaaaugg cucuccucaa gcguauucaa caaggggcug aaggaugccc       1560

agaagguacc ccauuguaug ggaucugauc uggggccucg gugcacaugc uuuacaugug       1620

uuuagucgag guuaaaaaaa cgucuaggcc ccccgaacca cggggacgug guuuuccuuu       1680

gaaaaacacg augauaccau gagcacaaau ccuaaaccuc aaagaaaaac caaaagaaac       1740

accaaccguc gcccagaaga cguuaaguuc ccgggcggcg gccagaucgu uggcggagua       1800

uacuuguugc cgcgcagggg ccccagguug ggugugcgca cgacaaggaa aacuucggag       1860

cggucccagc cacgugggag acgccagccc auccccaaag aucggcgcuc cacuggcaag       1920

gccuggggaa aaccaggucg ccccuggccc cuauauggga augagggacu cggcugggca       1980

ggauggcucc uguccccccg aggcucucgc cccuccuggg gccccacuga cccccggcau       2040

aggucgcgca acguggguaa agucaucgac acccuaacgu guggcuuugc cgaccucaug       2100

ggguacaucc ccgucguagg cgccccgcuu aguggcgccg ccagagcugu cgcgcacggc       2160

gugagagucc uggaggacgg gguuaauuau gcaacaggga accuacccgg uuuccccuuu       2220

ucuaucuucu ugcuggcccu guuguccugc aucaccguuc cggucucugc ugcccaggug       2280

aagaauacca guagcagcua cauggugacc aaugacugcu ccaaugacag caucacuugg       2340

cagcucgagg cugcgguucu ccacgucccc gggugcgucc cgugcgagag aguggggaau       2400

acgucacggu guugggugcc agucucgcca aacauggcug ugcggcagcc cggugcccuc       2460

acgcaggguc ugcggacgca caucgauaug guugugaugu ccgccaccuu cugcucugcu       2520

cucuacgugg gggaccucug uggcggggug augcucgcgg cccagguguu caucgucucg       2580

ccgcaguacc acugguuugu gcaagaaugc aauugcucca ucuacccugg caccaucacu       2640

ggacaccgca uggcauggga caugaugaug aacuggucgc ccacggccac caugauccug       2700

gcguacguga ugcgcguccc cgaggucauc auagacaucg uuagcggggc ucacuggggc       2760

gucauguucg gcuuggccua cuucucuaug cagggagcgu gggcgaaggu cauugucauc       2820

cuucugcugg ccgcuggggu ggacgcgggc accaccaccg uuggaggcgc uguugcacgu       2880

uccaccaacg ugauugccgg cguguucagc cauggcccuc agcagaacau ucagcucauu       2940

aacaccaacg gcaguuggca caucaaccgu acugccuuga auugcaauga cuccuugaac       3000

accggcuuuc ucgcggccuu guucuacacc aaccgcuuua acucgucagg guguccaggg       3060

cgccuguccg ccugccgcaa caucgaggcu uuccggauag gguggggcac ccuacaguac       3120

gaggauaaug ucaccaaucc agaggauaug aggccguacu gcuggcacua ccccccaaag       3180

ccguguggcg uaguccccgc gaggucugug uguggcccag uguacuguuu cacccccagc       3240

ccgguaguag ugggcacgac cgacagacgu ggagugccca ccuacacaug gggagagaau       3300

gagacagaug ucuuccuacu gaacagcacc cgaccgccgc agggcucaug guucggcugc       3360

acguggauga acuccacugg uuucaccaag acuuguggcg cgccaccuug ccgcaccaga       3420

gcugacuuca acgccagcac ggacuuguug ugcccuacgg auuguuuuag gaagcauccu       3480

gaugccacuu auauuaagug ugguucuggg cccuggcuca caccaaagug ccugguccac       3540

uacccuuaca gacucuggca uuaccccugc acagucaauu uuaccaucuu caagauaaga       3600

auguauguag gggggguuga gcacaggcuc acggccgcau gcaacuucac ucguggggau       3660

cgcugcgacu uggaggacag ggacaggagu cagcugucuc cucuguugca cucuaccacg       3720

gaaugggcca uccugcccug caccuacuca gacuuacccg cuuugucaac uggucuucuc       3780

caccuucacc agaacaucgu ggacguacaa uacauguaug gccucucacc ugcuaucaca       3840

aaauacgucg uucgauggga guggguggua cucuuauucc ugcucuuagc ggacgccaga       3900

gucugcgccu gcuuguggau gcucaucuug uugggccagg ccgaagcagc auuggagaag       3960

uuggucgucu ugcacgcugc gagugcggcu aacugccaug gccuccuaua uuuugccauc       4020

uucuucgugg cagcuuggca caucaggggu cggguggucc ccuugaccac cuauugccuc       4080

acuggccuau ggcccuucug ccuacugcuc auggcacugc cccggcaggc uuaugccuau       4140

gacgcaccug ugcacggaca gauaggcgug gguuuguuga uauugaucac ccucuucaca       4200

cucaccccgg gguauaagac ccuccucggc cagugucugu ggugguugug cuaucuccug       4260

acccuggggg aagccaugau ucaggagugg guaccaccca ugcaggugcg cggcggccgc       4320

gauggcaucg cgugggccgu cacuauauuc ugcccgggug ugguguuuga cauuaccaaa       4380

uggcuuuugg cguugcuugg gccugcuuac cucuuaaggg ccgcuuugac acaugugccg       4440

uacuucguca gagcucacgc ucugauaagg guaugcgcuu uggugaagca gcucgcgggg       4500

gguagguaug uucagguggc gcuauuggcc cuuggcaggu ggacuggcac cuacaucuau       4560

gaccaccuca caccuauguc ggacugggcc gcuagcggcc ugcgcgacuu agcggucgcc       4620

guggaaccca ucaucuucag uccgauggag aagaagguca ucgucugggg agcggagacg       4680

gcugcaugug gggacauucu acauggacuu cccguguccg cccgacucgg ccaggagauc       4740

cuccucggcc cagcugaugg cuacaccucc aaggggugga agcuccuugc ucccaucacu       4800

gcuuaugccc agcaaacacg aggccuccug ggcgccauag uggugaguau gacggggcgu       4860

gacaggacag aacaggccgg ggaaguccaa auccugucca cagucucuca guccuuccuc       4920

ggaacaaccaucucgggggu  uuuguggacu guuuaccacg gagcuggcaa caagacucua       4980

gccggcuuac gggguccggu cacgcagaug uacucgagug cugaggggga cuugguaggc       5040

uggcccagcc ccccugggac caagucuuug gagccgugca aguguggagc cgucgaccua       5100

uaucugguca cgcggaacgc ugaugucauc ccggcucgga gacgcgggga caagcgggga       5160

gcauugcucu ccccgagacc cauuucgacc uugaaggggu ccucgggggg gccggugcuc       5220

ugcccuaggg gccacgucgu ugggcucuuc cgagcagcug ugugcucucg gggcguggcc       5280

aaauccaucg auuucauccc cguugagaca cucgacguug uuacaagguc ucccacuuuc       5340

agugacaaca gcacgccacc ggcugugccc cagaccuauc aggucgggua cuugcaugcu       5400

ccaacuggca guggaaagag caccaagguc ccugucgcgu augccgccca gggguacaaa       5460

guacuagugc uuaaccccuc gguagcugcc acccuggggu uuggggcgua ccuauccaag       5520

gcacauggca ucaaucccaa cauuaggacu ggagucagga ccgugaugac cggggaggcc       5580

aucacguacu ccacauaugg caaauuucuc gccgaugggg gcugcgcuag cggcgccuau       5640

gacaucauca uaugcgauga augccacgcu guggaugcua ccuccauucu cggcaucgga       5700

