CN112011570B - 特异杀伤肿瘤细胞的溶瘤病毒系统及其应用 - Google Patents
特异杀伤肿瘤细胞的溶瘤病毒系统及其应用 Download PDFInfo
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Abstract
本发明提出了一种表达系统。根据本发明的实施例的表达系统在胃癌、胰腺癌细胞中的翻转能力高，具有该表达系统的表达载体，如腺病毒载体等，对胃癌、胰腺癌细胞的特异性杀伤效率高。
Description
技术领域
本发明涉及生物医药领域，具体地，本发明涉及特异杀伤肿瘤细胞的溶瘤病毒系统及其应用，更具体地，本发明涉及表达系统、重组病毒、重组细胞以及表达系统、重组病毒、重组细胞在制备药物中的用途以及药物组合物。
背景技术
溶瘤病毒是指一类具有复制与包装能力实现肿瘤杀伤作用的病毒。目前，多数研究通过改造一些自然界存在的致病力较弱的毒种，使其特异的在肿瘤细胞中表达包装继而实现溶瘤作用。利用溶瘤病毒识别肿瘤细胞的原理主要有两种：第一，利用靶细胞中抑癌基因的失活或缺陷从而选择性地感染肿瘤细胞；第二，选择利用肿瘤特异性的启动子调控病毒关键基因的表达使溶瘤病毒在肿瘤细胞内大量复制并表达毒性蛋白摧毁肿瘤细胞，并/或同时分泌细胞因子刺激自身免疫系统攻击肿瘤细胞。相应的溶瘤病毒无法在正常机体细胞内复制而不具有杀伤作用，因此溶瘤病毒具有更高的抗肿瘤效应和更低的副作用。近几十年来，溶瘤病毒治疗引起了广泛关注，相关研究取得了巨大进展。目前腺病毒(adenovirus)、单纯疱疹病毒-1(herpes simplex virus-1，HSV-1)、新城疫病毒等相继被改造成溶瘤病毒。2006年，溶瘤腺病毒产品(oncorine)在中国已经用于临床治疗，主要用于治疗头颈部肿瘤、鼻窦癌等。Gendicine和oncorine的原理类似，将人5型腺病毒E1B-55kD区删除，使该病毒可在p53基因突变的癌细胞中繁殖并杀伤宿主细胞，产生溶瘤治疗作用。但是，临床数据显示这两种基于单一p53基因突变的溶瘤腺病毒的治疗效果并不十分理想。美国生物治疗公司Jennerex的JX-594是一种经过修改的牛痘病毒。在2013年完成的二期临床试验中，发现原发性肝癌患者在注射了高剂量的病毒后，其生命延长时间的中间值可达到14.1个月，而接受低剂量注射的患者只有6.7个月的生命延长期。目前该药物已经处于肝癌治疗的三期临床阶段。生物技术公司BioVex研发的基因工程化的单纯疱疹病毒OncoVEXGM-CSF已于2015年10月通过了FDA的批准成为美国和欧洲首个上市的溶瘤病毒产品。OncoVex可选择性地杀灭肿瘤细胞同时表达分泌GM-CSF启动机体产生系统的免疫反应杀伤剩余肿瘤细胞及其转移位点。2009年BioVex公布的一项转移性黑色素瘤II期试验的结果显示50名患者中有26％对治疗有反应，有8名患者完全恢复健康。该公司于2011年被Agmen以10亿美元的价格收购，用于推进三期临床试验。2013年3月，安进(Amgen)公布了OncoVex的治疗数据，临床证明它能成功地让晚期患者的肿瘤缩小，并在超过400名试验患者的III期研究中，Amgen的该药物要比同类其他药物表现得更加出色。
综上所述，溶瘤病毒确实在肿瘤的靶向治疗方面具有巨大的应用前景。但目前传统的溶瘤病毒研发平台仍然存在调控单一，靶向性差，平台转化能力低的问题。发明人前期构建了一个响应多输入信号的互抑制闭合回路，该回路通过开关调控E1A的表达进而调控腺病毒在癌细胞中的表达、复制和包装，在闭合回路中，去除一些病毒包装非必要的基因减少病毒对非靶标细胞的毒性，同时提高病毒的包装容量，如E3，E4等，替换腺病毒的外壳蛋白，改变病毒对特定细胞和组织的靶向性。
但基于该闭合回路仍存在较大的改进空间，发明人在前期工作的基础上，对该闭合回路进行了更深入和全面的研究和改进，以使该闭合回路应用于溶瘤病毒后，溶瘤病毒对疾病的治疗更加安全和有效。
发明内容
本申请是基于发明人对以下事实和问题的发现和认识作出的：
发明人在对前期开发的闭合回路进一步改进的过程中，惊喜地发现，闭合回路中，某些肿瘤细胞特异性启动子在特定肿瘤细胞中对某些特异性肿瘤细胞microRNA的翻转能力显著高，而某些肿瘤细胞特异性启动子在某些肿瘤细胞中对其特异性microRNA的翻转能力却不高或没有翻转能力。基于此，发明人在前期研究平台的基础上，提出了特异性针对某种癌症的闭合回路，该闭合回路在特异癌症细胞中的翻转能力高，承载该闭合回路的溶瘤病毒对特异癌症细胞的杀伤效率高而有效，且对正常细胞没有杀伤，安全性也更高。
在本发明的第一方面，本发明提出了一种表达系统。根据本发明的实施例，所述系统包括：第一核酸分子，所述第一核酸分子含有细胞特异性启动子；第二核酸分子，所述第二核酸分子与所述第一核酸分子可操作地连接，所述第二核酸分子编码转录激活因子；第三核酸分子，所述第三核酸分子含有所述转录激活因子的第一识别序列；第四核酸分子，所述第四核酸分子与所述第三核酸分子可操作地连接，所述第四核酸分子含有第一启动子和第一调控元件；第五核酸分子，所述第五核酸分子与第四核酸分子可操作地连接，所述第五核酸分子编码第一调控蛋白和目的蛋白，并且所述目的蛋白包括选自病毒复制包装蛋白、免疫效应因子的至少之一；第六核酸分子，所述第六核酸分子含有所述转录激活因子的第二识别序列；第七核酸分子，所述第七核酸分子与所述第六核酸分子可操作地连接，所述第七核酸分子含有第二启动子和第二调控元件；第八核酸分子，所述第八核酸分子与第七核酸分子可操作地连接，并且所述第八核酸分子编码第二调控蛋白；第九核酸分子，所述第九核酸分子与所述第五核酸分子可操作地连接，所述第九核酸分子被配置为条件性抑制所述第一调控蛋白的表达；以及第十核酸分子，所述第十核酸分子与所述第八核酸分子可操作地连接，所述第十核酸分子被配置为条件性抑制所述第二调控蛋白的表达，其中，所述第一调控元件适于通过结合所述第二调控蛋白抑制所述第一启动子的功能，所述第二调控元件适于通过结合所述第一调控蛋白抑制所述第二启动子的功能；所述第九核酸分子与所述第十核酸分子分别独立地借助RNA干扰抑制所述第一调控蛋白或所述第二调控蛋白的表达；所述第九核酸分子含有被第一microRNA特异性识别的核酸序列，所述第十核酸分子含有被第二microRNA特异性识别的核酸序列，所述第一microRNA为正常细胞特异性microRNA，所述第二microRNA为异常细胞特异性microRNA；所述细胞特异性启动子为CEA205，所述第一microRNA为mir-135a-5p，所述第二microRNA为mir-21；或所述细胞特异性启动子为CEA368，所述第一microRNA为mir-135a-5p，所述第二microRNA为mir-21；或所述细胞特异性启动子为CXCR4，所述第一microRNA为mir-135a-5p，所述第二microRNA为mir-21；或所述细胞特异性启动子为Survivin1，所述第一microRNA为mir-135a-5p，所述第二microRNA为mir-21；或所述细胞特异性启动子为CEA368，所述第一microRNA为mir-143-3p，所述第二microRNA为mir-21；或所述细胞特异性启动子为hMuc1，所述第一microRNA为mir-199a-3p所述第二microRNA为mir-21。根据本发明的实施例的表达系统在胃癌、胰腺癌细胞中的翻转能力高，具有该表达系统的表达载体，如腺病毒载体等，对胃癌、胰腺癌细胞的特异性杀伤效率高。
根据本发明的实施例，所述表达系统还可以进一步包括如下附加技术特征至少之一：
根据本发明的实施例，所述第一识别序列与所述第二识别序列分别独立地选自UAS,tetO以及dCas9的靶标序列至少之一。其中，所述UAS或tetO片段的重复次数可根据需要进行调整，如可选择2×UAS、3×UAS、4×UAS或5×UAS，5×tetO、6×tetO或7×tetO。
根据本发明的实施例，所述第一识别序列与所述第二识别序列为5×UAS。发明人在实验中意外地发现，使用癌特异启动子CEA368与癌特异microRNA mir-21及mir-143-3p调控的开关线路控制的溶瘤腺病毒、使用癌特异启动子CEA205与癌特异microRNA mir-21及mir-135a-5p调控的开关线路控制的溶瘤腺病毒、使用癌特异启动子CEA368与癌特异microRNA mir-21及mir-135a-5p调控的开关线路控制的溶瘤腺病毒、使用癌特异启动子CXCR4与癌特异microRNA mir-21及mir-135a-5p调控的开关线路控制的溶瘤腺病毒、使用癌特异启动子Survivin1与癌特异microRNA mir-21及mir-135a-5p调控的开关线路控制的溶瘤腺病毒可以特异性地杀伤胃癌细胞系AGS，5×UAS开关线路相较于4×UAS开关线路对胃正常细胞系GES1影响更小。
根据本发明的实施例，所述转录激活因子为选自Gal4VP16、Gal4VP64、Gal4esn、dCas9-VP16、dCas9-VP64、dCas9-VPR、dCas9-VTR以及rtTA至少之一。
根据本发明的实施例，所述第一识别序列与所述第二识别序列分别独立地选自5×UAS,7×tetO和dCas9的靶标序列至少之一。
根据本发明的实施例，所述第一启动子与所述第二启动子分别独立地选自miniCMV、TATA box。
根据本发明的实施例，所述第一调控蛋白和第二调控蛋白分别独立地选自Lacl、tetR、zinc finger(锌指)、TALE、KRAB、tetR-KRAB、、TALE-KRAB、dCas9-KRAB、miniCas9-KRAB、分割dCas9-KRAB至少之一。
根据本发明的实施例，所述第一调控元件和所述第二调控元件分别独立地选自tetO、LacO、锌指蛋白目的序列(zinc finger target site)、TALE蛋白靶标序列、dCas9的靶标序列以及miniCas9的靶标序列至少之一。
根据本发明的实施例，所述第一调控蛋白是LacI，所述第二调控元件包括多个重复的LacO序列，所述多个重复的LacO序列的至少之一设置在所述第二启动子的下游。LacI表达后，可特异性结合LacO序列，进而抑制第二启动子的功能。根据本发明的实施案例的LacI/LacO抑制系统，通过实验表明该系统可以有效地抑制启动子下游基因的表达。
根据本发明的实施例，所述第二调控蛋白是tetR-KRAB所述第一调控元件包括多个重复的tetO序列，所述多个重复的tetO序列的至少之一设置在所述第一启动子的下游。根据本发明的实施案例的tetR-KRAB/tetO抑制系统，可以有效的抑制启动子下游基因的表达。
根据本发明的实施例，所述病毒复制包装蛋白、免疫效应因子可以以融合蛋白的形式存在。病毒复制包装蛋白可有效保证表达系统载体在宿主中的存活和复制；免疫效应因子的表达可有效激活机体的免疫系统，进而促进对胃癌细胞、胰腺癌细胞的免疫杀伤。
根据本发明的实施例，所述病毒复制包装相关蛋白包括选自腺病毒E1基因,E1A基因，E1B基因，E2基因，E4基因至少之一。
根据本发明的实施例，所述免疫效应因子包括选自拮抗PD-1基因的抑制序列、拮抗PD-L1基因的抑制序列、拮抗CTLA4基因的抑制序列、拮抗Tim-3基因的抑制序列、GM-CSF、IL-2、IL-12、IL-15至少之一。任选地，上述的免疫效应因子可以以融合蛋白的形式存在。
根据本发明的实施例，所述目的蛋白与所述第一调控蛋白是以融合蛋白的形式表达的，并且所述目的蛋白与所述第一调控蛋白之间通过可切割的连接肽连接的。目的蛋白与第一调控蛋白在相同启动子下被调控表达，其表达后在连接肽处被切割，目的蛋白与第一调控蛋白分开，目的蛋白和第一调控蛋白相互独立地发挥功能。
根据本发明的实施例，所述第一核酸分子与所述第二核酸分子负载在第一表达载体上，所述第三核酸分子、所述第四核酸分子、所述第五核酸分子以及所述第九核酸分子负载在第二表达载体上，所述第六核酸分子、所述第七核酸分子、所述第八核酸分子以及所述第十核酸分子负载在第三表达载体上。第一、第二和第三表达载体作为表达系统的负载载体，实现目的蛋白在胃癌细胞、胰腺癌细胞的特异性表达。
所述表达载体的选择不受特别限制，只要能够实现表达系统在胃癌细胞、胰腺癌细胞的特异性发挥功能即可。根据本发明的具体实施例，所述第一表达载体、第二表达载体和第三表达载体分别独立地选自下列的至少之一：
质粒、病毒、稳定细胞系以及其他材料载体如纳米材料，脂质体，分子耦联载体、裸露DNA、染色体载体、多聚物。
根据本发明的实施例，所述病毒包括选自腺病毒、牛痘病毒、疱疹病毒、逆转录病毒的至少之一。
根据本发明的实施例，所述第一表达载体、第二表达载体和第三表达载体构建负载在同一个载体上。需要说明的是，第一表达载体、第二表达载体和第三表达载体的连接顺序不受特别限制，只要不影响系统的生物学功能的实现即可。根据本发明的具体实施例，负载在同一表达载体上可以有效的解决多个大片段载体共转染效率极低的问题。
根据本发明的实施例，所述同一个载体为腺病毒载体。腺病毒作为基因治疗载体具有主范围广，对人致病性低、在增殖和非增殖细胞中感染和表达基因、滴度高、与人类基因同源、无插入致突变性、能在悬浮培养液中扩增、能同时表达多个基因的优点。
根据本发明的实施例，所述细胞特异性启动子为CEA205，所述第一microRNA为mir-135a-5p，所述第二microRNA为mir-21，所述第一识别序列与所述第二识别序列为5×UAS；或所述细胞特异性启动子为CEA368，所述第一microRNA为mir-135a-5p，所述第二microRNA为mir-21，所述第一识别序列与所述第二识别序列为5×UAS；或所述细胞特异性启动子为CXCR4，所述第一microRNA为mir-135a-5p，所述第二microRNA为mir-21，所述第一识别序列与所述第二识别序列为5×UAS；或所述细胞特异性启动子为Survivin1，所述第一microRNA为mir-135a-5p，所述第二microRNA为mir-21，所述第一识别序列与所述第二识别序列为5×UAS；或所述细胞特异性启动子为CEA368，所述第一microRNA为mir-143-3p，所述第二microRNA为mir-21，所述第一识别序列与所述第二识别序列为5×UAS；或所述细胞特异性启动子为CEA205，所述第一microRNA为mir-135a-5p，所述第二microRNA为mir-21，所述第一识别序列与所述第二识别序列为4×UAS；或所述细胞特异性启动子为CEA368，所述第一microRNA为mir-135a-5p，所述第二microRNA为mir-21，所述第一识别序列与所述第二识别序列为4×UAS；或所述细胞特异性启动子为CXCR4，所述第一microRNA为mir-135a-5p，所述第二microRNA为mir-21，所述第一识别序列与所述第二识别序列为4×UAS；或所述细胞特异性启动子为Survivin1，所述第一microRNA为mir-135a-5p，所述第二microRNA为mir-21，所述第一识别序列与所述第二识别序列为4×UAS；或所述细胞特异性启动子为CEA368，所述第一microRNA为mir-143-3p，所述第二microRNA为mir-21，所述第一识别序列与所述第二识别序列为4×UAS。根据本发明实施例的上述元件组成的表达系统控制的溶瘤腺病毒可以特异性地杀伤胃癌细胞，同时具有5×UAS的开关线路相较于具有4×UAS开关线路胃正常细胞的杀伤影响更小。
根据本发明的实施例，所述细胞特异性启动子为hMuc1，所述第一microRNA为mir-199a-3p，所述第二microRNA为mir-21，所述第一识别序列与所述第二识别序列为4×UAS。根据本发明实施例的上述元件组成的表达系统控制的溶瘤腺病毒可以特异性地杀伤胰腺癌细胞。
根据本发明的实施例，所述腺病毒载体携带具有SEQ ID NO：1～26任一所示核苷酸序列的核酸。
具有SEQ ID NO:1所述核苷酸序列的核酸编码由细胞特异性启动子CEA205、第一microRNA为mir-135a-5p、第二microRNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白分别为Lacl、tetR-KRAB、第一和第二识别序列为5×UAS的元件组成的表达系统。
CGGGGTGCCGCCCCCCGCGGGTGACCGCCCTCCGCGGGTGACAGCCCTCCGCGGAGTGCCGCACTCCGCGGAGTGCTGTACTCCGAGCGGAGACTCTAGACTCCCTATCAGTGATAGAGATAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCTCCCTATCAGTGATAGAGAGAATTCGACCGCCACCATGAGACATATTATCTGCCACGGAGGTGTTATTACCGAAGAAATGGCCGCCAGTCTTTTGGACCAGCTGATCGAAGAGGTACTGGCTGATAATCTTCCACCTCCTAGCCATTTTGAACCACCTACCCTTCACGAACTGTATGATTTAGACGTGACGGCCCCCGAAGATCCCAACGAGGAGGCGGTTTCGCAGATTTTTCCCGACTCTGTAATGTTGGCGGTGCAGGAAGGGATTGACTTACTCACTTTTCCGCCGGCGCCCGGTTCTCCGGAGCCGCCTCACCTTTCCCGGCAGCCCGAGCAGCCGGAGCAGAGAGCCTTGGGTCCGGTTTCTATGCCAAACCTTGTACCGGAGGTGATCGATCTTACCTGCCACGAGGCTGGCTTTCCACCCAGTGACGACGAGGATGAAGAGGGTGAGGAGTTTGTGTTAGATTATGTGGAGCACCCCGGGCACGGTTGCAGGTCTTGTCATTATCACCGGAGGAATACGGGGGACCCAGATATTATGTGTTCGCTTTGCTATATGAGGACCTGTGGCATGTTTGTCTACAGTAAGTGAAAATTATGGGCAGTGGGTGATAGAGTGGTGGGTTTGGTGTGGTAATTTTTTTTTTAATTTTTACAGTTTTGTGGTTTAAAGAATTTTGTATTGTGATTTTTTTAAAAGGTCCTGTGTCTGAACCTGAGCCTGAGCCCGAGCCAGAACCGGAGCCTGCAAGACCTACCCGCCGTCCTAAAATGGCGCCTGCTATCCTGAGGCGCCCGACATCACCTGTGTCTAGAGAATGCAATAGTAGTACGGATAGCTGTGACTCCGGTCCTTCTAACACACCTCCTGAGATACACCCGGTGGTCCCGCTGTGCCCCATTAAACCAGTTGCCGTGAGAGTTGGTGGGCGTCGCCAGGCTGTGGAATGTATCGAGGACTTGCTTAACGAGCCTGGGCAACCTTTGGACTTGAGCTGTAAACGCCCCAGGCCACTCGAGGGTACCGGGTCCGGAGCTACAAATTTTTCCCTCCTCAAACAGGCTGGAGATGTCGAAGAAAATCCTGGGCCTACCGGTATGTGGCTGCAGAGCCTCCTTCTACTGGGCACCGTGGCTTGCAGCATCTCGGCCCCCGCCAGAAGTCCCAGCCCCAGCACCCAGCCCTGGGAACACGTAAATGCAATCCAGGAGGCCCGCCGCCTCCTGAACTTGTCACGGGACACGGCAGCCGAGATGAATGAAACTGTGGAGGTCATCAGTGAAATGTTTGACTTGCAGGAGCCCACATGCCTGCAGACTCGGCTGGAGCTCTACAAACAGGGCCTGCGTGGCAGCCTCACCAAGCTGAAGGGCCCACTGACCATGATGGCTAGCCATTACAAGCAGCACTGCCCGCCGACCCCTGAGACATCCTGTGCCACTCAGATCATTACCTTTGAGTCTTTCAAGGAGAACCTCAAGGACTTTCTGCTGGTCATCCCATTTGACTGCTGGGAACCCGTCCAGGAAAGATCTGAGGGCCGCGGCAGCCTGCTGACCTGCGGCGACGTGGAGGAAAACCCCGGCCCCGGATCCATGAAACCAGTAACGTTATACGATGTCGCAGAGTATGCCGGTGTCTCTTATCAGACCGTTTCCCGCGTGGTGAACCAGGCCAGCCACGTTTCTGCGAAAACGCGGGAAAAAGTGGAAGCGGCGATGGCGGAGCTGAATTACATTCCCAACCGCGTGGCACAACAACTGGCGGGCAAACAGTCGTTGCTGATTGGCGTTGCCACCTCCAGTCTGGCCCTGCACGCGCCGTCGCAAATTGTCGCGGCGATTAAATCTCGCGCCGATCAACTGGGTGCCAGCGTGGTGGTGTCGATGGTAGAACGAAGCGGCGTCGAAGCCTGTAAAGCGGCGGTGCACAATCTTCTCGCGCAACGCGTCAGTGGGCTGATCATTAACTATCCGCTGGATGACCAGGATGCCATTGCTGTGGAAGCTGCCTGCACTAATGTTCCGGCGTTATTTCTTGATGTCTCTGACCAGACACCCATCAACAGTATTATTTTCTCCCATGAAGACGGTACGCGACTGGGCGTGGAGCATCTGGTCGCATTGGGTCACCAGCAAATCGCGCTGTTAGCGGGCCCATTAAGTTCTGTCTCGGCGCGTCTGCGTCTGGCTGGCTGGCATAAATATCTCACTCGCAATCAAATTCAGCCGATAGCGGAACGGGAAGGCGACTGGAGTGCCATGTCCGGTTTTCAACAAACCATGCAAATGCTGAATGAGGGCATCGTTCCCACTGCGATGCTGGTTGCCAACGATCAGATGGCGCTGGGCGCAATGCGCGCCATTACCGAGTCCGGGCTGCGCGTTGGTGCGGATATCTCGGTAGTGGGATACGACGATACCGAAGACAGCTCATGTTATATCCCGCCGTTAACCACCATCAAACAGGATTTTCGCCTGCTGGGGCAAACCAGCGTGGACCGCTTGCTGCAACTCTCTCAGGGCCAGGCGGTGAAGGGCAATCAGCTGTTGCCGGTCTCACTGGTGAAAAGAAAAACCACCCTGGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGAGCAGCCTGAGGCCTCCTAAGAAGAAGAGGAAGGTTGTTACAAACACCATTGTCACACTCCACAAACACCATTGTCACACTCCACAAACACCATTGTCACACTCCACAAACACCATTGTCACACTCCATCACATAGGAATAAAAAGCCATATCACATAGGAATAAAAAGCCATATCACATAGGAATAAAAAGCCATATCACATAGGAATAAAAAGCCATAGAAAGCGGACTACTGTAGTCCGCGGAGTACTGTCCTCCGCGGACGACTGTCGTCCGCGGAGCACTGTGCTCCGCGGATGACTGTCATCCGAGCGGAGACTCTAGAAATTGTGAGCGGATAACAATTGGGTATATAATGGGGGCCGTCAGCTACTACCAGAGCCGATCGAACACTCGAGCCGAGCAGACGTGCCTACAATTGTGAGCGCTCACAATTACTAGAAGATCTTGAATTCGCCACCATGGCTAGATTAGATAAAAGTAAAGTGATTAACAGCGCATTAGAGCTGCTTAATGAGGTCGGAATCGAAGGTTTAACAACCCGTAAACTCGCCCAGAAGCTAGGTGTAGAGCAGCCTACATTGTATTGGCATGTAAAAAATAAGCGGGCTTTGCTCGACGCCTTAGCCATTGAGATGTTAGATAGGCACCATACTCACTTTTGCCCTTTAGAAGGGGAAAGCTGGCAAGATTTTTTACGTAATAACGCTAAAAGTTTTAGATGTGCTTTACTAAGTCATCGCGATGGAGCAAAAGTACATTTAGGTACACGGCCTACAGAAAAACAGTATGAAACTCTCGAAAATCAATTAGCCTTTTTATGCCAACAAGGTTTTTCACTAGAGAATGCATTATATGCACTCAGCGCTGTGGGGCATTTTACTTTAGGTTGCGTATTGGAAGATCAAGAGCATCAAGTCGCTAAAGAAGAAAGGGAAACACCTACTACTGATAGTATGCCGCCATTATTACGACAAGCTATCGAATTATTTGATCACCAAGGTGCAGAGCCAGCCTTCTTATTCGGCCTTGAATTGATCATATGCGGATTAGAAAAACAACTTAAATGTGAAAGTGGGTCGCCAAAAAAGAAGAGAAAGGTGGATGGTGGCGGAGCACTGTCTCCCCAGCATTCAGCTGTGACCCAGGGATCAATCATTAAGAACAAGGAAGGTATGGATGCTAAGTCCCTGACCGCTTGGTCAAGAACTTTGGTAACTTTTAAGGACGTGTTCGTCGATTTTACTCGGGAGGAGTGGAAACTATTGGATACCGCCCAGCAAATCGTGTATAGAAACGTTATGCTCGAGAACTATAAGAATCTTGTGTCACTGGGCTACCAACTGACAAAGCCTGACGTTATCCTGAGGCTGGAAAAGGGTGAGGAGCCTTGGTTGGTCGAGCGGGAGATCCACCAGGAAACTCACCCAGACAGTGAGACTGCTTTCGAGATCAAGTCCTCTGTGTAATACAAGGCGGCCGCAAATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTAAAGCTTCGAATTCTGATAATCAGCCATACCACATTTGTAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCAGCTCGCCTTTTGCTCACATGTGAAAATAGAAGGGAAAAAAGAGGAGGGACAAAAGAGGCAGAAATGAGAGGGGAGGGGACAGAGGACACCTGAATAAAGACCACACCCATGACCCACGTGATGCTGAGAAGTACTCCTGCCCTAGGAAGAGACTCAGGGCAGAGGGAGGAAGGACAGCAGACCAGACAGTCACAGCAGCCTTGACAAAACGTTCCTGGAACTCAAGCTCGAGACCGGTATGGCCCCCCCGACCGATGTCAGCCTGGGGGACGAGCTCCACTTAGACGGCGAGGACGTGGCGATGGCGCATGCCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGGACGGGGATTCCCCGGGTCCGGGATTTACCCCCCACGACTCCGCCCCCTACGGCGCTCTGGATATGGCCGACTTCGAGTTTGAGCAGATGTTTACCGATGCCCTTGGAATTGACGAGTACGGTGGGACGCGTATGAAGCTACTGTCTTCTATCGAACAAGCATGCGATATTTGCCGACTTAAAAAGCTCAAGTGCTCCAAAGAAAAACCGAAGTGCGCCAAGTGTCTGAAGAACAACTGGGAGTGTCGCTACTCTCCCAAAACCAAAAGGTCTCCGCTGACTAGGGCACATCTGACAGAAGTGGAATCAAGGCTAGAAAGACTGGAACAGCTATTTCTACTGATTTTTCCTCGAGAAGACCTTGACATGATTTTGAAAATGGATTCTTTACAGGATATAAAAGCATTGTTAACAGGATTATTTGTACAAGATAATGTGAATAAAGATGCCGTCACAGATAGATTGGCTTCAGTGGAGACTGATATGCCTCTAACATTGAGACAGCATAGAATAAGTGCGACATCATCATCGGAAGAGAGTAGTAACAAAGGTCAAAGACAGTTGACTGTATAAAAGCTTCACTCCTCAGGTGCAGGCTGCCTATCAGAAGGTGGTGGCTGGTGTGGCCAATGCCCTGGCTCACAAATACCACTGAGATCTTTTTCCCTCTGCCAAAAATTATGGGGACATCATGAAGCCCCTTGAGCATCTGACTTCTGGCTAATAAAGGAAATTTATTTTCATTGCAATAGTGTGTTGGAATTTTTTGTGTCTCTCACTCGGAAGGACATATGGGAGGGCAAATCATTTAAAACATCAGAATGAGTATTTGGTTTAGAGTTTGGCAACATATGCCCATATGCTGGCTGCCATGAACAAAGGTTGGCTATAAAGAGGTCATCAGTATATGAAACAGCCCCCTGCTGTCCATTCCTTATTCCATAGAAAAGCCTTGACTTGAGGTTAGATTTTTTTTATATTTTGTTTTGTGTTATTTTTTTCTTTAACATCCCTAAAATTTTCCTTACATGTTTTACTAGCCAGATTTTTCCTCCTCTCCTGACTACTCCCAGTCATAGCTGTCCCTCTTCTCTTATGGAGATC(SEQ ID NO:1)。
具有SEQ ID NO:2所述核苷酸序列的核酸编码由细胞特异性启动子CEA205、第一microRNA为mir-143-3p、第二microRNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白分别为TALE蛋白、第一和第二识别序列为5×UAS的元件组成的表达系统。
CGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTTCTGTCCTCCGAGCGGAGACTCTAGATACCCCTCTCCGCTTCTGAATTCTAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCCTCGAGTACCCCTCTCCGCTTCTTACCCCTCTCCGCTTCTTACCCCTCTCCGCTTCTGAATTCGACCGCCACCATGAGACATATTATCTGCCACGGAGGTGTTATTACCGAAGAAATGGCCGCCAGTCTTTTGGACCAGCTGATCGAAGAGGTACTGGCTGATAATCTTCCACCTCCTAGCCATTTTGAACCACCTACCCTTCACGAACTGTATGATTTAGACGTGACGGCCCCCGAAGATCCCAACGAGGAGGCGGTTTCGCAGATTTTTCCCGACTCTGTAATGTTGGCGGTGCAGGAAGGGATTGACTTACTCACTTTTCCGCCGGCGCCCGGTTCTCCGGAGCCGCCTCACCTTTCCCGGCAGCCCGAGCAGCCGGAGCAGAGAGCCTTGGGTCCGGTTTCTATGCCAAACCTTGTACCGGAGGTGATCGATCTTACCTGCCACGAGGCTGGCTTTCCACCCAGTGACGACGAGGATGAAGAGGGTGAGGAGTTTGTGTTAGATTATGTGGAGCACCCCGGGCACGGTTGCAGGTCTTGTCATTATCACCGGAGGAATACGGGGGACCCAGATATTATGTGTTCGCTTTGCTATATGAGGACCTGTGGCATGTTTGTCTACAGTAAGTGAAAATTATGGGCAGTGGGTGATAGAGTGGTGGGTTTGGTGTGGTAATTTTTTTTTTAATTTTTACAGTTTTGTGGTTTAAAGAATTTTGTATTGTGATTTTTTTAAAAGGTCCTGTGTCTGAACCTGAGCCTGAGCCCGAGCCAGAACCGGAGCCTGCAAGACCTACCCGCCGTCCTAAAATGGCGCCTGCTATCCTGAGGCGCCCGACATCACCTGTGTCTAGAGAATGCAATAGTAGTACGGATAGCTGTGACTCCGGTCCTTCTAACACACCTCCTGAGATACACCCGGTGGTCCCGCTGTGCCCCATTAAACCAGTTGCCGTGAGAGTTGGTGGGCGTCGCCAGGCTGTGGAATGTATCGAGGACTTGCTTAACGAGCCTGGGCAACCTTTGGACTTGAGCTGTAAACGCCCCAGGCCAGGGTCCGGAGCTACAAATTTTTCCCTCCTCAAACAGGCTGGAGATGTCGAAGAAAATCCTGGGCCTATGGAGGCTTGGGAGTGTTTGGAAGATTTTTCTGCTGTGCGTAACTTGCTGGAACAGAGCTCTAACAGTACCTCTTGGTTTTGGAGGTTTCTGTGGGGCTCATCCCAGGCAAAGTTAGTCTGCAGAATTAAGGAGGATTACAAGTGGGAATTTGAAGAGCTTTTGAAATCCTGTGGTGAGCTGTTTGATTCTTTGAATCTGGGTCACCAGGCGCTTTTCCAAGAGAAGGTCATCAAGACTTTGGATTTTTCCACACCGGGGCGCGCTGCGGCTGCTGTTGCTTTTTTGAGTTTTATAAAGGATAAATGGAGCGAAGAAACCCATCTGAGCGGGGGGTACCTGCTGGATTTTCTGGCCATGCATCTGTGGAGAGCGGTTGTGAGACACAAGAATCGCCTGCTACTGTTGTCTTCCGTCCGCCCGGCGATAATACCGACGGAGGAGCAGCAGCAGCAGCAGGAGGAAGCCAGGCGGCGGCGGCAGGAGCAGAGCCCATGGAACCCGAGAGCCGGCCTGGACCCTCGGGAATGAATGTTGTACAGGTGGCTGAACTGTATCCAGAACTGAGACGCATTTTGACAATTACAGAGGATGGGCAGGGGCTAAAGGGGGTAAAGAGGGAGCGGGGGGCTTGTGAGGCTACAGAGGAGGCTAGGAATCTAGCTTTTAGCTTAATGACCAGACACCGTCCTGAGTGTATTACTTTTCAACAGATCAAGGATAATTGCGCTAATGAGCTTGATCTGCTGGCGCAGAAGTATTCCATAGAGCAGCTGACCACTTACTGGCTGCAGCCAGGGGATGATTTTGAGGAGGCTATTAGGGTATATGCAAAGGTGGCACTTAGGCCAGATTGCAAGTACAAGATCAGCAAACTTGTAAATATCAGGAATTGTTGCTACATTTCTGGGAACGGGGCCGAGGTGGAGATAGATACGGAGGATAGGGTGGCCTTTAGATGTAGCATGATAAATATGTGGCCGGGGGTGCTTGGCATGGACGGGGTGGTTATTATGAATGTAAGGTTTACTGGCCCCAATTTTAGCGGTACGGTTTTCCTGGCCAATACCAACCTTATCCTACACGGTGTAAGCTTCTATGGGTTTAACAATACCTGTGTGGAAGCCTGGACCGATGTAAGGGTTCGGGGCTGTGCCTTTTACTGCTGCTGGAAGGGGGTGGTGTGTCGCCCCAAAAGCAGGGCTTCAATTAAGAAATGCCTCTTTGAAAGGTGTACCTTGGGTATCCTGTCTGAGGGTAACTCCAGGGTGCGCCACAATGTGGCCTCCGACTGTGGTTGCTTCATGCTAGTGAAAAGCGTGGCTGTGATTAAGCATAACATGGTATGTGGCAACTGCGAGGACAGGGCCTCTCAGATGCTGACCTGCTCGGACGGCAACTGTCACCTGCTGAAGACCATTCACGTAGCCAGCCACTCTCGCAAGGCCTGGCCAGTGTTTGAGCATAACATACTGACCCGCTGTTCCTTGCATTTGGGTAACAGGAGGGGGGTGTTCCTACCTTACCAATGCAATTTGAGTCACACTAAGATATTGCTTGAGCCCGAGAGCATGTCCAAGGTGAACCTGAACGGGGTGTTTGACATGACCATGAAGATCTGGAAGGTGCTGAGGTACGATGAGACCCGCACCAGGTGCAGACCCTGCGAGTGTGGCGGTAAACATATTAGGAACCAGCCTGTGATGCTGGATGTGACCGAGGAGCTGAGGCCCGATCACTTGGTGCTGGCCTGCACCCGCGCTGAGTTTGGCTCTAGCGATGAAGATACAGATCTCGAGGGGTCCGGAGCTACAAATTTTTCCCTCCTCAAACAGGCTGGAGATGTCGAAGAAAATCCTGGGCCTACCGGTATGTATAGGATGCAGTTGCTCTCTTGCATCGCCCTGTCTCTAGCGTTGGTGACTAATAGCGCCCCTACTTCTTCTTCCACGAAGAAAACCCAGCTGCAACTGGAACACCTTCTGCTGGATCTCCAGATGATCCTTAACGGGATTAACAACTATAAGAACCCGAAGCTCACGGCTATGCTGACGGCGAAGTTCGCTATGCCTAAAAAGGCTACTGAGCTGAAACACTTACAGTGTCTCGAAGAGGCACTCAAACCCCTCGAGGAGGTACTCAATCTGGCGCAAAGCAAAAACTTCCATCTCCGCCCACGGGACCTAATCTCCAACATCAACGTGATTGTGCTGGAGCTCAAGGGGTCTGAAACCACGTTTATGTGCGAGTACGCAGATGAAACTGCTACCATAGTGGAATTCCTCAATCGTTGGATCACTTTCTGCCAGTCAATCATTTCAACCCTGACCAGATCTGAGGGCCGCGGCAGCCTGCTGACCTGCGGCGACGTGGAGGAAAACCCCGGCCCCGGATCCTTGTCGACCATTCGTCCGCGCAGGCCAAGTCCTGCCCGCGAGCTTCTGCCCGGACCCCAACCGGATAGGGTTCAGCCGACTGCAGATCGTGGGGTGTCTGCGCCTGCTGGCAGCCCTCTGGATGGCTTGCCCGCTCGGCGGACGGTGTCCCGGACCCGGCTGCCATCTCCCCCTGCGCCCTCACCTGCGTTCTCGGCGGGCAGCTTCAGCGATCTGCTCCGTCCGTTCGATCCGTCGCTTCTTGATACATCGCTTCTTGATTCGATGCCTGCCGTCGGCACGCCGCATACAGCGGCTGCCCCAGCAGAGTGGGATGAGGCGCAATCGGCTCTGCGTGCAGCCGATGACCCGCCACCCACCGTGCGTGTCGCTGTCACTGCCGCGCGGCCGCCGCGCGCCAAGCCGGCCCCGCGACGGCGTGCTGCGCAACCCTCCGACGCTTCGCCGGCCGCGCAGGTGGATCTACGCACGCTCGGCTACAGTCAGCAGCAGCAAGAGAAGATCAAACCGAAGGTGCGTTCGACAGTGGCGCAGCACCACGAGGCACTGGTGGGCCATGGGTTTACACACGCGCACATCGTTGCGCTCAGCCAACACCCGGCAGCGTTAGGGACCGTCGCTGTCACGTATCAGCACATAATCACGGCGTTGCCAGAGGCGACACACGAAGACATCGTTGGCGTCGGCAAACAGTGGTCCGGCGCACGCGCCCTGGAGGCCTTGCTCACGGATGCGGGGGAGTTGAGAGGTCCGCCGTTACAGTTGGACACAGGCCAACTTGTGAAGATTGCAAAACGTGGCGGCGTGACCGCAATGGAGGCAGTGCATGCATCGCGCAATGCACTGACGGGTGCCCCCCTGAACCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACAATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACAATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAAGCATTGTGGCCCAGCTGAGCCGGCCTGATCCGGCGTTGGCCGCGTTGACCAACGACCACCTCGTCGCCTTGGCCTGCCTCGGCGGACGTCCTGCCATGGATGCAGTGAAAAAGGGATTGCCGCACGCGCCGGAATTGATCAGAAGAGTCAATCGCCGTATTGGCGAACGCACGTCCCATCGCGTTGCCGACTACGCGCAAGTGGTTCGCGTGCTGGAGTTTTTCCAGTGCCACTCCCACCCAGCGTACGCATTTGATGAGGCCATGACGCAGTTCGGGATGAGCAGGAACGGGTTGGTACAGCTCTTTCGCAGAGTGGGCGTCACCGAACTCGAAGCCCGCGGTGGAACGCTCCCCCCAGCCTCGCAGCGTTGGGACCGTATCCTCCAGGCATCAGGGATGAAAAGGGCCAAACCGTCCCCTACTTCAGCTCAAACACCGGATCAGGCGTCTTTGCATGCATTCGCCGATTCGCTGGAGCGTGACCTTGATGCGCCCAGCCCAATGCACGAGGGAGATCAGACGCGGGCAAGCAGCCGTAAACGGTCCCGATCGGATCGTGCTGTCACCGGCCCCTCCGCACAGCAGGCTGTCGAGGTGCGCGTTCCCGAACAGCGCGATGCGCTGCATTTGCCCCTCAGCTGGAGGGTAAAACGCCCGCGTACCAGGATCTGGGGCGGCCTCCCGGTTAAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGGAGCTACAGTGCTTCATCTCAGAGCTACAGTGCTTCATCTCAGAGCTACAGTGCTTCATCTCAGAGCTACAGTGCTTCATCTCAGAAAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTTCTGTCCTCCGAGCGGAGACTCTAGATATAGAACCGATCCTCCCATTGAATTCTAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCCTCGAGTATAGAACCGATCCTCCCATTTATAGAACCGATCCTCCCATTTATAGAACCGATCCTCCCATTACTAGAAGATCTTGAATTCGCCACCTTGTCGACCATTCGTCCGCGCAGGCCAAGTCCTGCCCGCGAGCTTCTGCCCGGACCCCAACCGGATAGGGTTCAGCCGACTGCAGATCGTGGGGTGTCTGCGCCTGCTGGCAGCCCTCTGGATGGCTTGCCCGCTCGGCGGACGGTGTCCCGGACCCGGCTGCCATCTCCCCCTGCGCCCTCACCTGCGTTCTCGGCGGGCAGCTTCAGCGATCTGCTCCGTCCGTTCGATCCGTCGCTTCTTGATACATCGCTTCTTGATTCGATGCCTGCCGTCGGCACGCCGCATACAGCGGCTGCCCCAGCAGAGTGGGATGAGGCGCAATCGGCTCTGCGTGCAGCCGATGACCCGCCACCCACCGTGCGTGTCGCTGTCACTGCCGCGCGGCCGCCGCGCGCCAAGCCGGCCCCGCGACGGCGTGCTGCGCAACCCTCCGACGCTTCGCCGGCCGCGCAGGTGGATCTACGCACGCTCGGCTACAGTCAGCAGCAGCAAGAGAAGATCAAACCGAAGGTGCGTTCGACAGTGGCGCAGCACCACGAGGCACTGGTGGGCCATGGGTTTACACACGCGCACATCGTTGCGCTCAGCCAACACCCGGCAGCGTTAGGGACCGTCGCTGTCACGTATCAGCACATAATCACGGCGTTGCCAGAGGCGACACACGAAGACATCGTTGGCGTCGGCAAACAGTGGTCCGGCGCACGCGCCCTGGAGGCCTTGCTCACGGATGCGGGGGAGTTGAGAGGTCCGCCGTTACAGTTGGACACAGGCCAACTTGTGAAGATTGCAAAACGTGGCGGCGTGACCGCAATGGAGGCAGTGCATGCATCGCGCAATGCACTGACGGGTGCCCCCCTGAACCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACAATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAAGCATTGTGGCCCAGCTGAGCCGGCCTGATCCGGCGTTGGCCGCGTTGACCAACGACCACCTCGTCGCCTTGGCCTGCCTCGGCGGACGTCCTGCCATGGATGCAGTGAAAAAGGGATTGCCGCACGCGCCGGAATTGATCAGAAGAGTCAATCGCCGTATTGGCGAACGCACGTCCCATCGCGTTGCCGACTACGCGCAAGTGGTTCGCGTGCTGGAGTTTTTCCAGTGCCACTCCCACCCAGCGTACGCATTTGATGAGGCCATGACGCAGTTCGGGATGAGCAGGAACGGGTTGGTACAGCTCTTTCGCAGAGTGGGCGTCACCGAACTCGAAGCCCGCGGTGGAACGCTCCCCCCAGCCTCGCAGCGTTGGGACCGTATCCTCCAGGCATCAGGGATGAAAAGGGCCAAACCGTCCCCTACTTCAGCTCAAACACCGGATCAGGCGTCTTTGCATGCATTCGCCGATTCGCTGGAGCGTGACCTTGATGCGCCCAGCCCAATGCACGAGGGAGATCAGACGCGGGCAAGCAGCCGTAAACGGTCCCGATCGGATCGTGCTGTCACCGGCCCCTCCGCACAGCAGGCTGTCGAGGTGCGCGTTCCCGAACAGCGCGATGCGCTGCATTTGCCCCTCAGCTGGAGGGTAAAACGCCCGCGTACCAGGATCTGGGGCGGCCTCCCGTAATACAAGGCGGCCGCAAATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTAAAGCTTCGAATTCTGATAATCAGCCATACCACATTTGTAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCGAAAATAGAAGGGAAAAAAGAGGAGGGACAAAAGAGGCAGAAATGAGAGGGGAGGGGACAGAGGACACCTGAATAAAGACCACACCCATGACCCACGTGATGCTGAGAAGTACTCCTGCCCTAGGAAGAGACTCAGGGCAGAGGGAGGAAGGACAGCAGACCAGACAGTCACAGCAGCCTTGACAAAACGTTCCTGGAACTCAAGATGGCCCCCCCGACCGATGTCAGCCTGGGGGACGAGCTCCACTTAGACGGCGAGGACGTGGCGATGGCGCATGCCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGAGCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGAGCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGAGCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGGACGGGGATTCCCCGGGTCCGGGATTTACCCCCCACGACTCCGCCCCCTACGGCGCTCTGGATATGGCCGACTTCGAGTTTGAGCAGATGTTTACCGATGCCCTTGGAATTGACGAGTACGGTGGGACGCGTATGAAGCTACTGTCTTCTATCGAACAAGCATGCGATATTTGCCGACTTAAAAAGCTCAAGTGCTCCAAAGAAAAACCGAAGTGCGCCAAGTGTCTGAAGAACAACTGGGAGTGTCGCTACTCTCCCAAAACCAAAAGGTCTCCGCTGACTAGGGCACATCTGACAGAAGTGGAATCAAGGCTAGAAAGACTGGAACAGCTATTTCTACTGATTTTTCCTCGAGAAGACCTTGACATGATTTTGAAAATGGATTCTTTACAGGATATAAAAGCATTGTTAACAGGATTATTTGTACAAGATAATGTGAATAAAGATGCCGTCACAGATAGATTGGCTTCAGTGGAGACTGATATGCCTCTAACATTGAGACAGCATAGAATAAGTGCGACATCATCATCGGAAGAGAGTAGTAACAAAGGTCAAAGACAGTTGACTGTATAAAAGCTTCACTCCTCAGGTGCAGGCTGCCTATCAGAAGGTGGTGGCTGGTGTGGCCAATGCCCTGGCTCACAAATACCACTGAGATCTTTTTCCCTCTGCCAAAAATTATGGGGACATCATGAAGCCCCTTGAGCATCTGACTTCTGGCTAATAAAGGAAATTTATTTTCATTGCAATAGTGTGTTGGAATTTTTTGTGTCTCTCACTCGGAAGGACATATGGGAGGGCAAATCATTTAAAACATCAGAATGAGTATTTGGTTTAGAGTTTGGCAACATATGCCCATATGCTGGCTGCCATGAACAAAGGTTGGCTATAAAGAGGTCATCAGTATATGAAACAGCCCCCTGCTGTCCATTCCTTATTCCATAGAAAAGCCTTGACTTGAGGTTAGATTTTTTTTATATTTTGTTTTGTGTTATTTTTTTCTTTAACATCCCTAAAATTTTCCTTACATGTTTTACTAGCCAGATTTTTCCTCCTCTCCTGACTACTCCCAGTCATAGCTGTCCCTCTTCTCTTATGGAGATC(SEQ ID NO:2)。
具有SEQ ID NO:3所述核苷酸序列的核酸编码由细胞特异性启动子CEA368、第一microRNA为mir-135a-5p、第二microRNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白分别为LacI与tetR-KRAB、第一和第二识别序列为5×UAS的元件组成的表达系统。
CGGGGTGCCGCCCCCCGCGGGTGACCGCCCTCCGCGGGTGACAGCCCTCCGCGGAGTGCCGCACTCCGCGGAGTGCTGTACTCCGAGCGGAGACTCTAGACTCCCTATCAGTGATAGAGATAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCTCCCTATCAGTGATAGAGAGAATTCGACCGCCACCATGAGACATATTATCTGCCACGGAGGTGTTATTACCGAAGAAATGGCCGCCAGTCTTTTGGACCAGCTGATCGAAGAGGTACTGGCTGATAATCTTCCACCTCCTAGCCATTTTGAACCACCTACCCTTCACGAACTGTATGATTTAGACGTGACGGCCCCCGAAGATCCCAACGAGGAGGCGGTTTCGCAGATTTTTCCCGACTCTGTAATGTTGGCGGTGCAGGAAGGGATTGACTTACTCACTTTTCCGCCGGCGCCCGGTTCTCCGGAGCCGCCTCACCTTTCCCGGCAGCCCGAGCAGCCGGAGCAGAGAGCCTTGGGTCCGGTTTCTATGCCAAACCTTGTACCGGAGGTGATCGATCTTACCTGCCACGAGGCTGGCTTTCCACCCAGTGACGACGAGGATGAAGAGGGTGAGGAGTTTGTGTTAGATTATGTGGAGCACCCCGGGCACGGTTGCAGGTCTTGTCATTATCACCGGAGGAATACGGGGGACCCAGATATTATGTGTTCGCTTTGCTATATGAGGACCTGTGGCATGTTTGTCTACAGTAAGTGAAAATTATGGGCAGTGGGTGATAGAGTGGTGGGTTTGGTGTGGTAATTTTTTTTTTAATTTTTACAGTTTTGTGGTTTAAAGAATTTTGTATTGTGATTTTTTTAAAAGGTCCTGTGTCTGAACCTGAGCCTGAGCCCGAGCCAGAACCGGAGCCTGCAAGACCTACCCGCCGTCCTAAAATGGCGCCTGCTATCCTGAGGCGCCCGACATCACCTGTGTCTAGAGAATGCAATAGTAGTACGGATAGCTGTGACTCCGGTCCTTCTAACACACCTCCTGAGATACACCCGGTGGTCCCGCTGTGCCCCATTAAACCAGTTGCCGTGAGAGTTGGTGGGCGTCGCCAGGCTGTGGAATGTATCGAGGACTTGCTTAACGAGCCTGGGCAACCTTTGGACTTGAGCTGTAAACGCCCCAGGCCACTCGAGGGTACCGGGTCCGGAGCTACAAATTTTTCCCTCCTCAAACAGGCTGGAGATGTCGAAGAAAATCCTGGGCCTACCGGTATGTGGCTGCAGAGCCTCCTTCTACTGGGCACCGTGGCTTGCAGCATCTCGGCCCCCGCCAGAAGTCCCAGCCCCAGCACCCAGCCCTGGGAACACGTAAATGCAATCCAGGAGGCCCGCCGCCTCCTGAACTTGTCACGGGACACGGCAGCCGAGATGAATGAAACTGTGGAGGTCATCAGTGAAATGTTTGACTTGCAGGAGCCCACATGCCTGCAGACTCGGCTGGAGCTCTACAAACAGGGCCTGCGTGGCAGCCTCACCAAGCTGAAGGGCCCACTGACCATGATGGCTAGCCATTACAAGCAGCACTGCCCGCCGACCCCTGAGACATCCTGTGCCACTCAGATCATTACCTTTGAGTCTTTCAAGGAGAACCTCAAGGACTTTCTGCTGGTCATCCCATTTGACTGCTGGGAACCCGTCCAGGAAAGATCTGAGGGCCGCGGCAGCCTGCTGACCTGCGGCGACGTGGAGGAAAACCCCGGCCCCGGATCCATGAAACCAGTAACGTTATACGATGTCGCAGAGTATGCCGGTGTCTCTTATCAGACCGTTTCCCGCGTGGTGAACCAGGCCAGCCACGTTTCTGCGAAAACGCGGGAAAAAGTGGAAGCGGCGATGGCGGAGCTGAATTACATTCCCAACCGCGTGGCACAACAACTGGCGGGCAAACAGTCGTTGCTGATTGGCGTTGCCACCTCCAGTCTGGCCCTGCACGCGCCGTCGCAAATTGTCGCGGCGATTAAATCTCGCGCCGATCAACTGGGTGCCAGCGTGGTGGTGTCGATGGTAGAACGAAGCGGCGTCGAAGCCTGTAAAGCGGCGGTGCACAATCTTCTCGCGCAACGCGTCAGTGGGCTGATCATTAACTATCCGCTGGATGACCAGGATGCCATTGCTGTGGAAGCTGCCTGCACTAATGTTCCGGCGTTATTTCTTGATGTCTCTGACCAGACACCCATCAACAGTATTATTTTCTCCCATGAAGACGGTACGCGACTGGGCGTGGAGCATCTGGTCGCATTGGGTCACCAGCAAATCGCGCTGTTAGCGGGCCCATTAAGTTCTGTCTCGGCGCGTCTGCGTCTGGCTGGCTGGCATAAATATCTCACTCGCAATCAAATTCAGCCGATAGCGGAACGGGAAGGCGACTGGAGTGCCATGTCCGGTTTTCAACAAACCATGCAAATGCTGAATGAGGGCATCGTTCCCACTGCGATGCTGGTTGCCAACGATCAGATGGCGCTGGGCGCAATGCGCGCCATTACCGAGTCCGGGCTGCGCGTTGGTGCGGATATCTCGGTAGTGGGATACGACGATACCGAAGACAGCTCATGTTATATCCCGCCGTTAACCACCATCAAACAGGATTTTCGCCTGCTGGGGCAAACCAGCGTGGACCGCTTGCTGCAACTCTCTCAGGGCCAGGCGGTGAAGGGCAATCAGCTGTTGCCGGTCTCACTGGTGAAAAGAAAAACCACCCTGGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGAGCAGCCTGAGGCCTCCTAAGAAGAAGAGGAAGGTTGTTACAAACACCATTGTCACACTCCACAAACACCATTGTCACACTCCACAAACACCATTGTCACACTCCACAAACACCATTGTCACACTCCATCACATAGGAATAAAAAGCCATATCACATAGGAATAAAAAGCCATATCACATAGGAATAAAAAGCCATATCACATAGGAATAAAAAGCCATAGAAAGCGGACTACTGTAGTCCGCGGAGTACTGTCCTCCGCGGACGACTGTCGTCCGCGGAGCACTGTGCTCCGCGGATGACTGTCATCCGAGCGGAGACTCTAGAAATTGTGAGCGGATAACAATTGGGTATATAATGGGGGCCGTCAGCTACTACCAGAGCCGATCGAACACTCGAGCCGAGCAGACGTGCCTACAATTGTGAGCGCTCACAATTACTAGAAGATCTTGAATTCGCCACCATGGCTAGATTAGATAAAAGTAAAGTGATTAACAGCGCATTAGAGCTGCTTAATGAGGTCGGAATCGAAGGTTTAACAACCCGTAAACTCGCCCAGAAGCTAGGTGTAGAGCAGCCTACATTGTATTGGCATGTAAAAAATAAGCGGGCTTTGCTCGACGCCTTAGCCATTGAGATGTTAGATAGGCACCATACTCACTTTTGCCCTTTAGAAGGGGAAAGCTGGCAAGATTTTTTACGTAATAACGCTAAAAGTTTTAGATGTGCTTTACTAAGTCATCGCGATGGAGCAAAAGTACATTTAGGTACACGGCCTACAGAAAAACAGTATGAAACTCTCGAAAATCAATTAGCCTTTTTATGCCAACAAGGTTTTTCACTAGAGAATGCATTATATGCACTCAGCGCTGTGGGGCATTTTACTTTAGGTTGCGTATTGGAAGATCAAGAGCATCAAGTCGCTAAAGAAGAAAGGGAAACACCTACTACTGATAGTATGCCGCCATTATTACGACAAGCTATCGAATTATTTGATCACCAAGGTGCAGAGCCAGCCTTCTTATTCGGCCTTGAATTGATCATATGCGGATTAGAAAAACAACTTAAATGTGAAAGTGGGTCGCCAAAAAAGAAGAGAAAGGTGGATGGTGGCGGAGCACTGTCTCCCCAGCATTCAGCTGTGACCCAGGGATCAATCATTAAGAACAAGGAAGGTATGGATGCTAAGTCCCTGACCGCTTGGTCAAGAACTTTGGTAACTTTTAAGGACGTGTTCGTCGATTTTACTCGGGAGGAGTGGAAACTATTGGATACCGCCCAGCAAATCGTGTATAGAAACGTTATGCTCGAGAACTATAAGAATCTTGTGTCACTGGGCTACCAACTGACAAAGCCTGACGTTATCCTGAGGCTGGAAAAGGGTGAGGAGCCTTGGTTGGTCGAGCGGGAGATCCACCAGGAAACTCACCCAGACAGTGAGACTGCTTTCGAGATCAAGTCCTCTGTGTAATACAAGGCGGCCGCAAATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTAAAGCTTCGAATTCTGATAATCAGCCATACCACATTTGTAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCAGCTCGCCTTTTGCTCACATGTCTCCCAGGGGATGGGGGTCCATCCACCTTGCCGAAAAGATTTGTCTGAGGAACTGAAAATAGAAGGGAAAAAAGAGGAGGGACAAAAGAGGCAGAAATGAGAGGGGAGGGGACAGAGGACACCTGAATAAAGACCACACCCATGACCCACGTGATGCTGAGAAGTACTCCTGCCCTAGGAAGAGACTCAGGGCAGAGGGAGGAAGGACAGCAGACCAGACAGTCACAGCAGCCTTGACAAAACGTTCCTGGAACTCAAGCTCTTCTCCACAGAGGAGGACAGAGCAGACAGCAGAGACCATGGAGTCTCCCTCGGCCCCTCCCCACAGATGGTGCATCCCCTGGCAGAGGCTCCTGCTCACAGGTGAACTCGAGACCGGTATGGCCCCCCCGACCGATGTCAGCCTGGGGGACGAGCTCCACTTAGACGGCGAGGACGTGGCGATGGCGCATGCCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGGACGGGGATTCCCCGGGTCCGGGATTTACCCCCCACGACTCCGCCCCCTACGGCGCTCTGGATATGGCCGACTTCGAGTTTGAGCAGATGTTTACCGATGCCCTTGGAATTGACGAGTACGGTGGGACGCGTATGAAGCTACTGTCTTCTATCGAACAAGCATGCGATATTTGCCGACTTAAAAAGCTCAAGTGCTCCAAAGAAAAACCGAAGTGCGCCAAGTGTCTGAAGAACAACTGGGAGTGTCGCTACTCTCCCAAAACCAAAAGGTCTCCGCTGACTAGGGCACATCTGACAGAAGTGGAATCAAGGCTAGAAAGACTGGAACAGCTATTTCTACTGATTTTTCCTCGAGAAGACCTTGACATGATTTTGAAAATGGATTCTTTACAGGATATAAAAGCATTGTTAACAGGATTATTTGTACAAGATAATGTGAATAAAGATGCCGTCACAGATAGATTGGCTTCAGTGGAGACTGATATGCCTCTAACATTGAGACAGCATAGAATAAGTGCGACATCATCATCGGAAGAGAGTAGTAACAAAGGTCAAAGACAGTTGACTGTATAAAAGCTTCACTCCTCAGGTGCAGGCTGCCTATCAGAAGGTGGTGGCTGGTGTGGCCAATGCCCTGGCTCACAAATACCACTGAGATCTTTTTCCCTCTGCCAAAAATTATGGGGACATCATGAAGCCCCTTGAGCATCTGACTTCTGGCTAATAAAGGAAATTTATTTTCATTGCAATAGTGTGTTGGAATTTTTTGTGTCTCTCACTCGGAAGGACATATGGGAGGGCAAATCATTTAAAACATCAGAATGAGTATTTGGTTTAGAGTTTGGCAACATATGCCCATATGCTGGCTGCCATGAACAAAGGTTGGCTATAAAGAGGTCATCAGTATATGAAACAGCCCCCTGCTGTCCATTCCTTATTCCATAGAAAAGCCTTGACTTGAGGTTAGATTTTTTTTATATTTTGTTTTGTGTTATTTTTTTCTTTAACATCCCTAAAATTTTCCTTACATGTTTTACTAGCCAGATTTTTCCTCCTCTCCTGACTACTCCCAGTCATAGCTGTCCCTCTTCTCTTATGGAGATC(SEQ ID NO:3)。
具有SEQ ID NO:4所述核苷酸序列的核酸编码由细胞特异性启动子CEA368、第一microRNA为mir-143-3p、第二microRNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白分别为TALE蛋白、第一和第二识别序列为5×UAS的元件组成的表达系统。
CGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTTCTGTCCTCCGAGCGGAGACTCTAGATACCCCTCTCCGCTTCTGAATTCTAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCCTCGAGTACCCCTCTCCGCTTCTTACCCCTCTCCGCTTCTTACCCCTCTCCGCTTCTGAATTCGACCGCCACCATGAGACATATTATCTGCCACGGAGGTGTTATTACCGAAGAAATGGCCGCCAGTCTTTTGGACCAGCTGATCGAAGAGGTACTGGCTGATAATCTTCCACCTCCTAGCCATTTTGAACCACCTACCCTTCACGAACTGTATGATTTAGACGTGACGGCCCCCGAAGATCCCAACGAGGAGGCGGTTTCGCAGATTTTTCCCGACTCTGTAATGTTGGCGGTGCAGGAAGGGATTGACTTACTCACTTTTCCGCCGGCGCCCGGTTCTCCGGAGCCGCCTCACCTTTCCCGGCAGCCCGAGCAGCCGGAGCAGAGAGCCTTGGGTCCGGTTTCTATGCCAAACCTTGTACCGGAGGTGATCGATCTTACCTGCCACGAGGCTGGCTTTCCACCCAGTGACGACGAGGATGAAGAGGGTGAGGAGTTTGTGTTAGATTATGTGGAGCACCCCGGGCACGGTTGCAGGTCTTGTCATTATCACCGGAGGAATACGGGGGACCCAGATATTATGTGTTCGCTTTGCTATATGAGGACCTGTGGCATGTTTGTCTACAGTAAGTGAAAATTATGGGCAGTGGGTGATAGAGTGGTGGGTTTGGTGTGGTAATTTTTTTTTTAATTTTTACAGTTTTGTGGTTTAAAGAATTTTGTATTGTGATTTTTTTAAAAGGTCCTGTGTCTGAACCTGAGCCTGAGCCCGAGCCAGAACCGGAGCCTGCAAGACCTACCCGCCGTCCTAAAATGGCGCCTGCTATCCTGAGGCGCCCGACATCACCTGTGTCTAGAGAATGCAATAGTAGTACGGATAGCTGTGACTCCGGTCCTTCTAACACACCTCCTGAGATACACCCGGTGGTCCCGCTGTGCCCCATTAAACCAGTTGCCGTGAGAGTTGGTGGGCGTCGCCAGGCTGTGGAATGTATCGAGGACTTGCTTAACGAGCCTGGGCAACCTTTGGACTTGAGCTGTAAACGCCCCAGGCCAGGGTCCGGAGCTACAAATTTTTCCCTCCTCAAACAGGCTGGAGATGTCGAAGAAAATCCTGGGCCTATGGAGGCTTGGGAGTGTTTGGAAGATTTTTCTGCTGTGCGTAACTTGCTGGAACAGAGCTCTAACAGTACCTCTTGGTTTTGGAGGTTTCTGTGGGGCTCATCCCAGGCAAAGTTAGTCTGCAGAATTAAGGAGGATTACAAGTGGGAATTTGAAGAGCTTTTGAAATCCTGTGGTGAGCTGTTTGATTCTTTGAATCTGGGTCACCAGGCGCTTTTCCAAGAGAAGGTCATCAAGACTTTGGATTTTTCCACACCGGGGCGCGCTGCGGCTGCTGTTGCTTTTTTGAGTTTTATAAAGGATAAATGGAGCGAAGAAACCCATCTGAGCGGGGGGTACCTGCTGGATTTTCTGGCCATGCATCTGTGGAGAGCGGTTGTGAGACACAAGAATCGCCTGCTACTGTTGTCTTCCGTCCGCCCGGCGATAATACCGACGGAGGAGCAGCAGCAGCAGCAGGAGGAAGCCAGGCGGCGGCGGCAGGAGCAGAGCCCATGGAACCCGAGAGCCGGCCTGGACCCTCGGGAATGAATGTTGTACAGGTGGCTGAACTGTATCCAGAACTGAGACGCATTTTGACAATTACAGAGGATGGGCAGGGGCTAAAGGGGGTAAAGAGGGAGCGGGGGGCTTGTGAGGCTACAGAGGAGGCTAGGAATCTAGCTTTTAGCTTAATGACCAGACACCGTCCTGAGTGTATTACTTTTCAACAGATCAAGGATAATTGCGCTAATGAGCTTGATCTGCTGGCGCAGAAGTATTCCATAGAGCAGCTGACCACTTACTGGCTGCAGCCAGGGGATGATTTTGAGGAGGCTATTAGGGTATATGCAAAGGTGGCACTTAGGCCAGATTGCAAGTACAAGATCAGCAAACTTGTAAATATCAGGAATTGTTGCTACATTTCTGGGAACGGGGCCGAGGTGGAGATAGATACGGAGGATAGGGTGGCCTTTAGATGTAGCATGATAAATATGTGGCCGGGGGTGCTTGGCATGGACGGGGTGGTTATTATGAATGTAAGGTTTACTGGCCCCAATTTTAGCGGTACGGTTTTCCTGGCCAATACCAACCTTATCCTACACGGTGTAAGCTTCTATGGGTTTAACAATACCTGTGTGGAAGCCTGGACCGATGTAAGGGTTCGGGGCTGTGCCTTTTACTGCTGCTGGAAGGGGGTGGTGTGTCGCCCCAAAAGCAGGGCTTCAATTAAGAAATGCCTCTTTGAAAGGTGTACCTTGGGTATCCTGTCTGAGGGTAACTCCAGGGTGCGCCACAATGTGGCCTCCGACTGTGGTTGCTTCATGCTAGTGAAAAGCGTGGCTGTGATTAAGCATAACATGGTATGTGGCAACTGCGAGGACAGGGCCTCTCAGATGCTGACCTGCTCGGACGGCAACTGTCACCTGCTGAAGACCATTCACGTAGCCAGCCACTCTCGCAAGGCCTGGCCAGTGTTTGAGCATAACATACTGACCCGCTGTTCCTTGCATTTGGGTAACAGGAGGGGGGTGTTCCTACCTTACCAATGCAATTTGAGTCACACTAAGATATTGCTTGAGCCCGAGAGCATGTCCAAGGTGAACCTGAACGGGGTGTTTGACATGACCATGAAGATCTGGAAGGTGCTGAGGTACGATGAGACCCGCACCAGGTGCAGACCCTGCGAGTGTGGCGGTAAACATATTAGGAACCAGCCTGTGATGCTGGATGTGACCGAGGAGCTGAGGCCCGATCACTTGGTGCTGGCCTGCACCCGCGCTGAGTTTGGCTCTAGCGATGAAGATACAGATCTCGAGGGGTCCGGAGCTACAAATTTTTCCCTCCTCAAACAGGCTGGAGATGTCGAAGAAAATCCTGGGCCTACCGGTATGTATAGGATGCAGTTGCTCTCTTGCATCGCCCTGTCTCTAGCGTTGGTGACTAATAGCGCCCCTACTTCTTCTTCCACGAAGAAAACCCAGCTGCAACTGGAACACCTTCTGCTGGATCTCCAGATGATCCTTAACGGGATTAACAACTATAAGAACCCGAAGCTCACGGCTATGCTGACGGCGAAGTTCGCTATGCCTAAAAAGGCTACTGAGCTGAAACACTTACAGTGTCTCGAAGAGGCACTCAAACCCCTCGAGGAGGTACTCAATCTGGCGCAAAGCAAAAACTTCCATCTCCGCCCACGGGACCTAATCTCCAACATCAACGTGATTGTGCTGGAGCTCAAGGGGTCTGAAACCACGTTTATGTGCGAGTACGCAGATGAAACTGCTACCATAGTGGAATTCCTCAATCGTTGGATCACTTTCTGCCAGTCAATCATTTCAACCCTGACCAGATCTGAGGGCCGCGGCAGCCTGCTGACCTGCGGCGACGTGGAGGAAAACCCCGGCCCCGGATCCTTGTCGACCATTCGTCCGCGCAGGCCAAGTCCTGCCCGCGAGCTTCTGCCCGGACCCCAACCGGATAGGGTTCAGCCGACTGCAGATCGTGGGGTGTCTGCGCCTGCTGGCAGCCCTCTGGATGGCTTGCCCGCTCGGCGGACGGTGTCCCGGACCCGGCTGCCATCTCCCCCTGCGCCCTCACCTGCGTTCTCGGCGGGCAGCTTCAGCGATCTGCTCCGTCCGTTCGATCCGTCGCTTCTTGATACATCGCTTCTTGATTCGATGCCTGCCGTCGGCACGCCGCATACAGCGGCTGCCCCAGCAGAGTGGGATGAGGCGCAATCGGCTCTGCGTGCAGCCGATGACCCGCCACCCACCGTGCGTGTCGCTGTCACTGCCGCGCGGCCGCCGCGCGCCAAGCCGGCCCCGCGACGGCGTGCTGCGCAACCCTCCGACGCTTCGCCGGCCGCGCAGGTGGATCTACGCACGCTCGGCTACAGTCAGCAGCAGCAAGAGAAGATCAAACCGAAGGTGCGTTCGACAGTGGCGCAGCACCACGAGGCACTGGTGGGCCATGGGTTTACACACGCGCACATCGTTGCGCTCAGCCAACACCCGGCAGCGTTAGGGACCGTCGCTGTCACGTATCAGCACATAATCACGGCGTTGCCAGAGGCGACACACGAAGACATCGTTGGCGTCGGCAAACAGTGGTCCGGCGCACGCGCCCTGGAGGCCTTGCTCACGGATGCGGGGGAGTTGAGAGGTCCGCCGTTACAGTTGGACACAGGCCAACTTGTGAAGATTGCAAAACGTGGCGGCGTGACCGCAATGGAGGCAGTGCATGCATCGCGCAATGCACTGACGGGTGCCCCCCTGAACCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACAATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACAATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAAGCATTGTGGCCCAGCTGAGCCGGCCTGATCCGGCGTTGGCCGCGTTGACCAACGACCACCTCGTCGCCTTGGCCTGCCTCGGCGGACGTCCTGCCATGGATGCAGTGAAAAAGGGATTGCCGCACGCGCCGGAATTGATCAGAAGAGTCAATCGCCGTATTGGCGAACGCACGTCCCATCGCGTTGCCGACTACGCGCAAGTGGTTCGCGTGCTGGAGTTTTTCCAGTGCCACTCCCACCCAGCGTACGCATTTGATGAGGCCATGACGCAGTTCGGGATGAGCAGGAACGGGTTGGTACAGCTCTTTCGCAGAGTGGGCGTCACCGAACTCGAAGCCCGCGGTGGAACGCTCCCCCCAGCCTCGCAGCGTTGGGACCGTATCCTCCAGGCATCAGGGATGAAAAGGGCCAAACCGTCCCCTACTTCAGCTCAAACACCGGATCAGGCGTCTTTGCATGCATTCGCCGATTCGCTGGAGCGTGACCTTGATGCGCCCAGCCCAATGCACGAGGGAGATCAGACGCGGGCAAGCAGCCGTAAACGGTCCCGATCGGATCGTGCTGTCACCGGCCCCTCCGCACAGCAGGCTGTCGAGGTGCGCGTTCCCGAACAGCGCGATGCGCTGCATTTGCCCCTCAGCTGGAGGGTAAAACGCCCGCGTACCAGGATCTGGGGCGGCCTCCCGGTTAAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGGAGCTACAGTGCTTCATCTCAGAGCTACAGTGCTTCATCTCAGAGCTACAGTGCTTCATCTCAGAGCTACAGTGCTTCATCTCAGAAAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTTCTGTCCTCCGAGCGGAGACTCTAGATATAGAACCGATCCTCCCATTGAATTCTAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCCTCGAGTATAGAACCGATCCTCCCATTTATAGAACCGATCCTCCCATTTATAGAACCGATCCTCCCATTACTAGAAGATCTTGAATTCGCCACCTTGTCGACCATTCGTCCGCGCAGGCCAAGTCCTGCCCGCGAGCTTCTGCCCGGACCCCAACCGGATAGGGTTCAGCCGACTGCAGATCGTGGGGTGTCTGCGCCTGCTGGCAGCCCTCTGGATGGCTTGCCCGCTCGGCGGACGGTGTCCCGGACCCGGCTGCCATCTCCCCCTGCGCCCTCACCTGCGTTCTCGGCGGGCAGCTTCAGCGATCTGCTCCGTCCGTTCGATCCGTCGCTTCTTGATACATCGCTTCTTGATTCGATGCCTGCCGTCGGCACGCCGCATACAGCGGCTGCCCCAGCAGAGTGGGATGAGGCGCAATCGGCTCTGCGTGCAGCCGATGACCCGCCACCCACCGTGCGTGTCGCTGTCACTGCCGCGCGGCCGCCGCGCGCCAAGCCGGCCCCGCGACGGCGTGCTGCGCAACCCTCCGACGCTTCGCCGGCCGCGCAGGTGGATCTACGCACGCTCGGCTACAGTCAGCAGCAGCAAGAGAAGATCAAACCGAAGGTGCGTTCGACAGTGGCGCAGCACCACGAGGCACTGGTGGGCCATGGGTTTACACACGCGCACATCGTTGCGCTCAGCCAACACCCGGCAGCGTTAGGGACCGTCGCTGTCACGTATCAGCACATAATCACGGCGTTGCCAGAGGCGACACACGAAGACATCGTTGGCGTCGGCAAACAGTGGTCCGGCGCACGCGCCCTGGAGGCCTTGCTCACGGATGCGGGGGAGTTGAGAGGTCCGCCGTTACAGTTGGACACAGGCCAACTTGTGAAGATTGCAAAACGTGGCGGCGTGACCGCAATGGAGGCAGTGCATGCATCGCGCAATGCACTGACGGGTGCCCCCCTGAACCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACAATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAAGCATTGTGGCCCAGCTGAGCCGGCCTGATCCGGCGTTGGCCGCGTTGACCAACGACCACCTCGTCGCCTTGGCCTGCCTCGGCGGACGTCCTGCCATGGATGCAGTGAAAAAGGGATTGCCGCACGCGCCGGAATTGATCAGAAGAGTCAATCGCCGTATTGGCGAACGCACGTCCCATCGCGTTGCCGACTACGCGCAAGTGGTTCGCGTGCTGGAGTTTTTCCAGTGCCACTCCCACCCAGCGTACGCATTTGATGAGGCCATGACGCAGTTCGGGATGAGCAGGAACGGGTTGGTACAGCTCTTTCGCAGAGTGGGCGTCACCGAACTCGAAGCCCGCGGTGGAACGCTCCCCCCAGCCTCGCAGCGTTGGGACCGTATCCTCCAGGCATCAGGGATGAAAAGGGCCAAACCGTCCCCTACTTCAGCTCAAACACCGGATCAGGCGTCTTTGCATGCATTCGCCGATTCGCTGGAGCGTGACCTTGATGCGCCCAGCCCAATGCACGAGGGAGATCAGACGCGGGCAAGCAGCCGTAAACGGTCCCGATCGGATCGTGCTGTCACCGGCCCCTCCGCACAGCAGGCTGTCGAGGTGCGCGTTCCCGAACAGCGCGATGCGCTGCATTTGCCCCTCAGCTGGAGGGTAAAACGCCCGCGTACCAGGATCTGGGGCGGCCTCCCGTAATACAAGGCGGCCGCAAATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTAAAGCTTCGAATTCTGATAATCAGCCATACCACATTTGTAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCAGCTCGCCTTTTGCTCACATGTCTCCCAGGGGATGGGGGTCCATCCACCTTGCCGAAAAGATTTGTCTGAGGAACTGAAAATAGAAGGGAAAAAAGAGGAGGGACAAAAGAGGCAGAAATGAGAGGGGAGGGGACAGAGGACACCTGAATAAAGACCACACCCATGACCCACGTGATGCTGAGAAGTACTCCTGCCCTAGGAAGAGACTCAGGGCAGAGGGAGGAAGGACAGCAGACCAGACAGTCACAGCAGCCTTGACAAAACGTTCCTGGAACTCAAGCTCTTCTCCACAGAGGAGGACAGAGCAGACAGCAGAGACCATGGAGTCTCCCTCGGCCCCTCCCCACAGATGGTGCATCCCCTGGCAGAGGCTCCTGCTCACAGGTGAACTCGAGACCGGTATGGCCCCCCCGACCGATGTCAGCCTGGGGGACGAGCTCCACTTAGACGGCGAGGACGTGGCGATGGCGCATGCCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGAGCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGAGCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGAGCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGGACGGGGATTCCCCGGGTCCGGGATTTACCCCCCACGACTCCGCCCCCTACGGCGCTCTGGATATGGCCGACTTCGAGTTTGAGCAGATGTTTACCGATGCCCTTGGAATTGACGAGTACGGTGGGACGCGTATGAAGCTACTGTCTTCTATCGAACAAGCATGCGATATTTGCCGACTTAAAAAGCTCAAGTGCTCCAAAGAAAAACCGAAGTGCGCCAAGTGTCTGAAGAACAACTGGGAGTGTCGCTACTCTCCCAAAACCAAAAGGTCTCCGCTGACTAGGGCACATCTGACAGAAGTGGAATCAAGGCTAGAAAGACTGGAACAGCTATTTCTACTGATTTTTCCTCGAGAAGACCTTGACATGATTTTGAAAATGGATTCTTTACAGGATATAAAAGCATTGTTAACAGGATTATTTGTACAAGATAATGTGAATAAAGATGCCGTCACAGATAGATTGGCTTCAGTGGAGACTGATATGCCTCTAACATTGAGACAGCATAGAATAAGTGCGACATCATCATCGGAAGAGAGTAGTAACAAAGGTCAAAGACAGTTGACTGTATAAAAGCTTCACTCCTCAGGTGCAGGCTGCCTATCAGAAGGTGGTGGCTGGTGTGGCCAATGCCCTGGCTCACAAATACCACTGAGATCTTTTTCCCTCTGCCAAAAATTATGGGGACATCATGAAGCCCCTTGAGCATCTGACTTCTGGCTAATAAAGGAAATTTATTTTCATTGCAATAGTGTGTTGGAATTTTTTGTGTCTCTCACTCGGAAGGACATATGGGAGGGCAAATCATTTAAAACATCAGAATGAGTATTTGGTTTAGAGTTTGGCAACATATGCCCATATGCTGGCTGCCATGAACAAAGGTTGGCTATAAAGAGGTCATCAGTATATGAAACAGCCCCCTGCTGTCCATTCCTTATTCCATAGAAAAGCCTTGACTTGAGGTTAGATTTTTTTTATATTTTGTTTTGTGTTATTTTTTTCTTTAACATCCCTAAAATTTTCCTTACATGTTTTACTAGCCAGATTTTTCCTCCTCTCCTGACTACTCCCAGTCATAGCTGTCCCTCTTCTCTTATGGAGATC(SEQ IDNO:4)。
具有SEQ ID NO:5所述核苷酸序列的核酸编码由细胞特异性启动子CXCR4、第一microRNA为mir-135a-5p、第二microRNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白分别为Lacl、tetR-KRAB、第一和第二识别序列为5×UAS的元件组成的表达系统(SEQ ID NO:5)。
CGGGGTGCCGCCCCCCGCGGGTGACCGCCCTCCGCGGGTGACAGCCCTCCGCGGAGTGCCGCACTCCGCGGAGTGCTGTACTCCGAGCGGAGACTCTAGACTCCCTATCAGTGATAGAGATAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCTCCCTATCAGTGATAGAGAGAATTCGACCGCCACCATGAGACATATTATCTGCCACGGAGGTGTTATTACCGAAGAAATGGCCGCCAGTCTTTTGGACCAGCTGATCGAAGAGGTACTGGCTGATAATCTTCCACCTCCTAGCCATTTTGAACCACCTACCCTTCACGAACTGTATGATTTAGACGTGACGGCCCCCGAAGATCCCAACGAGGAGGCGGTTTCGCAGATTTTTCCCGACTCTGTAATGTTGGCGGTGCAGGAAGGGATTGACTTACTCACTTTTCCGCCGGCGCCCGGTTCTCCGGAGCCGCCTCACCTTTCCCGGCAGCCCGAGCAGCCGGAGCAGAGAGCCTTGGGTCCGGTTTCTATGCCAAACCTTGTACCGGAGGTGATCGATCTTACCTGCCACGAGGCTGGCTTTCCACCCAGTGACGACGAGGATGAAGAGGGTGAGGAGTTTGTGTTAGATTATGTGGAGCACCCCGGGCACGGTTGCAGGTCTTGTCATTATCACCGGAGGAATACGGGGGACCCAGATATTATGTGTTCGCTTTGCTATATGAGGACCTGTGGCATGTTTGTCTACAGTAAGTGAAAATTATGGGCAGTGGGTGATAGAGTGGTGGGTTTGGTGTGGTAATTTTTTTTTTAATTTTTACAGTTTTGTGGTTTAAAGAATTTTGTATTGTGATTTTTTTAAAAGGTCCTGTGTCTGAACCTGAGCCTGAGCCCGAGCCAGAACCGGAGCCTGCAAGACCTACCCGCCGTCCTAAAATGGCGCCTGCTATCCTGAGGCGCCCGACATCACCTGTGTCTAGAGAATGCAATAGTAGTACGGATAGCTGTGACTCCGGTCCTTCTAACACACCTCCTGAGATACACCCGGTGGTCCCGCTGTGCCCCATTAAACCAGTTGCCGTGAGAGTTGGTGGGCGTCGCCAGGCTGTGGAATGTATCGAGGACTTGCTTAACGAGCCTGGGCAACCTTTGGACTTGAGCTGTAAACGCCCCAGGCCACTCGAGGGTACCGGGTCCGGAGCTACAAATTTTTCCCTCCTCAAACAGGCTGGAGATGTCGAAGAAAATCCTGGGCCTACCGGTATGTGGCTGCAGAGCCTCCTTCTACTGGGCACCGTGGCTTGCAGCATCTCGGCCCCCGCCAGAAGTCCCAGCCCCAGCACCCAGCCCTGGGAACACGTAAATGCAATCCAGGAGGCCCGCCGCCTCCTGAACTTGTCACGGGACACGGCAGCCGAGATGAATGAAACTGTGGAGGTCATCAGTGAAATGTTTGACTTGCAGGAGCCCACATGCCTGCAGACTCGGCTGGAGCTCTACAAACAGGGCCTGCGTGGCAGCCTCACCAAGCTGAAGGGCCCACTGACCATGATGGCTAGCCATTACAAGCAGCACTGCCCGCCGACCCCTGAGACATCCTGTGCCACTCAGATCATTACCTTTGAGTCTTTCAAGGAGAACCTCAAGGACTTTCTGCTGGTCATCCCATTTGACTGCTGGGAACCCGTCCAGGAAAGATCTGAGGGCCGCGGCAGCCTGCTGACCTGCGGCGACGTGGAGGAAAACCCCGGCCCCGGATCCATGAAACCAGTAACGTTATACGATGTCGCAGAGTATGCCGGTGTCTCTTATCAGACCGTTTCCCGCGTGGTGAACCAGGCCAGCCACGTTTCTGCGAAAACGCGGGAAAAAGTGGAAGCGGCGATGGCGGAGCTGAATTACATTCCCAACCGCGTGGCACAACAACTGGCGGGCAAACAGTCGTTGCTGATTGGCGTTGCCACCTCCAGTCTGGCCCTGCACGCGCCGTCGCAAATTGTCGCGGCGATTAAATCTCGCGCCGATCAACTGGGTGCCAGCGTGGTGGTGTCGATGGTAGAACGAAGCGGCGTCGAAGCCTGTAAAGCGGCGGTGCACAATCTTCTCGCGCAACGCGTCAGTGGGCTGATCATTAACTATCCGCTGGATGACCAGGATGCCATTGCTGTGGAAGCTGCCTGCACTAATGTTCCGGCGTTATTTCTTGATGTCTCTGACCAGACACCCATCAACAGTATTATTTTCTCCCATGAAGACGGTACGCGACTGGGCGTGGAGCATCTGGTCGCATTGGGTCACCAGCAAATCGCGCTGTTAGCGGGCCCATTAAGTTCTGTCTCGGCGCGTCTGCGTCTGGCTGGCTGGCATAAATATCTCACTCGCAATCAAATTCAGCCGATAGCGGAACGGGAAGGCGACTGGAGTGCCATGTCCGGTTTTCAACAAACCATGCAAATGCTGAATGAGGGCATCGTTCCCACTGCGATGCTGGTTGCCAACGATCAGATGGCGCTGGGCGCAATGCGCGCCATTACCGAGTCCGGGCTGCGCGTTGGTGCGGATATCTCGGTAGTGGGATACGACGATACCGAAGACAGCTCATGTTATATCCCGCCGTTAACCACCATCAAACAGGATTTTCGCCTGCTGGGGCAAACCAGCGTGGACCGCTTGCTGCAACTCTCTCAGGGCCAGGCGGTGAAGGGCAATCAGCTGTTGCCGGTCTCACTGGTGAAAAGAAAAACCACCCTGGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGAGCAGCCTGAGGCCTCCTAAGAAGAAGAGGAAGGTTGTTACAAACACCATTGTCACACTCCACAAACACCATTGTCACACTCCACAAACACCATTGTCACACTCCACAAACACCATTGTCACACTCCATCACATAGGAATAAAAAGCCATATCACATAGGAATAAAAAGCCATATCACATAGGAATAAAAAGCCATATCACATAGGAATAAAAAGCCATAGAAAGCGGACTACTGTAGTCCGCGGAGTACTGTCCTCCGCGGACGACTGTCGTCCGCGGAGCACTGTGCTCCGCGGATGACTGTCATCCGAGCGGAGACTCTAGAAATTGTGAGCGGATAACAATTGGGTATATAATGGGGGCCGTCAGCTACTACCAGAGCCGATCGAACACTCGAGCCGAGCAGACGTGCCTACAATTGTGAGCGCTCACAATTACTAGAAGATCTTGAATTCGCCACCATGGCTAGATTAGATAAAAGTAAAGTGATTAACAGCGCATTAGAGCTGCTTAATGAGGTCGGAATCGAAGGTTTAACAACCCGTAAACTCGCCCAGAAGCTAGGTGTAGAGCAGCCTACATTGTATTGGCATGTAAAAAATAAGCGGGCTTTGCTCGACGCCTTAGCCATTGAGATGTTAGATAGGCACCATACTCACTTTTGCCCTTTAGAAGGGGAAAGCTGGCAAGATTTTTTACGTAATAACGCTAAAAGTTTTAGATGTGCTTTACTAAGTCATCGCGATGGAGCAAAAGTACATTTAGGTACACGGCCTACAGAAAAACAGTATGAAACTCTCGAAAATCAATTAGCCTTTTTATGCCAACAAGGTTTTTCACTAGAGAATGCATTATATGCACTCAGCGCTGTGGGGCATTTTACTTTAGGTTGCGTATTGGAAGATCAAGAGCATCAAGTCGCTAAAGAAGAAAGGGAAACACCTACTACTGATAGTATGCCGCCATTATTACGACAAGCTATCGAATTATTTGATCACCAAGGTGCAGAGCCAGCCTTCTTATTCGGCCTTGAATTGATCATATGCGGATTAGAAAAACAACTTAAATGTGAAAGTGGGTCGCCAAAAAAGAAGAGAAAGGTGGATGGTGGCGGAGCACTGTCTCCCCAGCATTCAGCTGTGACCCAGGGATCAATCATTAAGAACAAGGAAGGTATGGATGCTAAGTCCCTGACCGCTTGGTCAAGAACTTTGGTAACTTTTAAGGACGTGTTCGTCGATTTTACTCGGGAGGAGTGGAAACTATTGGATACCGCCCAGCAAATCGTGTATAGAAACGTTATGCTCGAGAACTATAAGAATCTTGTGTCACTGGGCTACCAACTGACAAAGCCTGACGTTATCCTGAGGCTGGAAAAGGGTGAGGAGCCTTGGTTGGTCGAGCGGGAGATCCACCAGGAAACTCACCCAGACAGTGAGACTGCTTTCGAGATCAAGTCCTCTGTGTAATACAAGGCGGCCGCAAATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTAAAGCTTCGAATTCTGATAATCAGCCATACCACATTTGTAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCAGCTCGCCTTTTGCTCACATGTTACCGACCACCCGCAAACAGCAGGGTCCCCTGGGCTTCCCAAGCCGCGCACCTCTCCGCCCCGCCCCTGCGCCCTCCTTCCTCGCGTCTGCCCCTCTCCCCCACCCCGCCTTCTCCCTCCCCGCCCCAGCGGCGCATGCGCCGCGCTCGGAGCGTGTTTTTATAAAAGTCCGGCCGCGGCCAGAAACTTCAGTTTGTTGGCTGCGGCAGCAGGTAGCAAAGTGACGCCGAGGGCCTGAGTGCTCCAGTAGCCACCGCATCTGGAGAACCAGCGGTTACCCTCGAGACCGGTATGGCCCCCCCGACCGATGTCAGCCTGGGGGACGAGCTCCACTTAGACGGCGAGGACGTGGCGATGGCGCATGCCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGGACGGGGATTCCCCGGGTCCGGGATTTACCCCCCACGACTCCGCCCCCTACGGCGCTCTGGATATGGCCGACTTCGAGTTTGAGCAGATGTTTACCGATGCCCTTGGAATTGACGAGTACGGTGGGACGCGTATGAAGCTACTGTCTTCTATCGAACAAGCATGCGATATTTGCCGACTTAAAAAGCTCAAGTGCTCCAAAGAAAAACCGAAGTGCGCCAAGTGTCTGAAGAACAACTGGGAGTGTCGCTACTCTCCCAAAACCAAAAGGTCTCCGCTGACTAGGGCACATCTGACAGAAGTGGAATCAAGGCTAGAAAGACTGGAACAGCTATTTCTACTGATTTTTCCTCGAGAAGACCTTGACATGATTTTGAAAATGGATTCTTTACAGGATATAAAAGCATTGTTAACAGGATTATTTGTACAAGATAATGTGAATAAAGATGCCGTCACAGATAGATTGGCTTCAGTGGAGACTGATATGCCTCTAACATTGAGACAGCATAGAATAAGTGCGACATCATCATCGGAAGAGAGTAGTAACAAAGGTCAAAGACAGTTGACTGTATAAAAGCTTCACTCCTCAGGTGCAGGCTGCCTATCAGAAGGTGGTGGCTGGTGTGGCCAATGCCCTGGCTCACAAATACCACTGAGATCTTTTTCCCTCTGCCAAAAATTATGGGGACATCATGAAGCCCCTTGAGCATCTGACTTCTGGCTAATAAAGGAAATTTATTTTCATTGCAATAGTGTGTTGGAATTTTTTGTGTCTCTCACTCGGAAGGACATATGGGAGGGCAAATCATTTAAAACATCAGAATGAGTATTTGGTTTAGAGTTTGGCAACATATGCCCATATGCTGGCTGCCATGAACAAAGGTTGGCTATAAAGAGGTCATCAGTATATGAAACAGCCCCCTGCTGTCCATTCCTTATTCCATAGAAAAGCCTTGACTTGAGGTTAGATTTTTTTTATATTTTGTTTTGTGTTATTTTTTTCTTTAACATCCCTAAAATTTTCCTTACATGTTTTACTAGCCAGATTTTTCCTCCTCTCCTGACTACTCCCAGTCATAGCTGTCCCTCTTCTCTTATGGAGATC(SEQ ID NO:5)。