acgguccuug aucaagcaga gacagccggg gucagacuaa cugugcuggc uacggccaca       5760

ccccccgggu cagugacaac cccccauccc gauauagaag agguaggccu cgggcgggag       5820

ggugagaucc ccuucuaugg gagggcgauu ccccuauccu gcaucaaggg agggagacac       5880

cugauuuucu gccacucaaa gaaaaagugu gacgagcucg cggcggcccu ucggggcaug       5940

ggcuugaaug ccguggcaua cuauagaggg uuggacgucu ccauaauacc agcucaggga       6000

gauguggugg ucgucgccac cgacgcccuc augacggggu acacuggaga cuuugacucc       6060

gugaucgacu gcaauguagc ggucacccaa gcugucgacu ucagccugga ccccaccuuc       6120

acuauaacca cacagacugu cccacaagac gcugucucac gcagucagcg ccgcgggcgc       6180

acagguagag gaagacaggg cacuuauagg uauguuucca cuggugaacg agccucagga       6240

auguuugaca guguagugcu uugugagugc uacgacgcag gggcugcgug guacgaucuc       6300

acaccagcgg agaccaccgu caggcuuaga gcguauuuca acacgcccgg ccuacccgug       6360

ugucaagacc aucuugaauu uugggaggca guuuucaccg gccucacaca cauagacgcc       6420

cacuuccucu cccaaacaaa gcaagcgggg gagaacuucg cguaccuagu agccuaccaa       6480

gcuacggugu gcgccagagc caaggccccu cccccguccu gggacgccau guggaagugc       6540

cuggcccgac ucaagccuac gcuugcgggc cccacaccuc uccuguaccg uuugggcccu       6600

auuaccaaug aggucacccu cacacacccu gggacgaagu acaucgccac augcaugcaa       6660

gcugaccuug aggucaugac cagcacgugg guccuagcug gaggaguccu ggcagccguc       6720

gccgcauauu gccuggcgac uggaugcguu uccaucaucg gccgcuugca cgucaaccag       6780

cgagucgucg uugcgccgga uaaggagguc cuguaugagg cuuuugauga gauggaggaa       6840

ugcgccucua gggcggcucu caucgaagag gggcagcgga uagccgagau guugaagucc       6900

aagauccaag gcuugcugca gcaggccucu aagcaggccc aggacauaca acccgcuaug       6960

caggcuucau ggcccaaagu ggaacaauuu ugggccagac acauguggaa cuucauuagc       7020

ggcauccaau accucgcagg auugucaaca cugccaggga accccgcggu ggcuuccaug       7080

auggcauuca gugccgcccu caccaguccg uugucgacca guaccaccau ccuucucaac       7140

aucaugggag gcugguuagc gucccagauc gcaccacccg cgggggccac cggcuuuguc       7200

gucaguggcc uggugggggc ugccgugggc agcauaggcc uggguaaggu gcugguggac       7260

auccuggcag gauauggugc gggcauuucg ggggcccucg ucgcauucaa gaucaugucu       7320

ggcgagaagc ccucuaugga agaugucauc aaucuacugc cugggauccu gucuccggga       7380

gcccuggugg ugggggucau cugcgcggcc auucugcgcc gccacguggg accgggggag       7440

ggcgcggucc aauggaugaa caggcuuauu gccuuugcuu ccagaggaaa ccacgucgcc       7500

ccuacucacu acgugacgga gucggaugcg ucgcagcgug ugacccaacu acuuggcucu       7560

cuuacuauaa ccagccuacu cagaagacuc cacaauugga uaacugagga cugccccauc       7620

ccaugcuccg gauccuggcu ccgcgacgug ugggacuggg uuugcaccau cuugacagac       7680

uucaaaaauu ggcugaccuc uaaauuguuc cccaagcugc ccggccuccc cuucaucucu       7740

ugucaaaagg gguacaaggg ugugugggcc ggcacuggca ucaugaccac gcgcugcccu       7800

ugcggcgcca acaucucugg caauguccgc cugggcucua ugaggaucac agggccuaaa       7860

accugcauga acaccuggca ggggaccuuu ccuaucaauu gcuacacgga gggccagugc       7920

gcgccgaaac cccccacgaa cuacaagacc gccaucugga ggguggcggc cucggaguac       7980

gcggagguga cgcagcaugg gucguacucc uauguaacag gacugaccac ugacaaucug       8040

aaaauuccuu gccaacuacc uucuccagag uuuuucuccu ggguggacgg ugugcagauc       8100

cauagguuug cacccacacc aaagccguuu uuccgggaug aggucucguu cugcguuggg       8160

cuuaauuccu augcugucgg gucccagcuu cccugugaac cugagcccga cgcagacgua       8220

uugaggucca ugcuaacaga uccgccccac aucacggcgg agacugcggc gcggcgcuug       8280

gcacggggau caccuccauc ugaggcgagc uccucaguga gccagcuauc agcaccgucg       8340

cugcgggcca ccugcaccac ccacagcaac accuaugacg uggacauggu cgaugccaac       8400

cugcucaugg agggcggugu ggcucagaca gagccugagu ccagggugcc cguucuggac       8460

uuucucgagc caauggccga ggaagagagc gaccuugagc ccucaauacc aucggagugc       8520

augcucccca ggagcggguu uccacgggcc uuaccggcuu gggcacggcc ugacuacaac       8580

ccgccgcucg uggaaucgug gaggaggcca gauuaccaac cgcccaccgu ugcugguugu       8640

gcucuccccc cccccaagaa ggccccgacg ccucccccaa ggagacgccg gacagugggu       8700

cugagcgaga gcaccauauc agaagcccuc cagcaacugg ccaucaagac cuuuggccag       8760

ccccccucga gcggugaugc aggcucgucc acgggggcgg gcgccgccga auccggcggu       8820

ccgacguccc cuggugagcc ggcccccuca gagacagguu ccgccuccuc uaugcccccc       8880

cucgaggggg agccuggaga uccggaccug gagucugauc agguagagcu ucaaccuccc       8940

ccccaggggg gggggguagc ucccgguucg ggcucggggu cuuggucuac uugcuccgag       9000

gaggacgaua ccaccgugug cugcuccaug ucauacuccu ggaccggggc ucuaauaacu       9060

cccuguagcc ccgaagagga aaaguugcca aucaacccuu ugaguaacuc gcuguugcga       9120

uaccauaaca agguguacug uacaacauca aagagcgccu cacagagggc uaaaaaggua       9180

acuuuugaca ggacgcaagu gcucgacgcc cauuaugacu cagucuuaaa ggacaucaag       9240

cuagcggcuu ccaaggucag cgcaaggcuc cucaccuugg aggaggcgug ccaguugacu       9300

ccaccccauu cugcaagauc caaguaugga uucggggcca aggagguccg cagcuugucc       9360

gggagggccg uuaaccacau caaguccgug uggaaggacc uccuggaaga cccacaaaca       9420

ccaauuccca caaccaucau ggccaaaaau gagguguucu gcguggaccc cgccaagggg       9480

gguaagaaac cagcucgccu caucguuuac ccugaccucg gcguccgggu cugcgagaaa       9540

auggcccucu augacauuac acaaaagcuu ccucaggcgg uaaugggagc uuccuauggc       9600

uuccaguacu ccccugccca acggguggag uaucucuuga aagcaugggc ggaaaagaag       9660

gaccccaugg guuuuucgua ugauacccga ugcuucgacu caaccgucac ugagagagac       9720

aucaggaccg aggaguccau auaccaggcc ugcucccugc ccgaggaggc ccgcacugcc       9780

auacacucgc ugacugagag acuuuacgua ggagggccca uguucaacag caagggucaa       9840

accugcgguu acagacguug ccgcgccagc ggggugcuaa ccacuagcau ggguaacacc       9900