具有SEQ ID NO:6所述核苷酸序列的核酸编码由细胞特异性启动子CXCR4、第一microRNA为mir-143-3p、第二microRNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白分别为TALE蛋白、第一和第二识别序列为5×UAS的元件组成的表达系统。
CGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTTCTGTCCTCCGAGCGGAGACTCTAGATACCCCTCTCCGCTTCTGAATTCTAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCCTCGAGTACCCCTCTCCGCTTCTTACCCCTCTCCGCTTCTTACCCCTCTCCGCTTCTGAATTCGACCGCCACCATGAGACATATTATCTGCCACGGAGGTGTTATTACCGAAGAAATGGCCGCCAGTCTTTTGGACCAGCTGATCGAAGAGGTACTGGCTGATAATCTTCCACCTCCTAGCCATTTTGAACCACCTACCCTTCACGAACTGTATGATTTAGACGTGACGGCCCCCGAAGATCCCAACGAGGAGGCGGTTTCGCAGATTTTTCCCGACTCTGTAATGTTGGCGGTGCAGGAAGGGATTGACTTACTCACTTTTCCGCCGGCGCCCGGTTCTCCGGAGCCGCCTCACCTTTCCCGGCAGCCCGAGCAGCCGGAGCAGAGAGCCTTGGGTCCGGTTTCTATGCCAAACCTTGTACCGGAGGTGATCGATCTTACCTGCCACGAGGCTGGCTTTCCACCCAGTGACGACGAGGATGAAGAGGGTGAGGAGTTTGTGTTAGATTATGTGGAGCACCCCGGGCACGGTTGCAGGTCTTGTCATTATCACCGGAGGAATACGGGGGACCCAGATATTATGTGTTCGCTTTGCTATATGAGGACCTGTGGCATGTTTGTCTACAGTAAGTGAAAATTATGGGCAGTGGGTGATAGAGTGGTGGGTTTGGTGTGGTAATTTTTTTTTTAATTTTTACAGTTTTGTGGTTTAAAGAATTTTGTATTGTGATTTTTTTAAAAGGTCCTGTGTCTGAACCTGAGCCTGAGCCCGAGCCAGAACCGGAGCCTGCAAGACCTACCCGCCGTCCTAAAATGGCGCCTGCTATCCTGAGGCGCCCGACATCACCTGTGTCTAGAGAATGCAATAGTAGTACGGATAGCTGTGACTCCGGTCCTTCTAACACACCTCCTGAGATACACCCGGTGGTCCCGCTGTGCCCCATTAAACCAGTTGCCGTGAGAGTTGGTGGGCGTCGCCAGGCTGTGGAATGTATCGAGGACTTGCTTAACGAGCCTGGGCAACCTTTGGACTTGAGCTGTAAACGCCCCAGGCCAGGGTCCGGAGCTACAAATTTTTCCCTCCTCAAACAGGCTGGAGATGTCGAAGAAAATCCTGGGCCTATGGAGGCTTGGGAGTGTTTGGAAGATTTTTCTGCTGTGCGTAACTTGCTGGAACAGAGCTCTAACAGTACCTCTTGGTTTTGGAGGTTTCTGTGGGGCTCATCCCAGGCAAAGTTAGTCTGCAGAATTAAGGAGGATTACAAGTGGGAATTTGAAGAGCTTTTGAAATCCTGTGGTGAGCTGTTTGATTCTTTGAATCTGGGTCACCAGGCGCTTTTCCAAGAGAAGGTCATCAAGACTTTGGATTTTTCCACACCGGGGCGCGCTGCGGCTGCTGTTGCTTTTTTGAGTTTTATAAAGGATAAATGGAGCGAAGAAACCCATCTGAGCGGGGGGTACCTGCTGGATTTTCTGGCCATGCATCTGTGGAGAGCGGTTGTGAGACACAAGAATCGCCTGCTACTGTTGTCTTCCGTCCGCCCGGCGATAATACCGACGGAGGAGCAGCAGCAGCAGCAGGAGGAAGCCAGGCGGCGGCGGCAGGAGCAGAGCCCATGGAACCCGAGAGCCGGCCTGGACCCTCGGGAATGAATGTTGTACAGGTGGCTGAACTGTATCCAGAACTGAGACGCATTTTGACAATTACAGAGGATGGGCAGGGGCTAAAGGGGGTAAAGAGGGAGCGGGGGGCTTGTGAGGCTACAGAGGAGGCTAGGAATCTAGCTTTTAGCTTAATGACCAGACACCGTCCTGAGTGTATTACTTTTCAACAGATCAAGGATAATTGCGCTAATGAGCTTGATCTGCTGGCGCAGAAGTATTCCATAGAGCAGCTGACCACTTACTGGCTGCAGCCAGGGGATGATTTTGAGGAGGCTATTAGGGTATATGCAAAGGTGGCACTTAGGCCAGATTGCAAGTACAAGATCAGCAAACTTGTAAATATCAGGAATTGTTGCTACATTTCTGGGAACGGGGCCGAGGTGGAGATAGATACGGAGGATAGGGTGGCCTTTAGATGTAGCATGATAAATATGTGGCCGGGGGTGCTTGGCATGGACGGGGTGGTTATTATGAATGTAAGGTTTACTGGCCCCAATTTTAGCGGTACGGTTTTCCTGGCCAATACCAACCTTATCCTACACGGTGTAAGCTTCTATGGGTTTAACAATACCTGTGTGGAAGCCTGGACCGATGTAAGGGTTCGGGGCTGTGCCTTTTACTGCTGCTGGAAGGGGGTGGTGTGTCGCCCCAAAAGCAGGGCTTCAATTAAGAAATGCCTCTTTGAAAGGTGTACCTTGGGTATCCTGTCTGAGGGTAACTCCAGGGTGCGCCACAATGTGGCCTCCGACTGTGGTTGCTTCATGCTAGTGAAAAGCGTGGCTGTGATTAAGCATAACATGGTATGTGGCAACTGCGAGGACAGGGCCTCTCAGATGCTGACCTGCTCGGACGGCAACTGTCACCTGCTGAAGACCATTCACGTAGCCAGCCACTCTCGCAAGGCCTGGCCAGTGTTTGAGCATAACATACTGACCCGCTGTTCCTTGCATTTGGGTAACAGGAGGGGGGTGTTCCTACCTTACCAATGCAATTTGAGTCACACTAAGATATTGCTTGAGCCCGAGAGCATGTCCAAGGTGAACCTGAACGGGGTGTTTGACATGACCATGAAGATCTGGAAGGTGCTGAGGTACGATGAGACCCGCACCAGGTGCAGACCCTGCGAGTGTGGCGGTAAACATATTAGGAACCAGCCTGTGATGCTGGATGTGACCGAGGAGCTGAGGCCCGATCACTTGGTGCTGGCCTGCACCCGCGCTGAGTTTGGCTCTAGCGATGAAGATACAGATCTCGAGGGGTCCGGAGCTACAAATTTTTCCCTCCTCAAACAGGCTGGAGATGTCGAAGAAAATCCTGGGCCTACCGGTATGTATAGGATGCAGTTGCTCTCTTGCATCGCCCTGTCTCTAGCGTTGGTGACTAATAGCGCCCCTACTTCTTCTTCCACGAAGAAAACCCAGCTGCAACTGGAACACCTTCTGCTGGATCTCCAGATGATCCTTAACGGGATTAACAACTATAAGAACCCGAAGCTCACGGCTATGCTGACGGCGAAGTTCGCTATGCCTAAAAAGGCTACTGAGCTGAAACACTTACAGTGTCTCGAAGAGGCACTCAAACCCCTCGAGGAGGTACTCAATCTGGCGCAAAGCAAAAACTTCCATCTCCGCCCACGGGACCTAATCTCCAACATCAACGTGATTGTGCTGGAGCTCAAGGGGTCTGAAACCACGTTTATGTGCGAGTACGCAGATGAAACTGCTACCATAGTGGAATTCCTCAATCGTTGGATCACTTTCTGCCAGTCAATCATTTCAACCCTGACCAGATCTGAGGGCCGCGGCAGCCTGCTGACCTGCGGCGACGTGGAGGAAAACCCCGGCCCCGGATCCTTGTCGACCATTCGTCCGCGCAGGCCAAGTCCTGCCCGCGAGCTTCTGCCCGGACCCCAACCGGATAGGGTTCAGCCGACTGCAGATCGTGGGGTGTCTGCGCCTGCTGGCAGCCCTCTGGATGGCTTGCCCGCTCGGCGGACGGTGTCCCGGACCCGGCTGCCATCTCCCCCTGCGCCCTCACCTGCGTTCTCGGCGGGCAGCTTCAGCGATCTGCTCCGTCCGTTCGATCCGTCGCTTCTTGATACATCGCTTCTTGATTCGATGCCTGCCGTCGGCACGCCGCATACAGCGGCTGCCCCAGCAGAGTGGGATGAGGCGCAATCGGCTCTGCGTGCAGCCGATGACCCGCCACCCACCGTGCGTGTCGCTGTCACTGCCGCGCGGCCGCCGCGCGCCAAGCCGGCCCCGCGACGGCGTGCTGCGCAACCCTCCGACGCTTCGCCGGCCGCGCAGGTGGATCTACGCACGCTCGGCTACAGTCAGCAGCAGCAAGAGAAGATCAAACCGAAGGTGCGTTCGACAGTGGCGCAGCACCACGAGGCACTGGTGGGCCATGGGTTTACACACGCGCACATCGTTGCGCTCAGCCAACACCCGGCAGCGTTAGGGACCGTCGCTGTCACGTATCAGCACATAATCACGGCGTTGCCAGAGGCGACACACGAAGACATCGTTGGCGTCGGCAAACAGTGGTCCGGCGCACGCGCCCTGGAGGCCTTGCTCACGGATGCGGGGGAGTTGAGAGGTCCGCCGTTACAGTTGGACACAGGCCAACTTGTGAAGATTGCAAAACGTGGCGGCGTGACCGCAATGGAGGCAGTGCATGCATCGCGCAATGCACTGACGGGTGCCCCCCTGAACCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACAATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACAATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAAGCATTGTGGCCCAGCTGAGCCGGCCTGATCCGGCGTTGGCCGCGTTGACCAACGACCACCTCGTCGCCTTGGCCTGCCTCGGCGGACGTCCTGCCATGGATGCAGTGAAAAAGGGATTGCCGCACGCGCCGGAATTGATCAGAAGAGTCAATCGCCGTATTGGCGAACGCACGTCCCATCGCGTTGCCGACTACGCGCAAGTGGTTCGCGTGCTGGAGTTTTTCCAGTGCCACTCCCACCCAGCGTACGCATTTGATGAGGCCATGACGCAGTTCGGGATGAGCAGGAACGGGTTGGTACAGCTCTTTCGCAGAGTGGGCGTCACCGAACTCGAAGCCCGCGGTGGAACGCTCCCCCCAGCCTCGCAGCGTTGGGACCGTATCCTCCAGGCATCAGGGATGAAAAGGGCCAAACCGTCCCCTACTTCAGCTCAAACACCGGATCAGGCGTCTTTGCATGCATTCGCCGATTCGCTGGAGCGTGACCTTGATGCGCCCAGCCCAATGCACGAGGGAGATCAGACGCGGGCAAGCAGCCGTAAACGGTCCCGATCGGATCGTGCTGTCACCGGCCCCTCCGCACAGCAGGCTGTCGAGGTGCGCGTTCCCGAACAGCGCGATGCGCTGCATTTGCCCCTCAGCTGGAGGGTAAAACGCCCGCGTACCAGGATCTGGGGCGGCCTCCCGGTTAAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGGAGCTACAGTGCTTCATCTCAGAGCTACAGTGCTTCATCTCAGAGCTACAGTGCTTCATCTCAGAGCTACAGTGCTTCATCTCAGAAAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTTCTGTCCTCCGAGCGGAGACTCTAGATATAGAACCGATCCTCCCATTGAATTCTAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCCTCGAGTATAGAACCGATCCTCCCATTTATAGAACCGATCCTCCCATTTATAGAACCGATCCTCCCATTACTAGAAGATCTTGAATTCGCCACCTTGTCGACCATTCGTCCGCGCAGGCCAAGTCCTGCCCGCGAGCTTCTGCCCGGACCCCAACCGGATAGGGTTCAGCCGACTGCAGATCGTGGGGTGTCTGCGCCTGCTGGCAGCCCTCTGGATGGCTTGCCCGCTCGGCGGACGGTGTCCCGGACCCGGCTGCCATCTCCCCCTGCGCCCTCACCTGCGTTCTCGGCGGGCAGCTTCAGCGATCTGCTCCGTCCGTTCGATCCGTCGCTTCTTGATACATCGCTTCTTGATTCGATGCCTGCCGTCGGCACGCCGCATACAGCGGCTGCCCCAGCAGAGTGGGATGAGGCGCAATCGGCTCTGCGTGCAGCCGATGACCCGCCACCCACCGTGCGTGTCGCTGTCACTGCCGCGCGGCCGCCGCGCGCCAAGCCGGCCCCGCGACGGCGTGCTGCGCAACCCTCCGACGCTTCGCCGGCCGCGCAGGTGGATCTACGCACGCTCGGCTACAGTCAGCAGCAGCAAGAGAAGATCAAACCGAAGGTGCGTTCGACAGTGGCGCAGCACCACGAGGCACTGGTGGGCCATGGGTTTACACACGCGCACATCGTTGCGCTCAGCCAACACCCGGCAGCGTTAGGGACCGTCGCTGTCACGTATCAGCACATAATCACGGCGTTGCCAGAGGCGACACACGAAGACATCGTTGGCGTCGGCAAACAGTGGTCCGGCGCACGCGCCCTGGAGGCCTTGCTCACGGATGCGGGGGAGTTGAGAGGTCCGCCGTTACAGTTGGACACAGGCCAACTTGTGAAGATTGCAAAACGTGGCGGCGTGACCGCAATGGAGGCAGTGCATGCATCGCGCAATGCACTGACGGGTGCCCCCCTGAACCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACAATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAAGCATTGTGGCCCAGCTGAGCCGGCCTGATCCGGCGTTGGCCGCGTTGACCAACGACCACCTCGTCGCCTTGGCCTGCCTCGGCGGACGTCCTGCCATGGATGCAGTGAAAAAGGGATTGCCGCACGCGCCGGAATTGATCAGAAGAGTCAATCGCCGTATTGGCGAACGCACGTCCCATCGCGTTGCCGACTACGCGCAAGTGGTTCGCGTGCTGGAGTTTTTCCAGTGCCACTCCCACCCAGCGTACGCATTTGATGAGGCCATGACGCAGTTCGGGATGAGCAGGAACGGGTTGGTACAGCTCTTTCGCAGAGTGGGCGTCACCGAACTCGAAGCCCGCGGTGGAACGCTCCCCCCAGCCTCGCAGCGTTGGGACCGTATCCTCCAGGCATCAGGGATGAAAAGGGCCAAACCGTCCCCTACTTCAGCTCAAACACCGGATCAGGCGTCTTTGCATGCATTCGCCGATTCGCTGGAGCGTGACCTTGATGCGCCCAGCCCAATGCACGAGGGAGATCAGACGCGGGCAAGCAGCCGTAAACGGTCCCGATCGGATCGTGCTGTCACCGGCCCCTCCGCACAGCAGGCTGTCGAGGTGCGCGTTCCCGAACAGCGCGATGCGCTGCATTTGCCCCTCAGCTGGAGGGTAAAACGCCCGCGTACCAGGATCTGGGGCGGCCTCCCGTAATACAAGGCGGCCGCAAATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTAAAGCTTCGAATTCTGATAATCAGCCATACCACATTTGTAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCTACCGACCACCCGCAAACAGCAGGGTCCCCTGGGCTTCCCAAGCCGCGCACCTCTCCGCCCCGCCCCTGCGCCCTCCTTCCTCGCGTCTGCCCCTCTCCCCCACCCCGCCTTCTCCCTCCCCGCCCCAGCGGCGCATGCGCCGCGCTCGGAGCGTGTTTTTATAAAAGTCCGGCCGCGGCCAGAAACTTCAGTTTGTTGGCTGCGGCAGCAGGTAGCAAAGTGACGCCGAGGGCCTGAGTGCTCCAGTAGCCACCGCATCTGGAGAACCAGCGGTTACCATGGCCCCCCCGACCGATGTCAGCCTGGGGGACGAGCTCCACTTAGACGGCGAGGACGTGGCGATGGCGCATGCCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGAGCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGAGCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGAGCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGGACGGGGATTCCCCGGGTCCGGGATTTACCCCCCACGACTCCGCCCCCTACGGCGCTCTGGATATGGCCGACTTCGAGTTTGAGCAGATGTTTACCGATGCCCTTGGAATTGACGAGTACGGTGGGACGCGTATGAAGCTACTGTCTTCTATCGAACAAGCATGCGATATTTGCCGACTTAAAAAGCTCAAGTGCTCCAAAGAAAAACCGAAGTGCGCCAAGTGTCTGAAGAACAACTGGGAGTGTCGCTACTCTCCCAAAACCAAAAGGTCTCCGCTGACTAGGGCACATCTGACAGAAGTGGAATCAAGGCTAGAAAGACTGGAACAGCTATTTCTACTGATTTTTCCTCGAGAAGACCTTGACATGATTTTGAAAATGGATTCTTTACAGGATATAAAAGCATTGTTAACAGGATTATTTGTACAAGATAATGTGAATAAAGATGCCGTCACAGATAGATTGGCTTCAGTGGAGACTGATATGCCTCTAACATTGAGACAGCATAGAATAAGTGCGACATCATCATCGGAAGAGAGTAGTAACAAAGGTCAAAGACAGTTGACTGTATAAAAGCTTCACTCCTCAGGTGCAGGCTGCCTATCAGAAGGTGGTGGCTGGTGTGGCCAATGCCCTGGCTCACAAATACCACTGAGATCTTTTTCCCTCTGCCAAAAATTATGGGGACATCATGAAGCCCCTTGAGCATCTGACTTCTGGCTAATAAAGGAAATTTATTTTCATTGCAATAGTGTGTTGGAATTTTTTGTGTCTCTCACTCGGAAGGACATATGGGAGGGCAAATCATTTAAAACATCAGAATGAGTATTTGGTTTAGAGTTTGGCAACATATGCCCATATGCTGGCTGCCATGAACAAAGGTTGGCTATAAAGAGGTCATCAGTATATGAAACAGCCCCCTGCTGTCCATTCCTTATTCCATAGAAAAGCCTTGACTTGAGGTTAGATTTTTTTTATATTTTGTTTTGTGTTATTTTTTTCTTTAACATCCCTAAAATTTTCCTTACATGTTTTACTAGCCAGATTTTTCCTCCTCTCCTGACTACTCCCAGTCATAGCTGTCCCTCTTCTCTTATGGAGATC(SEQ ID NO:6)。
具有SEQ ID NO:7所述核苷酸序列的核酸编码由细胞特异性启动子Survivin1、第一microRNA为mir-135a-5p、第二microRNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白分别为Lacl、tetR-KRAB、第一和第二识别序列为5×UAS的元件组成的表达系统。
CGGGGTGCCGCCCCCCGCGGGTGACCGCCCTCCGCGGGTGACAGCCCTCCGCGGAGTGCCGCACTCCGCGGAGTGCTGTACTCCGAGCGGAGACTCTAGACTCCCTATCAGTGATAGAGATAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCTCCCTATCAGTGATAGAGAGAATTCGACCGCCACCATGAGACATATTATCTGCCACGGAGGTGTTATTACCGAAGAAATGGCCGCCAGTCTTTTGGACCAGCTGATCGAAGAGGTACTGGCTGATAATCTTCCACCTCCTAGCCATTTTGAACCACCTACCCTTCACGAACTGTATGATTTAGACGTGACGGCCCCCGAAGATCCCAACGAGGAGGCGGTTTCGCAGATTTTTCCCGACTCTGTAATGTTGGCGGTGCAGGAAGGGATTGACTTACTCACTTTTCCGCCGGCGCCCGGTTCTCCGGAGCCGCCTCACCTTTCCCGGCAGCCCGAGCAGCCGGAGCAGAGAGCCTTGGGTCCGGTTTCTATGCCAAACCTTGTACCGGAGGTGATCGATCTTACCTGCCACGAGGCTGGCTTTCCACCCAGTGACGACGAGGATGAAGAGGGTGAGGAGTTTGTGTTAGATTATGTGGAGCACCCCGGGCACGGTTGCAGGTCTTGTCATTATCACCGGAGGAATACGGGGGACCCAGATATTATGTGTTCGCTTTGCTATATGAGGACCTGTGGCATGTTTGTCTACAGTAAGTGAAAATTATGGGCAGTGGGTGATAGAGTGGTGGGTTTGGTGTGGTAATTTTTTTTTTAATTTTTACAGTTTTGTGGTTTAAAGAATTTTGTATTGTGATTTTTTTAAAAGGTCCTGTGTCTGAACCTGAGCCTGAGCCCGAGCCAGAACCGGAGCCTGCAAGACCTACCCGCCGTCCTAAAATGGCGCCTGCTATCCTGAGGCGCCCGACATCACCTGTGTCTAGAGAATGCAATAGTAGTACGGATAGCTGTGACTCCGGTCCTTCTAACACACCTCCTGAGATACACCCGGTGGTCCCGCTGTGCCCCATTAAACCAGTTGCCGTGAGAGTTGGTGGGCGTCGCCAGGCTGTGGAATGTATCGAGGACTTGCTTAACGAGCCTGGGCAACCTTTGGACTTGAGCTGTAAACGCCCCAGGCCACTCGAGGGTACCGGGTCCGGAGCTACAAATTTTTCCCTCCTCAAACAGGCTGGAGATGTCGAAGAAAATCCTGGGCCTACCGGTATGTGGCTGCAGAGCCTCCTTCTACTGGGCACCGTGGCTTGCAGCATCTCGGCCCCCGCCAGAAGTCCCAGCCCCAGCACCCAGCCCTGGGAACACGTAAATGCAATCCAGGAGGCCCGCCGCCTCCTGAACTTGTCACGGGACACGGCAGCCGAGATGAATGAAACTGTGGAGGTCATCAGTGAAATGTTTGACTTGCAGGAGCCCACATGCCTGCAGACTCGGCTGGAGCTCTACAAACAGGGCCTGCGTGGCAGCCTCACCAAGCTGAAGGGCCCACTGACCATGATGGCTAGCCATTACAAGCAGCACTGCCCGCCGACCCCTGAGACATCCTGTGCCACTCAGATCATTACCTTTGAGTCTTTCAAGGAGAACCTCAAGGACTTTCTGCTGGTCATCCCATTTGACTGCTGGGAACCCGTCCAGGAAAGATCTGAGGGCCGCGGCAGCCTGCTGACCTGCGGCGACGTGGAGGAAAACCCCGGCCCCGGATCCATGAAACCAGTAACGTTATACGATGTCGCAGAGTATGCCGGTGTCTCTTATCAGACCGTTTCCCGCGTGGTGAACCAGGCCAGCCACGTTTCTGCGAAAACGCGGGAAAAAGTGGAAGCGGCGATGGCGGAGCTGAATTACATTCCCAACCGCGTGGCACAACAACTGGCGGGCAAACAGTCGTTGCTGATTGGCGTTGCCACCTCCAGTCTGGCCCTGCACGCGCCGTCGCAAATTGTCGCGGCGATTAAATCTCGCGCCGATCAACTGGGTGCCAGCGTGGTGGTGTCGATGGTAGAACGAAGCGGCGTCGAAGCCTGTAAAGCGGCGGTGCACAATCTTCTCGCGCAACGCGTCAGTGGGCTGATCATTAACTATCCGCTGGATGACCAGGATGCCATTGCTGTGGAAGCTGCCTGCACTAATGTTCCGGCGTTATTTCTTGATGTCTCTGACCAGACACCCATCAACAGTATTATTTTCTCCCATGAAGACGGTACGCGACTGGGCGTGGAGCATCTGGTCGCATTGGGTCACCAGCAAATCGCGCTGTTAGCGGGCCCATTAAGTTCTGTCTCGGCGCGTCTGCGTCTGGCTGGCTGGCATAAATATCTCACTCGCAATCAAATTCAGCCGATAGCGGAACGGGAAGGCGACTGGAGTGCCATGTCCGGTTTTCAACAAACCATGCAAATGCTGAATGAGGGCATCGTTCCCACTGCGATGCTGGTTGCCAACGATCAGATGGCGCTGGGCGCAATGCGCGCCATTACCGAGTCCGGGCTGCGCGTTGGTGCGGATATCTCGGTAGTGGGATACGACGATACCGAAGACAGCTCATGTTATATCCCGCCGTTAACCACCATCAAACAGGATTTTCGCCTGCTGGGGCAAACCAGCGTGGACCGCTTGCTGCAACTCTCTCAGGGCCAGGCGGTGAAGGGCAATCAGCTGTTGCCGGTCTCACTGGTGAAAAGAAAAACCACCCTGGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGAGCAGCCTGAGGCCTCCTAAGAAGAAGAGGAAGGTTGTTACAAACACCATTGTCACACTCCACAAACACCATTGTCACACTCCACAAACACCATTGTCACACTCCACAAACACCATTGTCACACTCCATCACATAGGAATAAAAAGCCATATCACATAGGAATAAAAAGCCATATCACATAGGAATAAAAAGCCATATCACATAGGAATAAAAAGCCATAGAAAGCGGACTACTGTAGTCCGCGGAGTACTGTCCTCCGCGGACGACTGTCGTCCGCGGAGCACTGTGCTCCGCGGATGACTGTCATCCGAGCGGAGACTCTAGAAATTGTGAGCGGATAACAATTGGGTATATAATGGGGGCCGTCAGCTACTACCAGAGCCGATCGAACACTCGAGCCGAGCAGACGTGCCTACAATTGTGAGCGCTCACAATTACTAGAAGATCTTGAATTCGCCACCATGGCTAGATTAGATAAAAGTAAAGTGATTAACAGCGCATTAGAGCTGCTTAATGAGGTCGGAATCGAAGGTTTAACAACCCGTAAACTCGCCCAGAAGCTAGGTGTAGAGCAGCCTACATTGTATTGGCATGTAAAAAATAAGCGGGCTTTGCTCGACGCCTTAGCCATTGAGATGTTAGATAGGCACCATACTCACTTTTGCCCTTTAGAAGGGGAAAGCTGGCAAGATTTTTTACGTAATAACGCTAAAAGTTTTAGATGTGCTTTACTAAGTCATCGCGATGGAGCAAAAGTACATTTAGGTACACGGCCTACAGAAAAACAGTATGAAACTCTCGAAAATCAATTAGCCTTTTTATGCCAACAAGGTTTTTCACTAGAGAATGCATTATATGCACTCAGCGCTGTGGGGCATTTTACTTTAGGTTGCGTATTGGAAGATCAAGAGCATCAAGTCGCTAAAGAAGAAAGGGAAACACCTACTACTGATAGTATGCCGCCATTATTACGACAAGCTATCGAATTATTTGATCACCAAGGTGCAGAGCCAGCCTTCTTATTCGGCCTTGAATTGATCATATGCGGATTAGAAAAACAACTTAAATGTGAAAGTGGGTCGCCAAAAAAGAAGAGAAAGGTGGATGGTGGCGGAGCACTGTCTCCCCAGCATTCAGCTGTGACCCAGGGATCAATCATTAAGAACAAGGAAGGTATGGATGCTAAGTCCCTGACCGCTTGGTCAAGAACTTTGGTAACTTTTAAGGACGTGTTCGTCGATTTTACTCGGGAGGAGTGGAAACTATTGGATACCGCCCAGCAAATCGTGTATAGAAACGTTATGCTCGAGAACTATAAGAATCTTGTGTCACTGGGCTACCAACTGACAAAGCCTGACGTTATCCTGAGGCTGGAAAAGGGTGAGGAGCCTTGGTTGGTCGAGCGGGAGATCCACCAGGAAACTCACCCAGACAGTGAGACTGCTTTCGAGATCAAGTCCTCTGTGTAATACAAGGCGGCCGCAAATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTAAAGCTTCGAATTCTGATAATCAGCCATACCACATTTGTAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCAGCTCGCCTTTTGCTCACATGTTAGGTGTGGGCAGGGACGAGCTGGCGCGGCGTCGCTGGGTGCACCGCGACCACGGGCAGAGCCACGCGGCGGGAGGACTACAACTCCCGGCACACCCCGCGCCGCCCCGCCTCTACTCCCAGAAGGCCGCGGGGGGTGGACCGCCTAAGAGGGCGTGCGCTCCCGACATGCCCCGCGGCGCGCCATTAACCGCCAGATTTGAATCGCGGGACCCGTTGGCAGAGGTGGCGGCGGCGGCATGGGTGCCCCGACGTTGCCCCCTGCCTGGCTCGAGACCGGTATGGCCCCCCCGACCGATGTCAGCCTGGGGGACGAGCTCCACTTAGACGGCGAGGACGTGGCGATGGCGCATGCCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGGACGGGGATTCCCCGGGTCCGGGATTTACCCCCCACGACTCCGCCCCCTACGGCGCTCTGGATATGGCCGACTTCGAGTTTGAGCAGATGTTTACCGATGCCCTTGGAATTGACGAGTACGGTGGGACGCGTATGAAGCTACTGTCTTCTATCGAACAAGCATGCGATATTTGCCGACTTAAAAAGCTCAAGTGCTCCAAAGAAAAACCGAAGTGCGCCAAGTGTCTGAAGAACAACTGGGAGTGTCGCTACTCTCCCAAAACCAAAAGGTCTCCGCTGACTAGGGCACATCTGACAGAAGTGGAATCAAGGCTAGAAAGACTGGAACAGCTATTTCTACTGATTTTTCCTCGAGAAGACCTTGACATGATTTTGAAAATGGATTCTTTACAGGATATAAAAGCATTGTTAACAGGATTATTTGTACAAGATAATGTGAATAAAGATGCCGTCACAGATAGATTGGCTTCAGTGGAGACTGATATGCCTCTAACATTGAGACAGCATAGAATAAGTGCGACATCATCATCGGAAGAGAGTAGTAACAAAGGTCAAAGACAGTTGACTGTATAAAAGCTTCACTCCTCAGGTGCAGGCTGCCTATCAGAAGGTGGTGGCTGGTGTGGCCAATGCCCTGGCTCACAAATACCACTGAGATCTTTTTCCCTCTGCCAAAAATTATGGGGACATCATGAAGCCCCTTGAGCATCTGACTTCTGGCTAATAAAGGAAATTTATTTTCATTGCAATAGTGTGTTGGAATTTTTTGTGTCTCTCACTCGGAAGGACATATGGGAGGGCAAATCATTTAAAACATCAGAATGAGTATTTGGTTTAGAGTTTGGCAACATATGCCCATATGCTGGCTGCCATGAACAAAGGTTGGCTATAAAGAGGTCATCAGTATATGAAACAGCCCCCTGCTGTCCATTCCTTATTCCATAGAAAAGCCTTGACTTGAGGTTAGATTTTTTTTATATTTTGTTTTGTGTTATTTTTTTCTTTAACATCCCTAAAATTTTCCTTACATGTTTTACTAGCCAGATTTTTCCTCCTCTCCTGACTACTCCCAGTCATAGCTGTCCCTCTTCTCTTATGGAGATC(SEQ ID NO:7)。
具有SEQ ID NO:8所述核苷酸序列的核酸编码由细胞特异性启动子Survivin1、第一microRNA为mir-143-3p、第二microRNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白分别为TALE蛋白、第一和第二识别序列为5×UAS的元件组成的表达系统。
CGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTTCTGTCCTCCGAGCGGAGACTCTAGATACCCCTCTCCGCTTCTGAATTCTAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCCTCGAGTACCCCTCTCCGCTTCTTACCCCTCTCCGCTTCTTACCCCTCTCCGCTTCTGAATTCGACCGCCACCATGAGACATATTATCTGCCACGGAGGTGTTATTACCGAAGAAATGGCCGCCAGTCTTTTGGACCAGCTGATCGAAGAGGTACTGGCTGATAATCTTCCACCTCCTAGCCATTTTGAACCACCTACCCTTCACGAACTGTATGATTTAGACGTGACGGCCCCCGAAGATCCCAACGAGGAGGCGGTTTCGCAGATTTTTCCCGACTCTGTAATGTTGGCGGTGCAGGAAGGGATTGACTTACTCACTTTTCCGCCGGCGCCCGGTTCTCCGGAGCCGCCTCACCTTTCCCGGCAGCCCGAGCAGCCGGAGCAGAGAGCCTTGGGTCCGGTTTCTATGCCAAACCTTGTACCGGAGGTGATCGATCTTACCTGCCACGAGGCTGGCTTTCCACCCAGTGACGACGAGGATGAAGAGGGTGAGGAGTTTGTGTTAGATTATGTGGAGCACCCCGGGCACGGTTGCAGGTCTTGTCATTATCACCGGAGGAATACGGGGGACCCAGATATTATGTGTTCGCTTTGCTATATGAGGACCTGTGGCATGTTTGTCTACAGTAAGTGAAAATTATGGGCAGTGGGTGATAGAGTGGTGGGTTTGGTGTGGTAATTTTTTTTTTAATTTTTACAGTTTTGTGGTTTAAAGAATTTTGTATTGTGATTTTTTTAAAAGGTCCTGTGTCTGAACCTGAGCCTGAGCCCGAGCCAGAACCGGAGCCTGCAAGACCTACCCGCCGTCCTAAAATGGCGCCTGCTATCCTGAGGCGCCCGACATCACCTGTGTCTAGAGAATGCAATAGTAGTACGGATAGCTGTGACTCCGGTCCTTCTAACACACCTCCTGAGATACACCCGGTGGTCCCGCTGTGCCCCATTAAACCAGTTGCCGTGAGAGTTGGTGGGCGTCGCCAGGCTGTGGAATGTATCGAGGACTTGCTTAACGAGCCTGGGCAACCTTTGGACTTGAGCTGTAAACGCCCCAGGCCAGGGTCCGGAGCTACAAATTTTTCCCTCCTCAAACAGGCTGGAGATGTCGAAGAAAATCCTGGGCCTATGGAGGCTTGGGAGTGTTTGGAAGATTTTTCTGCTGTGCGTAACTTGCTGGAACAGAGCTCTAACAGTACCTCTTGGTTTTGGAGGTTTCTGTGGGGCTCATCCCAGGCAAAGTTAGTCTGCAGAATTAAGGAGGATTACAAGTGGGAATTTGAAGAGCTTTTGAAATCCTGTGGTGAGCTGTTTGATTCTTTGAATCTGGGTCACCAGGCGCTTTTCCAAGAGAAGGTCATCAAGACTTTGGATTTTTCCACACCGGGGCGCGCTGCGGCTGCTGTTGCTTTTTTGAGTTTTATAAAGGATAAATGGAGCGAAGAAACCCATCTGAGCGGGGGGTACCTGCTGGATTTTCTGGCCATGCATCTGTGGAGAGCGGTTGTGAGACACAAGAATCGCCTGCTACTGTTGTCTTCCGTCCGCCCGGCGATAATACCGACGGAGGAGCAGCAGCAGCAGCAGGAGGAAGCCAGGCGGCGGCGGCAGGAGCAGAGCCCATGGAACCCGAGAGCCGGCCTGGACCCTCGGGAATGAATGTTGTACAGGTGGCTGAACTGTATCCAGAACTGAGACGCATTTTGACAATTACAGAGGATGGGCAGGGGCTAAAGGGGGTAAAGAGGGAGCGGGGGGCTTGTGAGGCTACAGAGGAGGCTAGGAATCTAGCTTTTAGCTTAATGACCAGACACCGTCCTGAGTGTATTACTTTTCAACAGATCAAGGATAATTGCGCTAATGAGCTTGATCTGCTGGCGCAGAAGTATTCCATAGAGCAGCTGACCACTTACTGGCTGCAGCCAGGGGATGATTTTGAGGAGGCTATTAGGGTATATGCAAAGGTGGCACTTAGGCCAGATTGCAAGTACAAGATCAGCAAACTTGTAAATATCAGGAATTGTTGCTACATTTCTGGGAACGGGGCCGAGGTGGAGATAGATACGGAGGATAGGGTGGCCTTTAGATGTAGCATGATAAATATGTGGCCGGGGGTGCTTGGCATGGACGGGGTGGTTATTATGAATGTAAGGTTTACTGGCCCCAATTTTAGCGGTACGGTTTTCCTGGCCAATACCAACCTTATCCTACACGGTGTAAGCTTCTATGGGTTTAACAATACCTGTGTGGAAGCCTGGACCGATGTAAGGGTTCGGGGCTGTGCCTTTTACTGCTGCTGGAAGGGGGTGGTGTGTCGCCCCAAAAGCAGGGCTTCAATTAAGAAATGCCTCTTTGAAAGGTGTACCTTGGGTATCCTGTCTGAGGGTAACTCCAGGGTGCGCCACAATGTGGCCTCCGACTGTGGTTGCTTCATGCTAGTGAAAAGCGTGGCTGTGATTAAGCATAACATGGTATGTGGCAACTGCGAGGACAGGGCCTCTCAGATGCTGACCTGCTCGGACGGCAACTGTCACCTGCTGAAGACCATTCACGTAGCCAGCCACTCTCGCAAGGCCTGGCCAGTGTTTGAGCATAACATACTGACCCGCTGTTCCTTGCATTTGGGTAACAGGAGGGGGGTGTTCCTACCTTACCAATGCAATTTGAGTCACACTAAGATATTGCTTGAGCCCGAGAGCATGTCCAAGGTGAACCTGAACGGGGTGTTTGACATGACCATGAAGATCTGGAAGGTGCTGAGGTACGATGAGACCCGCACCAGGTGCAGACCCTGCGAGTGTGGCGGTAAACATATTAGGAACCAGCCTGTGATGCTGGATGTGACCGAGGAGCTGAGGCCCGATCACTTGGTGCTGGCCTGCACCCGCGCTGAGTTTGGCTCTAGCGATGAAGATACAGATCTCGAGGGGTCCGGAGCTACAAATTTTTCCCTCCTCAAACAGGCTGGAGATGTCGAAGAAAATCCTGGGCCTACCGGTATGTATAGGATGCAGTTGCTCTCTTGCATCGCCCTGTCTCTAGCGTTGGTGACTAATAGCGCCCCTACTTCTTCTTCCACGAAGAAAACCCAGCTGCAACTGGAACACCTTCTGCTGGATCTCCAGATGATCCTTAACGGGATTAACAACTATAAGAACCCGAAGCTCACGGCTATGCTGACGGCGAAGTTCGCTATGCCTAAAAAGGCTACTGAGCTGAAACACTTACAGTGTCTCGAAGAGGCACTCAAACCCCTCGAGGAGGTACTCAATCTGGCGCAAAGCAAAAACTTCCATCTCCGCCCACGGGACCTAATCTCCAACATCAACGTGATTGTGCTGGAGCTCAAGGGGTCTGAAACCACGTTTATGTGCGAGTACGCAGATGAAACTGCTACCATAGTGGAATTCCTCAATCGTTGGATCACTTTCTGCCAGTCAATCATTTCAACCCTGACCAGATCTGAGGGCCGCGGCAGCCTGCTGACCTGCGGCGACGTGGAGGAAAACCCCGGCCCCGGATCCTTGTCGACCATTCGTCCGCGCAGGCCAAGTCCTGCCCGCGAGCTTCTGCCCGGACCCCAACCGGATAGGGTTCAGCCGACTGCAGATCGTGGGGTGTCTGCGCCTGCTGGCAGCCCTCTGGATGGCTTGCCCGCTCGGCGGACGGTGTCCCGGACCCGGCTGCCATCTCCCCCTGCGCCCTCACCTGCGTTCTCGGCGGGCAGCTTCAGCGATCTGCTCCGTCCGTTCGATCCGTCGCTTCTTGATACATCGCTTCTTGATTCGATGCCTGCCGTCGGCACGCCGCATACAGCGGCTGCCCCAGCAGAGTGGGATGAGGCGCAATCGGCTCTGCGTGCAGCCGATGACCCGCCACCCACCGTGCGTGTCGCTGTCACTGCCGCGCGGCCGCCGCGCGCCAAGCCGGCCCCGCGACGGCGTGCTGCGCAACCCTCCGACGCTTCGCCGGCCGCGCAGGTGGATCTACGCACGCTCGGCTACAGTCAGCAGCAGCAAGAGAAGATCAAACCGAAGGTGCGTTCGACAGTGGCGCAGCACCACGAGGCACTGGTGGGCCATGGGTTTACACACGCGCACATCGTTGCGCTCAGCCAACACCCGGCAGCGTTAGGGACCGTCGCTGTCACGTATCAGCACATAATCACGGCGTTGCCAGAGGCGACACACGAAGACATCGTTGGCGTCGGCAAACAGTGGTCCGGCGCACGCGCCCTGGAGGCCTTGCTCACGGATGCGGGGGAGTTGAGAGGTCCGCCGTTACAGTTGGACACAGGCCAACTTGTGAAGATTGCAAAACGTGGCGGCGTGACCGCAATGGAGGCAGTGCATGCATCGCGCAATGCACTGACGGGTGCCCCCCTGAACCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACAATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACAATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAAGCATTGTGGCCCAGCTGAGCCGGCCTGATCCGGCGTTGGCCGCGTTGACCAACGACCACCTCGTCGCCTTGGCCTGCCTCGGCGGACGTCCTGCCATGGATGCAGTGAAAAAGGGATTGCCGCACGCGCCGGAATTGATCAGAAGAGTCAATCGCCGTATTGGCGAACGCACGTCCCATCGCGTTGCCGACTACGCGCAAGTGGTTCGCGTGCTGGAGTTTTTCCAGTGCCACTCCCACCCAGCGTACGCATTTGATGAGGCCATGACGCAGTTCGGGATGAGCAGGAACGGGTTGGTACAGCTCTTTCGCAGAGTGGGCGTCACCGAACTCGAAGCCCGCGGTGGAACGCTCCCCCCAGCCTCGCAGCGTTGGGACCGTATCCTCCAGGCATCAGGGATGAAAAGGGCCAAACCGTCCCCTACTTCAGCTCAAACACCGGATCAGGCGTCTTTGCATGCATTCGCCGATTCGCTGGAGCGTGACCTTGATGCGCCCAGCCCAATGCACGAGGGAGATCAGACGCGGGCAAGCAGCCGTAAACGGTCCCGATCGGATCGTGCTGTCACCGGCCCCTCCGCACAGCAGGCTGTCGAGGTGCGCGTTCCCGAACAGCGCGATGCGCTGCATTTGCCCCTCAGCTGGAGGGTAAAACGCCCGCGTACCAGGATCTGGGGCGGCCTCCCGGTTAAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGGAGCTACAGTGCTTCATCTCAGAGCTACAGTGCTTCATCTCAGAGCTACAGTGCTTCATCTCAGAGCTACAGTGCTTCATCTCAGAAAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTTCTGTCCTCCGAGCGGAGACTCTAGATATAGAACCGATCCTCCCATTGAATTCTAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCCTCGAGTATAGAACCGATCCTCCCATTTATAGAACCGATCCTCCCATTTATAGAACCGATCCTCCCATTACTAGAAGATCTTGAATTCGCCACCTTGTCGACCATTCGTCCGCGCAGGCCAAGTCCTGCCCGCGAGCTTCTGCCCGGACCCCAACCGGATAGGGTTCAGCCGACTGCAGATCGTGGGGTGTCTGCGCCTGCTGGCAGCCCTCTGGATGGCTTGCCCGCTCGGCGGACGGTGTCCCGGACCCGGCTGCCATCTCCCCCTGCGCCCTCACCTGCGTTCTCGGCGGGCAGCTTCAGCGATCTGCTCCGTCCGTTCGATCCGTCGCTTCTTGATACATCGCTTCTTGATTCGATGCCTGCCGTCGGCACGCCGCATACAGCGGCTGCCCCAGCAGAGTGGGATGAGGCGCAATCGGCTCTGCGTGCAGCCGATGACCCGCCACCCACCGTGCGTGTCGCTGTCACTGCCGCGCGGCCGCCGCGCGCCAAGCCGGCCCCGCGACGGCGTGCTGCGCAACCCTCCGACGCTTCGCCGGCCGCGCAGGTGGATCTACGCACGCTCGGCTACAGTCAGCAGCAGCAAGAGAAGATCAAACCGAAGGTGCGTTCGACAGTGGCGCAGCACCACGAGGCACTGGTGGGCCATGGGTTTACACACGCGCACATCGTTGCGCTCAGCCAACACCCGGCAGCGTTAGGGACCGTCGCTGTCACGTATCAGCACATAATCACGGCGTTGCCAGAGGCGACACACGAAGACATCGTTGGCGTCGGCAAACAGTGGTCCGGCGCACGCGCCCTGGAGGCCTTGCTCACGGATGCGGGGGAGTTGAGAGGTCCGCCGTTACAGTTGGACACAGGCCAACTTGTGAAGATTGCAAAACGTGGCGGCGTGACCGCAATGGAGGCAGTGCATGCATCGCGCAATGCACTGACGGGTGCCCCCCTGAACCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACAATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAAGCATTGTGGCCCAGCTGAGCCGGCCTGATCCGGCGTTGGCCGCGTTGACCAACGACCACCTCGTCGCCTTGGCCTGCCTCGGCGGACGTCCTGCCATGGATGCAGTGAAAAAGGGATTGCCGCACGCGCCGGAATTGATCAGAAGAGTCAATCGCCGTATTGGCGAACGCACGTCCCATCGCGTTGCCGACTACGCGCAAGTGGTTCGCGTGCTGGAGTTTTTCCAGTGCCACTCCCACCCAGCGTACGCATTTGATGAGGCCATGACGCAGTTCGGGATGAGCAGGAACGGGTTGGTACAGCTCTTTCGCAGAGTGGGCGTCACCGAACTCGAAGCCCGCGGTGGAACGCTCCCCCCAGCCTCGCAGCGTTGGGACCGTATCCTCCAGGCATCAGGGATGAAAAGGGCCAAACCGTCCCCTACTTCAGCTCAAACACCGGATCAGGCGTCTTTGCATGCATTCGCCGATTCGCTGGAGCGTGACCTTGATGCGCCCAGCCCAATGCACGAGGGAGATCAGACGCGGGCAAGCAGCCGTAAACGGTCCCGATCGGATCGTGCTGTCACCGGCCCCTCCGCACAGCAGGCTGTCGAGGTGCGCGTTCCCGAACAGCGCGATGCGCTGCATTTGCCCCTCAGCTGGAGGGTAAAACGCCCGCGTACCAGGATCTGGGGCGGCCTCCCGTAATACAAGGCGGCCGCAAATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTAAAGCTTCGAATTCTGATAATCAGCCATACCACATTTGTAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCCATGTTAGGTGTGGGCAGGGACGAGCTGGCGCGGCGTCGCTGGGTGCACCGCGACCACGGGCAGAGCCACGCGGCGGGAGGACTACAACTCCCGGCACACCCCGCGCCGCCCCGCCTCTACTCCCAGAAGGCCGCGGGGGGTGGACCGCCTAAGAGGGCGTGCGCTCCCGACATGCCCCGCGGCGCGCCATTAACCGCCAGATTTGAATCGCGGGACCCGTTGGCAGAGGTGGCGGCGGCGGCATGGGTGCCCCGACGTTGCCCCCTGCCTGGCTCGAGACCGATGGCCCCCCCGACCGATGTCAGCCTGGGGGACGAGCTCCACTTAGACGGCGAGGACGTGGCGATGGCGCATGCCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGAGCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGAGCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGAGCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGGACGGGGATTCCCCGGGTCCGGGATTTACCCCCCACGACTCCGCCCCCTACGGCGCTCTGGATATGGCCGACTTCGAGTTTGAGCAGATGTTTACCGATGCCCTTGGAATTGACGAGTACGGTGGGACGCGTATGAAGCTACTGTCTTCTATCGAACAAGCATGCGATATTTGCCGACTTAAAAAGCTCAAGTGCTCCAAAGAAAAACCGAAGTGCGCCAAGTGTCTGAAGAACAACTGGGAGTGTCGCTACTCTCCCAAAACCAAAAGGTCTCCGCTGACTAGGGCACATCTGACAGAAGTGGAATCAAGGCTAGAAAGACTGGAACAGCTATTTCTACTGATTTTTCCTCGAGAAGACCTTGACATGATTTTGAAAATGGATTCTTTACAGGATATAAAAGCATTGTTAACAGGATTATTTGTACAAGATAATGTGAATAAAGATGCCGTCACAGATAGATTGGCTTCAGTGGAGACTGATATGCCTCTAACATTGAGACAGCATAGAATAAGTGCGACATCATCATCGGAAGAGAGTAGTAACAAAGGTCAAAGACAGTTGACTGTATAAAAGCTTCACTCCTCAGGTGCAGGCTGCCTATCAGAAGGTGGTGGCTGGTGTGGCCAATGCCCTGGCTCACAAATACCACTGAGATCTTTTTCCCTCTGCCAAAAATTATGGGGACATCATGAAGCCCCTTGAGCATCTGACTTCTGGCTAATAAAGGAAATTTATTTTCATTGCAATAGTGTGTTGGAATTTTTTGTGTCTCTCACTCGGAAGGACATATGGGAGGGCAAATCATTTAAAACATCAGAATGAGTATTTGGTTTAGAGTTTGGCAACATATGCCCATATGCTGGCTGCCATGAACAAAGGTTGGCTATAAAGAGGTCATCAGTATATGAAACAGCCCCCTGCTGTCCATTCCTTATTCCATAGAAAAGCCTTGACTTGAGGTTAGATTTTTTTTATATTTTGTTTTGTGTTATTTTTTTCTTTAACATCCCTAAAATTTTCCTTACATGTTTTACTAGCCAGATTTTTCCTCCTCTCCTGACTACTCCCAGTCATAGCTGTCCCTCTTCTCTTATGGAGATC(SEQ ID NO:8)。
具有SEQ ID NO:9所述核苷酸序列的核酸编码由细胞特异性启动子CEA368、第一microRNA为mir-143-3p、第二microRNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白分别为Lacl、tetR-KRAB、第一和第二识别序列为5×UAS的元件组成的表达系统。
CGGGGTGCCGCCCCCCGCGGGTGACCGCCCTCCGCGGGTGACAGCCCTCCGCGGAGTGCCGCACTCCGCGGAGTGCTGTACTCCGAGCGGAGACTCTAGACTCCCTATCAGTGATAGAGATAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCTCCCTATCAGTGATAGAGAGAATTCGACCGCCACCATGAGACATATTATCTGCCACGGAGGTGTTATTACCGAAGAAATGGCCGCCAGTCTTTTGGACCAGCTGATCGAAGAGGTACTGGCTGATAATCTTCCACCTCCTAGCCATTTTGAACCACCTACCCTTCACGAACTGTATGATTTAGACGTGACGGCCCCCGAAGATCCCAACGAGGAGGCGGTTTCGCAGATTTTTCCCGACTCTGTAATGTTGGCGGTGCAGGAAGGGATTGACTTACTCACTTTTCCGCCGGCGCCCGGTTCTCCGGAGCCGCCTCACCTTTCCCGGCAGCCCGAGCAGCCGGAGCAGAGAGCCTTGGGTCCGGTTTCTATGCCAAACCTTGTACCGGAGGTGATCGATCTTACCTGCCACGAGGCTGGCTTTCCACCCAGTGACGACGAGGATGAAGAGGGTGAGGAGTTTGTGTTAGATTATGTGGAGCACCCCGGGCACGGTTGCAGGTCTTGTCATTATCACCGGAGGAATACGGGGGACCCAGATATTATGTGTTCGCTTTGCTATATGAGGACCTGTGGCATGTTTGTCTACAGTAAGTGAAAATTATGGGCAGTGGGTGATAGAGTGGTGGGTTTGGTGTGGTAATTTTTTTTTTAATTTTTACAGTTTTGTGGTTTAAAGAATTTTGTATTGTGATTTTTTTAAAAGGTCCTGTGTCTGAACCTGAGCCTGAGCCCGAGCCAGAACCGGAGCCTGCAAGACCTACCCGCCGTCCTAAAATGGCGCCTGCTATCCTGAGGCGCCCGACATCACCTGTGTCTAGAGAATGCAATAGTAGTACGGATAGCTGTGACTCCGGTCCTTCTAACACACCTCCTGAGATACACCCGGTGGTCCCGCTGTGCCCCATTAAACCAGTTGCCGTGAGAGTTGGTGGGCGTCGCCAGGCTGTGGAATGTATCGAGGACTTGCTTAACGAGCCTGGGCAACCTTTGGACTTGAGCTGTAAACGCCCCAGGCCACTCGAGGGTACCGGGTCCGGAGCTACAAATTTTTCCCTCCTCAAACAGGCTGGAGATGTCGAAGAAAATCCTGGGCCTACCGGTATGTGGCTGCAGAGCCTCCTTCTACTGGGCACCGTGGCTTGCAGCATCTCGGCCCCCGCCAGAAGTCCCAGCCCCAGCACCCAGCCCTGGGAACACGTAAATGCAATCCAGGAGGCCCGCCGCCTCCTGAACTTGTCACGGGACACGGCAGCCGAGATGAATGAAACTGTGGAGGTCATCAGTGAAATGTTTGACTTGCAGGAGCCCACATGCCTGCAGACTCGGCTGGAGCTCTACAAACAGGGCCTGCGTGGCAGCCTCACCAAGCTGAAGGGCCCACTGACCATGATGGCTAGCCATTACAAGCAGCACTGCCCGCCGACCCCTGAGACATCCTGTGCCACTCAGATCATTACCTTTGAGTCTTTCAAGGAGAACCTCAAGGACTTTCTGCTGGTCATCCCATTTGACTGCTGGGAACCCGTCCAGGAAAGATCTGAGGGCCGCGGCAGCCTGCTGACCTGCGGCGACGTGGAGGAAAACCCCGGCCCCGGATCCATGAAACCAGTAACGTTATACGATGTCGCAGAGTATGCCGGTGTCTCTTATCAGACCGTTTCCCGCGTGGTGAACCAGGCCAGCCACGTTTCTGCGAAAACGCGGGAAAAAGTGGAAGCGGCGATGGCGGAGCTGAATTACATTCCCAACCGCGTGGCACAACAACTGGCGGGCAAACAGTCGTTGCTGATTGGCGTTGCCACCTCCAGTCTGGCCCTGCACGCGCCGTCGCAAATTGTCGCGGCGATTAAATCTCGCGCCGATCAACTGGGTGCCAGCGTGGTGGTGTCGATGGTAGAACGAAGCGGCGTCGAAGCCTGTAAAGCGGCGGTGCACAATCTTCTCGCGCAACGCGTCAGTGGGCTGATCATTAACTATCCGCTGGATGACCAGGATGCCATTGCTGTGGAAGCTGCCTGCACTAATGTTCCGGCGTTATTTCTTGATGTCTCTGACCAGACACCCATCAACAGTATTATTTTCTCCCATGAAGACGGTACGCGACTGGGCGTGGAGCATCTGGTCGCATTGGGTCACCAGCAAATCGCGCTGTTAGCGGGCCCATTAAGTTCTGTCTCGGCGCGTCTGCGTCTGGCTGGCTGGCATAAATATCTCACTCGCAATCAAATTCAGCCGATAGCGGAACGGGAAGGCGACTGGAGTGCCATGTCCGGTTTTCAACAAACCATGCAAATGCTGAATGAGGGCATCGTTCCCACTGCGATGCTGGTTGCCAACGATCAGATGGCGCTGGGCGCAATGCGCGCCATTACCGAGTCCGGGCTGCGCGTTGGTGCGGATATCTCGGTAGTGGGATACGACGATACCGAAGACAGCTCATGTTATATCCCGCCGTTAACCACCATCAAACAGGATTTTCGCCTGCTGGGGCAAACCAGCGTGGACCGCTTGCTGCAACTCTCTCAGGGCCAGGCGGTGAAGGGCAATCAGCTGTTGCCGGTCTCACTGGTGAAAAGAAAAACCACCCTGGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGAGCAGCCTGAGGCCTCCTAAGAAGAAGAGGAAGGTTGTTACAAACACCATTGTCACACTCCACAAACACCATTGTCACACTCCACAAACACCATTGTCACACTCCACAAACACCATTGTCACACTCCAGAGCTACAGTGCTTCATCTCAGAGCTACAGTGCTTCATCTCAGAGCTACAGTGCTTCATCTCAGAGCTACAGTGCTTCATCTCAGAAAGCGGACTACTGTAGTCCGCGGAGTACTGTCCTCCGCGGACGACTGTCGTCCGCGGAGCACTGTGCTCCGCGGATGACTGTCATCCGAGCGGAGACTCTAGAAATTGTGAGCGGATAACAATTGGGTATATAATGGGGGCCGTCAGCTACTACCAGAGCCGATCGAACACTCGAGCCGAGCAGACGTGCCTACAATTGTGAGCGCTCACAATTACTAGAAGATCTTGAATTCGCCACCATGGCTAGATTAGATAAAAGTAAAGTGATTAACAGCGCATTAGAGCTGCTTAATGAGGTCGGAATCGAAGGTTTAACAACCCGTAAACTCGCCCAGAAGCTAGGTGTAGAGCAGCCTACATTGTATTGGCATGTAAAAAATAAGCGGGCTTTGCTCGACGCCTTAGCCATTGAGATGTTAGATAGGCACCATACTCACTTTTGCCCTTTAGAAGGGGAAAGCTGGCAAGATTTTTTACGTAATAACGCTAAAAGTTTTAGATGTGCTTTACTAAGTCATCGCGATGGAGCAAAAGTACATTTAGGTACACGGCCTACAGAAAAACAGTATGAAACTCTCGAAAATCAATTAGCCTTTTTATGCCAACAAGGTTTTTCACTAGAGAATGCATTATATGCACTCAGCGCTGTGGGGCATTTTACTTTAGGTTGCGTATTGGAAGATCAAGAGCATCAAGTCGCTAAAGAAGAAAGGGAAACACCTACTACTGATAGTATGCCGCCATTATTACGACAAGCTATCGAATTATTTGATCACCAAGGTGCAGAGCCAGCCTTCTTATTCGGCCTTGAATTGATCATATGCGGATTAGAAAAACAACTTAAATGTGAAAGTGGGTCGCCAAAAAAGAAGAGAAAGGTGGATGGTGGCGGAGCACTGTCTCCCCAGCATTCAGCTGTGACCCAGGGATCAATCATTAAGAACAAGGAAGGTATGGATGCTAAGTCCCTGACCGCTTGGTCAAGAACTTTGGTAACTTTTAAGGACGTGTTCGTCGATTTTACTCGGGAGGAGTGGAAACTATTGGATACCGCCCAGCAAATCGTGTATAGAAACGTTATGCTCGAGAACTATAAGAATCTTGTGTCACTGGGCTACCAACTGACAAAGCCTGACGTTATCCTGAGGCTGGAAAAGGGTGAGGAGCCTTGGTTGGTCGAGCGGGAGATCCACCAGGAAACTCACCCAGACAGTGAGACTGCTTTCGAGATCAAGTCCTCTGTGTAATACAAGGCGGCCGCAAATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTAAAGCTTCGAATTCTGATAATCAGCCATACCACATTTGTAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCAGCTCGCCTTTTGCTCACATGTCTCCCAGGGGATGGGGGTCCATCCACCTTGCCGAAAAGATTTGTCTGAGGAACTGAAAATAGAAGGGAAAAAAGAGGAGGGACAAAAGAGGCAGAAATGAGAGGGGAGGGGACAGAGGACACCTGAATAAAGACCACACCCATGACCCACGTGATGCTGAGAAGTACTCCTGCCCTAGGAAGAGACTCAGGGCAGAGGGAGGAAGGACAGCAGACCAGACAGTCACAGCAGCCTTGACAAAACGTTCCTGGAACTCAAGCTCTTCTCCACAGAGGAGGACAGAGCAGACAGCAGAGACCATGGAGTCTCCCTCGGCCCCTCCCCACAGATGGTGCATCCCCTGGCAGAGGCTCCTGCTCACAGGTGAACTCGAGACCGGTATGGCCCCCCCGACCGATGTCAGCCTGGGGGACGAGCTCCACTTAGACGGCGAGGACGTGGCGATGGCGCATGCCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGGACGGGGATTCCCCGGGTCCGGGATTTACCCCCCACGACTCCGCCCCCTACGGCGCTCTGGATATGGCCGACTTCGAGTTTGAGCAGATGTTTACCGATGCCCTTGGAATTGACGAGTACGGTGGGACGCGTATGAAGCTACTGTCTTCTATCGAACAAGCATGCGATATTTGCCGACTTAAAAAGCTCAAGTGCTCCAAAGAAAAACCGAAGTGCGCCAAGTGTCTGAAGAACAACTGGGAGTGTCGCTACTCTCCCAAAACCAAAAGGTCTCCGCTGACTAGGGCACATCTGACAGAAGTGGAATCAAGGCTAGAAAGACTGGAACAGCTATTTCTACTGATTTTTCCTCGAGAAGACCTTGACATGATTTTGAAAATGGATTCTTTACAGGATATAAAAGCATTGTTAACAGGATTATTTGTACAAGATAATGTGAATAAAGATGCCGTCACAGATAGATTGGCTTCAGTGGAGACTGATATGCCTCTAACATTGAGACAGCATAGAATAAGTGCGACATCATCATCGGAAGAGAGTAGTAACAAAGGTCAAAGACAGTTGACTGTATAAAAGCTTCACTCCTCAGGTGCAGGCTGCCTATCAGAAGGTGGTGGCTGGTGTGGCCAATGCCCTGGCTCACAAATACCACTGAGATCTTTTTCCCTCTGCCAAAAATTATGGGGACATCATGAAGCCCCTTGAGCATCTGACTTCTGGCTAATAAAGGAAATTTATTTTCATTGCAATAGTGTGTTGGAATTTTTTGTGTCTCTCACTCGGAAGGACATATGGGAGGGCAAATCATTTAAAACATCAGAATGAGTATTTGGTTTAGAGTTTGGCAACATATGCCCATATGCTGGCTGCCATGAACAAAGGTTGGCTATAAAGAGGTCATCAGTATATGAAACAGCCCCCTGCTGTCCATTCCTTATTCCATAGAAAAGCCTTGACTTGAGGTTAGATTTTTTTTATATTTTGTTTTGTGTTATTTTTTTCTTTAACATCCCTAAAATTTTCCTTACATGTTTTACTAGCCAGATTTTTCCTCCTCTCCTGACTACTCCCAGTCATAGCTGTCCCTCTTCTCTTATGGAGATC(SEQ ID NO:9)。
具有SEQ ID NO:10所述核苷酸序列的核酸编码由细胞特异性启动子CEA368、第一microRNA为mir-135a-5p、第二microRNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白分别为TALE蛋白、第一和第二识别序列为5×UAS的元件组成的表达系统。
CGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTTCTGTCCTCCGAGCGGAGACTCTAGATACCCCTCTCCGCTTCTGAATTCTAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCCTCGAGTACCCCTCTCCGCTTCTTACCCCTCTCCGCTTCTTACCCCTCTCCGCTTCTGAATTCGACCGCCACCATGAGACATATTATCTGCCACGGAGGTGTTATTACCGAAGAAATGGCCGCCAGTCTTTTGGACCAGCTGATCGAAGAGGTACTGGCTGATAATCTTCCACCTCCTAGCCATTTTGAACCACCTACCCTTCACGAACTGTATGATTTAGACGTGACGGCCCCCGAAGATCCCAACGAGGAGGCGGTTTCGCAGATTTTTCCCGACTCTGTAATGTTGGCGGTGCAGGAAGGGATTGACTTACTCACTTTTCCGCCGGCGCCCGGTTCTCCGGAGCCGCCTCACCTTTCCCGGCAGCCCGAGCAGCCGGAGCAGAGAGCCTTGGGTCCGGTTTCTATGCCAAACCTTGTACCGGAGGTGATCGATCTTACCTGCCACGAGGCTGGCTTTCCACCCAGTGACGACGAGGATGAAGAGGGTGAGGAGTTTGTGTTAGATTATGTGGAGCACCCCGGGCACGGTTGCAGGTCTTGTCATTATCACCGGAGGAATACGGGGGACCCAGATATTATGTGTTCGCTTTGCTATATGAGGACCTGTGGCATGTTTGTCTACAGTAAGTGAAAATTATGGGCAGTGGGTGATAGAGTGGTGGGTTTGGTGTGGTAATTTTTTTTTTAATTTTTACAGTTTTGTGGTTTAAAGAATTTTGTATTGTGATTTTTTTAAAAGGTCCTGTGTCTGAACCTGAGCCTGAGCCCGAGCCAGAACCGGAGCCTGCAAGACCTACCCGCCGTCCTAAAATGGCGCCTGCTATCCTGAGGCGCCCGACATCACCTGTGTCTAGAGAATGCAATAGTAGTACGGATAGCTGTGACTCCGGTCCTTCTAACACACCTCCTGAGATACACCCGGTGGTCCCGCTGTGCCCCATTAAACCAGTTGCCGTGAGAGTTGGTGGGCGTCGCCAGGCTGTGGAATGTATCGAGGACTTGCTTAACGAGCCTGGGCAACCTTTGGACTTGAGCTGTAAACGCCCCAGGCCAGGGTCCGGAGCTACAAATTTTTCCCTCCTCAAACAGGCTGGAGATGTCGAAGAAAATCCTGGGCCTATGGAGGCTTGGGAGTGTTTGGAAGATTTTTCTGCTGTGCGTAACTTGCTGGAACAGAGCTCTAACAGTACCTCTTGGTTTTGGAGGTTTCTGTGGGGCTCATCCCAGGCAAAGTTAGTCTGCAGAATTAAGGAGGATTACAAGTGGGAATTTGAAGAGCTTTTGAAATCCTGTGGTGAGCTGTTTGATTCTTTGAATCTGGGTCACCAGGCGCTTTTCCAAGAGAAGGTCATCAAGACTTTGGATTTTTCCACACCGGGGCGCGCTGCGGCTGCTGTTGCTTTTTTGAGTTTTATAAAGGATAAATGGAGCGAAGAAACCCATCTGAGCGGGGGGTACCTGCTGGATTTTCTGGCCATGCATCTGTGGAGAGCGGTTGTGAGACACAAGAATCGCCTGCTACTGTTGTCTTCCGTCCGCCCGGCGATAATACCGACGGAGGAGCAGCAGCAGCAGCAGGAGGAAGCCAGGCGGCGGCGGCAGGAGCAGAGCCCATGGAACCCGAGAGCCGGCCTGGACCCTCGGGAATGAATGTTGTACAGGTGGCTGAACTGTATCCAGAACTGAGACGCATTTTGACAATTACAGAGGATGGGCAGGGGCTAAAGGGGGTAAAGAGGGAGCGGGGGGCTTGTGAGGCTACAGAGGAGGCTAGGAATCTAGCTTTTAGCTTAATGACCAGACACCGTCCTGAGTGTATTACTTTTCAACAGATCAAGGATAATTGCGCTAATGAGCTTGATCTGCTGGCGCAGAAGTATTCCATAGAGCAGCTGACCACTTACTGGCTGCAGCCAGGGGATGATTTTGAGGAGGCTATTAGGGTATATGCAAAGGTGGCACTTAGGCCAGATTGCAAGTACAAGATCAGCAAACTTGTAAATATCAGGAATTGTTGCTACATTTCTGGGAACGGGGCCGAGGTGGAGATAGATACGGAGGATAGGGTGGCCTTTAGATGTAGCATGATAAATATGTGGCCGGGGGTGCTTGGCATGGACGGGGTGGTTATTATGAATGTAAGGTTTACTGGCCCCAATTTTAGCGGTACGGTTTTCCTGGCCAATACCAACCTTATCCTACACGGTGTAAGCTTCTATGGGTTTAACAATACCTGTGTGGAAGCCTGGACCGATGTAAGGGTTCGGGGCTGTGCCTTTTACTGCTGCTGGAAGGGGGTGGTGTGTCGCCCCAAAAGCAGGGCTTCAATTAAGAAATGCCTCTTTGAAAGGTGTACCTTGGGTATCCTGTCTGAGGGTAACTCCAGGGTGCGCCACAATGTGGCCTCCGACTGTGGTTGCTTCATGCTAGTGAAAAGCGTGGCTGTGATTAAGCATAACATGGTATGTGGCAACTGCGAGGACAGGGCCTCTCAGATGCTGACCTGCTCGGACGGCAACTGTCACCTGCTGAAGACCATTCACGTAGCCAGCCACTCTCGCAAGGCCTGGCCAGTGTTTGAGCATAACATACTGACCCGCTGTTCCTTGCATTTGGGTAACAGGAGGGGGGTGTTCCTACCTTACCAATGCAATTTGAGTCACACTAAGATATTGCTTGAGCCCGAGAGCATGTCCAAGGTGAACCTGAACGGGGTGTTTGACATGACCATGAAGATCTGGAAGGTGCTGAGGTACGATGAGACCCGCACCAGGTGCAGACCCTGCGAGTGTGGCGGTAAACATATTAGGAACCAGCCTGTGATGCTGGATGTGACCGAGGAGCTGAGGCCCGATCACTTGGTGCTGGCCTGCACCCGCGCTGAGTTTGGCTCTAGCGATGAAGATACAGATCTCGAGGGGTCCGGAGCTACAAATTTTTCCCTCCTCAAACAGGCTGGAGATGTCGAAGAAAATCCTGGGCCTACCGGTATGTATAGGATGCAGTTGCTCTCTTGCATCGCCCTGTCTCTAGCGTTGGTGACTAATAGCGCCCCTACTTCTTCTTCCACGAAGAAAACCCAGCTGCAACTGGAACACCTTCTGCTGGATCTCCAGATGATCCTTAACGGGATTAACAACTATAAGAACCCGAAGCTCACGGCTATGCTGACGGCGAAGTTCGCTATGCCTAAAAAGGCTACTGAGCTGAAACACTTACAGTGTCTCGAAGAGGCACTCAAACCCCTCGAGGAGGTACTCAATCTGGCGCAAAGCAAAAACTTCCATCTCCGCCCACGGGACCTAATCTCCAACATCAACGTGATTGTGCTGGAGCTCAAGGGGTCTGAAACCACGTTTATGTGCGAGTACGCAGATGAAACTGCTACCATAGTGGAATTCCTCAATCGTTGGATCACTTTCTGCCAGTCAATCATTTCAACCCTGACCAGATCTGAGGGCCGCGGCAGCCTGCTGACCTGCGGCGACGTGGAGGAAAACCCCGGCCCCGGATCCTTGTCGACCATTCGTCCGCGCAGGCCAAGTCCTGCCCGCGAGCTTCTGCCCGGACCCCAACCGGATAGGGTTCAGCCGACTGCAGATCGTGGGGTGTCTGCGCCTGCTGGCAGCCCTCTGGATGGCTTGCCCGCTCGGCGGACGGTGTCCCGGACCCGGCTGCCATCTCCCCCTGCGCCCTCACCTGCGTTCTCGGCGGGCAGCTTCAGCGATCTGCTCCGTCCGTTCGATCCGTCGCTTCTTGATACATCGCTTCTTGATTCGATGCCTGCCGTCGGCACGCCGCATACAGCGGCTGCCCCAGCAGAGTGGGATGAGGCGCAATCGGCTCTGCGTGCAGCCGATGACCCGCCACCCACCGTGCGTGTCGCTGTCACTGCCGCGCGGCCGCCGCGCGCCAAGCCGGCCCCGCGACGGCGTGCTGCGCAACCCTCCGACGCTTCGCCGGCCGCGCAGGTGGATCTACGCACGCTCGGCTACAGTCAGCAGCAGCAAGAGAAGATCAAACCGAAGGTGCGTTCGACAGTGGCGCAGCACCACGAGGCACTGGTGGGCCATGGGTTTACACACGCGCACATCGTTGCGCTCAGCCAACACCCGGCAGCGTTAGGGACCGTCGCTGTCACGTATCAGCACATAATCACGGCGTTGCCAGAGGCGACACACGAAGACATCGTTGGCGTCGGCAAACAGTGGTCCGGCGCACGCGCCCTGGAGGCCTTGCTCACGGATGCGGGGGAGTTGAGAGGTCCGCCGTTACAGTTGGACACAGGCCAACTTGTGAAGATTGCAAAACGTGGCGGCGTGACCGCAATGGAGGCAGTGCATGCATCGCGCAATGCACTGACGGGTGCCCCCCTGAACCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACAATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACAATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAAGCATTGTGGCCCAGCTGAGCCGGCCTGATCCGGCGTTGGCCGCGTTGACCAACGACCACCTCGTCGCCTTGGCCTGCCTCGGCGGACGTCCTGCCATGGATGCAGTGAAAAAGGGATTGCCGCACGCGCCGGAATTGATCAGAAGAGTCAATCGCCGTATTGGCGAACGCACGTCCCATCGCGTTGCCGACTACGCGCAAGTGGTTCGCGTGCTGGAGTTTTTCCAGTGCCACTCCCACCCAGCGTACGCATTTGATGAGGCCATGACGCAGTTCGGGATGAGCAGGAACGGGTTGGTACAGCTCTTTCGCAGAGTGGGCGTCACCGAACTCGAAGCCCGCGGTGGAACGCTCCCCCCAGCCTCGCAGCGTTGGGACCGTATCCTCCAGGCATCAGGGATGAAAAGGGCCAAACCGTCCCCTACTTCAGCTCAAACACCGGATCAGGCGTCTTTGCATGCATTCGCCGATTCGCTGGAGCGTGACCTTGATGCGCCCAGCCCAATGCACGAGGGAGATCAGACGCGGGCAAGCAGCCGTAAACGGTCCCGATCGGATCGTGCTGTCACCGGCCCCTCCGCACAGCAGGCTGTCGAGGTGCGCGTTCCCGAACAGCGCGATGCGCTGCATTTGCCCCTCAGCTGGAGGGTAAAACGCCCGCGTACCAGGATCTGGGGCGGCCTCCCGGTTAAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGTCACATAGGAATAAAAAGCCATATCACATAGGAATAAAAAGCCATATCACATAGGAATAAAAAGCCATATCACATAGGAATAAAAAGCCATAGAAAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTTCTGTCCTCCGAGCGGAGACTCTAGATATAGAACCGATCCTCCCATTGAATTCTAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCCTCGAGTATAGAACCGATCCTCCCATTTATAGAACCGATCCTCCCATTTATAGAACCGATCCTCCCATTACTAGAAGATCTTGAATTCGCCACCTTGTCGACCATTCGTCCGCGCAGGCCAAGTCCTGCCCGCGAGCTTCTGCCCGGACCCCAACCGGATAGGGTTCAGCCGACTGCAGATCGTGGGGTGTCTGCGCCTGCTGGCAGCCCTCTGGATGGCTTGCCCGCTCGGCGGACGGTGTCCCGGACCCGGCTGCCATCTCCCCCTGCGCCCTCACCTGCGTTCTCGGCGGGCAGCTTCAGCGATCTGCTCCGTCCGTTCGATCCGTCGCTTCTTGATACATCGCTTCTTGATTCGATGCCTGCCGTCGGCACGCCGCATACAGCGGCTGCCCCAGCAGAGTGGGATGAGGCGCAATCGGCTCTGCGTGCAGCCGATGACCCGCCACCCACCGTGCGTGTCGCTGTCACTGCCGCGCGGCCGCCGCGCGCCAAGCCGGCCCCGCGACGGCGTGCTGCGCAACCCTCCGACGCTTCGCCGGCCGCGCAGGTGGATCTACGCACGCTCGGCTACAGTCAGCAGCAGCAAGAGAAGATCAAACCGAAGGTGCGTTCGACAGTGGCGCAGCACCACGAGGCACTGGTGGGCCATGGGTTTACACACGCGCACATCGTTGCGCTCAGCCAACACCCGGCAGCGTTAGGGACCGTCGCTGTCACGTATCAGCACATAATCACGGCGTTGCCAGAGGCGACACACGAAGACATCGTTGGCGTCGGCAAACAGTGGTCCGGCGCACGCGCCCTGGAGGCCTTGCTCACGGATGCGGGGGAGTTGAGAGGTCCGCCGTTACAGTTGGACACAGGCCAACTTGTGAAGATTGCAAAACGTGGCGGCGTGACCGCAATGGAGGCAGTGCATGCATCGCGCAATGCACTGACGGGTGCCCCCCTGAACCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACAATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAAGCATTGTGGCCCAGCTGAGCCGGCCTGATCCGGCGTTGGCCGCGTTGACCAACGACCACCTCGTCGCCTTGGCCTGCCTCGGCGGACGTCCTGCCATGGATGCAGTGAAAAAGGGATTGCCGCACGCGCCGGAATTGATCAGAAGAGTCAATCGCCGTATTGGCGAACGCACGTCCCATCGCGTTGCCGACTACGCGCAAGTGGTTCGCGTGCTGGAGTTTTTCCAGTGCCACTCCCACCCAGCGTACGCATTTGATGAGGCCATGACGCAGTTCGGGATGAGCAGGAACGGGTTGGTACAGCTCTTTCGCAGAGTGGGCGTCACCGAACTCGAAGCCCGCGGTGGAACGCTCCCCCCAGCCTCGCAGCGTTGGGACCGTATCCTCCAGGCATCAGGGATGAAAAGGGCCAAACCGTCCCCTACTTCAGCTCAAACACCGGATCAGGCGTCTTTGCATGCATTCGCCGATTCGCTGGAGCGTGACCTTGATGCGCCCAGCCCAATGCACGAGGGAGATCAGACGCGGGCAAGCAGCCGTAAACGGTCCCGATCGGATCGTGCTGTCACCGGCCCCTCCGCACAGCAGGCTGTCGAGGTGCGCGTTCCCGAACAGCGCGATGCGCTGCATTTGCCCCTCAGCTGGAGGGTAAAACGCCCGCGTACCAGGATCTGGGGCGGCCTCCCGTAATACAAGGCGGCCGCAAATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTAAAGCTTCGAATTCTGATAATCAGCCATACCACATTTGTAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCAGCTCGCCTTTTGCTCACATGTCTCCCAGGGGATGGGGGTCCATCCACCTTGCCGAAAAGATTTGTCTGAGGAACTGAAAATAGAAGGGAAAAAAGAGGAGGGACAAAAGAGGCAGAAATGAGAGGGGAGGGGACAGAGGACACCTGAATAAAGACCACACCCATGACCCACGTGATGCTGAGAAGTACTCCTGCCCTAGGAAGAGACTCAGGGCAGAGGGAGGAAGGACAGCAGACCAGACAGTCACAGCAGCCTTGACAAAACGTTCCTGGAACTCAAGCTCTTCTCCACAGAGGAGGACAGAGCAGACAGCAGAGACCATGGAGTCTCCCTCGGCCCCTCCCCACAGATGGTGCATCCCCTGGCAGAGGCTCCTGCTCACAGGTGAACTCGAGACCGGTATGGCCCCCCCGACCGATGTCAGCCTGGGGGACGAGCTCCACTTAGACGGCGAGGACGTGGCGATGGCGCATGCCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGAGCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGAGCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGAGCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGGACGGGGATTCCCCGGGTCCGGGATTTACCCCCCACGACTCCGCCCCCTACGGCGCTCTGGATATGGCCGACTTCGAGTTTGAGCAGATGTTTACCGATGCCCTTGGAATTGACGAGTACGGTGGGACGCGTATGAAGCTACTGTCTTCTATCGAACAAGCATGCGATATTTGCCGACTTAAAAAGCTCAAGTGCTCCAAAGAAAAACCGAAGTGCGCCAAGTGTCTGAAGAACAACTGGGAGTGTCGCTACTCTCCCAAAACCAAAAGGTCTCCGCTGACTAGGGCACATCTGACAGAAGTGGAATCAAGGCTAGAAAGACTGGAACAGCTATTTCTACTGATTTTTCCTCGAGAAGACCTTGACATGATTTTGAAAATGGATTCTTTACAGGATATAAAAGCATTGTTAACAGGATTATTTGTACAAGATAATGTGAATAAAGATGCCGTCACAGATAGATTGGCTTCAGTGGAGACTGATATGCCTCTAACATTGAGACAGCATAGAATAAGTGCGACATCATCATCGGAAGAGAGTAGTAACAAAGGTCAAAGACAGTTGACTGTATAAAAGCTTCACTCCTCAGGTGCAGGCTGCCTATCAGAAGGTGGTGGCTGGTGTGGCCAATGCCCTGGCTCACAAATACCACTGAGATCTTTTTCCCTCTGCCAAAAATTATGGGGACATCATGAAGCCCCTTGAGCATCTGACTTCTGGCTAATAAAGGAAATTTATTTTCATTGCAATAGTGTGTTGGAATTTTTTGTGTCTCTCACTCGGAAGGACATATGGGAGGGCAAATCATTTAAAACATCAGAATGAGTATTTGGTTTAGAGTTTGGCAACATATGCCCATATGCTGGCTGCCATGAACAAAGGTTGGCTATAAAGAGGTCATCAGTATATGAAACAGCCCCCTGCTGTCCATTCCTTATTCCATAGAAAAGCCTTGACTTGAGGTTAGATTTTTTTTATATTTTGTTTTGTGTTATTTTTTTCTTTAACATCCCTAAAATTTTCCTTACATGTTTTACTAGCCAGATTTTTCCTCCTCTCCTGACTACTCCCAGTCATAGCTGTCCCTCTTCTCTTATGGAGATC(SEQ ID NO:10)。