aucacaugcu augugaaagc ccuagcggcc ugcaaggcug cggggauagu ugcgcccaca       9960

augcugguau gcggcaauga ccuaguaguc aucucagaaa gccaggggac ugaggaggac       10020

gagcggaacc ugagagccuu cacggaggcc augaccaggu acucugcccc uccuggugau       10080

ccccccagac cggaauauga ccuggagcua auaacauccu guuccucaaa ugugucugug       10140

gcguugggcc cgcggggccg ccgcagauac uaccugacca gagacccaac cacuccacuc       10200

gcccgggcug ccugggaaac aguuagacac uccccuauca auucauggcu gggaaacauc       10260

auccaguaug cuccaaccau auggguucgc augguccuaa ugacacacuu cuucuccauu       10320

cucauggucc aagacacccu ggaccagaac cucaacuuug agauguaugg aucaguauac       10380

uccgugaauc cuuuggaccu uccagccaua auugagaggu uacacgggcu ugacgccuuu       10440

ucuaugcaca cauacucuca ccacgaacug acgcgggugg cuucagcccu cagaaaacuu       10500

ggggcgccac cccucagggu guggaagagu cgggcucgcg cagucagggc gucccucauc       10560

ucccguggag ggaaagcggc cguuugcggc cgauaucucu ucaauugggc ggugaagacc       10620

aagcucaaac ucacuccauu gccggaggcg cgccuacugg acuuauccag uugguucacc       10680

gucggcgccg gcgggggcga cauuuuucac agcgugucgc gcgcccgacc ccgcucauua       10740

cucuucggcc uacuccuacu uuucguaggg guaggccucu uccuacuccc cgcucgguag       10800

agcggcacac acuagguaca cuccauagcu aacuguuccu uuuuuuuuuu uuuuuuuuuu       10860

uuuuuuuuuu uuuuuuuuuu uucuuuuuuu uuuuuuuccc ucuuucuucc cuucucaucu       10920

uauucuacuu ucuuucuugg uggcuccauc uuagcccuag ucacggcuag cugugaaagg       10980

uccgugagcc gcaugacugc agagagugcc guaacugguc ucucugcagaucaugu            11036

<210>25

<211>11876

<212>RNA

<213>人工序列

<220>

<223>人工序列的描述：来源于表达载体rFGR-JFH1/SEAP的复制子RNA

<400>25

accugccccu aauaggggcg acacuccgcc augaaucacu ccccugugag gaacuacugu       60

cuucacgcag aaagcgccua gccauggcgu uaguaugagu gucguacagc cuccaggccc       120

cccccucccg ggagagccau aguggucugc ggaaccggug aguacaccgg aauugccggg       180

aagacugggu ccuuucuugg auaaacccac ucuaugcccg gccauuuggg cgugcccccg       240

caagacugcu agccgaguag cguuggguug cgaaaggccu ugugguacug ccugauaggg       300

cgcuugcgag ugccccggga ggucucguag accgugcacc augagcacaa auccuaaacc       360

ucaaagaaaa accaaaagaa acaccaaccg acgcguaaug cugcugcugc ugcugcugcu       420

gggccugagg cuacagcucu cccugggcau caucccaguu gaggaggaga acccggacuu       480

cuggaaccgc gaggcagccg aggcccuggg ugccgccaag aagcugcagc cugcacagac       540

agccgccaag aaccucauca ucuuccuggg cgaugggaug ggggugucua cggugacagc       600

ugccaggauc cuaaaagggc agaagaagga caaacugggg ccugagauac cccuggccau       660

ggaccgcuuc ccauaugugg cucuguccaa gacauacaau guagacaaac augugccaga       720

caguggagcc acagccacgg ccuaccugug cggggucaag ggcaacuucc agaccauugg       780

cuugagugca gccgcccgcu uuaaccagug caacacgaca cgcggcaacg aggucaucuc       840

cgugaugaau cgggccaaga aagcagggaa gucaguggga gugguaacca ccacacgagu       900

gcagcacgcc ucgccagccg gcaccuacgc ccacacggug aaccgcaacu gguacucgga       960

cgccgacgug ccugccucgg cccgccagga ggggugccag gacaucgcua cgcagcucau       1020

cuccaacaug gacauugacg ugauccuagg uggaggccga aaguacaugu uucgcauggg       1080

aaccccagac ccugaguacc cagaugacua cagccaaggu gggaccaggc uggacgggaa       1140

gaaucuggu gcaggaauggc uggcgaagcg ccagggugcc cgguaugugu ggaaccgcac       1200

ugagcucaug caggcuuccc uggacccguc ugugacccau cucauggguc ucuuugagcc       1260

uggagacaug aaauacgaga uccaccgaga cuccacacug gaccccuccc ugauggagau       1320

gacagaggcu gcccugcgcc ugcugagcag gaacccccgc ggcuucuucc ucuucgugga       1380

ggguggucgc aucgaccaug gucaucauga aagcagggcu uaccgggcac ugacugagac       1440

gaucauguuc gacgacgcca uugagagggc gggccagcuc accagcgagg aggacacgcu       1500

gagccucguc acugccgacc acucccacgu cuucuccuuc ggaggcuacc cccugcgagg       1560

gagcuccauc uucgggcugg ccccuggcaa ggcccgggac aggaaggccu acacgguccu       1620

ccuauacgga aacgguccag gcuaugugcu caaggacggc gcccggccgg auguuaccga       1680

gagcgagagc gggagccccg aguaucggca gcagucagca gugccccugg acgaagagac       1740

ccacgcaggc gaggacgugg cgguguucgc gcgcggcccg caggcgcacc ugguucacgg       1800

cgugcaggag cagaccuuca uagcgcacgu cauggccuuc gccgccugcc uggagcccua       1860

caccgccugc gaccuggcgc cccccgccgg caccaccgac gccgcgcacc cggguuacuc       1920

uagagucggg gcggccggcc gcuucgagca gacaugaguu uaaacccucu cccucccccc       1980

ccccuaacgu uacuggccga agccgcuugg aauaaggccg gugugcguuu gucuauaugu       2040

uauuuuccac cauauugccg ucuuuuggca augugagggc ccggaaaccu ggcccugucu       2100

ucuugacgag cauuccuagg ggucuuuccc cucucgccaa aggaaugcaa ggucuguuga       2160

augucgugaa ggaagcaguu ccucuggaag cuucuugaag acaaacaacg ucuguagcga       2220

cccuuugcag gcagcggaac cccccaccug gcgacaggug ccucugcggc caaaagccac       2280

guguauaaga uacaccugca aaggcggcac aaccccagug ccacguugug aguuggauag       2340

uuguggaaag agucaaaugg cucuccucaa gcguauucaa caaggggcug aaggaugccc       2400

agaagguacc ccauuguaug ggaucugauc uggggccucg gugcacaugc uuuacaugug       2460

uuuagucgag guuaaaaaaa cgucuaggcc ccccgaacca cggggacgug guuuuccuuu       2520

gaaaaacacg augauaccau gagcacaaau ccuaaaccuc aaagaaaaac caaaagaaac       2580

accaaccguc gcccagaaga cguuaaguuc ccgggcggcg gccagaucgu uggcggagua       2640

uacuuguugc cgcgcagggg ccccagguug ggugugcgca cgacaaggaa aacuucggag       2700

cggucccagc cacgugggag acgccagccc auccccaaag aucggcgcuc cacuggcaag       2760

gccuggggaa aaccaggucg ccccuggccc cuauauggga augagggacu cggcugggca       2820

ggauggcucc uguccccccg aggcucucgc cccuccuggg gccccacuga cccccggcau       2880

aggucgcgca acguggguaa agucaucgac acccuaacgu guggcuuugc cgaccucaug       2940