具有SEQ ID NO:11所述核苷酸序列的核酸编码由细胞特异性启动子CEA205、第一microRNA为mir-135a-5p、第二microRNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白分别为Lacl、tetR-KRAB、第一和第二识别序列为4×UAS的元件组成的表达系统。
CGGGTGACCGCCCTCCGCGGAGTACTGTCCTCCGCGGGTGACAGCCCTCCGCGGAGTGCCGCACTCCGAGCGGAGACTCTAGACTCCCTATCAGTGATAGAGATAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCTCCCTATCAGTGATAGAGAGAATTCGACCGCCACCATGAGACATATTATCTGCCACGGAGGTGTTATTACCGAAGAAATGGCCGCCAGTCTTTTGGACCAGCTGATCGAAGAGGTACTGGCTGATAATCTTCCACCTCCTAGCCATTTTGAACCACCTACCCTTCACGAACTGTATGATTTAGACGTGACGGCCCCCGAAGATCCCAACGAGGAGGCGGTTTCGCAGATTTTTCCCGACTCTGTAATGTTGGCGGTGCAGGAAGGGATTGACTTACTCACTTTTCCGCCGGCGCCCGGTTCTCCGGAGCCGCCTCACCTTTCCCGGCAGCCCGAGCAGCCGGAGCAGAGAGCCTTGGGTCCGGTTTCTATGCCAAACCTTGTACCGGAGGTGATCGATCTTACCTGCCACGAGGCTGGCTTTCCACCCAGTGACGACGAGGATGAAGAGGGTGAGGAGTTTGTGTTAGATTATGTGGAGCACCCCGGGCACGGTTGCAGGTCTTGTCATTATCACCGGAGGAATACGGGGGACCCAGATATTATGTGTTCGCTTTGCTATATGAGGACCTGTGGCATGTTTGTCTACAGTAAGTGAAAATTATGGGCAGTGGGTGATAGAGTGGTGGGTTTGGTGTGGTAATTTTTTTTTTAATTTTTACAGTTTTGTGGTTTAAAGAATTTTGTATTGTGATTTTTTTAAAAGGTCCTGTGTCTGAACCTGAGCCTGAGCCCGAGCCAGAACCGGAGCCTGCAAGACCTACCCGCCGTCCTAAAATGGCGCCTGCTATCCTGAGGCGCCCGACATCACCTGTGTCTAGAGAATGCAATAGTAGTACGGATAGCTGTGACTCCGGTCCTTCTAACACACCTCCTGAGATACACCCGGTGGTCCCGCTGTGCCCCATTAAACCAGTTGCCGTGAGAGTTGGTGGGCGTCGCCAGGCTGTGGAATGTATCGAGGACTTGCTTAACGAGCCTGGGCAACCTTTGGACTTGAGCTGTAAACGCCCCAGGCCACTCGAGGGTACCGGGTCCGGAGCTACAAATTTTTCCCTCCTCAAACAGGCTGGAGATGTCGAAGAAAATCCTGGGCCTACCGGTATGTGGCTGCAGAGCCTCCTTCTACTGGGCACCGTGGCTTGCAGCATCTCGGCCCCCGCCAGAAGTCCCAGCCCCAGCACCCAGCCCTGGGAACACGTAAATGCAATCCAGGAGGCCCGCCGCCTCCTGAACTTGTCACGGGACACGGCAGCCGAGATGAATGAAACTGTGGAGGTCATCAGTGAAATGTTTGACTTGCAGGAGCCCACATGCCTGCAGACTCGGCTGGAGCTCTACAAACAGGGCCTGCGTGGCAGCCTCACCAAGCTGAAGGGCCCACTGACCATGATGGCTAGCCATTACAAGCAGCACTGCCCGCCGACCCCTGAGACATCCTGTGCCACTCAGATCATTACCTTTGAGTCTTTCAAGGAGAACCTCAAGGACTTTCTGCTGGTCATCCCATTTGACTGCTGGGAACCCGTCCAGGAAAGATCTGAGGGCCGCGGCAGCCTGCTGACCTGCGGCGACGTGGAGGAAAACCCCGGCCCCGGATCCATGAAACCAGTAACGTTATACGATGTCGCAGAGTATGCCGGTGTCTCTTATCAGACCGTTTCCCGCGTGGTGAACCAGGCCAGCCACGTTTCTGCGAAAACGCGGGAAAAAGTGGAAGCGGCGATGGCGGAGCTGAATTACATTCCCAACCGCGTGGCACAACAACTGGCGGGCAAACAGTCGTTGCTGATTGGCGTTGCCACCTCCAGTCTGGCCCTGCACGCGCCGTCGCAAATTGTCGCGGCGATTAAATCTCGCGCCGATCAACTGGGTGCCAGCGTGGTGGTGTCGATGGTAGAACGAAGCGGCGTCGAAGCCTGTAAAGCGGCGGTGCACAATCTTCTCGCGCAACGCGTCAGTGGGCTGATCATTAACTATCCGCTGGATGACCAGGATGCCATTGCTGTGGAAGCTGCCTGCACTAATGTTCCGGCGTTATTTCTTGATGTCTCTGACCAGACACCCATCAACAGTATTATTTTCTCCCATGAAGACGGTACGCGACTGGGCGTGGAGCATCTGGTCGCATTGGGTCACCAGCAAATCGCGCTGTTAGCGGGCCCATTAAGTTCTGTCTCGGCGCGTCTGCGTCTGGCTGGCTGGCATAAATATCTCACTCGCAATCAAATTCAGCCGATAGCGGAACGGGAAGGCGACTGGAGTGCCATGTCCGGTTTTCAACAAACCATGCAAATGCTGAATGAGGGCATCGTTCCCACTGCGATGCTGGTTGCCAACGATCAGATGGCGCTGGGCGCAATGCGCGCCATTACCGAGTCCGGGCTGCGCGTTGGTGCGGATATCTCGGTAGTGGGATACGACGATACCGAAGACAGCTCATGTTATATCCCGCCGTTAACCACCATCAAACAGGATTTTCGCCTGCTGGGGCAAACCAGCGTGGACCGCTTGCTGCAACTCTCTCAGGGCCAGGCGGTGAAGGGCAATCAGCTGTTGCCGGTCTCACTGGTGAAAAGAAAAACCACCCTGGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGAGCAGCCTGAGGCCTCCTAAGAAGAAGAGGAAGGTTGTTACAAACACCATTGTCACACTCCACAAACACCATTGTCACACTCCACAAACACCATTGTCACACTCCACAAACACCATTGTCACACTCCATCACATAGGAATAAAAAGCCATATCACATAGGAATAAAAAGCCATATCACATAGGAATAAAAAGCCATATCACATAGGAATAAAAAGCCATAGAAAGCGGGGTGCCGCCCCCCGCGGACTACTGTAGTCCGCGGACGACTGTCGTCCGCGGAGCACTGTGCTCCGAGCGGAGACTCTAGAAATTGTGAGCGGATAACAATTGGGTATATAATGGGGGCCGTCAGCTACTACCAGAGCCGATCGAACACTCGAGCCGAGCAGACGTGCCTACAATTGTGAGCGCTCACAATTACTAGAAGATCTTGAATTCGCCACCATGGCTAGATTAGATAAAAGTAAAGTGATTAACAGCGCATTAGAGCTGCTTAATGAGGTCGGAATCGAAGGTTTAACAACCCGTAAACTCGCCCAGAAGCTAGGTGTAGAGCAGCCTACATTGTATTGGCATGTAAAAAATAAGCGGGCTTTGCTCGACGCCTTAGCCATTGAGATGTTAGATAGGCACCATACTCACTTTTGCCCTTTAGAAGGGGAAAGCTGGCAAGATTTTTTACGTAATAACGCTAAAAGTTTTAGATGTGCTTTACTAAGTCATCGCGATGGAGCAAAAGTACATTTAGGTACACGGCCTACAGAAAAACAGTATGAAACTCTCGAAAATCAATTAGCCTTTTTATGCCAACAAGGTTTTTCACTAGAGAATGCATTATATGCACTCAGCGCTGTGGGGCATTTTACTTTAGGTTGCGTATTGGAAGATCAAGAGCATCAAGTCGCTAAAGAAGAAAGGGAAACACCTACTACTGATAGTATGCCGCCATTATTACGACAAGCTATCGAATTATTTGATCACCAAGGTGCAGAGCCAGCCTTCTTATTCGGCCTTGAATTGATCATATGCGGATTAGAAAAACAACTTAAATGTGAAAGTGGGTCGCCAAAAAAGAAGAGAAAGGTGGATGGTGGCGGAGCACTGTCTCCCCAGCATTCAGCTGTGACCCAGGGATCAATCATTAAGAACAAGGAAGGTATGGATGCTAAGTCCCTGACCGCTTGGTCAAGAACTTTGGTAACTTTTAAGGACGTGTTCGTCGATTTTACTCGGGAGGAGTGGAAACTATTGGATACCGCCCAGCAAATCGTGTATAGAAACGTTATGCTCGAGAACTATAAGAATCTTGTGTCACTGGGCTACCAACTGACAAAGCCTGACGTTATCCTGAGGCTGGAAAAGGGTGAGGAGCCTTGGTTGGTCGAGCGGGAGATCCACCAGGAAACTCACCCAGACAGTGAGACTGCTTTCGAGATCAAGTCCTCTGTGTAATACAAGGCGGCCGCAAATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTAAAGCTTCGAATTCTGATAATCAGCCATACCACATTTGTAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCAGCTCGCCTTTTGCTCACATGTGAAAATAGAAGGGAAAAAAGAGGAGGGACAAAAGAGGCAGAAATGAGAGGGGAGGGGACAGAGGACACCTGAATAAAGACCACACCCATGACCCACGTGATGCTGAGAAGTACTCCTGCCCTAGGAAGAGACTCAGGGCAGAGGGAGGAAGGACAGCAGACCAGACAGTCACAGCAGCCTTGACAAAACGTTCCTGGAACTCAAGCTCGAGACCGGTATGGCCCCCCCGACCGATGTCAGCCTGGGGGACGAGCTCCACTTAGACGGCGAGGACGTGGCGATGGCGCATGCCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGGACGGGGATTCCCCGGGTCCGGGATTTACCCCCCACGACTCCGCCCCCTACGGCGCTCTGGATATGGCCGACTTCGAGTTTGAGCAGATGTTTACCGATGCCCTTGGAATTGACGAGTACGGTGGGACGCGTATGAAGCTACTGTCTTCTATCGAACAAGCATGCGATATTTGCCGACTTAAAAAGCTCAAGTGCTCCAAAGAAAAACCGAAGTGCGCCAAGTGTCTGAAGAACAACTGGGAGTGTCGCTACTCTCCCAAAACCAAAAGGTCTCCGCTGACTAGGGCACATCTGACAGAAGTGGAATCAAGGCTAGAAAGACTGGAACAGCTATTTCTACTGATTTTTCCTCGAGAAGACCTTGACATGATTTTGAAAATGGATTCTTTACAGGATATAAAAGCATTGTTAACAGGATTATTTGTACAAGATAATGTGAATAAAGATGCCGTCACAGATAGATTGGCTTCAGTGGAGACTGATATGCCTCTAACATTGAGACAGCATAGAATAAGTGCGACATCATCATCGGAAGAGAGTAGTAACAAAGGTCAAAGACAGTTGACTGTATAAAAGCTTCACTCCTCAGGTGCAGGCTGCCTATCAGAAGGTGGTGGCTGGTGTGGCCAATGCCCTGGCTCACAAATACCACTGAGATCTTTTTCCCTCTGCCAAAAATTATGGGGACATCATGAAGCCCCTTGAGCATCTGACTTCTGGCTAATAAAGGAAATTTATTTTCATTGCAATAGTGTGTTGGAATTTTTTGTGTCTCTCACTCGGAAGGACATATGGGAGGGCAAATCATTTAAAACATCAGAATGAGTATTTGGTTTAGAGTTTGGCAACATATGCCCATATGCTGGCTGCCATGAACAAAGGTTGGCTATAAAGAGGTCATCAGTATATGAAACAGCCCCCTGCTGTCCATTCCTTATTCCATAGAAAAGCCTTGACTTGAGGTTAGATTTTTTTTATATTTTGTTTTGTGTTATTTTTTTCTTTAACATCCCTAAAATTTTCCTTACATGTTTTACTAGCCAGATTTTTCCTCCTCTCCTGACTACTCCCAGTCATAGCTGTCCCTCTTCTCTTATGGAGATC(SEQ ID NO:11)。
具有SEQ ID NO:12所述核苷酸序列的核酸编码由细胞特异性启动子CEA205、第一microRNA为mir-143-3p、第二microRNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白分别为TALE蛋白、第一和第二识别序列为4×UAS的元件组成的表达系统。
CGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTTCTGTCCTCCGAGCGGAGACTCTAGATACCCCTCTCCGCTTCTGAATTCTAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCCTCGAGTACCCCTCTCCGCTTCTTACCCCTCTCCGCTTCTTACCCCTCTCCGCTTCTGAATTCGACCGCCACCATGAGACATATTATCTGCCACGGAGGTGTTATTACCGAAGAAATGGCCGCCAGTCTTTTGGACCAGCTGATCGAAGAGGTACTGGCTGATAATCTTCCACCTCCTAGCCATTTTGAACCACCTACCCTTCACGAACTGTATGATTTAGACGTGACGGCCCCCGAAGATCCCAACGAGGAGGCGGTTTCGCAGATTTTTCCCGACTCTGTAATGTTGGCGGTGCAGGAAGGGATTGACTTACTCACTTTTCCGCCGGCGCCCGGTTCTCCGGAGCCGCCTCACCTTTCCCGGCAGCCCGAGCAGCCGGAGCAGAGAGCCTTGGGTCCGGTTTCTATGCCAAACCTTGTACCGGAGGTGATCGATCTTACCTGCCACGAGGCTGGCTTTCCACCCAGTGACGACGAGGATGAAGAGGGTGAGGAGTTTGTGTTAGATTATGTGGAGCACCCCGGGCACGGTTGCAGGTCTTGTCATTATCACCGGAGGAATACGGGGGACCCAGATATTATGTGTTCGCTTTGCTATATGAGGACCTGTGGCATGTTTGTCTACAGTAAGTGAAAATTATGGGCAGTGGGTGATAGAGTGGTGGGTTTGGTGTGGTAATTTTTTTTTTAATTTTTACAGTTTTGTGGTTTAAAGAATTTTGTATTGTGATTTTTTTAAAAGGTCCTGTGTCTGAACCTGAGCCTGAGCCCGAGCCAGAACCGGAGCCTGCAAGACCTACCCGCCGTCCTAAAATGGCGCCTGCTATCCTGAGGCGCCCGACATCACCTGTGTCTAGAGAATGCAATAGTAGTACGGATAGCTGTGACTCCGGTCCTTCTAACACACCTCCTGAGATACACCCGGTGGTCCCGCTGTGCCCCATTAAACCAGTTGCCGTGAGAGTTGGTGGGCGTCGCCAGGCTGTGGAATGTATCGAGGACTTGCTTAACGAGCCTGGGCAACCTTTGGACTTGAGCTGTAAACGCCCCAGGCCAGGGTCCGGAGCTACAAATTTTTCCCTCCTCAAACAGGCTGGAGATGTCGAAGAAAATCCTGGGCCTATGGAGGCTTGGGAGTGTTTGGAAGATTTTTCTGCTGTGCGTAACTTGCTGGAACAGAGCTCTAACAGTACCTCTTGGTTTTGGAGGTTTCTGTGGGGCTCATCCCAGGCAAAGTTAGTCTGCAGAATTAAGGAGGATTACAAGTGGGAATTTGAAGAGCTTTTGAAATCCTGTGGTGAGCTGTTTGATTCTTTGAATCTGGGTCACCAGGCGCTTTTCCAAGAGAAGGTCATCAAGACTTTGGATTTTTCCACACCGGGGCGCGCTGCGGCTGCTGTTGCTTTTTTGAGTTTTATAAAGGATAAATGGAGCGAAGAAACCCATCTGAGCGGGGGGTACCTGCTGGATTTTCTGGCCATGCATCTGTGGAGAGCGGTTGTGAGACACAAGAATCGCCTGCTACTGTTGTCTTCCGTCCGCCCGGCGATAATACCGACGGAGGAGCAGCAGCAGCAGCAGGAGGAAGCCAGGCGGCGGCGGCAGGAGCAGAGCCCATGGAACCCGAGAGCCGGCCTGGACCCTCGGGAATGAATGTTGTACAGGTGGCTGAACTGTATCCAGAACTGAGACGCATTTTGACAATTACAGAGGATGGGCAGGGGCTAAAGGGGGTAAAGAGGGAGCGGGGGGCTTGTGAGGCTACAGAGGAGGCTAGGAATCTAGCTTTTAGCTTAATGACCAGACACCGTCCTGAGTGTATTACTTTTCAACAGATCAAGGATAATTGCGCTAATGAGCTTGATCTGCTGGCGCAGAAGTATTCCATAGAGCAGCTGACCACTTACTGGCTGCAGCCAGGGGATGATTTTGAGGAGGCTATTAGGGTATATGCAAAGGTGGCACTTAGGCCAGATTGCAAGTACAAGATCAGCAAACTTGTAAATATCAGGAATTGTTGCTACATTTCTGGGAACGGGGCCGAGGTGGAGATAGATACGGAGGATAGGGTGGCCTTTAGATGTAGCATGATAAATATGTGGCCGGGGGTGCTTGGCATGGACGGGGTGGTTATTATGAATGTAAGGTTTACTGGCCCCAATTTTAGCGGTACGGTTTTCCTGGCCAATACCAACCTTATCCTACACGGTGTAAGCTTCTATGGGTTTAACAATACCTGTGTGGAAGCCTGGACCGATGTAAGGGTTCGGGGCTGTGCCTTTTACTGCTGCTGGAAGGGGGTGGTGTGTCGCCCCAAAAGCAGGGCTTCAATTAAGAAATGCCTCTTTGAAAGGTGTACCTTGGGTATCCTGTCTGAGGGTAACTCCAGGGTGCGCCACAATGTGGCCTCCGACTGTGGTTGCTTCATGCTAGTGAAAAGCGTGGCTGTGATTAAGCATAACATGGTATGTGGCAACTGCGAGGACAGGGCCTCTCAGATGCTGACCTGCTCGGACGGCAACTGTCACCTGCTGAAGACCATTCACGTAGCCAGCCACTCTCGCAAGGCCTGGCCAGTGTTTGAGCATAACATACTGACCCGCTGTTCCTTGCATTTGGGTAACAGGAGGGGGGTGTTCCTACCTTACCAATGCAATTTGAGTCACACTAAGATATTGCTTGAGCCCGAGAGCATGTCCAAGGTGAACCTGAACGGGGTGTTTGACATGACCATGAAGATCTGGAAGGTGCTGAGGTACGATGAGACCCGCACCAGGTGCAGACCCTGCGAGTGTGGCGGTAAACATATTAGGAACCAGCCTGTGATGCTGGATGTGACCGAGGAGCTGAGGCCCGATCACTTGGTGCTGGCCTGCACCCGCGCTGAGTTTGGCTCTAGCGATGAAGATACAGATCTCGAGGGGTCCGGAGCTACAAATTTTTCCCTCCTCAAACAGGCTGGAGATGTCGAAGAAAATCCTGGGCCTACCGGTATGTATAGGATGCAGTTGCTCTCTTGCATCGCCCTGTCTCTAGCGTTGGTGACTAATAGCGCCCCTACTTCTTCTTCCACGAAGAAAACCCAGCTGCAACTGGAACACCTTCTGCTGGATCTCCAGATGATCCTTAACGGGATTAACAACTATAAGAACCCGAAGCTCACGGCTATGCTGACGGCGAAGTTCGCTATGCCTAAAAAGGCTACTGAGCTGAAACACTTACAGTGTCTCGAAGAGGCACTCAAACCCCTCGAGGAGGTACTCAATCTGGCGCAAAGCAAAAACTTCCATCTCCGCCCACGGGACCTAATCTCCAACATCAACGTGATTGTGCTGGAGCTCAAGGGGTCTGAAACCACGTTTATGTGCGAGTACGCAGATGAAACTGCTACCATAGTGGAATTCCTCAATCGTTGGATCACTTTCTGCCAGTCAATCATTTCAACCCTGACCAGATCTGAGGGCCGCGGCAGCCTGCTGACCTGCGGCGACGTGGAGGAAAACCCCGGCCCCGGATCCTTGTCGACCATTCGTCCGCGCAGGCCAAGTCCTGCCCGCGAGCTTCTGCCCGGACCCCAACCGGATAGGGTTCAGCCGACTGCAGATCGTGGGGTGTCTGCGCCTGCTGGCAGCCCTCTGGATGGCTTGCCCGCTCGGCGGACGGTGTCCCGGACCCGGCTGCCATCTCCCCCTGCGCCCTCACCTGCGTTCTCGGCGGGCAGCTTCAGCGATCTGCTCCGTCCGTTCGATCCGTCGCTTCTTGATACATCGCTTCTTGATTCGATGCCTGCCGTCGGCACGCCGCATACAGCGGCTGCCCCAGCAGAGTGGGATGAGGCGCAATCGGCTCTGCGTGCAGCCGATGACCCGCCACCCACCGTGCGTGTCGCTGTCACTGCCGCGCGGCCGCCGCGCGCCAAGCCGGCCCCGCGACGGCGTGCTGCGCAACCCTCCGACGCTTCGCCGGCCGCGCAGGTGGATCTACGCACGCTCGGCTACAGTCAGCAGCAGCAAGAGAAGATCAAACCGAAGGTGCGTTCGACAGTGGCGCAGCACCACGAGGCACTGGTGGGCCATGGGTTTACACACGCGCACATCGTTGCGCTCAGCCAACACCCGGCAGCGTTAGGGACCGTCGCTGTCACGTATCAGCACATAATCACGGCGTTGCCAGAGGCGACACACGAAGACATCGTTGGCGTCGGCAAACAGTGGTCCGGCGCACGCGCCCTGGAGGCCTTGCTCACGGATGCGGGGGAGTTGAGAGGTCCGCCGTTACAGTTGGACACAGGCCAACTTGTGAAGATTGCAAAACGTGGCGGCGTGACCGCAATGGAGGCAGTGCATGCATCGCGCAATGCACTGACGGGTGCCCCCCTGAACCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACAATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACAATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAAGCATTGTGGCCCAGCTGAGCCGGCCTGATCCGGCGTTGGCCGCGTTGACCAACGACCACCTCGTCGCCTTGGCCTGCCTCGGCGGACGTCCTGCCATGGATGCAGTGAAAAAGGGATTGCCGCACGCGCCGGAATTGATCAGAAGAGTCAATCGCCGTATTGGCGAACGCACGTCCCATCGCGTTGCCGACTACGCGCAAGTGGTTCGCGTGCTGGAGTTTTTCCAGTGCCACTCCCACCCAGCGTACGCATTTGATGAGGCCATGACGCAGTTCGGGATGAGCAGGAACGGGTTGGTACAGCTCTTTCGCAGAGTGGGCGTCACCGAACTCGAAGCCCGCGGTGGAACGCTCCCCCCAGCCTCGCAGCGTTGGGACCGTATCCTCCAGGCATCAGGGATGAAAAGGGCCAAACCGTCCCCTACTTCAGCTCAAACACCGGATCAGGCGTCTTTGCATGCATTCGCCGATTCGCTGGAGCGTGACCTTGATGCGCCCAGCCCAATGCACGAGGGAGATCAGACGCGGGCAAGCAGCCGTAAACGGTCCCGATCGGATCGTGCTGTCACCGGCCCCTCCGCACAGCAGGCTGTCGAGGTGCGCGTTCCCGAACAGCGCGATGCGCTGCATTTGCCCCTCAGCTGGAGGGTAAAACGCCCGCGTACCAGGATCTGGGGCGGCCTCCCGGTTAAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGGAGCTACAGTGCTTCATCTCAGAGCTACAGTGCTTCATCTCAGAGCTACAGTGCTTCATCTCAGAGCTACAGTGCTTCATCTCAGAAAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTTCTGTCCTCCGAGCGGAGACTCTAGATATAGAACCGATCCTCCCATTGAATTCTAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCCTCGAGTATAGAACCGATCCTCCCATTTATAGAACCGATCCTCCCATTTATAGAACCGATCCTCCCATTACTAGAAGATCTTGAATTCGCCACCTTGTCGACCATTCGTCCGCGCAGGCCAAGTCCTGCCCGCGAGCTTCTGCCCGGACCCCAACCGGATAGGGTTCAGCCGACTGCAGATCGTGGGGTGTCTGCGCCTGCTGGCAGCCCTCTGGATGGCTTGCCCGCTCGGCGGACGGTGTCCCGGACCCGGCTGCCATCTCCCCCTGCGCCCTCACCTGCGTTCTCGGCGGGCAGCTTCAGCGATCTGCTCCGTCCGTTCGATCCGTCGCTTCTTGATACATCGCTTCTTGATTCGATGCCTGCCGTCGGCACGCCGCATACAGCGGCTGCCCCAGCAGAGTGGGATGAGGCGCAATCGGCTCTGCGTGCAGCCGATGACCCGCCACCCACCGTGCGTGTCGCTGTCACTGCCGCGCGGCCGCCGCGCGCCAAGCCGGCCCCGCGACGGCGTGCTGCGCAACCCTCCGACGCTTCGCCGGCCGCGCAGGTGGATCTACGCACGCTCGGCTACAGTCAGCAGCAGCAAGAGAAGATCAAACCGAAGGTGCGTTCGACAGTGGCGCAGCACCACGAGGCACTGGTGGGCCATGGGTTTACACACGCGCACATCGTTGCGCTCAGCCAACACCCGGCAGCGTTAGGGACCGTCGCTGTCACGTATCAGCACATAATCACGGCGTTGCCAGAGGCGACACACGAAGACATCGTTGGCGTCGGCAAACAGTGGTCCGGCGCACGCGCCCTGGAGGCCTTGCTCACGGATGCGGGGGAGTTGAGAGGTCCGCCGTTACAGTTGGACACAGGCCAACTTGTGAAGATTGCAAAACGTGGCGGCGTGACCGCAATGGAGGCAGTGCATGCATCGCGCAATGCACTGACGGGTGCCCCCCTGAACCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACAATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAAGCATTGTGGCCCAGCTGAGCCGGCCTGATCCGGCGTTGGCCGCGTTGACCAACGACCACCTCGTCGCCTTGGCCTGCCTCGGCGGACGTCCTGCCATGGATGCAGTGAAAAAGGGATTGCCGCACGCGCCGGAATTGATCAGAAGAGTCAATCGCCGTATTGGCGAACGCACGTCCCATCGCGTTGCCGACTACGCGCAAGTGGTTCGCGTGCTGGAGTTTTTCCAGTGCCACTCCCACCCAGCGTACGCATTTGATGAGGCCATGACGCAGTTCGGGATGAGCAGGAACGGGTTGGTACAGCTCTTTCGCAGAGTGGGCGTCACCGAACTCGAAGCCCGCGGTGGAACGCTCCCCCCAGCCTCGCAGCGTTGGGACCGTATCCTCCAGGCATCAGGGATGAAAAGGGCCAAACCGTCCCCTACTTCAGCTCAAACACCGGATCAGGCGTCTTTGCATGCATTCGCCGATTCGCTGGAGCGTGACCTTGATGCGCCCAGCCCAATGCACGAGGGAGATCAGACGCGGGCAAGCAGCCGTAAACGGTCCCGATCGGATCGTGCTGTCACCGGCCCCTCCGCACAGCAGGCTGTCGAGGTGCGCGTTCCCGAACAGCGCGATGCGCTGCATTTGCCCCTCAGCTGGAGGGTAAAACGCCCGCGTACCAGGATCTGGGGCGGCCTCCCGTAATACAAGGCGGCCGCAAATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTAAAGCTTCGAATTCTGATAATCAGCCATACCACATTTGTAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCGAAAATAGAAGGGAAAAAAGAGGAGGGACAAAAGAGGCAGAAATGAGAGGGGAGGGGACAGAGGACACCTGAATAAAGACCACACCCATGACCCACGTGATGCTGAGAAGTACTCCTGCCCTAGGAAGAGACTCAGGGCAGAGGGAGGAAGGACAGCAGACCAGACAGTCACAGCAGCCTTGACAAAACGTTCCTGGAACTCAAGATGGCCCCCCCGACCGATGTCAGCCTGGGGGACGAGCTCCACTTAGACGGCGAGGACGTGGCGATGGCGCATGCCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGAGCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGAGCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGAGCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGGACGGGGATTCCCCGGGTCCGGGATTTACCCCCCACGACTCCGCCCCCTACGGCGCTCTGGATATGGCCGACTTCGAGTTTGAGCAGATGTTTACCGATGCCCTTGGAATTGACGAGTACGGTGGGACGCGTATGAAGCTACTGTCTTCTATCGAACAAGCATGCGATATTTGCCGACTTAAAAAGCTCAAGTGCTCCAAAGAAAAACCGAAGTGCGCCAAGTGTCTGAAGAACAACTGGGAGTGTCGCTACTCTCCCAAAACCAAAAGGTCTCCGCTGACTAGGGCACATCTGACAGAAGTGGAATCAAGGCTAGAAAGACTGGAACAGCTATTTCTACTGATTTTTCCTCGAGAAGACCTTGACATGATTTTGAAAATGGATTCTTTACAGGATATAAAAGCATTGTTAACAGGATTATTTGTACAAGATAATGTGAATAAAGATGCCGTCACAGATAGATTGGCTTCAGTGGAGACTGATATGCCTCTAACATTGAGACAGCATAGAATAAGTGCGACATCATCATCGGAAGAGAGTAGTAACAAAGGTCAAAGACAGTTGACTGTATAAAAGCTTCACTCCTCAGGTGCAGGCTGCCTATCAGAAGGTGGTGGCTGGTGTGGCCAATGCCCTGGCTCACAAATACCACTGAGATCTTTTTCCCTCTGCCAAAAATTATGGGGACATCATGAAGCCCCTTGAGCATCTGACTTCTGGCTAATAAAGGAAATTTATTTTCATTGCAATAGTGTGTTGGAATTTTTTGTGTCTCTCACTCGGAAGGACATATGGGAGGGCAAATCATTTAAAACATCAGAATGAGTATTTGGTTTAGAGTTTGGCAACATATGCCCATATGCTGGCTGCCATGAACAAAGGTTGGCTATAAAGAGGTCATCAGTATATGAAACAGCCCCCTGCTGTCCATTCCTTATTCCATAGAAAAGCCTTGACTTGAGGTTAGATTTTTTTTATATTTTGTTTTGTGTTATTTTTTTCTTTAACATCCCTAAAATTTTCCTTACATGTTTTACTAGCCAGATTTTTCCTCCTCTCCTGACTACTCCCAGTCATAGCTGTCCCTCTTCTCTTATGGAGATC(SEQ ID NO:12)。
具有SEQ ID NO:13所述核苷酸序列的核酸编码由细胞特异性启动子CEA368、第一microRNA为mir-135a-5p、第二microRNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白分别为Lacl、tetR-KRAB、第一和第二识别序列为4×UAS的元件组成的表达系统。
CGGGTGACCGCCCTCCGCGGAGTACTGTCCTCCGCGGGTGACAGCCCTCCGCGGAGTGCCGCACTCCGAGCGGAGACTCTAGACTCCCTATCAGTGATAGAGATAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCTCCCTATCAGTGATAGAGAGAATTCGACCGCCACCATGAGACATATTATCTGCCACGGAGGTGTTATTACCGAAGAAATGGCCGCCAGTCTTTTGGACCAGCTGATCGAAGAGGTACTGGCTGATAATCTTCCACCTCCTAGCCATTTTGAACCACCTACCCTTCACGAACTGTATGATTTAGACGTGACGGCCCCCGAAGATCCCAACGAGGAGGCGGTTTCGCAGATTTTTCCCGACTCTGTAATGTTGGCGGTGCAGGAAGGGATTGACTTACTCACTTTTCCGCCGGCGCCCGGTTCTCCGGAGCCGCCTCACCTTTCCCGGCAGCCCGAGCAGCCGGAGCAGAGAGCCTTGGGTCCGGTTTCTATGCCAAACCTTGTACCGGAGGTGATCGATCTTACCTGCCACGAGGCTGGCTTTCCACCCAGTGACGACGAGGATGAAGAGGGTGAGGAGTTTGTGTTAGATTATGTGGAGCACCCCGGGCACGGTTGCAGGTCTTGTCATTATCACCGGAGGAATACGGGGGACCCAGATATTATGTGTTCGCTTTGCTATATGAGGACCTGTGGCATGTTTGTCTACAGTAAGTGAAAATTATGGGCAGTGGGTGATAGAGTGGTGGGTTTGGTGTGGTAATTTTTTTTTTAATTTTTACAGTTTTGTGGTTTAAAGAATTTTGTATTGTGATTTTTTTAAAAGGTCCTGTGTCTGAACCTGAGCCTGAGCCCGAGCCAGAACCGGAGCCTGCAAGACCTACCCGCCGTCCTAAAATGGCGCCTGCTATCCTGAGGCGCCCGACATCACCTGTGTCTAGAGAATGCAATAGTAGTACGGATAGCTGTGACTCCGGTCCTTCTAACACACCTCCTGAGATACACCCGGTGGTCCCGCTGTGCCCCATTAAACCAGTTGCCGTGAGAGTTGGTGGGCGTCGCCAGGCTGTGGAATGTATCGAGGACTTGCTTAACGAGCCTGGGCAACCTTTGGACTTGAGCTGTAAACGCCCCAGGCCACTCGAGGGTACCGGGTCCGGAGCTACAAATTTTTCCCTCCTCAAACAGGCTGGAGATGTCGAAGAAAATCCTGGGCCTACCGGTATGTGGCTGCAGAGCCTCCTTCTACTGGGCACCGTGGCTTGCAGCATCTCGGCCCCCGCCAGAAGTCCCAGCCCCAGCACCCAGCCCTGGGAACACGTAAATGCAATCCAGGAGGCCCGCCGCCTCCTGAACTTGTCACGGGACACGGCAGCCGAGATGAATGAAACTGTGGAGGTCATCAGTGAAATGTTTGACTTGCAGGAGCCCACATGCCTGCAGACTCGGCTGGAGCTCTACAAACAGGGCCTGCGTGGCAGCCTCACCAAGCTGAAGGGCCCACTGACCATGATGGCTAGCCATTACAAGCAGCACTGCCCGCCGACCCCTGAGACATCCTGTGCCACTCAGATCATTACCTTTGAGTCTTTCAAGGAGAACCTCAAGGACTTTCTGCTGGTCATCCCATTTGACTGCTGGGAACCCGTCCAGGAAAGATCTGAGGGCCGCGGCAGCCTGCTGACCTGCGGCGACGTGGAGGAAAACCCCGGCCCCGGATCCATGAAACCAGTAACGTTATACGATGTCGCAGAGTATGCCGGTGTCTCTTATCAGACCGTTTCCCGCGTGGTGAACCAGGCCAGCCACGTTTCTGCGAAAACGCGGGAAAAAGTGGAAGCGGCGATGGCGGAGCTGAATTACATTCCCAACCGCGTGGCACAACAACTGGCGGGCAAACAGTCGTTGCTGATTGGCGTTGCCACCTCCAGTCTGGCCCTGCACGCGCCGTCGCAAATTGTCGCGGCGATTAAATCTCGCGCCGATCAACTGGGTGCCAGCGTGGTGGTGTCGATGGTAGAACGAAGCGGCGTCGAAGCCTGTAAAGCGGCGGTGCACAATCTTCTCGCGCAACGCGTCAGTGGGCTGATCATTAACTATCCGCTGGATGACCAGGATGCCATTGCTGTGGAAGCTGCCTGCACTAATGTTCCGGCGTTATTTCTTGATGTCTCTGACCAGACACCCATCAACAGTATTATTTTCTCCCATGAAGACGGTACGCGACTGGGCGTGGAGCATCTGGTCGCATTGGGTCACCAGCAAATCGCGCTGTTAGCGGGCCCATTAAGTTCTGTCTCGGCGCGTCTGCGTCTGGCTGGCTGGCATAAATATCTCACTCGCAATCAAATTCAGCCGATAGCGGAACGGGAAGGCGACTGGAGTGCCATGTCCGGTTTTCAACAAACCATGCAAATGCTGAATGAGGGCATCGTTCCCACTGCGATGCTGGTTGCCAACGATCAGATGGCGCTGGGCGCAATGCGCGCCATTACCGAGTCCGGGCTGCGCGTTGGTGCGGATATCTCGGTAGTGGGATACGACGATACCGAAGACAGCTCATGTTATATCCCGCCGTTAACCACCATCAAACAGGATTTTCGCCTGCTGGGGCAAACCAGCGTGGACCGCTTGCTGCAACTCTCTCAGGGCCAGGCGGTGAAGGGCAATCAGCTGTTGCCGGTCTCACTGGTGAAAAGAAAAACCACCCTGGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGAGCAGCCTGAGGCCTCCTAAGAAGAAGAGGAAGGTTGTTACAAACACCATTGTCACACTCCACAAACACCATTGTCACACTCCACAAACACCATTGTCACACTCCACAAACACCATTGTCACACTCCATCACATAGGAATAAAAAGCCATATCACATAGGAATAAAAAGCCATATCACATAGGAATAAAAAGCCATATCACATAGGAATAAAAAGCCATAGAAAGCGGGGTGCCGCCCCCCGCGGACTACTGTAGTCCGCGGACGACTGTCGTCCGCGGAGCACTGTGCTCCGAGCGGAGACTCTAGAAATTGTGAGCGGATAACAATTGGGTATATAATGGGGGCCGTCAGCTACTACCAGAGCCGATCGAACACTCGAGCCGAGCAGACGTGCCTACAATTGTGAGCGCTCACAATTACTAGAAGATCTTGAATTCGCCACCATGGCTAGATTAGATAAAAGTAAAGTGATTAACAGCGCATTAGAGCTGCTTAATGAGGTCGGAATCGAAGGTTTAACAACCCGTAAACTCGCCCAGAAGCTAGGTGTAGAGCAGCCTACATTGTATTGGCATGTAAAAAATAAGCGGGCTTTGCTCGACGCCTTAGCCATTGAGATGTTAGATAGGCACCATACTCACTTTTGCCCTTTAGAAGGGGAAAGCTGGCAAGATTTTTTACGTAATAACGCTAAAAGTTTTAGATGTGCTTTACTAAGTCATCGCGATGGAGCAAAAGTACATTTAGGTACACGGCCTACAGAAAAACAGTATGAAACTCTCGAAAATCAATTAGCCTTTTTATGCCAACAAGGTTTTTCACTAGAGAATGCATTATATGCACTCAGCGCTGTGGGGCATTTTACTTTAGGTTGCGTATTGGAAGATCAAGAGCATCAAGTCGCTAAAGAAGAAAGGGAAACACCTACTACTGATAGTATGCCGCCATTATTACGACAAGCTATCGAATTATTTGATCACCAAGGTGCAGAGCCAGCCTTCTTATTCGGCCTTGAATTGATCATATGCGGATTAGAAAAACAACTTAAATGTGAAAGTGGGTCGCCAAAAAAGAAGAGAAAGGTGGATGGTGGCGGAGCACTGTCTCCCCAGCATTCAGCTGTGACCCAGGGATCAATCATTAAGAACAAGGAAGGTATGGATGCTAAGTCCCTGACCGCTTGGTCAAGAACTTTGGTAACTTTTAAGGACGTGTTCGTCGATTTTACTCGGGAGGAGTGGAAACTATTGGATACCGCCCAGCAAATCGTGTATAGAAACGTTATGCTCGAGAACTATAAGAATCTTGTGTCACTGGGCTACCAACTGACAAAGCCTGACGTTATCCTGAGGCTGGAAAAGGGTGAGGAGCCTTGGTTGGTCGAGCGGGAGATCCACCAGGAAACTCACCCAGACAGTGAGACTGCTTTCGAGATCAAGTCCTCTGTGTAATACAAGGCGGCCGCAAATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTAAAGCTTCGAATTCTGATAATCAGCCATACCACATTTGTAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCAGCTCGCCTTTTGCTCACATGTCTCCCAGGGGATGGGGGTCCATCCACCTTGCCGAAAAGATTTGTCTGAGGAACTGAAAATAGAAGGGAAAAAAGAGGAGGGACAAAAGAGGCAGAAATGAGAGGGGAGGGGACAGAGGACACCTGAATAAAGACCACACCCATGACCCACGTGATGCTGAGAAGTACTCCTGCCCTAGGAAGAGACTCAGGGCAGAGGGAGGAAGGACAGCAGACCAGACAGTCACAGCAGCCTTGACAAAACGTTCCTGGAACTCAAGCTCTTCTCCACAGAGGAGGACAGAGCAGACAGCAGAGACCATGGAGTCTCCCTCGGCCCCTCCCCACAGATGGTGCATCCCCTGGCAGAGGCTCCTGCTCACAGGTGAACTCGAGACCGGTATGGCCCCCCCGACCGATGTCAGCCTGGGGGACGAGCTCCACTTAGACGGCGAGGACGTGGCGATGGCGCATGCCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGGACGGGGATTCCCCGGGTCCGGGATTTACCCCCCACGACTCCGCCCCCTACGGCGCTCTGGATATGGCCGACTTCGAGTTTGAGCAGATGTTTACCGATGCCCTTGGAATTGACGAGTACGGTGGGACGCGTATGAAGCTACTGTCTTCTATCGAACAAGCATGCGATATTTGCCGACTTAAAAAGCTCAAGTGCTCCAAAGAAAAACCGAAGTGCGCCAAGTGTCTGAAGAACAACTGGGAGTGTCGCTACTCTCCCAAAACCAAAAGGTCTCCGCTGACTAGGGCACATCTGACAGAAGTGGAATCAAGGCTAGAAAGACTGGAACAGCTATTTCTACTGATTTTTCCTCGAGAAGACCTTGACATGATTTTGAAAATGGATTCTTTACAGGATATAAAAGCATTGTTAACAGGATTATTTGTACAAGATAATGTGAATAAAGATGCCGTCACAGATAGATTGGCTTCAGTGGAGACTGATATGCCTCTAACATTGAGACAGCATAGAATAAGTGCGACATCATCATCGGAAGAGAGTAGTAACAAAGGTCAAAGACAGTTGACTGTATAAAAGCTTCACTCCTCAGGTGCAGGCTGCCTATCAGAAGGTGGTGGCTGGTGTGGCCAATGCCCTGGCTCACAAATACCACTGAGATCTTTTTCCCTCTGCCAAAAATTATGGGGACATCATGAAGCCCCTTGAGCATCTGACTTCTGGCTAATAAAGGAAATTTATTTTCATTGCAATAGTGTGTTGGAATTTTTTGTGTCTCTCACTCGGAAGGACATATGGGAGGGCAAATCATTTAAAACATCAGAATGAGTATTTGGTTTAGAGTTTGGCAACATATGCCCATATGCTGGCTGCCATGAACAAAGGTTGGCTATAAAGAGGTCATCAGTATATGAAACAGCCCCCTGCTGTCCATTCCTTATTCCATAGAAAAGCCTTGACTTGAGGTTAGATTTTTTTTATATTTTGTTTTGTGTTATTTTTTTCTTTAACATCCCTAAAATTTTCCTTACATGTTTTACTAGCCAGATTTTTCCTCCTCTCCTGACTACTCCCAGTCATAGCTGTCCCTCTTCTCTTATGGAGATC(SEQ IDNO:13)。
具有SEQ ID NO:14所述核苷酸序列的核酸编码由细胞特异性启动子CEA368、第一microRNA为mir-143-3p、第二microRNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白分别为TALE蛋白、第一和第二识别序列为4×UAS的元件组成的表达系统。
CGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTTCTGTCCTCCGAGCGGAGACTCTAGATACCCCTCTCCGCTTCTGAATTCTAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCCTCGAGTACCCCTCTCCGCTTCTTACCCCTCTCCGCTTCTTACCCCTCTCCGCTTCTGAATTCGACCGCCACCATGAGACATATTATCTGCCACGGAGGTGTTATTACCGAAGAAATGGCCGCCAGTCTTTTGGACCAGCTGATCGAAGAGGTACTGGCTGATAATCTTCCACCTCCTAGCCATTTTGAACCACCTACCCTTCACGAACTGTATGATTTAGACGTGACGGCCCCCGAAGATCCCAACGAGGAGGCGGTTTCGCAGATTTTTCCCGACTCTGTAATGTTGGCGGTGCAGGAAGGGATTGACTTACTCACTTTTCCGCCGGCGCCCGGTTCTCCGGAGCCGCCTCACCTTTCCCGGCAGCCCGAGCAGCCGGAGCAGAGAGCCTTGGGTCCGGTTTCTATGCCAAACCTTGTACCGGAGGTGATCGATCTTACCTGCCACGAGGCTGGCTTTCCACCCAGTGACGACGAGGATGAAGAGGGTGAGGAGTTTGTGTTAGATTATGTGGAGCACCCCGGGCACGGTTGCAGGTCTTGTCATTATCACCGGAGGAATACGGGGGACCCAGATATTATGTGTTCGCTTTGCTATATGAGGACCTGTGGCATGTTTGTCTACAGTAAGTGAAAATTATGGGCAGTGGGTGATAGAGTGGTGGGTTTGGTGTGGTAATTTTTTTTTTAATTTTTACAGTTTTGTGGTTTAAAGAATTTTGTATTGTGATTTTTTTAAAAGGTCCTGTGTCTGAACCTGAGCCTGAGCCCGAGCCAGAACCGGAGCCTGCAAGACCTACCCGCCGTCCTAAAATGGCGCCTGCTATCCTGAGGCGCCCGACATCACCTGTGTCTAGAGAATGCAATAGTAGTACGGATAGCTGTGACTCCGGTCCTTCTAACACACCTCCTGAGATACACCCGGTGGTCCCGCTGTGCCCCATTAAACCAGTTGCCGTGAGAGTTGGTGGGCGTCGCCAGGCTGTGGAATGTATCGAGGACTTGCTTAACGAGCCTGGGCAACCTTTGGACTTGAGCTGTAAACGCCCCAGGCCAGGGTCCGGAGCTACAAATTTTTCCCTCCTCAAACAGGCTGGAGATGTCGAAGAAAATCCTGGGCCTATGGAGGCTTGGGAGTGTTTGGAAGATTTTTCTGCTGTGCGTAACTTGCTGGAACAGAGCTCTAACAGTACCTCTTGGTTTTGGAGGTTTCTGTGGGGCTCATCCCAGGCAAAGTTAGTCTGCAGAATTAAGGAGGATTACAAGTGGGAATTTGAAGAGCTTTTGAAATCCTGTGGTGAGCTGTTTGATTCTTTGAATCTGGGTCACCAGGCGCTTTTCCAAGAGAAGGTCATCAAGACTTTGGATTTTTCCACACCGGGGCGCGCTGCGGCTGCTGTTGCTTTTTTGAGTTTTATAAAGGATAAATGGAGCGAAGAAACCCATCTGAGCGGGGGGTACCTGCTGGATTTTCTGGCCATGCATCTGTGGAGAGCGGTTGTGAGACACAAGAATCGCCTGCTACTGTTGTCTTCCGTCCGCCCGGCGATAATACCGACGGAGGAGCAGCAGCAGCAGCAGGAGGAAGCCAGGCGGCGGCGGCAGGAGCAGAGCCCATGGAACCCGAGAGCCGGCCTGGACCCTCGGGAATGAATGTTGTACAGGTGGCTGAACTGTATCCAGAACTGAGACGCATTTTGACAATTACAGAGGATGGGCAGGGGCTAAAGGGGGTAAAGAGGGAGCGGGGGGCTTGTGAGGCTACAGAGGAGGCTAGGAATCTAGCTTTTAGCTTAATGACCAGACACCGTCCTGAGTGTATTACTTTTCAACAGATCAAGGATAATTGCGCTAATGAGCTTGATCTGCTGGCGCAGAAGTATTCCATAGAGCAGCTGACCACTTACTGGCTGCAGCCAGGGGATGATTTTGAGGAGGCTATTAGGGTATATGCAAAGGTGGCACTTAGGCCAGATTGCAAGTACAAGATCAGCAAACTTGTAAATATCAGGAATTGTTGCTACATTTCTGGGAACGGGGCCGAGGTGGAGATAGATACGGAGGATAGGGTGGCCTTTAGATGTAGCATGATAAATATGTGGCCGGGGGTGCTTGGCATGGACGGGGTGGTTATTATGAATGTAAGGTTTACTGGCCCCAATTTTAGCGGTACGGTTTTCCTGGCCAATACCAACCTTATCCTACACGGTGTAAGCTTCTATGGGTTTAACAATACCTGTGTGGAAGCCTGGACCGATGTAAGGGTTCGGGGCTGTGCCTTTTACTGCTGCTGGAAGGGGGTGGTGTGTCGCCCCAAAAGCAGGGCTTCAATTAAGAAATGCCTCTTTGAAAGGTGTACCTTGGGTATCCTGTCTGAGGGTAACTCCAGGGTGCGCCACAATGTGGCCTCCGACTGTGGTTGCTTCATGCTAGTGAAAAGCGTGGCTGTGATTAAGCATAACATGGTATGTGGCAACTGCGAGGACAGGGCCTCTCAGATGCTGACCTGCTCGGACGGCAACTGTCACCTGCTGAAGACCATTCACGTAGCCAGCCACTCTCGCAAGGCCTGGCCAGTGTTTGAGCATAACATACTGACCCGCTGTTCCTTGCATTTGGGTAACAGGAGGGGGGTGTTCCTACCTTACCAATGCAATTTGAGTCACACTAAGATATTGCTTGAGCCCGAGAGCATGTCCAAGGTGAACCTGAACGGGGTGTTTGACATGACCATGAAGATCTGGAAGGTGCTGAGGTACGATGAGACCCGCACCAGGTGCAGACCCTGCGAGTGTGGCGGTAAACATATTAGGAACCAGCCTGTGATGCTGGATGTGACCGAGGAGCTGAGGCCCGATCACTTGGTGCTGGCCTGCACCCGCGCTGAGTTTGGCTCTAGCGATGAAGATACAGATCTCGAGGGGTCCGGAGCTACAAATTTTTCCCTCCTCAAACAGGCTGGAGATGTCGAAGAAAATCCTGGGCCTACCGGTATGTATAGGATGCAGTTGCTCTCTTGCATCGCCCTGTCTCTAGCGTTGGTGACTAATAGCGCCCCTACTTCTTCTTCCACGAAGAAAACCCAGCTGCAACTGGAACACCTTCTGCTGGATCTCCAGATGATCCTTAACGGGATTAACAACTATAAGAACCCGAAGCTCACGGCTATGCTGACGGCGAAGTTCGCTATGCCTAAAAAGGCTACTGAGCTGAAACACTTACAGTGTCTCGAAGAGGCACTCAAACCCCTCGAGGAGGTACTCAATCTGGCGCAAAGCAAAAACTTCCATCTCCGCCCACGGGACCTAATCTCCAACATCAACGTGATTGTGCTGGAGCTCAAGGGGTCTGAAACCACGTTTATGTGCGAGTACGCAGATGAAACTGCTACCATAGTGGAATTCCTCAATCGTTGGATCACTTTCTGCCAGTCAATCATTTCAACCCTGACCAGATCTGAGGGCCGCGGCAGCCTGCTGACCTGCGGCGACGTGGAGGAAAACCCCGGCCCCGGATCCTTGTCGACCATTCGTCCGCGCAGGCCAAGTCCTGCCCGCGAGCTTCTGCCCGGACCCCAACCGGATAGGGTTCAGCCGACTGCAGATCGTGGGGTGTCTGCGCCTGCTGGCAGCCCTCTGGATGGCTTGCCCGCTCGGCGGACGGTGTCCCGGACCCGGCTGCCATCTCCCCCTGCGCCCTCACCTGCGTTCTCGGCGGGCAGCTTCAGCGATCTGCTCCGTCCGTTCGATCCGTCGCTTCTTGATACATCGCTTCTTGATTCGATGCCTGCCGTCGGCACGCCGCATACAGCGGCTGCCCCAGCAGAGTGGGATGAGGCGCAATCGGCTCTGCGTGCAGCCGATGACCCGCCACCCACCGTGCGTGTCGCTGTCACTGCCGCGCGGCCGCCGCGCGCCAAGCCGGCCCCGCGACGGCGTGCTGCGCAACCCTCCGACGCTTCGCCGGCCGCGCAGGTGGATCTACGCACGCTCGGCTACAGTCAGCAGCAGCAAGAGAAGATCAAACCGAAGGTGCGTTCGACAGTGGCGCAGCACCACGAGGCACTGGTGGGCCATGGGTTTACACACGCGCACATCGTTGCGCTCAGCCAACACCCGGCAGCGTTAGGGACCGTCGCTGTCACGTATCAGCACATAATCACGGCGTTGCCAGAGGCGACACACGAAGACATCGTTGGCGTCGGCAAACAGTGGTCCGGCGCACGCGCCCTGGAGGCCTTGCTCACGGATGCGGGGGAGTTGAGAGGTCCGCCGTTACAGTTGGACACAGGCCAACTTGTGAAGATTGCAAAACGTGGCGGCGTGACCGCAATGGAGGCAGTGCATGCATCGCGCAATGCACTGACGGGTGCCCCCCTGAACCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACAATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACAATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAAGCATTGTGGCCCAGCTGAGCCGGCCTGATCCGGCGTTGGCCGCGTTGACCAACGACCACCTCGTCGCCTTGGCCTGCCTCGGCGGACGTCCTGCCATGGATGCAGTGAAAAAGGGATTGCCGCACGCGCCGGAATTGATCAGAAGAGTCAATCGCCGTATTGGCGAACGCACGTCCCATCGCGTTGCCGACTACGCGCAAGTGGTTCGCGTGCTGGAGTTTTTCCAGTGCCACTCCCACCCAGCGTACGCATTTGATGAGGCCATGACGCAGTTCGGGATGAGCAGGAACGGGTTGGTACAGCTCTTTCGCAGAGTGGGCGTCACCGAACTCGAAGCCCGCGGTGGAACGCTCCCCCCAGCCTCGCAGCGTTGGGACCGTATCCTCCAGGCATCAGGGATGAAAAGGGCCAAACCGTCCCCTACTTCAGCTCAAACACCGGATCAGGCGTCTTTGCATGCATTCGCCGATTCGCTGGAGCGTGACCTTGATGCGCCCAGCCCAATGCACGAGGGAGATCAGACGCGGGCAAGCAGCCGTAAACGGTCCCGATCGGATCGTGCTGTCACCGGCCCCTCCGCACAGCAGGCTGTCGAGGTGCGCGTTCCCGAACAGCGCGATGCGCTGCATTTGCCCCTCAGCTGGAGGGTAAAACGCCCGCGTACCAGGATCTGGGGCGGCCTCCCGGTTAAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGGAGCTACAGTGCTTCATCTCAGAGCTACAGTGCTTCATCTCAGAGCTACAGTGCTTCATCTCAGAGCTACAGTGCTTCATCTCAGAAAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTTCTGTCCTCCGAGCGGAGACTCTAGATATAGAACCGATCCTCCCATTGAATTCTAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCCTCGAGTATAGAACCGATCCTCCCATTTATAGAACCGATCCTCCCATTTATAGAACCGATCCTCCCATTACTAGAAGATCTTGAATTCGCCACCTTGTCGACCATTCGTCCGCGCAGGCCAAGTCCTGCCCGCGAGCTTCTGCCCGGACCCCAACCGGATAGGGTTCAGCCGACTGCAGATCGTGGGGTGTCTGCGCCTGCTGGCAGCCCTCTGGATGGCTTGCCCGCTCGGCGGACGGTGTCCCGGACCCGGCTGCCATCTCCCCCTGCGCCCTCACCTGCGTTCTCGGCGGGCAGCTTCAGCGATCTGCTCCGTCCGTTCGATCCGTCGCTTCTTGATACATCGCTTCTTGATTCGATGCCTGCCGTCGGCACGCCGCATACAGCGGCTGCCCCAGCAGAGTGGGATGAGGCGCAATCGGCTCTGCGTGCAGCCGATGACCCGCCACCCACCGTGCGTGTCGCTGTCACTGCCGCGCGGCCGCCGCGCGCCAAGCCGGCCCCGCGACGGCGTGCTGCGCAACCCTCCGACGCTTCGCCGGCCGCGCAGGTGGATCTACGCACGCTCGGCTACAGTCAGCAGCAGCAAGAGAAGATCAAACCGAAGGTGCGTTCGACAGTGGCGCAGCACCACGAGGCACTGGTGGGCCATGGGTTTACACACGCGCACATCGTTGCGCTCAGCCAACACCCGGCAGCGTTAGGGACCGTCGCTGTCACGTATCAGCACATAATCACGGCGTTGCCAGAGGCGACACACGAAGACATCGTTGGCGTCGGCAAACAGTGGTCCGGCGCACGCGCCCTGGAGGCCTTGCTCACGGATGCGGGGGAGTTGAGAGGTCCGCCGTTACAGTTGGACACAGGCCAACTTGTGAAGATTGCAAAACGTGGCGGCGTGACCGCAATGGAGGCAGTGCATGCATCGCGCAATGCACTGACGGGTGCCCCCCTGAACCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACAATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAAGCATTGTGGCCCAGCTGAGCCGGCCTGATCCGGCGTTGGCCGCGTTGACCAACGACCACCTCGTCGCCTTGGCCTGCCTCGGCGGACGTCCTGCCATGGATGCAGTGAAAAAGGGATTGCCGCACGCGCCGGAATTGATCAGAAGAGTCAATCGCCGTATTGGCGAACGCACGTCCCATCGCGTTGCCGACTACGCGCAAGTGGTTCGCGTGCTGGAGTTTTTCCAGTGCCACTCCCACCCAGCGTACGCATTTGATGAGGCCATGACGCAGTTCGGGATGAGCAGGAACGGGTTGGTACAGCTCTTTCGCAGAGTGGGCGTCACCGAACTCGAAGCCCGCGGTGGAACGCTCCCCCCAGCCTCGCAGCGTTGGGACCGTATCCTCCAGGCATCAGGGATGAAAAGGGCCAAACCGTCCCCTACTTCAGCTCAAACACCGGATCAGGCGTCTTTGCATGCATTCGCCGATTCGCTGGAGCGTGACCTTGATGCGCCCAGCCCAATGCACGAGGGAGATCAGACGCGGGCAAGCAGCCGTAAACGGTCCCGATCGGATCGTGCTGTCACCGGCCCCTCCGCACAGCAGGCTGTCGAGGTGCGCGTTCCCGAACAGCGCGATGCGCTGCATTTGCCCCTCAGCTGGAGGGTAAAACGCCCGCGTACCAGGATCTGGGGCGGCCTCCCGTAATACAAGGCGGCCGCAAATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTAAAGCTTCGAATTCTGATAATCAGCCATACCACATTTGTAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCAGCTCGCCTTTTGCTCACATGTCTCCCAGGGGATGGGGGTCCATCCACCTTGCCGAAAAGATTTGTCTGAGGAACTGAAAATAGAAGGGAAAAAAGAGGAGGGACAAAAGAGGCAGAAATGAGAGGGGAGGGGACAGAGGACACCTGAATAAAGACCACACCCATGACCCACGTGATGCTGAGAAGTACTCCTGCCCTAGGAAGAGACTCAGGGCAGAGGGAGGAAGGACAGCAGACCAGACAGTCACAGCAGCCTTGACAAAACGTTCCTGGAACTCAAGCTCTTCTCCACAGAGGAGGACAGAGCAGACAGCAGAGACCATGGAGTCTCCCTCGGCCCCTCCCCACAGATGGTGCATCCCCTGGCAGAGGCTCCTGCTCACAGGTGAACTCGAGACCGGTATGGCCCCCCCGACCGATGTCAGCCTGGGGGACGAGCTCCACTTAGACGGCGAGGACGTGGCGATGGCGCATGCCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGAGCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGAGCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGAGCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGGACGGGGATTCCCCGGGTCCGGGATTTACCCCCCACGACTCCGCCCCCTACGGCGCTCTGGATATGGCCGACTTCGAGTTTGAGCAGATGTTTACCGATGCCCTTGGAATTGACGAGTACGGTGGGACGCGTATGAAGCTACTGTCTTCTATCGAACAAGCATGCGATATTTGCCGACTTAAAAAGCTCAAGTGCTCCAAAGAAAAACCGAAGTGCGCCAAGTGTCTGAAGAACAACTGGGAGTGTCGCTACTCTCCCAAAACCAAAAGGTCTCCGCTGACTAGGGCACATCTGACAGAAGTGGAATCAAGGCTAGAAAGACTGGAACAGCTATTTCTACTGATTTTTCCTCGAGAAGACCTTGACATGATTTTGAAAATGGATTCTTTACAGGATATAAAAGCATTGTTAACAGGATTATTTGTACAAGATAATGTGAATAAAGATGCCGTCACAGATAGATTGGCTTCAGTGGAGACTGATATGCCTCTAACATTGAGACAGCATAGAATAAGTGCGACATCATCATCGGAAGAGAGTAGTAACAAAGGTCAAAGACAGTTGACTGTATAAAAGCTTCACTCCTCAGGTGCAGGCTGCCTATCAGAAGGTGGTGGCTGGTGTGGCCAATGCCCTGGCTCACAAATACCACTGAGATCTTTTTCCCTCTGCCAAAAATTATGGGGACATCATGAAGCCCCTTGAGCATCTGACTTCTGGCTAATAAAGGAAATTTATTTTCATTGCAATAGTGTGTTGGAATTTTTTGTGTCTCTCACTCGGAAGGACATATGGGAGGGCAAATCATTTAAAACATCAGAATGAGTATTTGGTTTAGAGTTTGGCAACATATGCCCATATGCTGGCTGCCATGAACAAAGGTTGGCTATAAAGAGGTCATCAGTATATGAAACAGCCCCCTGCTGTCCATTCCTTATTCCATAGAAAAGCCTTGACTTGAGGTTAGATTTTTTTTATATTTTGTTTTGTGTTATTTTTTTCTTTAACATCCCTAAAATTTTCCTTACATGTTTTACTAGCCAGATTTTTCCTCCTCTCCTGACTACTCCCAGTCATAGCTGTCCCTCTTCTCTTATGGAGATC(SEQ ID NO:14)。
具有SEQ ID NO:15所述核苷酸序列的核酸编码由细胞特异性启动子CXCR4、第一microRNA为mir-135a-5p、第二microRNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白分别为Lacl、tetR-KRAB、第一和第二识别序列为4×UAS的元件组成的表达系统。
CGGGTGACCGCCCTCCGCGGAGTACTGTCCTCCGCGGGTGACAGCCCTCCGCGGAGTGCCGCACTCCGAGCGGAGACTCTAGACTCCCTATCAGTGATAGAGATAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCTCCCTATCAGTGATAGAGAGAATTCGACCGCCACCATGAGACATATTATCTGCCACGGAGGTGTTATTACCGAAGAAATGGCCGCCAGTCTTTTGGACCAGCTGATCGAAGAGGTACTGGCTGATAATCTTCCACCTCCTAGCCATTTTGAACCACCTACCCTTCACGAACTGTATGATTTAGACGTGACGGCCCCCGAAGATCCCAACGAGGAGGCGGTTTCGCAGATTTTTCCCGACTCTGTAATGTTGGCGGTGCAGGAAGGGATTGACTTACTCACTTTTCCGCCGGCGCCCGGTTCTCCGGAGCCGCCTCACCTTTCCCGGCAGCCCGAGCAGCCGGAGCAGAGAGCCTTGGGTCCGGTTTCTATGCCAAACCTTGTACCGGAGGTGATCGATCTTACCTGCCACGAGGCTGGCTTTCCACCCAGTGACGACGAGGATGAAGAGGGTGAGGAGTTTGTGTTAGATTATGTGGAGCACCCCGGGCACGGTTGCAGGTCTTGTCATTATCACCGGAGGAATACGGGGGACCCAGATATTATGTGTTCGCTTTGCTATATGAGGACCTGTGGCATGTTTGTCTACAGTAAGTGAAAATTATGGGCAGTGGGTGATAGAGTGGTGGGTTTGGTGTGGTAATTTTTTTTTTAATTTTTACAGTTTTGTGGTTTAAAGAATTTTGTATTGTGATTTTTTTAAAAGGTCCTGTGTCTGAACCTGAGCCTGAGCCCGAGCCAGAACCGGAGCCTGCAAGACCTACCCGCCGTCCTAAAATGGCGCCTGCTATCCTGAGGCGCCCGACATCACCTGTGTCTAGAGAATGCAATAGTAGTACGGATAGCTGTGACTCCGGTCCTTCTAACACACCTCCTGAGATACACCCGGTGGTCCCGCTGTGCCCCATTAAACCAGTTGCCGTGAGAGTTGGTGGGCGTCGCCAGGCTGTGGAATGTATCGAGGACTTGCTTAACGAGCCTGGGCAACCTTTGGACTTGAGCTGTAAACGCCCCAGGCCACTCGAGGGTACCGGGTCCGGAGCTACAAATTTTTCCCTCCTCAAACAGGCTGGAGATGTCGAAGAAAATCCTGGGCCTACCGGTATGTGGCTGCAGAGCCTCCTTCTACTGGGCACCGTGGCTTGCAGCATCTCGGCCCCCGCCAGAAGTCCCAGCCCCAGCACCCAGCCCTGGGAACACGTAAATGCAATCCAGGAGGCCCGCCGCCTCCTGAACTTGTCACGGGACACGGCAGCCGAGATGAATGAAACTGTGGAGGTCATCAGTGAAATGTTTGACTTGCAGGAGCCCACATGCCTGCAGACTCGGCTGGAGCTCTACAAACAGGGCCTGCGTGGCAGCCTCACCAAGCTGAAGGGCCCACTGACCATGATGGCTAGCCATTACAAGCAGCACTGCCCGCCGACCCCTGAGACATCCTGTGCCACTCAGATCATTACCTTTGAGTCTTTCAAGGAGAACCTCAAGGACTTTCTGCTGGTCATCCCATTTGACTGCTGGGAACCCGTCCAGGAAAGATCTGAGGGCCGCGGCAGCCTGCTGACCTGCGGCGACGTGGAGGAAAACCCCGGCCCCGGATCCATGAAACCAGTAACGTTATACGATGTCGCAGAGTATGCCGGTGTCTCTTATCAGACCGTTTCCCGCGTGGTGAACCAGGCCAGCCACGTTTCTGCGAAAACGCGGGAAAAAGTGGAAGCGGCGATGGCGGAGCTGAATTACATTCCCAACCGCGTGGCACAACAACTGGCGGGCAAACAGTCGTTGCTGATTGGCGTTGCCACCTCCAGTCTGGCCCTGCACGCGCCGTCGCAAATTGTCGCGGCGATTAAATCTCGCGCCGATCAACTGGGTGCCAGCGTGGTGGTGTCGATGGTAGAACGAAGCGGCGTCGAAGCCTGTAAAGCGGCGGTGCACAATCTTCTCGCGCAACGCGTCAGTGGGCTGATCATTAACTATCCGCTGGATGACCAGGATGCCATTGCTGTGGAAGCTGCCTGCACTAATGTTCCGGCGTTATTTCTTGATGTCTCTGACCAGACACCCATCAACAGTATTATTTTCTCCCATGAAGACGGTACGCGACTGGGCGTGGAGCATCTGGTCGCATTGGGTCACCAGCAAATCGCGCTGTTAGCGGGCCCATTAAGTTCTGTCTCGGCGCGTCTGCGTCTGGCTGGCTGGCATAAATATCTCACTCGCAATCAAATTCAGCCGATAGCGGAACGGGAAGGCGACTGGAGTGCCATGTCCGGTTTTCAACAAACCATGCAAATGCTGAATGAGGGCATCGTTCCCACTGCGATGCTGGTTGCCAACGATCAGATGGCGCTGGGCGCAATGCGCGCCATTACCGAGTCCGGGCTGCGCGTTGGTGCGGATATCTCGGTAGTGGGATACGACGATACCGAAGACAGCTCATGTTATATCCCGCCGTTAACCACCATCAAACAGGATTTTCGCCTGCTGGGGCAAACCAGCGTGGACCGCTTGCTGCAACTCTCTCAGGGCCAGGCGGTGAAGGGCAATCAGCTGTTGCCGGTCTCACTGGTGAAAAGAAAAACCACCCTGGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGAGCAGCCTGAGGCCTCCTAAGAAGAAGAGGAAGGTTGTTACAAACACCATTGTCACACTCCACAAACACCATTGTCACACTCCACAAACACCATTGTCACACTCCACAAACACCATTGTCACACTCCATCACATAGGAATAAAAAGCCATATCACATAGGAATAAAAAGCCATATCACATAGGAATAAAAAGCCATATCACATAGGAATAAAAAGCCATAGAAAGCGGGGTGCCGCCCCCCGCGGACTACTGTAGTCCGCGGACGACTGTCGTCCGCGGAGCACTGTGCTCCGAGCGGAGACTCTAGAAATTGTGAGCGGATAACAATTGGGTATATAATGGGGGCCGTCAGCTACTACCAGAGCCGATCGAACACTCGAGCCGAGCAGACGTGCCTACAATTGTGAGCGCTCACAATTACTAGAAGATCTTGAATTCGCCACCATGGCTAGATTAGATAAAAGTAAAGTGATTAACAGCGCATTAGAGCTGCTTAATGAGGTCGGAATCGAAGGTTTAACAACCCGTAAACTCGCCCAGAAGCTAGGTGTAGAGCAGCCTACATTGTATTGGCATGTAAAAAATAAGCGGGCTTTGCTCGACGCCTTAGCCATTGAGATGTTAGATAGGCACCATACTCACTTTTGCCCTTTAGAAGGGGAAAGCTGGCAAGATTTTTTACGTAATAACGCTAAAAGTTTTAGATGTGCTTTACTAAGTCATCGCGATGGAGCAAAAGTACATTTAGGTACACGGCCTACAGAAAAACAGTATGAAACTCTCGAAAATCAATTAGCCTTTTTATGCCAACAAGGTTTTTCACTAGAGAATGCATTATATGCACTCAGCGCTGTGGGGCATTTTACTTTAGGTTGCGTATTGGAAGATCAAGAGCATCAAGTCGCTAAAGAAGAAAGGGAAACACCTACTACTGATAGTATGCCGCCATTATTACGACAAGCTATCGAATTATTTGATCACCAAGGTGCAGAGCCAGCCTTCTTATTCGGCCTTGAATTGATCATATGCGGATTAGAAAAACAACTTAAATGTGAAAGTGGGTCGCCAAAAAAGAAGAGAAAGGTGGATGGTGGCGGAGCACTGTCTCCCCAGCATTCAGCTGTGACCCAGGGATCAATCATTAAGAACAAGGAAGGTATGGATGCTAAGTCCCTGACCGCTTGGTCAAGAACTTTGGTAACTTTTAAGGACGTGTTCGTCGATTTTACTCGGGAGGAGTGGAAACTATTGGATACCGCCCAGCAAATCGTGTATAGAAACGTTATGCTCGAGAACTATAAGAATCTTGTGTCACTGGGCTACCAACTGACAAAGCCTGACGTTATCCTGAGGCTGGAAAAGGGTGAGGAGCCTTGGTTGGTCGAGCGGGAGATCCACCAGGAAACTCACCCAGACAGTGAGACTGCTTTCGAGATCAAGTCCTCTGTGTAATACAAGGCGGCCGCAAATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTAAAGCTTCGAATTCTGATAATCAGCCATACCACATTTGTAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCAGCTCGCCTTTTGCTCACATGTTACCGACCACCCGCAAACAGCAGGGTCCCCTGGGCTTCCCAAGCCGCGCACCTCTCCGCCCCGCCCCTGCGCCCTCCTTCCTCGCGTCTGCCCCTCTCCCCCACCCCGCCTTCTCCCTCCCCGCCCCAGCGGCGCATGCGCCGCGCTCGGAGCGTGTTTTTATAAAAGTCCGGCCGCGGCCAGAAACTTCAGTTTGTTGGCTGCGGCAGCAGGTAGCAAAGTGACGCCGAGGGCCTGAGTGCTCCAGTAGCCACCGCATCTGGAGAACCAGCGGTTACCCTCGAGACCGGTATGGCCCCCCCGACCGATGTCAGCCTGGGGGACGAGCTCCACTTAGACGGCGAGGACGTGGCGATGGCGCATGCCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGGACGGGGATTCCCCGGGTCCGGGATTTACCCCCCACGACTCCGCCCCCTACGGCGCTCTGGATATGGCCGACTTCGAGTTTGAGCAGATGTTTACCGATGCCCTTGGAATTGACGAGTACGGTGGGACGCGTATGAAGCTACTGTCTTCTATCGAACAAGCATGCGATATTTGCCGACTTAAAAAGCTCAAGTGCTCCAAAGAAAAACCGAAGTGCGCCAAGTGTCTGAAGAACAACTGGGAGTGTCGCTACTCTCCCAAAACCAAAAGGTCTCCGCTGACTAGGGCACATCTGACAGAAGTGGAATCAAGGCTAGAAAGACTGGAACAGCTATTTCTACTGATTTTTCCTCGAGAAGACCTTGACATGATTTTGAAAATGGATTCTTTACAGGATATAAAAGCATTGTTAACAGGATTATTTGTACAAGATAATGTGAATAAAGATGCCGTCACAGATAGATTGGCTTCAGTGGAGACTGATATGCCTCTAACATTGAGACAGCATAGAATAAGTGCGACATCATCATCGGAAGAGAGTAGTAACAAAGGTCAAAGACAGTTGACTGTATAAAAGCTTCACTCCTCAGGTGCAGGCTGCCTATCAGAAGGTGGTGGCTGGTGTGGCCAATGCCCTGGCTCACAAATACCACTGAGATCTTTTTCCCTCTGCCAAAAATTATGGGGACATCATGAAGCCCCTTGAGCATCTGACTTCTGGCTAATAAAGGAAATTTATTTTCATTGCAATAGTGTGTTGGAATTTTTTGTGTCTCTCACTCGGAAGGACATATGGGAGGGCAAATCATTTAAAACATCAGAATGAGTATTTGGTTTAGAGTTTGGCAACATATGCCCATATGCTGGCTGCCATGAACAAAGGTTGGCTATAAAGAGGTCATCAGTATATGAAACAGCCCCCTGCTGTCCATTCCTTATTCCATAGAAAAGCCTTGACTTGAGGTTAGATTTTTTTTATATTTTGTTTTGTGTTATTTTTTTCTTTAACATCCCTAAAATTTTCCTTACATGTTTTACTAGCCAGATTTTTCCTCCTCTCCTGACTACTCCCAGTCATAGCTGTCCCTCTTCTCTTATGGAGATC(SEQ ID NO:15)。
具有SEQ ID NO:16所述核苷酸序列的核酸编码由细胞特异性启动子CXCR4、第一microRNA为mir-143-3p、第二microRNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白分别为TALE蛋白、第一和第二识别序列为4×UAS的元件组成的表达系统。
CGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTTCTGTCCTCCGAGCGGAGACTCTAGATACCCCTCTCCGCTTCTGAATTCTAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCCTCGAGTACCCCTCTCCGCTTCTTACCCCTCTCCGCTTCTTACCCCTCTCCGCTTCTGAATTCGACCGCCACCATGAGACATATTATCTGCCACGGAGGTGTTATTACCGAAGAAATGGCCGCCAGTCTTTTGGACCAGCTGATCGAAGAGGTACTGGCTGATAATCTTCCACCTCCTAGCCATTTTGAACCACCTACCCTTCACGAACTGTATGATTTAGACGTGACGGCCCCCGAAGATCCCAACGAGGAGGCGGTTTCGCAGATTTTTCCCGACTCTGTAATGTTGGCGGTGCAGGAAGGGATTGACTTACTCACTTTTCCGCCGGCGCCCGGTTCTCCGGAGCCGCCTCACCTTTCCCGGCAGCCCGAGCAGCCGGAGCAGAGAGCCTTGGGTCCGGTTTCTATGCCAAACCTTGTACCGGAGGTGATCGATCTTACCTGCCACGAGGCTGGCTTTCCACCCAGTGACGACGAGGATGAAGAGGGTGAGGAGTTTGTGTTAGATTATGTGGAGCACCCCGGGCACGGTTGCAGGTCTTGTCATTATCACCGGAGGAATACGGGGGACCCAGATATTATGTGTTCGCTTTGCTATATGAGGACCTGTGGCATGTTTGTCTACAGTAAGTGAAAATTATGGGCAGTGGGTGATAGAGTGGTGGGTTTGGTGTGGTAATTTTTTTTTTAATTTTTACAGTTTTGTGGTTTAAAGAATTTTGTATTGTGATTTTTTTAAAAGGTCCTGTGTCTGAACCTGAGCCTGAGCCCGAGCCAGAACCGGAGCCTGCAAGACCTACCCGCCGTCCTAAAATGGCGCCTGCTATCCTGAGGCGCCCGACATCACCTGTGTCTAGAGAATGCAATAGTAGTACGGATAGCTGTGACTCCGGTCCTTCTAACACACCTCCTGAGATACACCCGGTGGTCCCGCTGTGCCCCATTAAACCAGTTGCCGTGAGAGTTGGTGGGCGTCGCCAGGCTGTGGAATGTATCGAGGACTTGCTTAACGAGCCTGGGCAACCTTTGGACTTGAGCTGTAAACGCCCCAGGCCAGGGTCCGGAGCTACAAATTTTTCCCTCCTCAAACAGGCTGGAGATGTCGAAGAAAATCCTGGGCCTATGGAGGCTTGGGAGTGTTTGGAAGATTTTTCTGCTGTGCGTAACTTGCTGGAACAGAGCTCTAACAGTACCTCTTGGTTTTGGAGGTTTCTGTGGGGCTCATCCCAGGCAAAGTTAGTCTGCAGAATTAAGGAGGATTACAAGTGGGAATTTGAAGAGCTTTTGAAATCCTGTGGTGAGCTGTTTGATTCTTTGAATCTGGGTCACCAGGCGCTTTTCCAAGAGAAGGTCATCAAGACTTTGGATTTTTCCACACCGGGGCGCGCTGCGGCTGCTGTTGCTTTTTTGAGTTTTATAAAGGATAAATGGAGCGAAGAAACCCATCTGAGCGGGGGGTACCTGCTGGATTTTCTGGCCATGCATCTGTGGAGAGCGGTTGTGAGACACAAGAATCGCCTGCTACTGTTGTCTTCCGTCCGCCCGGCGATAATACCGACGGAGGAGCAGCAGCAGCAGCAGGAGGAAGCCAGGCGGCGGCGGCAGGAGCAGAGCCCATGGAACCCGAGAGCCGGCCTGGACCCTCGGGAATGAATGTTGTACAGGTGGCTGAACTGTATCCAGAACTGAGACGCATTTTGACAATTACAGAGGATGGGCAGGGGCTAAAGGGGGTAAAGAGGGAGCGGGGGGCTTGTGAGGCTACAGAGGAGGCTAGGAATCTAGCTTTTAGCTTAATGACCAGACACCGTCCTGAGTGTATTACTTTTCAACAGATCAAGGATAATTGCGCTAATGAGCTTGATCTGCTGGCGCAGAAGTATTCCATAGAGCAGCTGACCACTTACTGGCTGCAGCCAGGGGATGATTTTGAGGAGGCTATTAGGGTATATGCAAAGGTGGCACTTAGGCCAGATTGCAAGTACAAGATCAGCAAACTTGTAAATATCAGGAATTGTTGCTACATTTCTGGGAACGGGGCCGAGGTGGAGATAGATACGGAGGATAGGGTGGCCTTTAGATGTAGCATGATAAATATGTGGCCGGGGGTGCTTGGCATGGACGGGGTGGTTATTATGAATGTAAGGTTTACTGGCCCCAATTTTAGCGGTACGGTTTTCCTGGCCAATACCAACCTTATCCTACACGGTGTAAGCTTCTATGGGTTTAACAATACCTGTGTGGAAGCCTGGACCGATGTAAGGGTTCGGGGCTGTGCCTTTTACTGCTGCTGGAAGGGGGTGGTGTGTCGCCCCAAAAGCAGGGCTTCAATTAAGAAATGCCTCTTTGAAAGGTGTACCTTGGGTATCCTGTCTGAGGGTAACTCCAGGGTGCGCCACAATGTGGCCTCCGACTGTGGTTGCTTCATGCTAGTGAAAAGCGTGGCTGTGATTAAGCATAACATGGTATGTGGCAACTGCGAGGACAGGGCCTCTCAGATGCTGACCTGCTCGGACGGCAACTGTCACCTGCTGAAGACCATTCACGTAGCCAGCCACTCTCGCAAGGCCTGGCCAGTGTTTGAGCATAACATACTGACCCGCTGTTCCTTGCATTTGGGTAACAGGAGGGGGGTGTTCCTACCTTACCAATGCAATTTGAGTCACACTAAGATATTGCTTGAGCCCGAGAGCATGTCCAAGGTGAACCTGAACGGGGTGTTTGACATGACCATGAAGATCTGGAAGGTGCTGAGGTACGATGAGACCCGCACCAGGTGCAGACCCTGCGAGTGTGGCGGTAAACATATTAGGAACCAGCCTGTGATGCTGGATGTGACCGAGGAGCTGAGGCCCGATCACTTGGTGCTGGCCTGCACCCGCGCTGAGTTTGGCTCTAGCGATGAAGATACAGATCTCGAGGGGTCCGGAGCTACAAATTTTTCCCTCCTCAAACAGGCTGGAGATGTCGAAGAAAATCCTGGGCCTACCGGTATGTATAGGATGCAGTTGCTCTCTTGCATCGCCCTGTCTCTAGCGTTGGTGACTAATAGCGCCCCTACTTCTTCTTCCACGAAGAAAACCCAGCTGCAACTGGAACACCTTCTGCTGGATCTCCAGATGATCCTTAACGGGATTAACAACTATAAGAACCCGAAGCTCACGGCTATGCTGACGGCGAAGTTCGCTATGCCTAAAAAGGCTACTGAGCTGAAACACTTACAGTGTCTCGAAGAGGCACTCAAACCCCTCGAGGAGGTACTCAATCTGGCGCAAAGCAAAAACTTCCATCTCCGCCCACGGGACCTAATCTCCAACATCAACGTGATTGTGCTGGAGCTCAAGGGGTCTGAAACCACGTTTATGTGCGAGTACGCAGATGAAACTGCTACCATAGTGGAATTCCTCAATCGTTGGATCACTTTCTGCCAGTCAATCATTTCAACCCTGACCAGATCTGAGGGCCGCGGCAGCCTGCTGACCTGCGGCGACGTGGAGGAAAACCCCGGCCCCGGATCCTTGTCGACCATTCGTCCGCGCAGGCCAAGTCCTGCCCGCGAGCTTCTGCCCGGACCCCAACCGGATAGGGTTCAGCCGACTGCAGATCGTGGGGTGTCTGCGCCTGCTGGCAGCCCTCTGGATGGCTTGCCCGCTCGGCGGACGGTGTCCCGGACCCGGCTGCCATCTCCCCCTGCGCCCTCACCTGCGTTCTCGGCGGGCAGCTTCAGCGATCTGCTCCGTCCGTTCGATCCGTCGCTTCTTGATACATCGCTTCTTGATTCGATGCCTGCCGTCGGCACGCCGCATACAGCGGCTGCCCCAGCAGAGTGGGATGAGGCGCAATCGGCTCTGCGTGCAGCCGATGACCCGCCACCCACCGTGCGTGTCGCTGTCACTGCCGCGCGGCCGCCGCGCGCCAAGCCGGCCCCGCGACGGCGTGCTGCGCAACCCTCCGACGCTTCGCCGGCCGCGCAGGTGGATCTACGCACGCTCGGCTACAGTCAGCAGCAGCAAGAGAAGATCAAACCGAAGGTGCGTTCGACAGTGGCGCAGCACCACGAGGCACTGGTGGGCCATGGGTTTACACACGCGCACATCGTTGCGCTCAGCCAACACCCGGCAGCGTTAGGGACCGTCGCTGTCACGTATCAGCACATAATCACGGCGTTGCCAGAGGCGACACACGAAGACATCGTTGGCGTCGGCAAACAGTGGTCCGGCGCACGCGCCCTGGAGGCCTTGCTCACGGATGCGGGGGAGTTGAGAGGTCCGCCGTTACAGTTGGACACAGGCCAACTTGTGAAGATTGCAAAACGTGGCGGCGTGACCGCAATGGAGGCAGTGCATGCATCGCGCAATGCACTGACGGGTGCCCCCCTGAACCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACAATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACAATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAAGCATTGTGGCCCAGCTGAGCCGGCCTGATCCGGCGTTGGCCGCGTTGACCAACGACCACCTCGTCGCCTTGGCCTGCCTCGGCGGACGTCCTGCCATGGATGCAGTGAAAAAGGGATTGCCGCACGCGCCGGAATTGATCAGAAGAGTCAATCGCCGTATTGGCGAACGCACGTCCCATCGCGTTGCCGACTACGCGCAAGTGGTTCGCGTGCTGGAGTTTTTCCAGTGCCACTCCCACCCAGCGTACGCATTTGATGAGGCCATGACGCAGTTCGGGATGAGCAGGAACGGGTTGGTACAGCTCTTTCGCAGAGTGGGCGTCACCGAACTCGAAGCCCGCGGTGGAACGCTCCCCCCAGCCTCGCAGCGTTGGGACCGTATCCTCCAGGCATCAGGGATGAAAAGGGCCAAACCGTCCCCTACTTCAGCTCAAACACCGGATCAGGCGTCTTTGCATGCATTCGCCGATTCGCTGGAGCGTGACCTTGATGCGCCCAGCCCAATGCACGAGGGAGATCAGACGCGGGCAAGCAGCCGTAAACGGTCCCGATCGGATCGTGCTGTCACCGGCCCCTCCGCACAGCAGGCTGTCGAGGTGCGCGTTCCCGAACAGCGCGATGCGCTGCATTTGCCCCTCAGCTGGAGGGTAAAACGCCCGCGTACCAGGATCTGGGGCGGCCTCCCGGTTAAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGGAGCTACAGTGCTTCATCTCAGAGCTACAGTGCTTCATCTCAGAGCTACAGTGCTTCATCTCAGAGCTACAGTGCTTCATCTCAGAAAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTTCTGTCCTCCGAGCGGAGACTCTAGATATAGAACCGATCCTCCCATTGAATTCTAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCCTCGAGTATAGAACCGATCCTCCCATTTATAGAACCGATCCTCCCATTTATAGAACCGATCCTCCCATTACTAGAAGATCTTGAATTCGCCACCTTGTCGACCATTCGTCCGCGCAGGCCAAGTCCTGCCCGCGAGCTTCTGCCCGGACCCCAACCGGATAGGGTTCAGCCGACTGCAGATCGTGGGGTGTCTGCGCCTGCTGGCAGCCCTCTGGATGGCTTGCCCGCTCGGCGGACGGTGTCCCGGACCCGGCTGCCATCTCCCCCTGCGCCCTCACCTGCGTTCTCGGCGGGCAGCTTCAGCGATCTGCTCCGTCCGTTCGATCCGTCGCTTCTTGATACATCGCTTCTTGATTCGATGCCTGCCGTCGGCACGCCGCATACAGCGGCTGCCCCAGCAGAGTGGGATGAGGCGCAATCGGCTCTGCGTGCAGCCGATGACCCGCCACCCACCGTGCGTGTCGCTGTCACTGCCGCGCGGCCGCCGCGCGCCAAGCCGGCCCCGCGACGGCGTGCTGCGCAACCCTCCGACGCTTCGCCGGCCGCGCAGGTGGATCTACGCACGCTCGGCTACAGTCAGCAGCAGCAAGAGAAGATCAAACCGAAGGTGCGTTCGACAGTGGCGCAGCACCACGAGGCACTGGTGGGCCATGGGTTTACACACGCGCACATCGTTGCGCTCAGCCAACACCCGGCAGCGTTAGGGACCGTCGCTGTCACGTATCAGCACATAATCACGGCGTTGCCAGAGGCGACACACGAAGACATCGTTGGCGTCGGCAAACAGTGGTCCGGCGCACGCGCCCTGGAGGCCTTGCTCACGGATGCGGGGGAGTTGAGAGGTCCGCCGTTACAGTTGGACACAGGCCAACTTGTGAAGATTGCAAAACGTGGCGGCGTGACCGCAATGGAGGCAGTGCATGCATCGCGCAATGCACTGACGGGTGCCCCCCTGAACCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACAATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAAGCATTGTGGCCCAGCTGAGCCGGCCTGATCCGGCGTTGGCCGCGTTGACCAACGACCACCTCGTCGCCTTGGCCTGCCTCGGCGGACGTCCTGCCATGGATGCAGTGAAAAAGGGATTGCCGCACGCGCCGGAATTGATCAGAAGAGTCAATCGCCGTATTGGCGAACGCACGTCCCATCGCGTTGCCGACTACGCGCAAGTGGTTCGCGTGCTGGAGTTTTTCCAGTGCCACTCCCACCCAGCGTACGCATTTGATGAGGCCATGACGCAGTTCGGGATGAGCAGGAACGGGTTGGTACAGCTCTTTCGCAGAGTGGGCGTCACCGAACTCGAAGCCCGCGGTGGAACGCTCCCCCCAGCCTCGCAGCGTTGGGACCGTATCCTCCAGGCATCAGGGATGAAAAGGGCCAAACCGTCCCCTACTTCAGCTCAAACACCGGATCAGGCGTCTTTGCATGCATTCGCCGATTCGCTGGAGCGTGACCTTGATGCGCCCAGCCCAATGCACGAGGGAGATCAGACGCGGGCAAGCAGCCGTAAACGGTCCCGATCGGATCGTGCTGTCACCGGCCCCTCCGCACAGCAGGCTGTCGAGGTGCGCGTTCCCGAACAGCGCGATGCGCTGCATTTGCCCCTCAGCTGGAGGGTAAAACGCCCGCGTACCAGGATCTGGGGCGGCCTCCCGTAATACAAGGCGGCCGCAAATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTAAAGCTTCGAATTCTGATAATCAGCCATACCACATTTGTAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCTACCGACCACCCGCAAACAGCAGGGTCCCCTGGGCTTCCCAAGCCGCGCACCTCTCCGCCCCGCCCCTGCGCCCTCCTTCCTCGCGTCTGCCCCTCTCCCCCACCCCGCCTTCTCCCTCCCCGCCCCAGCGGCGCATGCGCCGCGCTCGGAGCGTGTTTTTATAAAAGTCCGGCCGCGGCCAGAAACTTCAGTTTGTTGGCTGCGGCAGCAGGTAGCAAAGTGACGCCGAGGGCCTGAGTGCTCCAGTAGCCACCGCATCTGGAGAACCAGCGGTTACCATGGCCCCCCCGACCGATGTCAGCCTGGGGGACGAGCTCCACTTAGACGGCGAGGACGTGGCGATGGCGCATGCCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGAGCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGAGCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGAGCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGGACGGGGATTCCCCGGGTCCGGGATTTACCCCCCACGACTCCGCCCCCTACGGCGCTCTGGATATGGCCGACTTCGAGTTTGAGCAGATGTTTACCGATGCCCTTGGAATTGACGAGTACGGTGGGACGCGTATGAAGCTACTGTCTTCTATCGAACAAGCATGCGATATTTGCCGACTTAAAAAGCTCAAGTGCTCCAAAGAAAAACCGAAGTGCGCCAAGTGTCTGAAGAACAACTGGGAGTGTCGCTACTCTCCCAAAACCAAAAGGTCTCCGCTGACTAGGGCACATCTGACAGAAGTGGAATCAAGGCTAGAAAGACTGGAACAGCTATTTCTACTGATTTTTCCTCGAGAAGACCTTGACATGATTTTGAAAATGGATTCTTTACAGGATATAAAAGCATTGTTAACAGGATTATTTGTACAAGATAATGTGAATAAAGATGCCGTCACAGATAGATTGGCTTCAGTGGAGACTGATATGCCTCTAACATTGAGACAGCATAGAATAAGTGCGACATCATCATCGGAAGAGAGTAGTAACAAAGGTCAAAGACAGTTGACTGTATAAAAGCTTCACTCCTCAGGTGCAGGCTGCCTATCAGAAGGTGGTGGCTGGTGTGGCCAATGCCCTGGCTCACAAATACCACTGAGATCTTTTTCCCTCTGCCAAAAATTATGGGGACATCATGAAGCCCCTTGAGCATCTGACTTCTGGCTAATAAAGGAAATTTATTTTCATTGCAATAGTGTGTTGGAATTTTTTGTGTCTCTCACTCGGAAGGACATATGGGAGGGCAAATCATTTAAAACATCAGAATGAGTATTTGGTTTAGAGTTTGGCAACATATGCCCATATGCTGGCTGCCATGAACAAAGGTTGGCTATAAAGAGGTCATCAGTATATGAAACAGCCCCCTGCTGTCCATTCCTTATTCCATAGAAAAGCCTTGACTTGAGGTTAGATTTTTTTTATATTTTGTTTTGTGTTATTTTTTTCTTTAACATCCCTAAAATTTTCCTTACATGTTTTACTAGCCAGATTTTTCCTCCTCTCCTGACTACTCCCAGTCATAGCTGTCCCTCTTCTCTTATGGAGATC(SEQ ID NO:16)。
具有SEQ ID NO:17所述核苷酸序列的核酸编码由细胞特异性启动子Survivin1、第一microRNA为mir-135a-5p、第二microRNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白分别为Lacl、tetR-KRAB、第一和第二识别序列为4×UAS的元件组成的表达系统。
CGGGTGACCGCCCTCCGCGGAGTACTGTCCTCCGCGGGTGACAGCCCTCCGCGGAGTGCCGCACTCCGAGCGGAGACTCTAGACTCCCTATCAGTGATAGAGATAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCTCCCTATCAGTGATAGAGAGAATTCGACCGCCACCATGAGACATATTATCTGCCACGGAGGTGTTATTACCGAAGAAATGGCCGCCAGTCTTTTGGACCAGCTGATCGAAGAGGTACTGGCTGATAATCTTCCACCTCCTAGCCATTTTGAACCACCTACCCTTCACGAACTGTATGATTTAGACGTGACGGCCCCCGAAGATCCCAACGAGGAGGCGGTTTCGCAGATTTTTCCCGACTCTGTAATGTTGGCGGTGCAGGAAGGGATTGACTTACTCACTTTTCCGCCGGCGCCCGGTTCTCCGGAGCCGCCTCACCTTTCCCGGCAGCCCGAGCAGCCGGAGCAGAGAGCCTTGGGTCCGGTTTCTATGCCAAACCTTGTACCGGAGGTGATCGATCTTACCTGCCACGAGGCTGGCTTTCCACCCAGTGACGACGAGGATGAAGAGGGTGAGGAGTTTGTGTTAGATTATGTGGAGCACCCCGGGCACGGTTGCAGGTCTTGTCATTATCACCGGAGGAATACGGGGGACCCAGATATTATGTGTTCGCTTTGCTATATGAGGACCTGTGGCATGTTTGTCTACAGTAAGTGAAAATTATGGGCAGTGGGTGATAGAGTGGTGGGTTTGGTGTGGTAATTTTTTTTTTAATTTTTACAGTTTTGTGGTTTAAAGAATTTTGTATTGTGATTTTTTTAAAAGGTCCTGTGTCTGAACCTGAGCCTGAGCCCGAGCCAGAACCGGAGCCTGCAAGACCTACCCGCCGTCCTAAAATGGCGCCTGCTATCCTGAGGCGCCCGACATCACCTGTGTCTAGAGAATGCAATAGTAGTACGGATAGCTGTGACTCCGGTCCTTCTAACACACCTCCTGAGATACACCCGGTGGTCCCGCTGTGCCCCATTAAACCAGTTGCCGTGAGAGTTGGTGGGCGTCGCCAGGCTGTGGAATGTATCGAGGACTTGCTTAACGAGCCTGGGCAACCTTTGGACTTGAGCTGTAAACGCCCCAGGCCACTCGAGGGTACCGGGTCCGGAGCTACAAATTTTTCCCTCCTCAAACAGGCTGGAGATGTCGAAGAAAATCCTGGGCCTACCGGTATGTGGCTGCAGAGCCTCCTTCTACTGGGCACCGTGGCTTGCAGCATCTCGGCCCCCGCCAGAAGTCCCAGCCCCAGCACCCAGCCCTGGGAACACGTAAATGCAATCCAGGAGGCCCGCCGCCTCCTGAACTTGTCACGGGACACGGCAGCCGAGATGAATGAAACTGTGGAGGTCATCAGTGAAATGTTTGACTTGCAGGAGCCCACATGCCTGCAGACTCGGCTGGAGCTCTACAAACAGGGCCTGCGTGGCAGCCTCACCAAGCTGAAGGGCCCACTGACCATGATGGCTAGCCATTACAAGCAGCACTGCCCGCCGACCCCTGAGACATCCTGTGCCACTCAGATCATTACCTTTGAGTCTTTCAAGGAGAACCTCAAGGACTTTCTGCTGGTCATCCCATTTGACTGCTGGGAACCCGTCCAGGAAAGATCTGAGGGCCGCGGCAGCCTGCTGACCTGCGGCGACGTGGAGGAAAACCCCGGCCCCGGATCCATGAAACCAGTAACGTTATACGATGTCGCAGAGTATGCCGGTGTCTCTTATCAGACCGTTTCCCGCGTGGTGAACCAGGCCAGCCACGTTTCTGCGAAAACGCGGGAAAAAGTGGAAGCGGCGATGGCGGAGCTGAATTACATTCCCAACCGCGTGGCACAACAACTGGCGGGCAAACAGTCGTTGCTGATTGGCGTTGCCACCTCCAGTCTGGCCCTGCACGCGCCGTCGCAAATTGTCGCGGCGATTAAATCTCGCGCCGATCAACTGGGTGCCAGCGTGGTGGTGTCGATGGTAGAACGAAGCGGCGTCGAAGCCTGTAAAGCGGCGGTGCACAATCTTCTCGCGCAACGCGTCAGTGGGCTGATCATTAACTATCCGCTGGATGACCAGGATGCCATTGCTGTGGAAGCTGCCTGCACTAATGTTCCGGCGTTATTTCTTGATGTCTCTGACCAGACACCCATCAACAGTATTATTTTCTCCCATGAAGACGGTACGCGACTGGGCGTGGAGCATCTGGTCGCATTGGGTCACCAGCAAATCGCGCTGTTAGCGGGCCCATTAAGTTCTGTCTCGGCGCGTCTGCGTCTGGCTGGCTGGCATAAATATCTCACTCGCAATCAAATTCAGCCGATAGCGGAACGGGAAGGCGACTGGAGTGCCATGTCCGGTTTTCAACAAACCATGCAAATGCTGAATGAGGGCATCGTTCCCACTGCGATGCTGGTTGCCAACGATCAGATGGCGCTGGGCGCAATGCGCGCCATTACCGAGTCCGGGCTGCGCGTTGGTGCGGATATCTCGGTAGTGGGATACGACGATACCGAAGACAGCTCATGTTATATCCCGCCGTTAACCACCATCAAACAGGATTTTCGCCTGCTGGGGCAAACCAGCGTGGACCGCTTGCTGCAACTCTCTCAGGGCCAGGCGGTGAAGGGCAATCAGCTGTTGCCGGTCTCACTGGTGAAAAGAAAAACCACCCTGGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGAGCAGCCTGAGGCCTCCTAAGAAGAAGAGGAAGGTTGTTACAAACACCATTGTCACACTCCACAAACACCATTGTCACACTCCACAAACACCATTGTCACACTCCACAAACACCATTGTCACACTCCATCACATAGGAATAAAAAGCCATATCACATAGGAATAAAAAGCCATATCACATAGGAATAAAAAGCCATATCACATAGGAATAAAAAGCCATAGAAAGCGGGGTGCCGCCCCCCGCGGACTACTGTAGTCCGCGGACGACTGTCGTCCGCGGAGCACTGTGCTCCGAGCGGAGACTCTAGAAATTGTGAGCGGATAACAATTGGGTATATAATGGGGGCCGTCAGCTACTACCAGAGCCGATCGAACACTCGAGCCGAGCAGACGTGCCTACAATTGTGAGCGCTCACAATTACTAGAAGATCTTGAATTCGCCACCATGGCTAGATTAGATAAAAGTAAAGTGATTAACAGCGCATTAGAGCTGCTTAATGAGGTCGGAATCGAAGGTTTAACAACCCGTAAACTCGCCCAGAAGCTAGGTGTAGAGCAGCCTACATTGTATTGGCATGTAAAAAATAAGCGGGCTTTGCTCGACGCCTTAGCCATTGAGATGTTAGATAGGCACCATACTCACTTTTGCCCTTTAGAAGGGGAAAGCTGGCAAGATTTTTTACGTAATAACGCTAAAAGTTTTAGATGTGCTTTACTAAGTCATCGCGATGGAGCAAAAGTACATTTAGGTACACGGCCTACAGAAAAACAGTATGAAACTCTCGAAAATCAATTAGCCTTTTTATGCCAACAAGGTTTTTCACTAGAGAATGCATTATATGCACTCAGCGCTGTGGGGCATTTTACTTTAGGTTGCGTATTGGAAGATCAAGAGCATCAAGTCGCTAAAGAAGAAAGGGAAACACCTACTACTGATAGTATGCCGCCATTATTACGACAAGCTATCGAATTATTTGATCACCAAGGTGCAGAGCCAGCCTTCTTATTCGGCCTTGAATTGATCATATGCGGATTAGAAAAACAACTTAAATGTGAAAGTGGGTCGCCAAAAAAGAAGAGAAAGGTGGATGGTGGCGGAGCACTGTCTCCCCAGCATTCAGCTGTGACCCAGGGATCAATCATTAAGAACAAGGAAGGTATGGATGCTAAGTCCCTGACCGCTTGGTCAAGAACTTTGGTAACTTTTAAGGACGTGTTCGTCGATTTTACTCGGGAGGAGTGGAAACTATTGGATACCGCCCAGCAAATCGTGTATAGAAACGTTATGCTCGAGAACTATAAGAATCTTGTGTCACTGGGCTACCAACTGACAAAGCCTGACGTTATCCTGAGGCTGGAAAAGGGTGAGGAGCCTTGGTTGGTCGAGCGGGAGATCCACCAGGAAACTCACCCAGACAGTGAGACTGCTTTCGAGATCAAGTCCTCTGTGTAATACAAGGCGGCCGCAAATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTAAAGCTTCGAATTCTGATAATCAGCCATACCACATTTGTAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCAGCTCGCCTTTTGCTCACATGTTAGGTGTGGGCAGGGACGAGCTGGCGCGGCGTCGCTGGGTGCACCGCGACCACGGGCAGAGCCACGCGGCGGGAGGACTACAACTCCCGGCACACCCCGCGCCGCCCCGCCTCTACTCCCAGAAGGCCGCGGGGGGTGGACCGCCTAAGAGGGCGTGCGCTCCCGACATGCCCCGCGGCGCGCCATTAACCGCCAGATTTGAATCGCGGGACCCGTTGGCAGAGGTGGCGGCGGCGGCATGGGTGCCCCGACGTTGCCCCCTGCCTGGCTCGAGACCGGTATGGCCCCCCCGACCGATGTCAGCCTGGGGGACGAGCTCCACTTAGACGGCGAGGACGTGGCGATGGCGCATGCCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGGACGGGGATTCCCCGGGTCCGGGATTTACCCCCCACGACTCCGCCCCCTACGGCGCTCTGGATATGGCCGACTTCGAGTTTGAGCAGATGTTTACCGATGCCCTTGGAATTGACGAGTACGGTGGGACGCGTATGAAGCTACTGTCTTCTATCGAACAAGCATGCGATATTTGCCGACTTAAAAAGCTCAAGTGCTCCAAAGAAAAACCGAAGTGCGCCAAGTGTCTGAAGAACAACTGGGAGTGTCGCTACTCTCCCAAAACCAAAAGGTCTCCGCTGACTAGGGCACATCTGACAGAAGTGGAATCAAGGCTAGAAAGACTGGAACAGCTATTTCTACTGATTTTTCCTCGAGAAGACCTTGACATGATTTTGAAAATGGATTCTTTACAGGATATAAAAGCATTGTTAACAGGATTATTTGTACAAGATAATGTGAATAAAGATGCCGTCACAGATAGATTGGCTTCAGTGGAGACTGATATGCCTCTAACATTGAGACAGCATAGAATAAGTGCGACATCATCATCGGAAGAGAGTAGTAACAAAGGTCAAAGACAGTTGACTGTATAAAAGCTTCACTCCTCAGGTGCAGGCTGCCTATCAGAAGGTGGTGGCTGGTGTGGCCAATGCCCTGGCTCACAAATACCACTGAGATCTTTTTCCCTCTGCCAAAAATTATGGGGACATCATGAAGCCCCTTGAGCATCTGACTTCTGGCTAATAAAGGAAATTTATTTTCATTGCAATAGTGTGTTGGAATTTTTTGTGTCTCTCACTCGGAAGGACATATGGGAGGGCAAATCATTTAAAACATCAGAATGAGTATTTGGTTTAGAGTTTGGCAACATATGCCCATATGCTGGCTGCCATGAACAAAGGTTGGCTATAAAGAGGTCATCAGTATATGAAACAGCCCCCTGCTGTCCATTCCTTATTCCATAGAAAAGCCTTGACTTGAGGTTAGATTTTTTTTATATTTTGTTTTGTGTTATTTTTTTCTTTAACATCCCTAAAATTTTCCTTACATGTTTTACTAGCCAGATTTTTCCTCCTCTCCTGACTACTCCCAGTCATAGCTGTCCCTCTTCTCTTATGGAGATC(SEQ ID NO:17)。
具有SEQ ID NO:18所述核苷酸序列的核酸编码由细胞特异性启动子Survivin1、第一microRNA为mir-143-3p、第二microRNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白分别为TALE蛋白、第一和第二识别序列为4×UAS的元件组成的表达系统。
CGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTTCTGTCCTCCGAGCGGAGACTCTAGATACCCCTCTCCGCTTCTGAATTCTAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCCTCGAGTACCCCTCTCCGCTTCTTACCCCTCTCCGCTTCTTACCCCTCTCCGCTTCTGAATTCGACCGCCACCATGAGACATATTATCTGCCACGGAGGTGTTATTACCGAAGAAATGGCCGCCAGTCTTTTGGACCAGCTGATCGAAGAGGTACTGGCTGATAATCTTCCACCTCCTAGCCATTTTGAACCACCTACCCTTCACGAACTGTATGATTTAGACGTGACGGCCCCCGAAGATCCCAACGAGGAGGCGGTTTCGCAGATTTTTCCCGACTCTGTAATGTTGGCGGTGCAGGAAGGGATTGACTTACTCACTTTTCCGCCGGCGCCCGGTTCTCCGGAGCCGCCTCACCTTTCCCGGCAGCCCGAGCAGCCGGAGCAGAGAGCCTTGGGTCCGGTTTCTATGCCAAACCTTGTACCGGAGGTGATCGATCTTACCTGCCACGAGGCTGGCTTTCCACCCAGTGACGACGAGGATGAAGAGGGTGAGGAGTTTGTGTTAGATTATGTGGAGCACCCCGGGCACGGTTGCAGGTCTTGTCATTATCACCGGAGGAATACGGGGGACCCAGATATTATGTGTTCGCTTTGCTATATGAGGACCTGTGGCATGTTTGTCTACAGTAAGTGAAAATTATGGGCAGTGGGTGATAGAGTGGTGGGTTTGGTGTGGTAATTTTTTTTTTAATTTTTACAGTTTTGTGGTTTAAAGAATTTTGTATTGTGATTTTTTTAAAAGGTCCTGTGTCTGAACCTGAGCCTGAGCCCGAGCCAGAACCGGAGCCTGCAAGACCTACCCGCCGTCCTAAAATGGCGCCTGCTATCCTGAGGCGCCCGACATCACCTGTGTCTAGAGAATGCAATAGTAGTACGGATAGCTGTGACTCCGGTCCTTCTAACACACCTCCTGAGATACACCCGGTGGTCCCGCTGTGCCCCATTAAACCAGTTGCCGTGAGAGTTGGTGGGCGTCGCCAGGCTGTGGAATGTATCGAGGACTTGCTTAACGAGCCTGGGCAACCTTTGGACTTGAGCTGTAAACGCCCCAGGCCAGGGTCCGGAGCTACAAATTTTTCCCTCCTCAAACAGGCTGGAGATGTCGAAGAAAATCCTGGGCCTATGGAGGCTTGGGAGTGTTTGGAAGATTTTTCTGCTGTGCGTAACTTGCTGGAACAGAGCTCTAACAGTACCTCTTGGTTTTGGAGGTTTCTGTGGGGCTCATCCCAGGCAAAGTTAGTCTGCAGAATTAAGGAGGATTACAAGTGGGAATTTGAAGAGCTTTTGAAATCCTGTGGTGAGCTGTTTGATTCTTTGAATCTGGGTCACCAGGCGCTTTTCCAAGAGAAGGTCATCAAGACTTTGGATTTTTCCACACCGGGGCGCGCTGCGGCTGCTGTTGCTTTTTTGAGTTTTATAAAGGATAAATGGAGCGAAGAAACCCATCTGAGCGGGGGGTACCTGCTGGATTTTCTGGCCATGCATCTGTGGAGAGCGGTTGTGAGACACAAGAATCGCCTGCTACTGTTGTCTTCCGTCCGCCCGGCGATAATACCGACGGAGGAGCAGCAGCAGCAGCAGGAGGAAGCCAGGCGGCGGCGGCAGGAGCAGAGCCCATGGAACCCGAGAGCCGGCCTGGACCCTCGGGAATGAATGTTGTACAGGTGGCTGAACTGTATCCAGAACTGAGACGCATTTTGACAATTACAGAGGATGGGCAGGGGCTAAAGGGGGTAAAGAGGGAGCGGGGGGCTTGTGAGGCTACAGAGGAGGCTAGGAATCTAGCTTTTAGCTTAATGACCAGACACCGTCCTGAGTGTATTACTTTTCAACAGATCAAGGATAATTGCGCTAATGAGCTTGATCTGCTGGCGCAGAAGTATTCCATAGAGCAGCTGACCACTTACTGGCTGCAGCCAGGGGATGATTTTGAGGAGGCTATTAGGGTATATGCAAAGGTGGCACTTAGGCCAGATTGCAAGTACAAGATCAGCAAACTTGTAAATATCAGGAATTGTTGCTACATTTCTGGGAACGGGGCCGAGGTGGAGATAGATACGGAGGATAGGGTGGCCTTTAGATGTAGCATGATAAATATGTGGCCGGGGGTGCTTGGCATGGACGGGGTGGTTATTATGAATGTAAGGTTTACTGGCCCCAATTTTAGCGGTACGGTTTTCCTGGCCAATACCAACCTTATCCTACACGGTGTAAGCTTCTATGGGTTTAACAATACCTGTGTGGAAGCCTGGACCGATGTAAGGGTTCGGGGCTGTGCCTTTTACTGCTGCTGGAAGGGGGTGGTGTGTCGCCCCAAAAGCAGGGCTTCAATTAAGAAATGCCTCTTTGAAAGGTGTACCTTGGGTATCCTGTCTGAGGGTAACTCCAGGGTGCGCCACAATGTGGCCTCCGACTGTGGTTGCTTCATGCTAGTGAAAAGCGTGGCTGTGATTAAGCATAACATGGTATGTGGCAACTGCGAGGACAGGGCCTCTCAGATGCTGACCTGCTCGGACGGCAACTGTCACCTGCTGAAGACCATTCACGTAGCCAGCCACTCTCGCAAGGCCTGGCCAGTGTTTGAGCATAACATACTGACCCGCTGTTCCTTGCATTTGGGTAACAGGAGGGGGGTGTTCCTACCTTACCAATGCAATTTGAGTCACACTAAGATATTGCTTGAGCCCGAGAGCATGTCCAAGGTGAACCTGAACGGGGTGTTTGACATGACCATGAAGATCTGGAAGGTGCTGAGGTACGATGAGACCCGCACCAGGTGCAGACCCTGCGAGTGTGGCGGTAAACATATTAGGAACCAGCCTGTGATGCTGGATGTGACCGAGGAGCTGAGGCCCGATCACTTGGTGCTGGCCTGCACCCGCGCTGAGTTTGGCTCTAGCGATGAAGATACAGATCTCGAGGGGTCCGGAGCTACAAATTTTTCCCTCCTCAAACAGGCTGGAGATGTCGAAGAAAATCCTGGGCCTACCGGTATGTATAGGATGCAGTTGCTCTCTTGCATCGCCCTGTCTCTAGCGTTGGTGACTAATAGCGCCCCTACTTCTTCTTCCACGAAGAAAACCCAGCTGCAACTGGAACACCTTCTGCTGGATCTCCAGATGATCCTTAACGGGATTAACAACTATAAGAACCCGAAGCTCACGGCTATGCTGACGGCGAAGTTCGCTATGCCTAAAAAGGCTACTGAGCTGAAACACTTACAGTGTCTCGAAGAGGCACTCAAACCCCTCGAGGAGGTACTCAATCTGGCGCAAAGCAAAAACTTCCATCTCCGCCCACGGGACCTAATCTCCAACATCAACGTGATTGTGCTGGAGCTCAAGGGGTCTGAAACCACGTTTATGTGCGAGTACGCAGATGAAACTGCTACCATAGTGGAATTCCTCAATCGTTGGATCACTTTCTGCCAGTCAATCATTTCAACCCTGACCAGATCTGAGGGCCGCGGCAGCCTGCTGACCTGCGGCGACGTGGAGGAAAACCCCGGCCCCGGATCCTTGTCGACCATTCGTCCGCGCAGGCCAAGTCCTGCCCGCGAGCTTCTGCCCGGACCCCAACCGGATAGGGTTCAGCCGACTGCAGATCGTGGGGTGTCTGCGCCTGCTGGCAGCCCTCTGGATGGCTTGCCCGCTCGGCGGACGGTGTCCCGGACCCGGCTGCCATCTCCCCCTGCGCCCTCACCTGCGTTCTCGGCGGGCAGCTTCAGCGATCTGCTCCGTCCGTTCGATCCGTCGCTTCTTGATACATCGCTTCTTGATTCGATGCCTGCCGTCGGCACGCCGCATACAGCGGCTGCCCCAGCAGAGTGGGATGAGGCGCAATCGGCTCTGCGTGCAGCCGATGACCCGCCACCCACCGTGCGTGTCGCTGTCACTGCCGCGCGGCCGCCGCGCGCCAAGCCGGCCCCGCGACGGCGTGCTGCGCAACCCTCCGACGCTTCGCCGGCCGCGCAGGTGGATCTACGCACGCTCGGCTACAGTCAGCAGCAGCAAGAGAAGATCAAACCGAAGGTGCGTTCGACAGTGGCGCAGCACCACGAGGCACTGGTGGGCCATGGGTTTACACACGCGCACATCGTTGCGCTCAGCCAACACCCGGCAGCGTTAGGGACCGTCGCTGTCACGTATCAGCACATAATCACGGCGTTGCCAGAGGCGACACACGAAGACATCGTTGGCGTCGGCAAACAGTGGTCCGGCGCACGCGCCCTGGAGGCCTTGCTCACGGATGCGGGGGAGTTGAGAGGTCCGCCGTTACAGTTGGACACAGGCCAACTTGTGAAGATTGCAAAACGTGGCGGCGTGACCGCAATGGAGGCAGTGCATGCATCGCGCAATGCACTGACGGGTGCCCCCCTGAACCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACAATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACAATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAAGCATTGTGGCCCAGCTGAGCCGGCCTGATCCGGCGTTGGCCGCGTTGACCAACGACCACCTCGTCGCCTTGGCCTGCCTCGGCGGACGTCCTGCCATGGATGCAGTGAAAAAGGGATTGCCGCACGCGCCGGAATTGATCAGAAGAGTCAATCGCCGTATTGGCGAACGCACGTCCCATCGCGTTGCCGACTACGCGCAAGTGGTTCGCGTGCTGGAGTTTTTCCAGTGCCACTCCCACCCAGCGTACGCATTTGATGAGGCCATGACGCAGTTCGGGATGAGCAGGAACGGGTTGGTACAGCTCTTTCGCAGAGTGGGCGTCACCGAACTCGAAGCCCGCGGTGGAACGCTCCCCCCAGCCTCGCAGCGTTGGGACCGTATCCTCCAGGCATCAGGGATGAAAAGGGCCAAACCGTCCCCTACTTCAGCTCAAACACCGGATCAGGCGTCTTTGCATGCATTCGCCGATTCGCTGGAGCGTGACCTTGATGCGCCCAGCCCAATGCACGAGGGAGATCAGACGCGGGCAAGCAGCCGTAAACGGTCCCGATCGGATCGTGCTGTCACCGGCCCCTCCGCACAGCAGGCTGTCGAGGTGCGCGTTCCCGAACAGCGCGATGCGCTGCATTTGCCCCTCAGCTGGAGGGTAAAACGCCCGCGTACCAGGATCTGGGGCGGCCTCCCGGTTAAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGGAGCTACAGTGCTTCATCTCAGAGCTACAGTGCTTCATCTCAGAGCTACAGTGCTTCATCTCAGAGCTACAGTGCTTCATCTCAGAAAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTTCTGTCCTCCGAGCGGAGACTCTAGATATAGAACCGATCCTCCCATTGAATTCTAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCCTCGAGTATAGAACCGATCCTCCCATTTATAGAACCGATCCTCCCATTTATAGAACCGATCCTCCCATTACTAGAAGATCTTGAATTCGCCACCTTGTCGACCATTCGTCCGCGCAGGCCAAGTCCTGCCCGCGAGCTTCTGCCCGGACCCCAACCGGATAGGGTTCAGCCGACTGCAGATCGTGGGGTGTCTGCGCCTGCTGGCAGCCCTCTGGATGGCTTGCCCGCTCGGCGGACGGTGTCCCGGACCCGGCTGCCATCTCCCCCTGCGCCCTCACCTGCGTTCTCGGCGGGCAGCTTCAGCGATCTGCTCCGTCCGTTCGATCCGTCGCTTCTTGATACATCGCTTCTTGATTCGATGCCTGCCGTCGGCACGCCGCATACAGCGGCTGCCCCAGCAGAGTGGGATGAGGCGCAATCGGCTCTGCGTGCAGCCGATGACCCGCCACCCACCGTGCGTGTCGCTGTCACTGCCGCGCGGCCGCCGCGCGCCAAGCCGGCCCCGCGACGGCGTGCTGCGCAACCCTCCGACGCTTCGCCGGCCGCGCAGGTGGATCTACGCACGCTCGGCTACAGTCAGCAGCAGCAAGAGAAGATCAAACCGAAGGTGCGTTCGACAGTGGCGCAGCACCACGAGGCACTGGTGGGCCATGGGTTTACACACGCGCACATCGTTGCGCTCAGCCAACACCCGGCAGCGTTAGGGACCGTCGCTGTCACGTATCAGCACATAATCACGGCGTTGCCAGAGGCGACACACGAAGACATCGTTGGCGTCGGCAAACAGTGGTCCGGCGCACGCGCCCTGGAGGCCTTGCTCACGGATGCGGGGGAGTTGAGAGGTCCGCCGTTACAGTTGGACACAGGCCAACTTGTGAAGATTGCAAAACGTGGCGGCGTGACCGCAATGGAGGCAGTGCATGCATCGCGCAATGCACTGACGGGTGCCCCCCTGAACCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACAATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAAGCATTGTGGCCCAGCTGAGCCGGCCTGATCCGGCGTTGGCCGCGTTGACCAACGACCACCTCGTCGCCTTGGCCTGCCTCGGCGGACGTCCTGCCATGGATGCAGTGAAAAAGGGATTGCCGCACGCGCCGGAATTGATCAGAAGAGTCAATCGCCGTATTGGCGAACGCACGTCCCATCGCGTTGCCGACTACGCGCAAGTGGTTCGCGTGCTGGAGTTTTTCCAGTGCCACTCCCACCCAGCGTACGCATTTGATGAGGCCATGACGCAGTTCGGGATGAGCAGGAACGGGTTGGTACAGCTCTTTCGCAGAGTGGGCGTCACCGAACTCGAAGCCCGCGGTGGAACGCTCCCCCCAGCCTCGCAGCGTTGGGACCGTATCCTCCAGGCATCAGGGATGAAAAGGGCCAAACCGTCCCCTACTTCAGCTCAAACACCGGATCAGGCGTCTTTGCATGCATTCGCCGATTCGCTGGAGCGTGACCTTGATGCGCCCAGCCCAATGCACGAGGGAGATCAGACGCGGGCAAGCAGCCGTAAACGGTCCCGATCGGATCGTGCTGTCACCGGCCCCTCCGCACAGCAGGCTGTCGAGGTGCGCGTTCCCGAACAGCGCGATGCGCTGCATTTGCCCCTCAGCTGGAGGGTAAAACGCCCGCGTACCAGGATCTGGGGCGGCCTCCCGTAATACAAGGCGGCCGCAAATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTAAAGCTTCGAATTCTGATAATCAGCCATACCACATTTGTAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCCATGTTAGGTGTGGGCAGGGACGAGCTGGCGCGGCGTCGCTGGGTGCACCGCGACCACGGGCAGAGCCACGCGGCGGGAGGACTACAACTCCCGGCACACCCCGCGCCGCCCCGCCTCTACTCCCAGAAGGCCGCGGGGGGTGGACCGCCTAAGAGGGCGTGCGCTCCCGACATGCCCCGCGGCGCGCCATTAACCGCCAGATTTGAATCGCGGGACCCGTTGGCAGAGGTGGCGGCGGCGGCATGGGTGCCCCGACGTTGCCCCCTGCCTGGCTCGAGACCGATGGCCCCCCCGACCGATGTCAGCCTGGGGGACGAGCTCCACTTAGACGGCGAGGACGTGGCGATGGCGCATGCCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGAGCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGAGCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGAGCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGGACGGGGATTCCCCGGGTCCGGGATTTACCCCCCACGACTCCGCCCCCTACGGCGCTCTGGATATGGCCGACTTCGAGTTTGAGCAGATGTTTACCGATGCCCTTGGAATTGACGAGTACGGTGGGACGCGTATGAAGCTACTGTCTTCTATCGAACAAGCATGCGATATTTGCCGACTTAAAAAGCTCAAGTGCTCCAAAGAAAAACCGAAGTGCGCCAAGTGTCTGAAGAACAACTGGGAGTGTCGCTACTCTCCCAAAACCAAAAGGTCTCCGCTGACTAGGGCACATCTGACAGAAGTGGAATCAAGGCTAGAAAGACTGGAACAGCTATTTCTACTGATTTTTCCTCGAGAAGACCTTGACATGATTTTGAAAATGGATTCTTTACAGGATATAAAAGCATTGTTAACAGGATTATTTGTACAAGATAATGTGAATAAAGATGCCGTCACAGATAGATTGGCTTCAGTGGAGACTGATATGCCTCTAACATTGAGACAGCATAGAATAAGTGCGACATCATCATCGGAAGAGAGTAGTAACAAAGGTCAAAGACAGTTGACTGTATAAAAGCTTCACTCCTCAGGTGCAGGCTGCCTATCAGAAGGTGGTGGCTGGTGTGGCCAATGCCCTGGCTCACAAATACCACTGAGATCTTTTTCCCTCTGCCAAAAATTATGGGGACATCATGAAGCCCCTTGAGCATCTGACTTCTGGCTAATAAAGGAAATTTATTTTCATTGCAATAGTGTGTTGGAATTTTTTGTGTCTCTCACTCGGAAGGACATATGGGAGGGCAAATCATTTAAAACATCAGAATGAGTATTTGGTTTAGAGTTTGGCAACATATGCCCATATGCTGGCTGCCATGAACAAAGGTTGGCTATAAAGAGGTCATCAGTATATGAAACAGCCCCCTGCTGTCCATTCCTTATTCCATAGAAAAGCCTTGACTTGAGGTTAGATTTTTTTTATATTTTGTTTTGTGTTATTTTTTTCTTTAACATCCCTAAAATTTTCCTTACATGTTTTACTAGCCAGATTTTTCCTCCTCTCCTGACTACTCCCAGTCATAGCTGTCCCTCTTCTCTTATGGAGATC(SEQ IDNO:18)。
具有SEQ ID NO:19所述核苷酸序列的核酸编码由细胞特异性启动子CEA368、第一microRNA为mir-143-3p、第二microRNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白分别为Lacl、tetR-KRAB、第一和第二识别序列为4×UAS的元件组成的表达系统。