ggguacaucc ccgucguagg cgccccgcuu aguggcgccg ccagagcugu cgcgcacggc       3000

gugagagucc uggaggacgg gguuaauuau gcaacaggga accuacccgg uuuccccuuu       3060

ucuaucuucu ugcuggcccu guuguccugc aucaccguuc cggucucugc ugcccaggug       3120

aagaauacca guagcagcua cauggugacc aaugacugcu ccaaugacag caucacuugg       3180

cagcucgagg cugcgguucu ccacgucccc gggugcgucc cgugcgagag aguggggaau       3240

acgucacggu guugggugcc agucucgcca aacauggcug ugcggcagcc cggugcccuc       3300

acgcaggguc ugcggacgca caucgauaug guugugaugu ccgccaccuu cugcucugcu       3360

cucuacgugg gggaccucug uggcggggug augcucgcgg cccagguguu caucgucucg       3420

ccgcaguacc acugguuugu gcaagaaugc aauugcucca ucuacccugg caccaucacu       3480

ggacaccgca uggcauggga caugaugaug aacuggucgc ccacggccac caugauccug       3540

gcguacguga ugcgcguccc cgaggucauc auagacaucg uuagcggggc ucacuggggc       3600

gucauguucg gcuuggccua cuucucuaug cagggagcgu gggcgaaggu cauugucauc       3660

cuucugcugg ccgcuggggu ggacgcgggc accaccaccg uuggaggcgc uguugcacgu       3720

uccaccaacg ugauugccgg cguguucagc cauggcccuc agcagaacau ucagcucauu       3780

aacaccaacg gcaguuggca caucaaccgu acugccuuga auugcaauga cuccuugaac       3840

accggcuuuc ucgcggccuu guucuacacc aaccgcuuua acucgucagg guguccaggg       3900

cgccuguccg ccugccgcaa caucgaggcu uuccggauag gguggggcac ccuacaguac       3960

gaggauaaug ucaccaaucc agaggauaug aggccguacu gcuggcacua ccccccaaag       4020

ccguguggcg uaguccccgc gaggucugug uguggcccag uguacuguuu cacccccagc       4080

ccgguaguag ugggcacgac cgacagacgu ggagugccca ccuacacaug gggagagaau       4140

gagacagaug ucuuccuacu gaacagcacc cgaccgccgc agggcucaug guucggcugc       4200

acguggauga acuccacugg uuucaccaag acuuguggcg cgccaccuug ccgcaccaga       4260

gcugacuuca acgccagcac ggacuuguug ugcccuacgg auuguuuuag gaagcauccu       4320

gaugccacuu auauuaagug ugguucuggg cccuggcuca caccaaagug ccugguccac       4380

uacccuuaca gacucuggca uuaccccugc acagucaauu uuaccaucuu caagauaaga       4440

auguauguag gggggguuga gcacaggcuc acggccgcau gcaacuucac ucguggggau       4500

cgcugcgacu uggaggacag ggacaggagu cagcugucuc cucuguugca cucuaccacg       4560

gaaugggcca uccugcccug caccuacuca gacuuacccg cuuugucaac uggucuucuc       4620

caccuucacc agaacaucgu ggacguacaa uacauguaug gccucucacc ugcuaucaca       4680

aaauacgucg uucgauggga guggguggua cucuuauucc ugcucuuagc ggacgccaga       4740

gucugcgccu gcuuguggau gcucaucuug uugggccagg ccgaagcagc auuggagaag       4800

uuggucgucu ugcacgcugc gagugcggcu aacugccaug gccuccuaua uuuugccauc       4860

uucuucgugg cagcuuggca caucaggggu cggguggucc ccuugaccac cuauugccuc       4920

acuggccuau ggcccuucug ccuacugcuc auggcacugc cccggcaggc uuaugccuau       4980

gacgcaccug ugcacggaca gauaggcgug gguuuguuga uauugaucac ccucuucaca       5040

cucaccccgg gguauaagac ccuccucggc cagugucugu ggugguugug cuaucuccug       5100

acccuggggg aagccaugau ucaggagugg guaccaccca ugcaggugcg cggcggccgc       5160

gauggcaucg cgugggccgu cacuauauuc ugcccgggug ugguguuuga cauuaccaaa       5220

uggcuuuugg cguugcuugg gccugcuuac cucuuaaggg ccgcuuugac acaugugccg       5280

uacuucguca gagcucacgc ucugauaagg guaugcgcuu uggugaagca gcucgcgggg       5340

gguagguaug uucagguggc gcuauuggcc cuuggcaggu ggacuggcac cuacaucuau       5400

gaccaccuca caccuauguc ggacugggcc gcuagcggcc ugcgcgacuu agcggucgcc       5460

guggaaccca ucaucuucag uccgauggag aagaagguca ucgucugggg agcggagacg       5520

gcugcaugug gggacauucu acauggacuu cccguguccg cccgacucgg ccaggagauc       5580

cuccucggcc cagcugaugg cuacaccucc aaggggugga agcuccuugc ucccaucacu       5640

gcuuaugccc agcaaacacg aggccuccug ggcgccauag uggugaguau gacggggcgu       5700

gacaggacag aacaggccgg ggaaguccaa auccugucca cagucucuca guccuuccuc       5760

ggaacaacca ucucgggggu uuuguggacu guuuaccacg gagcuggcaa caagacucua       5820

gccggcuuac gggguccggu cacgcagaug uacucgagug cugaggggga cuugguaggc       5880

uggcccagcc ccccugggac caagucuuug gagccgugca aguguggagc cgucgaccua       5940

uaucugguca cgcggaacgc ugaugucauc ccggcucgga gacgcgggga caagcgggga       6000

gcauugcucu ccccgagacc cauuucgacc uugaaggggu ccucgggggg gccggugcuc       6060

ugcccuaggg gccacgucgu ugggcucuuc cgagcagcug ugugcucucg gggcguggcc       6120

aaauccaucg auuucauccc cguugagaca cucgacguug uuacaagguc ucccacuuuc       6180

agugacaaca gcacgccacc ggcugugccc cagaccuauc aggucgggua cuugcaugcu       6240

ccaacuggca guggaaagag caccaagguc ccugucgcgu augccgccca gggguacaaa       6300

guacuagugc uuaaccccuc gguagcugcc acccuggggu uuggggcgua ccuauccaag       6360

gcacauggca ucaaucccaa cauuaggacu ggagucagga ccgugaugac cggggaggcc       6420

aucacguacu ccacauaugg caaauuucuc gccgaugggg gcugcgcuag cggcgccuau       6480

gacaucauca uaugcgauga augccacgcu guggaugcua ccuccauucu cggcaucgga       6540

acgguccuug aucaagcaga gacagccggg gucagacuaa cugugcuggc uacggccaca       6600

ccccccgggu cagugacaac cccccauccc gauauagaag agguaggccu cgggcgggag       6660

ggugagaucc ccuucuaugg gagggcgauu ccccuauccu gcaucaaggg agggagacac       6720

cugauuuucu gccacucaaa gaaaaagugu gacgagcucg cggcggcccu ucggggcaug       6780

ggcuugaaug ccguggcaua cuauagaggg uuggacgucu ccauaauacc agcucaggga       6840

gauguggugg ucgucgccac cgacgcccuc augacggggu acacuggaga cuuugacucc       6900

gugaucgacu gcaauguagc ggucacccaa gcugucgacu ucagccugga ccccaccuuc       6960

acuauaacca cacagacugu cccacaagac gcugucucac gcagucagcg ccgcgggcgc       7020

acagguagag gaagacaggg cacuuauagg uauguuucca cuggugaacg agccucagga       7080

auguuugaca guguagugcu uugugagugc uacgacgcag gggcugcgug guacgaucuc       7140

acaccagcgg agaccaccgu caggcuuaga gcguauuuca acacgcccgg ccuacccgug       7200

ugucaagacc aucuugaauu uugggaggca guuuucaccg gccucacaca cauagacgcc       7260

cacuuccucu cccaaacaaa gcaagcgggg gagaacuucg cguaccuagu agccuaccaa       7320

gcuacggugu gcgccagagc caaggccccu cccccguccu gggacgccau guggaagugc       7380

cuggcccgac ucaagccuac gcuugcgggc cccacaccuc uccuguaccg uuugggcccu       7440

auuaccaaug aggucacccu cacacacccu gggacgaagu acaucgccac augcaugcaa       7500

gcugaccuug aggucaugac cagcacgugg guccuagcug gaggaguccu ggcagccguc       7560

gccgcauauu gccuggcgac uggaugcguu uccaucaucg gccgcuugca cgucaaccag       7620

cgagucgucg uugcgccgga uaaggagguc cuguaugagg cuuuugauga gauggaggaa       7680

ugcgccucua gggcggcucu caucgaagag gggcagcgga uagccgagau guugaagucc       7740

aagauccaag gcuugcugca gcaggccucu aagcaggccc aggacauaca acccgcuaug       7800

caggcuucau ggcccaaagu ggaacaauuu ugggccagac acauguggaa cuucauuagc       7860

ggcauccaau accucgcagg auugucaaca cugccaggga accccgcggu ggcuuccaug       7920

auggcauuca gugccgcccu caccaguccg uugucgacca guaccaccau ccuucucaac       7980

aucaugggag gcugguuagc gucccagauc gcaccacccg cgggggccac cggcuuuguc       8040

gucaguggcc uggugggggc ugccgugggc agcauaggcc uggguaaggu gcugguggac       8100

auccuggcag gauauggugc gggcauuucg ggggcccucg ucgcauucaa gaucaugucu       8160

ggcgagaagc ccucuaugga agaugucauc aaucuacugc cugggauccu gucuccggga       8220

gcccuggugg ugggggucau cugcgcggcc auucugcgcc gccacguggg accgggggag       8280

ggcgcggucc aauggaugaa caggcuuauu gccuuugcuu ccagaggaaa ccacgucgcc       8340

ccuacucacu acgugacgga gucggaugcg ucgcagcgug ugacccaacu acuuggcucu       8400

cuuacuauaa ccagccuacu cagaagacuc cacaauugga uaacugagga cugccccauc       8460

ccaugcuccg gauccuggcu ccgcgacgug ugggacuggg uuugcaccau cuugacagac       8520

uucaaaaauu ggcugaccuc uaaauuguuc cccaagcugc ccggccuccc cuucaucucu       8580

ugucaaaagg gguacaaggg ugugugggcc ggcacuggca ucaugaccac gcgcugcccu       8640

ugcggcgcca acaucucugg caauguccgc cugggcucua ugaggaucac agggccuaaa       8700

accugcauga acaccuggca ggggaccuuu ccuaucaauu gcuacacgga gggccagugc       8760

gcgccgaaac cccccacgaa cuacaagacc gccaucugga ggguggcggc cucggaguac       8820

gcggagguga cgcagcaugg gucguacucc uauguaacag gacugaccac ugacaaucug       8880

aaaauuccuu gccaacuacc uucuccagag uuuuucuccu ggguggacgg ugugcagauc       8940

cauagguuug cacccacacc aaagccguuu uuccgggaug aggucucguu cugcguuggg       9000

cuuaauuccu augcugucgg gucccagcuu cccugugaac cugagcccga cgcagacgua       9060

uugaggucca ugcuaacaga uccgccccac aucacggcgg agacugcggc gcggcgcuug       9120

gcacggggau caccuccauc ugaggcgagc uccucaguga gccagcuauc agcaccgucg       9180

cugcgggcca ccugcaccac ccacagcaac accuaugacg uggacauggu cgaugccaac       9240

cugcucaugg agggcggugu ggcucagaca gagccugagu ccagggugcc cguucuggac       9300

uuucucgagc caauggccga ggaagagagc gaccuugagc ccucaauacc aucggagugc       9360

augcucccca ggagcggguu uccacgggcc uuaccggcuu gggcacggcc ugacuacaac       9420

ccgccgcucg uggaaucgug gaggaggcca gauuaccaac cgcccaccgu ugcugguugu       9480

gcucuccccc cccccaagaa ggccccgacg ccucccccaa ggagacgccg gacagugggu       9540

cugagcgaga gcaccauauc agaagcccuc cagcaacugg ccaucaagac cuuuggccag       9600

ccccccucga gcggugaugc aggcucgucc acgggggcgg gcgccgccga auccggcggu       9660

ccgacguccc cuggugagcc ggcccccuca gagacagguu ccgccuccuc uaugcccccc       9720

cucgaggggg agccuggaga uccggaccug gagucugauc agguagagcu ucaaccuccc       9780

ccccaggggg gggggguagc ucccgguucg ggcucggggu cuuggucuac uugcuccgag       9840

gaggacgaua ccaccgugug cugcuccaug ucauacuccu ggaccggggc ucuaauaacu       9900

cccuguagcc ccgaagagga aaaguugcca aucaacccuu ugaguaacuc gcuguugcga       9960

uaccauaaca agguguacug uacaacauca aagagcgccu cacagagggc uaaaaaggua       10020

acuuuugaca ggacgcaagu gcucgacgcc cauuaugacu cagucuuaaa ggacaucaag       10080

cuagcggcuu ccaaggucag cgcaaggcuc cucaccuugg aggaggcgug ccaguugacu       10140

ccaccccauu cugcaagauc caaguaugga uucggggcca aggagguccg cagcuugucc       10200

gggagggccg uuaaccacau caaguccgug uggaaggacc uccuggaaga cccacaaaca       10260

ccaauuccca caaccaucau ggccaaaaau gagguguucu gcguggaccc cgccaagggg       10320

gguaagaaac cagcucgccu caucguuuac ccugaccucg gcguccgggu cugcgagaaa       10380

auggcccucu augacauuac acaaaagcuu ccucaggcgg uaaugggagc uuccuauggc       10440

uuccaguacu ccccugccca acggguggag uaucucuuga aagcaugggc ggaaaagaag       10500

gaccccaugg guuuuucgua ugauacccga ugcuucgacu caaccgucac ugagagagac       10560

aucaggaccg aggaguccau auaccaggcc ugcucccugc ccgaggaggc ccgcacugcc       10620

auacacucgc ugacugagag acuuuacgua ggagggccca uguucaacag caagggucaa       10680

accugcgguu acagacguug ccgcgccagc ggggugcuaa ccacuagcau ggguaacacc       10740

aucacaugcu augugaaagc ccuagcggcc ugcaaggcug cggggauagu ugcgcccaca       10800

augcugguau gcggcgauga ccuaguaguc aucucagaaa gccaggggac ugaggaggac       10860

gagcggaacc ugagagccuu cacggaggcc augaccaggu acucugcccc uccuggugau       10920

ccccccagac cggaauauga ccuggagcua auaacauccu guuccucaaa ugugucugug       10980

gcguugggcc cgcggggccg ccgcagauac uaccugacca gagacccaac cacuccacuc       11040

gcccgggcug ccugggaaac aguuagacac uccccuauca auucauggcu gggaaacauc       11100

auccaguaug cuccaaccau auggguucgc augguccuaa ugacacacuu cuucuccauu       11160

cucauggucc aagacacccu ggaccagaac cucaacuuug agauguaugg aucaguauac       11220

uccgugaauc cuuuggaccu uccagccaua auugagaggu uacacgggcu ugacgccuuu       11280

ucuaugcaca cauacucuca ccacgaacug acgcgggugg cuucagcccu cagaaaacuu       11340

ggggcgccac cccucagggu guggaagagu cgggcucgcg cagucagggc gucccucauc       11400

ucccguggag ggaaagcggc cguuugcggc cgauaucucu ucaauugggc ggugaagacc       11460

aagcucaaac ucacuccauu gccggaggcg cgccuacugg acuuauccag uugguucacc       11520

gucggcgccg gcgggggcga cauuuuucac agcgugucgc gcgcccgacc ccgcucauua       11580

cucuucggcc uacuccuacu uuucguaggg guaggccucu uccuacuccc cgcucgguag       11640

agcggcacac acuagguaca cuccauagcu aacuguuccu uuuuuuuuuu uuuuuuuuuu       11700

uuuuuuuuuu uuuuuuuuuu uucuuuuuuu uuuuuuuccc ucuuucuucc cuucucaucu       11760

uauucuacuu ucuuucuugg uggcuccauc uuagcccuag ucacggcuag cugugaaagg       11820

uccgugagcc gcaugacugc agagagugcc guaacugguc ucucugcaga ucaugu           11876

<210>26

<211>11876

<212>RNA

<213>人工序列

<220>

<223>人工序列的描述：来源于表达载体rFGR-JFH1/sEAP/GND的复制子RNA

<400>26

accugccccu aauaggggcg acacuccgcc augaaucacu ccccugugag gaacuacugu       60

cuucacgcag aaagcgccua gccauggcgu uaguaugagu gucguacagc cuccaggccc       120

cccccucccg ggagagccau aguggucugc ggaaccggug aguacaccgg aauugccggg       180

aagacugggu ccuuucuugg auaaacccac ucuaugcccg gccauuuggg cgugcccccg       240

caagacugcu agccgaguag cguuggguug cgaaaggccu ugugguacug ccugauaggg       300

cgcuugcgag ugccccggga ggucucguag accgugcacc augagcacaa auccuaaacc       360

ucaaagaaaa accaaaagaa acaccaaccg acgcguaaug cugcugcugc ugcugcugcu       420

gggccugagg cuacagcucu cccugggcau caucccaguu gaggaggaga acccggacuu       480

cuggaaccgc gaggcagccg aggcccuggg ugccgccaag aagcugcagc cugcacagac       540

agccgccaag aaccucauca ucuuccuggg cgaugggaug ggggugucua cggugacagc       600

ugccaggauc cuaaaagggc agaagaagga caaacugggg ccugagauac cccuggccau       660

ggaccgcuuc ccauaugugg cucuguccaa gacauacaau guagacaaac augugccaga       720

caguggagcc acagccacgg ccuaccugug cggggucaag ggcaacuucc agaccauugg       780

cuugagugca gccgcccgcu uuaaccagug caacacgaca cgcggcaacg aggucaucuc       840

cgugaugaau cgggccaaga aagcagggaa gucaguggga gugguaacca ccacacgagu       900

gcagcacgcc ucgccagccg gcaccuacgc ccacacggug aaccgcaacu gguacucgga       960

cgccgacgug ccugccucgg cccgccagga ggggugccag gacaucgcua cgcagcucau       1020

cuccaacaug gacauugacg ugauccuagg uggaggccga aaguacaugu uucgcauggg       1080

aaccccagac ccugaguacc cagaugacua cagccaaggu gggaccaggc uggacgggaa       1140

gaaucuggug caggaauggc uggcgaagcg ccagggugcc cgguaugugu ggaaccgcac       1200

ugagcucaug caggcuuccc uggacccguc ugugacccau cucauggguc ucuuugagcc       1260

uggagacaug aaauacgaga uccaccgaga cuccacacug gaccccuccc ugauggagau       1320

gacagaggcu gcccugcgcc ugcugagcag gaacccccgc ggcuucuucc ucuucgugga       1380

ggguggucgc aucgaccaug gucaucauga aagcagggcu uaccgggcac ugacugagac       1440

gaucauguuc gacgacgcca uugagagggc gggccagcuc accagcgagg aggacacgcu       1500

gagccucguc acugccgacc acucccacgu cuucuccuuc ggaggcuacc cccugcgagg       1560

gagcuccauc uucgggcugg ccccuggcaa ggcccgggac aggaaggccu acacgguccu       1620

ccuauacgga aacgguccag gcuaugugcu caaggacggc gcccggccgg auguuaccga       1680

gagcgagagc gggagccccg aguaucggca gcagucagca gugccccugg acgaagagac       1740

ccacgcaggc gaggacgugg cgguguucgc gcgcggcccg caggcgcacc ugguucacgg       1800

cgugcaggag cagaccuuca uagcgcacgu cauggccuuc gccgccugcc uggagcccua       1860

caccgccugc gaccuggcgc cccccgccgg caccaccgac gccgcgcacc cggguuacuc       1920

uagagucggg gcggccggcc gcuucgagca gacaugaguu uaaacccucu cccucccccc       1980

ccccuaacgu uacuggccga agccgcuugg aauaaggccg gugugcguuu gucuauaugu       2040

uauuuuccac cauauugccg ucuuuuggca augugagggc ccggaaaccu ggcccugucu       2100

ucuugacgag cauuccuagg ggucuuuccc cucucgccaa aggaaugcaa ggucuguuga       2160

augucgugaa ggaagcaguu ccucuggaag cuucuugaag acaaacaacg ucuguagcga       2220

cccuuugcag gcagcggaac cccccaccug gcgacaggug ccucugcggc caaaagccac       2280

guguauaaga uacaccugca aaggcggcac aaccccagug ccacguugug aguuggauag       2340

uuguggaaag agucaaaugg cucuccucaa gcguauucaa caaggggcug aaggaugccc       2400

agaagguacc ccauuguaug ggaucugauc uggggccucg gugcacaugc uuuacaugug       2460

uuuagucgag guuaaaaaaa cgucuaggcc ccccgaacca cggggacgug guuuuccuuu       2520

gaaaaacacg augauaccau gagcacaaau ccuaaaccuc aaagaaaaac caaaagaaac       2580

accaaccguc gcccagaaga cguuaaguuc ccgggcggcg gccagaucgu uggcggagua       2640

uacuuguugc cgcgcagggg ccccagguug ggugugcgca cgacaaggaa aacuucggag       2700

cggucccagc cacgugggag acgccagccc auccccaaag aucggcgcuc cacuggcaag       2760

gccuggggaa aaccaggucg ccccuggccc cuauauggga augagggacu cggcugggca       2820

ggauggcucc uguccccccg aggcucucgc cccuccuggg gccccacuga cccccggcau       2880

aggucgcgca acguggguaa agucaucgac acccuaacgu guggcuuugc cgaccucaug       2940

ggguacaucc ccgucguagg cgccccgcuu aguggcgccg ccagagcugu cgcgcacggc       3000

gugagagucc uggaggacgg gguuaauuau gcaacaggga accuacccgg uuuccccuuu       3060

ucuaucuucu ugcuggcccu guuguccugc aucaccguuc cggucucugc ugcccaggug       3120

aagaauacca guagcagcua cauggugacc aaugacugcu ccaaugacag caucacuugg       3180

cagcucgagg cugcgguucu ccacgucccc gggugcgucc cgugcgagag aguggggaau       3240

acgucacggu guugggugcc agucucgcca aacauggcug ugcggcagcc cggugcccuc       3300

acgcaggguc ugcggacgca caucgauaug guugugaugu ccgccaccuu cugcucugcu       3360

cucuacgugg gggaccucug uggcggggug augcucgcgg cccagguguu caucgucucg       3420

ccgcaguacc acugguuugu gcaagaaugc aauugcucca ucuacccugg caccaucacu       3480

ggacaccgca uggcauggga caugaugaug aacuggucgc ccacggccac caugauccug       3540

gcguacguga ugcgcguccc cgaggucauc auagacaucg uuagcggggc ucacuggggc       3600

gucauguucg gcuuggccua cuucucuaug cagggagcgu gggcgaaggu cauugucauc       3660

cuucugcugg ccgcuggggu ggacgcgggc accaccaccg uuggaggcgc uguugcacgu       3720

uccaccaacg ugauugccgg cguguucagc cauggcccuc agcagaacau ucagcucauu       3780

aacaccaacg gcaguuggca caucaaccgu acugccuuga auugcaauga cuccuugaac       3840

accggcuuuc ucgcggccuu guucuacacc aaccgcuuua acucgucagg guguccaggg       3900

cgccuguccg ccugccgcaa caucgaggcu uuccggauag gguggggcac ccuacaguac       3960

gaggauaaug ucaccaaucc agaggauaug aggccguacu gcuggcacua ccccccaaag       4020

ccguguggcg uaguccccgc gaggucugug uguggcccag uguacuguuu cacccccagc       4080

ccgguaguag ugggcacgac cgacagacgu ggagugccca ccuacacaug gggagagaau       4140

gagacagaug ucuuccuacu gaacagcacc cgaccgccgc agggcucaug guucggcugc       4200

acguggauga acuccacugg uuucaccaag acuuguggcg cgccaccuug ccgcaccaga       4260

gcugacuuca acgccagcac ggacuuguug ugcccuacgg auuguuuuag gaagcauccu       4320

gaugccacuu auauuaagug ugguucuggg cccuggcuca caccaaagug ccugguccac       4380

uacccuuaca gacucuggca uuaccccugc acagucaauu uuaccaucuu caagauaaga       4440

auguauguag gggggguuga gcacaggcuc acggccgcau gcaacuucac ucguggggau       4500

cgcugcgacu uggaggacag ggacaggagu cagcugucuc cucuguugca cucuaccacg       4560

gaaugggcca uccugcccug caccuacuca gacuuacccg cuuugucaac uggucuucuc       4620

caccuucacc agaacaucgu ggacguacaa uacauguaug gccucucacc ugcuaucaca       4680

aaauacgucg uucgauggga guggguggua cucuuauucc ugcucuuagc ggacgccaga       4740

gucugcgccu gcuuguggau gcucaucuug uugggccagg ccgaagcagc auuggagaag       4800

uuggucgucu ugcacgcugc gagugcggcu aacugccaug gccuccuaua uuuugccauc       4860

uucuucgugg cagcuuggca caucaggggu cggguggucc ccuugaccac cuauugccuc       4920

acuggccuau ggcccuucug ccuacugcuc auggcacugc cccggcaggc uuaugccuau       4980

gacgcaccug ugcacggaca gauaggcgug gguuuguuga uauugaucac ccucuucaca       5040

cucaccccgg gguauaagac ccuccucggc cagugucugu ggugguugug cuaucuccug       5100

acccuggggg aagccaugau ucaggagugg guaccaccca ugcaggugcg cggcggccgc       5160

gauggcaucg cgugggccgu cacuauauuc ugcccgggug ugguguuuga cauuaccaaa       5220

uggcuuuugg cguugcuugg gccugcuuac cucuuaaggg ccgcuuugac acaugugccg       5280

uacuucguca gagcucacgc ucugauaagg guaugcgcuu uggugaagca gcucgcgggg       5340

gguagguaug uucagguggc gcuauuggcc cuuggcaggu ggacuggcac cuacaucuau       5400

gaccaccuca caccuauguc ggacugggcc gcuagcggcc ugcgcgacuu agcggucgcc       5460

guggaaccca ucaucuucag uccgauggag aagaagguca ucgucugggg agcggagacg       5520

gcugcaugug gggacauucu acauggacuu cccguguccg cccgacucgg ccaggagauc       5580

cuccucggcc cagcugaugg cuacaccucc aaggggugga agcuccuugc ucccaucacu       5640

gcuuaugccc agcaaacacg aggccuccug ggcgccauag uggugaguau gacggggcgu       5700

gacaggacag aacaggccgg ggaaguccaa auccugucca cagucucuca guccuuccuc       5760

ggaacaacca ucucgggggu uuuguggacu guuuaccacg gagcuggcaa caagacucua       5820

gccggcuuac gggguccggu cacgcagaug uacucgagug cugaggggga cuugguaggc       5880

uggcccagcc ccccugggac caagucuuug gagccgugca aguguggagc cgucgaccua       5940

uaucugguca cgcggaacgc ugaugucauc ccggcucgga gacgcgggga caagcgggga       6000

gcauugcucu ccccgagacc cauuucgacc uugaaggggu ccucgggggg gccggugcuc       6060

ugcccuaggg gccacgucgu ugggcucuuc cgagcagcug ugugcucucg gggcguggcc       6120

aaauccaucg auuucauccc cguugagaca cucgacguug uuacaagguc ucccacuuuc       6180

agugacaaca gcacgccacc ggcugugccc cagaccuauc aggucgggua cuugcaugcu       6240

ccaacuggca guggaaagag caccaagguc ccugucgcgu augccgccca gggguacaaa       6300

guacuagugc uuaaccccuc gguagcugcc acccuggggu uuggggcgua ccuauccaag       6360

gcacauggca ucaaucccaa cauuaggacu ggagucagga ccgugaugac cggggaggcc       6420

aucacguacu ccacauaugg caaauuucuc gccgaugggg gcugcgcuag cggcgccuau       6480

gacaucauca uaugcgauga augccacgcu guggaugcua ccuccauucu cggcaucgga       6540

acgguccuug aucaagcaga gacagccggg gucagacuaa cugugcuggc uacggccaca       6600

ccccccgggu cagugacaac cccccauccc gauauagaag agguaggccu cgggcgggag       6660

ggugagaucc ccuucuaugg gagggcgauu ccccuauccu gcaucaaggg agggagacac       6720

cugauuuucu gccacucaaa gaaaaagugu gacgagcucg cggcggcccu ucggggcaug       6780

ggcuugaaug ccguggcaua cuauagaggg uuggacgucu ccauaauacc agcucaggga       6840

gauguggugg ucgucgccac cgacgcccuc augacggggu acacuggaga cuuugacucc       6900

gugaucgacu gcaauguagc ggucacccaa gcugucgacu ucagccugga ccccaccuuc       6960

acuauaacca cacagacugu cccacaagac gcugucucac gcagucagcg ccgcgggcgc       7020

acagguagag gaagacaggg cacuuauagg uauguuucca cuggugaacg agccucagga       7080

auguuugaca guguagugcu uugugagugc uacgacgcag gggcugcgug guacgaucuc       7140

acaccagcgg agaccaccgu caggcuuaga gcguauuuca acacgcccgg ccuacccgug       7200

ugucaagacc aucuugaauu uugggaggca guuuucaccg gccucacaca cauagacgcc       7260

cacuuccucu cccaaacaaa gcaagcgggg gagaacuucg cguaccuagu agccuaccaa       7320

gcuacggugu gcgccagagc caaggccccu cccccguccu gggacgccau guggaagugc       7380

cuggcccgac ucaagccuac gcuugcgggc cccacaccuc uccuguaccg uuugggcccu       7440

auuaccaaug aggucacccu cacacacccu gggacgaagu acaucgccac augcaugcaa       7500

gcugaccuug aggucaugac cagcacgugg guccuagcug gaggaguccu ggcagccguc       7560

gccgcauauu gccuggcgac uggaugcguu uccaucaucg gccgcuugca cgucaaccag       7620

cgagucgucg uugcgccgga uaaggagguc cuguaugagg cuuuugauga gauggaggaa       7680

ugcgccucua gggcggcucu caucgaagag gggcagcgga uagccgagau guugaagucc       7740

aagauccaag gcuugcugca gcaggccucu aagcaggccc aggacauaca acccgcuaug       7800

caggcuucau ggcccaaagu ggaacaauuu ugggccagac acauguggaa cuucauuagc       7860

ggcauccaau accucgcagg auugucaaca cugccaggga accccgcggu ggcuuccaug       7920

auggcauuca gugccgcccu caccaguccg uugucgacca guaccaccau ccuucucaac       7980

aucaugggag gcugguuagc gucccagauc gcaccacccg cgggggccac cggcuuuguc       8040

gucaguggcc uggugggggc ugccgugggc agcauaggcc uggguaaggu gcugguggac       8100

auccuggcag gauauggugc gggcauuucg ggggcccucg ucgcauucaa gaucaugucu       8160

ggcgagaagc ccucuaugga agaugucauc aaucuacugc cugggauccu gucuccggga       8220

gcccuggugg ugggggucau cugcgcggcc auucugcgcc gccacguggg accgggggag       8280

ggcgcggucc aauggaugaa caggcuuauu gccuuugcuu ccagaggaaa ccacgucgcc       8340

ccuacucacu acgugacgga gucggaugcg ucgcagcgug ugacccaacu acuuggcucu       8400

cuuacuauaa ccagccuacu cagaagacuc cacaauugga uaacugagga cugccccauc       8460

ccaugcuccg gauccuggcu ccgcgacgug ugggacuggg uuugcaccau cuugacagac       8520

uucaaaaauu ggcugaccuc uaaauuguuc cccaagcugc ccggccuccc cuucaucucu       8580

ugucaaaagg gguacaaggg ugugugggcc ggcacuggca ucaugaccac gcgcugcccu       8640

ugcggcgcca acaucucugg caauguccgc cugggcucua ugaggaucac agggccuaaa       8700

accugcauga acaccuggca ggggaccuuu ccuaucaauu gcuacacgga gggccagugc       8760

gcgccgaaac cccccacgaa cuacaagacc gccaucugga ggguggcggc cucggaguac       8820

gcggagguga cgcagcaugg gucguacucc uauguaacag gacugaccac ugacaaucug       8880

aaaauuccuu gccaacuacc uucuccagag uuuuucuccu ggguggacgg ugugcagauc       8940

cauagguuug cacccacacc aaagccguuu uuccgggaug aggucucguu cugcguuggg       9000

cuuaauuccu augcugucgg gucccagcuu cccugugaac cugagcccga cgcagacgua       9060

uugaggucca ugcuaacaga uccgccccac aucacggcgg agacugcggc gcggcgcuug       9120

gcacggggau caccuccauc ugaggcgagc uccucaguga gccagcuauc agcaccgucg       9180

cugcgggcca ccugcaccac ccacagcaac accuaugacg uggacauggu cgaugccaac       9240

cugcucaugg agggcggugu ggcucagaca gagccugagu ccagggugcc cguucuggac       9300

uuucucgagc caauggccga ggaagagagc gaccuugagc ccucaauacc aucggagugc       9360

augcucccca ggagcggguu uccacgggcc uuaccggcuu gggcacggcc ugacuacaac       9420

ccgccgcucg uggaaucgug gaggaggcca gauuaccaac cgcccaccgu ugcugguugu       9480

gcucuccccc cccccaagaa ggccccgacg ccucccccaa ggagacgccg gacagugggu       9540

cugagcgaga gcaccauauc agaagcccuc cagcaacugg ccaucaagac cuuuggccag       9600

ccccccucga gcggugaugc aggcucgucc acgggggcgg gcgccgccga auccggcggu       9660

ccgacguccc cuggugagcc ggcccccuca gagacagguu ccgccuccuc uaugcccccc       9720

cucgaggggg agccuggaga uccggaccug gagucugauc agguagagcu ucaaccuccc       9780

ccccaggggg gggggguagc ucccgguucg ggcucggggu cuuggucuac uugcuccgag       9840

gaggacgaua ccaccgugug cugcuccaug ucauacuccu ggaccggggc ucuaauaacu       9900

cccuguagcc ccgaagagga aaaguugcca aucaacccuu ugaguaacuc gcuguugcga       9960

uaccauaaca agguguacug uacaacauca aagagcgccu cacagagggc uaaaaaggua       10020

acuuuugaca ggacgcaagu gcucgacgcc cauuaugacu cagucuuaaa ggacaucaag       10080

cuagcggcuu ccaaggucag cgcaaggcuc cucaccuugg aggaggcgug ccaguugacu       10140

ccaccccauu cugcaagauc caaguaugga uucggggcca aggagguccg cagcuugucc       10200

gggagggccg uuaaccacau caaguccgug uggaaggacc uccuggaaga cccacaaaca       10260

ccaauuccca caaccaucau ggccaaaaau gagguguucu gcguggaccc cgccaagggg       10320

gguaagaaac cagcucgccu caucguuuac ccugaccucg gcguccgggu cugcgagaaa       10380

auggcccucu augacauuac acaaaagcuu ccucaggcgg uaaugggagc uuccuauggc       10440

uuccaguacu ccccugccca acggguggag uaucucuuga aagcaugggc ggaaaagaag       10500

gaccccaugg guuuuucgua ugauacccga ugcuucgacu caaccgucac ugagagagac       10560

aucaggaccg aggaguccau auaccaggcc ugcucccugc ccgaggaggc ccgcacugcc       10620

auacacucgc ugacugagag acuuuacgua ggagggccca uguucaacag caagggucaa       10680

accugcgguu acagacguug ccgcgccagc ggggugcuaa ccacuagcau ggguaacacc       10740

aucacaugcu augugaaagc ccuagcggcc ugcaaggcug cggggauagu ugcgcccaca       10800

augcugguau gcggcaauga ccuaguaguc aucucagaaa gccaggggac ugaggaggac       10860

gagcggaacc ugagagccuu cacggaggcc augaccaggu acucugcccc uccuggugau       10920

ccccccagac cggaauauga ccuggagcua auaacauccu guuccucaaa ugugucugug       10980

gcguugggcc cgcggggccg ccgcagauac uaccugacca gagacccaac cacuccacuc       11040

gcccgggcug ccugggaaac aguuagacac uccccuauca auucauggcu gggaaacauc       11100

auccaguaug cuccaaccau auggguucgc augguccuaa ugacacacuu cuucuccauu       11160

cucauggucc aagacacccu ggaccagaac cucaacuuug agauguaugg aucaguauac       11220

uccgugaauc cuuuggaccu uccagccaua auugagaggu uacacgggcu ugacgccuuu       11280

ucuaugcaca cauacucuca ccacgaacug acgcgggugg cuucagcccu cagaaaacuu       11340

ggggcgccac cccucagggu guggaagagu cgggcucgcg cagucagggc gucccucauc       11400

ucccguggag ggaaagcggc cguuugcggc cgauaucucu ucaauugggc ggugaagacc       11460

aagcucaaac ucacuccauu gccggaggcg cgccuacugg acuuauccag uugguucacc       11520

gucggcgccg gcgggggcga cauuuuucac agcgugucgc gcgcccgacc ccgcucauua       11580

cucuucggcc uacuccuacu uuucguaggg guaggccucu uccuacuccc cgcucgguag       11640

agcggcacac acuagguaca cuccauagcu aacuguuccu uuuuuuuuuu uuuuuuuuuu       11700

uuuuuuuuuu uuuuuuuuuu uucuuuuuuu uuuuuuuccc ucuuucuucc cuucucaucu       11760

uauucuacuu ucuuucuugg uggcuccauc uuagcccuag ucacggcuag cugugaaagg       11820

uccgugagcc gcaugacugc agagagugcc guaacugguc ucucugcaga ucaugu           11876