CGGGTGACCGCCCTCCGCGGAGTACTGTCCTCCGCGGGTGACAGCCCTCCGCGGAGTGCCGCACTCCGAGCGGAGACTCTAGACTCCCTATCAGTGATAGAGATAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCTCCCTATCAGTGATAGAGAGAATTCGACCGCCACCATGAGACATATTATCTGCCACGGAGGTGTTATTACCGAAGAAATGGCCGCCAGTCTTTTGGACCAGCTGATCGAAGAGGTACTGGCTGATAATCTTCCACCTCCTAGCCATTTTGAACCACCTACCCTTCACGAACTGTATGATTTAGACGTGACGGCCCCCGAAGATCCCAACGAGGAGGCGGTTTCGCAGATTTTTCCCGACTCTGTAATGTTGGCGGTGCAGGAAGGGATTGACTTACTCACTTTTCCGCCGGCGCCCGGTTCTCCGGAGCCGCCTCACCTTTCCCGGCAGCCCGAGCAGCCGGAGCAGAGAGCCTTGGGTCCGGTTTCTATGCCAAACCTTGTACCGGAGGTGATCGATCTTACCTGCCACGAGGCTGGCTTTCCACCCAGTGACGACGAGGATGAAGAGGGTGAGGAGTTTGTGTTAGATTATGTGGAGCACCCCGGGCACGGTTGCAGGTCTTGTCATTATCACCGGAGGAATACGGGGGACCCAGATATTATGTGTTCGCTTTGCTATATGAGGACCTGTGGCATGTTTGTCTACAGTAAGTGAAAATTATGGGCAGTGGGTGATAGAGTGGTGGGTTTGGTGTGGTAATTTTTTTTTTAATTTTTACAGTTTTGTGGTTTAAAGAATTTTGTATTGTGATTTTTTTAAAAGGTCCTGTGTCTGAACCTGAGCCTGAGCCCGAGCCAGAACCGGAGCCTGCAAGACCTACCCGCCGTCCTAAAATGGCGCCTGCTATCCTGAGGCGCCCGACATCACCTGTGTCTAGAGAATGCAATAGTAGTACGGATAGCTGTGACTCCGGTCCTTCTAACACACCTCCTGAGATACACCCGGTGGTCCCGCTGTGCCCCATTAAACCAGTTGCCGTGAGAGTTGGTGGGCGTCGCCAGGCTGTGGAATGTATCGAGGACTTGCTTAACGAGCCTGGGCAACCTTTGGACTTGAGCTGTAAACGCCCCAGGCCACTCGAGGGTACCGGGTCCGGAGCTACAAATTTTTCCCTCCTCAAACAGGCTGGAGATGTCGAAGAAAATCCTGGGCCTACCGGTATGTGGCTGCAGAGCCTCCTTCTACTGGGCACCGTGGCTTGCAGCATCTCGGCCCCCGCCAGAAGTCCCAGCCCCAGCACCCAGCCCTGGGAACACGTAAATGCAATCCAGGAGGCCCGCCGCCTCCTGAACTTGTCACGGGACACGGCAGCCGAGATGAATGAAACTGTGGAGGTCATCAGTGAAATGTTTGACTTGCAGGAGCCCACATGCCTGCAGACTCGGCTGGAGCTCTACAAACAGGGCCTGCGTGGCAGCCTCACCAAGCTGAAGGGCCCACTGACCATGATGGCTAGCCATTACAAGCAGCACTGCCCGCCGACCCCTGAGACATCCTGTGCCACTCAGATCATTACCTTTGAGTCTTTCAAGGAGAACCTCAAGGACTTTCTGCTGGTCATCCCATTTGACTGCTGGGAACCCGTCCAGGAAAGATCTGAGGGCCGCGGCAGCCTGCTGACCTGCGGCGACGTGGAGGAAAACCCCGGCCCCGGATCCATGAAACCAGTAACGTTATACGATGTCGCAGAGTATGCCGGTGTCTCTTATCAGACCGTTTCCCGCGTGGTGAACCAGGCCAGCCACGTTTCTGCGAAAACGCGGGAAAAAGTGGAAGCGGCGATGGCGGAGCTGAATTACATTCCCAACCGCGTGGCACAACAACTGGCGGGCAAACAGTCGTTGCTGATTGGCGTTGCCACCTCCAGTCTGGCCCTGCACGCGCCGTCGCAAATTGTCGCGGCGATTAAATCTCGCGCCGATCAACTGGGTGCCAGCGTGGTGGTGTCGATGGTAGAACGAAGCGGCGTCGAAGCCTGTAAAGCGGCGGTGCACAATCTTCTCGCGCAACGCGTCAGTGGGCTGATCATTAACTATCCGCTGGATGACCAGGATGCCATTGCTGTGGAAGCTGCCTGCACTAATGTTCCGGCGTTATTTCTTGATGTCTCTGACCAGACACCCATCAACAGTATTATTTTCTCCCATGAAGACGGTACGCGACTGGGCGTGGAGCATCTGGTCGCATTGGGTCACCAGCAAATCGCGCTGTTAGCGGGCCCATTAAGTTCTGTCTCGGCGCGTCTGCGTCTGGCTGGCTGGCATAAATATCTCACTCGCAATCAAATTCAGCCGATAGCGGAACGGGAAGGCGACTGGAGTGCCATGTCCGGTTTTCAACAAACCATGCAAATGCTGAATGAGGGCATCGTTCCCACTGCGATGCTGGTTGCCAACGATCAGATGGCGCTGGGCGCAATGCGCGCCATTACCGAGTCCGGGCTGCGCGTTGGTGCGGATATCTCGGTAGTGGGATACGACGATACCGAAGACAGCTCATGTTATATCCCGCCGTTAACCACCATCAAACAGGATTTTCGCCTGCTGGGGCAAACCAGCGTGGACCGCTTGCTGCAACTCTCTCAGGGCCAGGCGGTGAAGGGCAATCAGCTGTTGCCGGTCTCACTGGTGAAAAGAAAAACCACCCTGGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGAGCAGCCTGAGGCCTCCTAAGAAGAAGAGGAAGGTTGTTACAAACACCATTGTCACACTCCACAAACACCATTGTCACACTCCACAAACACCATTGTCACACTCCACAAACACCATTGTCACACTCCAGAGCTACAGTGCTTCATCTCAGAGCTACAGTGCTTCATCTCAGAGCTACAGTGCTTCATCTCAGAGCTACAGTGCTTCATCTCAGAAAGCGGGGTGCCGCCCCCCGCGGACTACTGTAGTCCGCGGACGACTGTCGTCCGCGGAGCACTGTGCTCCGAGCGGAGACTCTAGAAATTGTGAGCGGATAACAATTGGGTATATAATGGGGGCCGTCAGCTACTACCAGAGCCGATCGAACACTCGAGCCGAGCAGACGTGCCTACAATTGTGAGCGCTCACAATTACTAGAAGATCTTGAATTCGCCACCATGGCTAGATTAGATAAAAGTAAAGTGATTAACAGCGCATTAGAGCTGCTTAATGAGGTCGGAATCGAAGGTTTAACAACCCGTAAACTCGCCCAGAAGCTAGGTGTAGAGCAGCCTACATTGTATTGGCATGTAAAAAATAAGCGGGCTTTGCTCGACGCCTTAGCCATTGAGATGTTAGATAGGCACCATACTCACTTTTGCCCTTTAGAAGGGGAAAGCTGGCAAGATTTTTTACGTAATAACGCTAAAAGTTTTAGATGTGCTTTACTAAGTCATCGCGATGGAGCAAAAGTACATTTAGGTACACGGCCTACAGAAAAACAGTATGAAACTCTCGAAAATCAATTAGCCTTTTTATGCCAACAAGGTTTTTCACTAGAGAATGCATTATATGCACTCAGCGCTGTGGGGCATTTTACTTTAGGTTGCGTATTGGAAGATCAAGAGCATCAAGTCGCTAAAGAAGAAAGGGAAACACCTACTACTGATAGTATGCCGCCATTATTACGACAAGCTATCGAATTATTTGATCACCAAGGTGCAGAGCCAGCCTTCTTATTCGGCCTTGAATTGATCATATGCGGATTAGAAAAACAACTTAAATGTGAAAGTGGGTCGCCAAAAAAGAAGAGAAAGGTGGATGGTGGCGGAGCACTGTCTCCCCAGCATTCAGCTGTGACCCAGGGATCAATCATTAAGAACAAGGAAGGTATGGATGCTAAGTCCCTGACCGCTTGGTCAAGAACTTTGGTAACTTTTAAGGACGTGTTCGTCGATTTTACTCGGGAGGAGTGGAAACTATTGGATACCGCCCAGCAAATCGTGTATAGAAACGTTATGCTCGAGAACTATAAGAATCTTGTGTCACTGGGCTACCAACTGACAAAGCCTGACGTTATCCTGAGGCTGGAAAAGGGTGAGGAGCCTTGGTTGGTCGAGCGGGAGATCCACCAGGAAACTCACCCAGACAGTGAGACTGCTTTCGAGATCAAGTCCTCTGTGTAATACAAGGCGGCCGCAAATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTAAAGCTTCGAATTCTGATAATCAGCCATACCACATTTGTAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCAGCTCGCCTTTTGCTCACATGTCTCCCAGGGGATGGGGGTCCATCCACCTTGCCGAAAAGATTTGTCTGAGGAACTGAAAATAGAAGGGAAAAAAGAGGAGGGACAAAAGAGGCAGAAATGAGAGGGGAGGGGACAGAGGACACCTGAATAAAGACCACACCCATGACCCACGTGATGCTGAGAAGTACTCCTGCCCTAGGAAGAGACTCAGGGCAGAGGGAGGAAGGACAGCAGACCAGACAGTCACAGCAGCCTTGACAAAACGTTCCTGGAACTCAAGCTCTTCTCCACAGAGGAGGACAGAGCAGACAGCAGAGACCATGGAGTCTCCCTCGGCCCCTCCCCACAGATGGTGCATCCCCTGGCAGAGGCTCCTGCTCACAGGTGAACTCGAGACCGGTATGGCCCCCCCGACCGATGTCAGCCTGGGGGACGAGCTCCACTTAGACGGCGAGGACGTGGCGATGGCGCATGCCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGGACGGGGATTCCCCGGGTCCGGGATTTACCCCCCACGACTCCGCCCCCTACGGCGCTCTGGATATGGCCGACTTCGAGTTTGAGCAGATGTTTACCGATGCCCTTGGAATTGACGAGTACGGTGGGACGCGTATGAAGCTACTGTCTTCTATCGAACAAGCATGCGATATTTGCCGACTTAAAAAGCTCAAGTGCTCCAAAGAAAAACCGAAGTGCGCCAAGTGTCTGAAGAACAACTGGGAGTGTCGCTACTCTCCCAAAACCAAAAGGTCTCCGCTGACTAGGGCACATCTGACAGAAGTGGAATCAAGGCTAGAAAGACTGGAACAGCTATTTCTACTGATTTTTCCTCGAGAAGACCTTGACATGATTTTGAAAATGGATTCTTTACAGGATATAAAAGCATTGTTAACAGGATTATTTGTACAAGATAATGTGAATAAAGATGCCGTCACAGATAGATTGGCTTCAGTGGAGACTGATATGCCTCTAACATTGAGACAGCATAGAATAAGTGCGACATCATCATCGGAAGAGAGTAGTAACAAAGGTCAAAGACAGTTGACTGTATAAAAGCTTCACTCCTCAGGTGCAGGCTGCCTATCAGAAGGTGGTGGCTGGTGTGGCCAATGCCCTGGCTCACAAATACCACTGAGATCTTTTTCCCTCTGCCAAAAATTATGGGGACATCATGAAGCCCCTTGAGCATCTGACTTCTGGCTAATAAAGGAAATTTATTTTCATTGCAATAGTGTGTTGGAATTTTTTGTGTCTCTCACTCGGAAGGACATATGGGAGGGCAAATCATTTAAAACATCAGAATGAGTATTTGGTTTAGAGTTTGGCAACATATGCCCATATGCTGGCTGCCATGAACAAAGGTTGGCTATAAAGAGGTCATCAGTATATGAAACAGCCCCCTGCTGTCCATTCCTTATTCCATAGAAAAGCCTTGACTTGAGGTTAGATTTTTTTTATATTTTGTTTTGTGTTATTTTTTTCTTTAACATCCCTAAAATTTTCCTTACATGTTTTACTAGCCAGATTTTTCCTCCTCTCCTGACTACTCCCAGTCATAGCTGTCCCTCTTCTCTTATGGAGATC(SEQ ID NO:19)。
具有SEQ ID NO:20所述核苷酸序列的核酸编码由细胞特异性启动子CEA368、第一microRNA为mir-135a-5p、第二microRNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白分别为TALE蛋白、第一和第二识别序列为4×UAS的元件组成的表达系统。
CGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTTCTGTCCTCCGAGCGGAGACTCTAGATACCCCTCTCCGCTTCTGAATTCTAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCCTCGAGTACCCCTCTCCGCTTCTTACCCCTCTCCGCTTCTTACCCCTCTCCGCTTCTGAATTCGACCGCCACCATGAGACATATTATCTGCCACGGAGGTGTTATTACCGAAGAAATGGCCGCCAGTCTTTTGGACCAGCTGATCGAAGAGGTACTGGCTGATAATCTTCCACCTCCTAGCCATTTTGAACCACCTACCCTTCACGAACTGTATGATTTAGACGTGACGGCCCCCGAAGATCCCAACGAGGAGGCGGTTTCGCAGATTTTTCCCGACTCTGTAATGTTGGCGGTGCAGGAAGGGATTGACTTACTCACTTTTCCGCCGGCGCCCGGTTCTCCGGAGCCGCCTCACCTTTCCCGGCAGCCCGAGCAGCCGGAGCAGAGAGCCTTGGGTCCGGTTTCTATGCCAAACCTTGTACCGGAGGTGATCGATCTTACCTGCCACGAGGCTGGCTTTCCACCCAGTGACGACGAGGATGAAGAGGGTGAGGAGTTTGTGTTAGATTATGTGGAGCACCCCGGGCACGGTTGCAGGTCTTGTCATTATCACCGGAGGAATACGGGGGACCCAGATATTATGTGTTCGCTTTGCTATATGAGGACCTGTGGCATGTTTGTCTACAGTAAGTGAAAATTATGGGCAGTGGGTGATAGAGTGGTGGGTTTGGTGTGGTAATTTTTTTTTTAATTTTTACAGTTTTGTGGTTTAAAGAATTTTGTATTGTGATTTTTTTAAAAGGTCCTGTGTCTGAACCTGAGCCTGAGCCCGAGCCAGAACCGGAGCCTGCAAGACCTACCCGCCGTCCTAAAATGGCGCCTGCTATCCTGAGGCGCCCGACATCACCTGTGTCTAGAGAATGCAATAGTAGTACGGATAGCTGTGACTCCGGTCCTTCTAACACACCTCCTGAGATACACCCGGTGGTCCCGCTGTGCCCCATTAAACCAGTTGCCGTGAGAGTTGGTGGGCGTCGCCAGGCTGTGGAATGTATCGAGGACTTGCTTAACGAGCCTGGGCAACCTTTGGACTTGAGCTGTAAACGCCCCAGGCCAGGGTCCGGAGCTACAAATTTTTCCCTCCTCAAACAGGCTGGAGATGTCGAAGAAAATCCTGGGCCTATGGAGGCTTGGGAGTGTTTGGAAGATTTTTCTGCTGTGCGTAACTTGCTGGAACAGAGCTCTAACAGTACCTCTTGGTTTTGGAGGTTTCTGTGGGGCTCATCCCAGGCAAAGTTAGTCTGCAGAATTAAGGAGGATTACAAGTGGGAATTTGAAGAGCTTTTGAAATCCTGTGGTGAGCTGTTTGATTCTTTGAATCTGGGTCACCAGGCGCTTTTCCAAGAGAAGGTCATCAAGACTTTGGATTTTTCCACACCGGGGCGCGCTGCGGCTGCTGTTGCTTTTTTGAGTTTTATAAAGGATAAATGGAGCGAAGAAACCCATCTGAGCGGGGGGTACCTGCTGGATTTTCTGGCCATGCATCTGTGGAGAGCGGTTGTGAGACACAAGAATCGCCTGCTACTGTTGTCTTCCGTCCGCCCGGCGATAATACCGACGGAGGAGCAGCAGCAGCAGCAGGAGGAAGCCAGGCGGCGGCGGCAGGAGCAGAGCCCATGGAACCCGAGAGCCGGCCTGGACCCTCGGGAATGAATGTTGTACAGGTGGCTGAACTGTATCCAGAACTGAGACGCATTTTGACAATTACAGAGGATGGGCAGGGGCTAAAGGGGGTAAAGAGGGAGCGGGGGGCTTGTGAGGCTACAGAGGAGGCTAGGAATCTAGCTTTTAGCTTAATGACCAGACACCGTCCTGAGTGTATTACTTTTCAACAGATCAAGGATAATTGCGCTAATGAGCTTGATCTGCTGGCGCAGAAGTATTCCATAGAGCAGCTGACCACTTACTGGCTGCAGCCAGGGGATGATTTTGAGGAGGCTATTAGGGTATATGCAAAGGTGGCACTTAGGCCAGATTGCAAGTACAAGATCAGCAAACTTGTAAATATCAGGAATTGTTGCTACATTTCTGGGAACGGGGCCGAGGTGGAGATAGATACGGAGGATAGGGTGGCCTTTAGATGTAGCATGATAAATATGTGGCCGGGGGTGCTTGGCATGGACGGGGTGGTTATTATGAATGTAAGGTTTACTGGCCCCAATTTTAGCGGTACGGTTTTCCTGGCCAATACCAACCTTATCCTACACGGTGTAAGCTTCTATGGGTTTAACAATACCTGTGTGGAAGCCTGGACCGATGTAAGGGTTCGGGGCTGTGCCTTTTACTGCTGCTGGAAGGGGGTGGTGTGTCGCCCCAAAAGCAGGGCTTCAATTAAGAAATGCCTCTTTGAAAGGTGTACCTTGGGTATCCTGTCTGAGGGTAACTCCAGGGTGCGCCACAATGTGGCCTCCGACTGTGGTTGCTTCATGCTAGTGAAAAGCGTGGCTGTGATTAAGCATAACATGGTATGTGGCAACTGCGAGGACAGGGCCTCTCAGATGCTGACCTGCTCGGACGGCAACTGTCACCTGCTGAAGACCATTCACGTAGCCAGCCACTCTCGCAAGGCCTGGCCAGTGTTTGAGCATAACATACTGACCCGCTGTTCCTTGCATTTGGGTAACAGGAGGGGGGTGTTCCTACCTTACCAATGCAATTTGAGTCACACTAAGATATTGCTTGAGCCCGAGAGCATGTCCAAGGTGAACCTGAACGGGGTGTTTGACATGACCATGAAGATCTGGAAGGTGCTGAGGTACGATGAGACCCGCACCAGGTGCAGACCCTGCGAGTGTGGCGGTAAACATATTAGGAACCAGCCTGTGATGCTGGATGTGACCGAGGAGCTGAGGCCCGATCACTTGGTGCTGGCCTGCACCCGCGCTGAGTTTGGCTCTAGCGATGAAGATACAGATCTCGAGGGGTCCGGAGCTACAAATTTTTCCCTCCTCAAACAGGCTGGAGATGTCGAAGAAAATCCTGGGCCTACCGGTATGTATAGGATGCAGTTGCTCTCTTGCATCGCCCTGTCTCTAGCGTTGGTGACTAATAGCGCCCCTACTTCTTCTTCCACGAAGAAAACCCAGCTGCAACTGGAACACCTTCTGCTGGATCTCCAGATGATCCTTAACGGGATTAACAACTATAAGAACCCGAAGCTCACGGCTATGCTGACGGCGAAGTTCGCTATGCCTAAAAAGGCTACTGAGCTGAAACACTTACAGTGTCTCGAAGAGGCACTCAAACCCCTCGAGGAGGTACTCAATCTGGCGCAAAGCAAAAACTTCCATCTCCGCCCACGGGACCTAATCTCCAACATCAACGTGATTGTGCTGGAGCTCAAGGGGTCTGAAACCACGTTTATGTGCGAGTACGCAGATGAAACTGCTACCATAGTGGAATTCCTCAATCGTTGGATCACTTTCTGCCAGTCAATCATTTCAACCCTGACCAGATCTGAGGGCCGCGGCAGCCTGCTGACCTGCGGCGACGTGGAGGAAAACCCCGGCCCCGGATCCTTGTCGACCATTCGTCCGCGCAGGCCAAGTCCTGCCCGCGAGCTTCTGCCCGGACCCCAACCGGATAGGGTTCAGCCGACTGCAGATCGTGGGGTGTCTGCGCCTGCTGGCAGCCCTCTGGATGGCTTGCCCGCTCGGCGGACGGTGTCCCGGACCCGGCTGCCATCTCCCCCTGCGCCCTCACCTGCGTTCTCGGCGGGCAGCTTCAGCGATCTGCTCCGTCCGTTCGATCCGTCGCTTCTTGATACATCGCTTCTTGATTCGATGCCTGCCGTCGGCACGCCGCATACAGCGGCTGCCCCAGCAGAGTGGGATGAGGCGCAATCGGCTCTGCGTGCAGCCGATGACCCGCCACCCACCGTGCGTGTCGCTGTCACTGCCGCGCGGCCGCCGCGCGCCAAGCCGGCCCCGCGACGGCGTGCTGCGCAACCCTCCGACGCTTCGCCGGCCGCGCAGGTGGATCTACGCACGCTCGGCTACAGTCAGCAGCAGCAAGAGAAGATCAAACCGAAGGTGCGTTCGACAGTGGCGCAGCACCACGAGGCACTGGTGGGCCATGGGTTTACACACGCGCACATCGTTGCGCTCAGCCAACACCCGGCAGCGTTAGGGACCGTCGCTGTCACGTATCAGCACATAATCACGGCGTTGCCAGAGGCGACACACGAAGACATCGTTGGCGTCGGCAAACAGTGGTCCGGCGCACGCGCCCTGGAGGCCTTGCTCACGGATGCGGGGGAGTTGAGAGGTCCGCCGTTACAGTTGGACACAGGCCAACTTGTGAAGATTGCAAAACGTGGCGGCGTGACCGCAATGGAGGCAGTGCATGCATCGCGCAATGCACTGACGGGTGCCCCCCTGAACCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACAATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACAATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAAGCATTGTGGCCCAGCTGAGCCGGCCTGATCCGGCGTTGGCCGCGTTGACCAACGACCACCTCGTCGCCTTGGCCTGCCTCGGCGGACGTCCTGCCATGGATGCAGTGAAAAAGGGATTGCCGCACGCGCCGGAATTGATCAGAAGAGTCAATCGCCGTATTGGCGAACGCACGTCCCATCGCGTTGCCGACTACGCGCAAGTGGTTCGCGTGCTGGAGTTTTTCCAGTGCCACTCCCACCCAGCGTACGCATTTGATGAGGCCATGACGCAGTTCGGGATGAGCAGGAACGGGTTGGTACAGCTCTTTCGCAGAGTGGGCGTCACCGAACTCGAAGCCCGCGGTGGAACGCTCCCCCCAGCCTCGCAGCGTTGGGACCGTATCCTCCAGGCATCAGGGATGAAAAGGGCCAAACCGTCCCCTACTTCAGCTCAAACACCGGATCAGGCGTCTTTGCATGCATTCGCCGATTCGCTGGAGCGTGACCTTGATGCGCCCAGCCCAATGCACGAGGGAGATCAGACGCGGGCAAGCAGCCGTAAACGGTCCCGATCGGATCGTGCTGTCACCGGCCCCTCCGCACAGCAGGCTGTCGAGGTGCGCGTTCCCGAACAGCGCGATGCGCTGCATTTGCCCCTCAGCTGGAGGGTAAAACGCCCGCGTACCAGGATCTGGGGCGGCCTCCCGGTTAAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGTCACATAGGAATAAAAAGCCATATCACATAGGAATAAAAAGCCATATCACATAGGAATAAAAAGCCATATCACATAGGAATAAAAAGCCATAGAAAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTTCTGTCCTCCGAGCGGAGACTCTAGATATAGAACCGATCCTCCCATTGAATTCTAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCCTCGAGTATAGAACCGATCCTCCCATTTATAGAACCGATCCTCCCATTTATAGAACCGATCCTCCCATTACTAGAAGATCTTGAATTCGCCACCTTGTCGACCATTCGTCCGCGCAGGCCAAGTCCTGCCCGCGAGCTTCTGCCCGGACCCCAACCGGATAGGGTTCAGCCGACTGCAGATCGTGGGGTGTCTGCGCCTGCTGGCAGCCCTCTGGATGGCTTGCCCGCTCGGCGGACGGTGTCCCGGACCCGGCTGCCATCTCCCCCTGCGCCCTCACCTGCGTTCTCGGCGGGCAGCTTCAGCGATCTGCTCCGTCCGTTCGATCCGTCGCTTCTTGATACATCGCTTCTTGATTCGATGCCTGCCGTCGGCACGCCGCATACAGCGGCTGCCCCAGCAGAGTGGGATGAGGCGCAATCGGCTCTGCGTGCAGCCGATGACCCGCCACCCACCGTGCGTGTCGCTGTCACTGCCGCGCGGCCGCCGCGCGCCAAGCCGGCCCCGCGACGGCGTGCTGCGCAACCCTCCGACGCTTCGCCGGCCGCGCAGGTGGATCTACGCACGCTCGGCTACAGTCAGCAGCAGCAAGAGAAGATCAAACCGAAGGTGCGTTCGACAGTGGCGCAGCACCACGAGGCACTGGTGGGCCATGGGTTTACACACGCGCACATCGTTGCGCTCAGCCAACACCCGGCAGCGTTAGGGACCGTCGCTGTCACGTATCAGCACATAATCACGGCGTTGCCAGAGGCGACACACGAAGACATCGTTGGCGTCGGCAAACAGTGGTCCGGCGCACGCGCCCTGGAGGCCTTGCTCACGGATGCGGGGGAGTTGAGAGGTCCGCCGTTACAGTTGGACACAGGCCAACTTGTGAAGATTGCAAAACGTGGCGGCGTGACCGCAATGGAGGCAGTGCATGCATCGCGCAATGCACTGACGGGTGCCCCCCTGAACCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACAATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAAGCATTGTGGCCCAGCTGAGCCGGCCTGATCCGGCGTTGGCCGCGTTGACCAACGACCACCTCGTCGCCTTGGCCTGCCTCGGCGGACGTCCTGCCATGGATGCAGTGAAAAAGGGATTGCCGCACGCGCCGGAATTGATCAGAAGAGTCAATCGCCGTATTGGCGAACGCACGTCCCATCGCGTTGCCGACTACGCGCAAGTGGTTCGCGTGCTGGAGTTTTTCCAGTGCCACTCCCACCCAGCGTACGCATTTGATGAGGCCATGACGCAGTTCGGGATGAGCAGGAACGGGTTGGTACAGCTCTTTCGCAGAGTGGGCGTCACCGAACTCGAAGCCCGCGGTGGAACGCTCCCCCCAGCCTCGCAGCGTTGGGACCGTATCCTCCAGGCATCAGGGATGAAAAGGGCCAAACCGTCCCCTACTTCAGCTCAAACACCGGATCAGGCGTCTTTGCATGCATTCGCCGATTCGCTGGAGCGTGACCTTGATGCGCCCAGCCCAATGCACGAGGGAGATCAGACGCGGGCAAGCAGCCGTAAACGGTCCCGATCGGATCGTGCTGTCACCGGCCCCTCCGCACAGCAGGCTGTCGAGGTGCGCGTTCCCGAACAGCGCGATGCGCTGCATTTGCCCCTCAGCTGGAGGGTAAAACGCCCGCGTACCAGGATCTGGGGCGGCCTCCCGTAATACAAGGCGGCCGCAAATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTAAAGCTTCGAATTCTGATAATCAGCCATACCACATTTGTAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCAGCTCGCCTTTTGCTCACATGTCTCCCAGGGGATGGGGGTCCATCCACCTTGCCGAAAAGATTTGTCTGAGGAACTGAAAATAGAAGGGAAAAAAGAGGAGGGACAAAAGAGGCAGAAATGAGAGGGGAGGGGACAGAGGACACCTGAATAAAGACCACACCCATGACCCACGTGATGCTGAGAAGTACTCCTGCCCTAGGAAGAGACTCAGGGCAGAGGGAGGAAGGACAGCAGACCAGACAGTCACAGCAGCCTTGACAAAACGTTCCTGGAACTCAAGCTCTTCTCCACAGAGGAGGACAGAGCAGACAGCAGAGACCATGGAGTCTCCCTCGGCCCCTCCCCACAGATGGTGCATCCCCTGGCAGAGGCTCCTGCTCACAGGTGAACTCGAGACCGGTATGGCCCCCCCGACCGATGTCAGCCTGGGGGACGAGCTCCACTTAGACGGCGAGGACGTGGCGATGGCGCATGCCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGAGCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGAGCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGAGCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGGACGGGGATTCCCCGGGTCCGGGATTTACCCCCCACGACTCCGCCCCCTACGGCGCTCTGGATATGGCCGACTTCGAGTTTGAGCAGATGTTTACCGATGCCCTTGGAATTGACGAGTACGGTGGGACGCGTATGAAGCTACTGTCTTCTATCGAACAAGCATGCGATATTTGCCGACTTAAAAAGCTCAAGTGCTCCAAAGAAAAACCGAAGTGCGCCAAGTGTCTGAAGAACAACTGGGAGTGTCGCTACTCTCCCAAAACCAAAAGGTCTCCGCTGACTAGGGCACATCTGACAGAAGTGGAATCAAGGCTAGAAAGACTGGAACAGCTATTTCTACTGATTTTTCCTCGAGAAGACCTTGACATGATTTTGAAAATGGATTCTTTACAGGATATAAAAGCATTGTTAACAGGATTATTTGTACAAGATAATGTGAATAAAGATGCCGTCACAGATAGATTGGCTTCAGTGGAGACTGATATGCCTCTAACATTGAGACAGCATAGAATAAGTGCGACATCATCATCGGAAGAGAGTAGTAACAAAGGTCAAAGACAGTTGACTGTATAAAAGCTTCACTCCTCAGGTGCAGGCTGCCTATCAGAAGGTGGTGGCTGGTGTGGCCAATGCCCTGGCTCACAAATACCACTGAGATCTTTTTCCCTCTGCCAAAAATTATGGGGACATCATGAAGCCCCTTGAGCATCTGACTTCTGGCTAATAAAGGAAATTTATTTTCATTGCAATAGTGTGTTGGAATTTTTTGTGTCTCTCACTCGGAAGGACATATGGGAGGGCAAATCATTTAAAACATCAGAATGAGTATTTGGTTTAGAGTTTGGCAACATATGCCCATATGCTGGCTGCCATGAACAAAGGTTGGCTATAAAGAGGTCATCAGTATATGAAACAGCCCCCTGCTGTCCATTCCTTATTCCATAGAAAAGCCTTGACTTGAGGTTAGATTTTTTTTATATTTTGTTTTGTGTTATTTTTTTCTTTAACATCCCTAAAATTTTCCTTACATGTTTTACTAGCCAGATTTTTCCTCCTCTCCTGACTACTCCCAGTCATAGCTGTCCCTCTTCTCTTATGGAGATC(SEQ ID NO:20)。
具有SEQ ID NO:21所述核苷酸序列的核酸编码由细胞特异性启动子hMUC1、第一microRNA为mir-199a-3p、第二microRNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白分别为Lacl、tetR-KRAB、第一和第二识别序列为4×UAS的元件组成的表达系统。
CGGCCATATGTCTTCCGCGGAGTGCTGTACTCCGCGGAGTACTGTCCTCCGCGGAGTGCCGCACTCCGAGCGGAGACTCTAGACTCCCTATCAGTGATAGAGATAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCTCCCTATCAGTGATAGAGAGAATTCGACCGCCACCATGAGACATATTATCTGCCACGGAGGTGTTATTACCGAAGAAATGGCCGCCAGTCTTTTGGACCAGCTGATCGAAGAGGTACTGGCTGATAATCTTCCACCTCCTAGCCATTTTGAACCACCTACCCTTCACGAACTGTATGATTTAGACGTGACGGCCCCCGAAGATCCCAACGAGGAGGCGGTTTCGCAGATTTTTCCCGACTCTGTAATGTTGGCGGTGCAGGAAGGGATTGACTTACTCACTTTTCCGCCGGCGCCCGGTTCTCCGGAGCCGCCTCACCTTTCCCGGCAGCCCGAGCAGCCGGAGCAGAGAGCCTTGGGTCCGGTTTCTATGCCAAACCTTGTACCGGAGGTGATCGATCTTACCTGCCACGAGGCTGGCTTTCCACCCAGTGACGACGAGGATGAAGAGGGTGAGGAGTTTGTGTTAGATTATGTGGAGCACCCCGGGCACGGTTGCAGGTCTTGTCATTATCACCGGAGGAATACGGGGGACCCAGATATTATGTGTTCGCTTTGCTATATGAGGACCTGTGGCATGTTTGTCTACAGTAAGTGAAAATTATGGGCAGTGGGTGATAGAGTGGTGGGTTTGGTGTGGTAATTTTTTTTTTAATTTTTACAGTTTTGTGGTTTAAAGAATTTTGTATTGTGATTTTTTTAAAAGGTCCTGTGTCTGAACCTGAGCCTGAGCCCGAGCCAGAACCGGAGCCTGCAAGACCTACCCGCCGTCCTAAAATGGCGCCTGCTATCCTGAGGCGCCCGACATCACCTGTGTCTAGAGAATGCAATAGTAGTACGGATAGCTGTGACTCCGGTCCTTCTAACACACCTCCTGAGATACACCCGGTGGTCCCGCTGTGCCCCATTAAACCAGTTGCCGTGAGAGTTGGTGGGCGTCGCCAGGCTGTGGAATGTATCGAGGACTTGCTTAACGAGCCTGGGCAACCTTTGGACTTGAGCTGTAAACGCCCCAGGCCACTCGAGGGTACCGGGTCCGGAGCTACAAATTTTTCCCTCCTCAAACAGGCTGGAGATGTCGAAGAAAATCCTGGGCCTACCGGTATGTGGCTGCAGAGCCTCCTTCTACTGGGCACCGTGGCTTGCAGCATCTCGGCCCCCGCCAGAAGTCCCAGCCCCAGCACCCAGCCCTGGGAACACGTAAATGCAATCCAGGAGGCCCGCCGCCTCCTGAACTTGTCACGGGACACGGCAGCCGAGATGAATGAAACTGTGGAGGTCATCAGTGAAATGTTTGACTTGCAGGAGCCCACATGCCTGCAGACTCGGCTGGAGCTCTACAAACAGGGCCTGCGTGGCAGCCTCACCAAGCTGAAGGGCCCACTGACCATGATGGCTAGCCATTACAAGCAGCACTGCCCGCCGACCCCTGAGACATCCTGTGCCACTCAGATCATTACCTTTGAGTCTTTCAAGGAGAACCTCAAGGACTTTCTGCTGGTCATCCCATTTGACTGCTGGGAACCCGTCCAGGAAAGATCTGAGGGCCGCGGCAGCCTGCTGACCTGCGGCGACGTGGAGGAAAACCCCGGCCCCGGATCCATGAAACCAGTAACGTTATACGATGTCGCAGAGTATGCCGGTGTCTCTTATCAGACCGTTTCCCGCGTGGTGAACCAGGCCAGCCACGTTTCTGCGAAAACGCGGGAAAAAGTGGAAGCGGCGATGGCGGAGCTGAATTACATTCCCAACCGCGTGGCACAACAACTGGCGGGCAAACAGTCGTTGCTGATTGGCGTTGCCACCTCCAGTCTGGCCCTGCACGCGCCGTCGCAAATTGTCGCGGCGATTAAATCTCGCGCCGATCAACTGGGTGCCAGCGTGGTGGTGTCGATGGTAGAACGAAGCGGCGTCGAAGCCTGTAAAGCGGCGGTGCACAATCTTCTCGCGCAACGCGTCAGTGGGCTGATCATTAACTATCCGCTGGATGACCAGGATGCCATTGCTGTGGAAGCTGCCTGCACTAATGTTCCGGCGTTATTTCTTGATGTCTCTGACCAGACACCCATCAACAGTATTATTTTCTCCCATGAAGACGGTACGCGACTGGGCGTGGAGCATCTGGTCGCATTGGGTCACCAGCAAATCGCGCTGTTAGCGGGCCCATTAAGTTCTGTCTCGGCGCGTCTGCGTCTGGCTGGCTGGCATAAATATCTCACTCGCAATCAAATTCAGCCGATAGCGGAACGGGAAGGCGACTGGAGTGCCATGTCCGGTTTTCAACAAACCATGCAAATGCTGAATGAGGGCATCGTTCCCACTGCGATGCTGGTTGCCAACGATCAGATGGCGCTGGGCGCAATGCGCGCCATTACCGAGTCCGGGCTGCGCGTTGGTGCGGATATCTCGGTAGTGGGATACGACGATACCGAAGACAGCTCATGTTATATCCCGCCGTTAACCACCATCAAACAGGATTTTCGCCTGCTGGGGCAAACCAGCGTGGACCGCTTGCTGCAACTCTCTCAGGGCCAGGCGGTGAAGGGCAATCAGCTGTTGCCGGTCTCACTGGTGAAAAGAAAAACCACCCTGGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGAGCAGCCTGAGGCCTCCTAAGAAGAAGAGGAAGGTTGCGGCCGCTAACCAATGTGCAGACTACTGTTAACCAATGTGCAGACTACTGTGAATTCTAACCAATGTGCAGACTACTGTTAACCAATGTGCAGACTACTGTAAGCTTCGAATTGAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGGTCGACAATACTAGTTAAGAAAGCGGTCCACTGTGTGCCGCGGGTGACAGCCCTCCGCGGCGGTCTTTCGTCCGCGGGTGACCGCCCTCCGAGCGGAGACTCTAGAAATTGTGAGCGGATAACAATTGGGTATATAATGGGGGCCGTCAGCTACTACCAGAGCCGATCGAACACTCGAGCCGAGCAGACGTGCCTACAATTGTGAGCGCTCACAATTACTAGAAGATCTTGAATTCGCCACCATGGCTAGATTAGATAAAAGTAAAGTGATTAACAGCGCATTAGAGCTGCTTAATGAGGTCGGAATCGAAGGTTTAACAACCCGTAAACTCGCCCAGAAGCTAGGTGTAGAGCAGCCTACATTGTATTGGCATGTAAAAAATAAGCGGGCTTTGCTCGACGCCTTAGCCATTGAGATGTTAGATAGGCACCATACTCACTTTTGCCCTTTAGAAGGGGAAAGCTGGCAAGATTTTTTACGTAATAACGCTAAAAGTTTTAGATGTGCTTTACTAAGTCATCGCGATGGAGCAAAAGTACATTTAGGTACACGGCCTACAGAAAAACAGTATGAAACTCTCGAAAATCAATTAGCCTTTTTATGCCAACAAGGTTTTTCACTAGAGAATGCATTATATGCACTCAGCGCTGTGGGGCATTTTACTTTAGGTTGCGTATTGGAAGATCAAGAGCATCAAGTCGCTAAAGAAGAAAGGGAAACACCTACTACTGATAGTATGCCGCCATTATTACGACAAGCTATCGAATTATTTGATCACCAAGGTGCAGAGCCAGCCTTCTTATTCGGCCTTGAATTGATCATATGCGGATTAGAAAAACAACTTAAATGTGAAAGTGGGTCGCCAAAAAAGAAGAGAAAGGTGGATGGTGGCGGAGCACTGTCTCCCCAGCATTCAGCTGTGACCCAGGGATCAATCATTAAGAACAAGGAAGGTATGGATGCTAAGTCCCTGACCGCTTGGTCAAGAACTTTGGTAACTTTTAAGGACGTGTTCGTCGATTTTACTCGGGAGGAGTGGAAACTATTGGATACCGCCCAGCAAATCGTGTATAGAAACGTTATGCTCGAGAACTATAAGAATCTTGTGTCACTGGGCTACCAACTGACAAAGCCTGACGTTATCCTGAGGCTGGAAAAGGGTGAGGAGCCTTGGTTGGTCGAGCGGGAGATCCACCAGGAAACTCACCCAGACAGTGAGACTGCTTTCGAGATCAAGTCCTCTGTGTAATACAAGGCGGCCGCAAATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTAAAGCTTCGAATTCTGATAATCAGCCATACCACATTTGTAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCAGCTCGACTAGTGTTCATCGGAGCCCAGGTTTACTCCCTTAAGTGGAAATTTCTTCCCCCACTCCCTCCTTGGCTTTCTCCAAGGAGGGAACCCAGGCTACTGGAAAGTCCGGCTGGGGCGGGGACTGTGGGTTTCAGGGTAGAACTGCGTGTGGAACGGGACAGGGAGCGGTTAGAAGGGTGGGGCTATTCCGGGAAGTGGTGGGGGGAGGGAGCCCAAAACTAGCACCTAGTCCACTCATTATCCAGCCCTCTTATTTCTCGGCCCCGCTCTGCTTCAGTGGACCCGGGGAGGGCGGGGAAGTGGAGTGGGAGACCTAGGGGTGGGCTTCCCGACCTTGCTGTACAGGACCTCGACCTAGCTGGCTTTGTTCCCCATCCCCACGTTAGTTGTTGCCCTGAGGCTAAAACTAGAGCCCAGGGGCCCCAAGTTCCAGACTGCCCCTCCCCCCTCCCCCGGAGCCAGGGAGTGGTTGGTGAAAGGGGGAGGCCAGCTGGAGAACAAACGGGTAGTCAGGGGGTTGAGCGATTAGAGCCCTTGTACCCTACCCAGGAATGGTTGGGGAGGAGGAGGAAGAGGTAGGAGGTAGGGGAGGGGGCGGGGTTTTGTCACCTGTCACCTGCTCCGGCTGTGCCTAGGGCGGGCGGGCGGGGAGTGGGGGGACCGGTATAAAGCGGTAGGCGCCTGTGCCCGCTCCACCTCTCAAGCAGCCAGCGCCTGCCTGAATCTGTTCTGCCCCCTCCCCACCCATTTCACCACCACCCTCGAGGGATCCGGCCACCATGGCCCCCCCGACCGATGTCAGCCTGGGGGACGAGCTCCACTTAGACGGCGAGGACGTGGCGATGGCGCATGCCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGGACGGGGATTCCCCGGGTCCGGGATTTACCCCCCACGACTCCGCCCCCTACGGCGCTCTGGATATGGCCGACTTCGAGTTTGAGCAGATGTTTACCGATGCCCTTGGAATTGACGAGTACGGTGGGACGCGTATGAAGCTACTGTCTTCTATCGAACAAGCATGCGATATTTGCCGACTTAAAAAGCTCAAGTGCTCCAAAGAAAAACCGAAGTGCGCCAAGTGTCTGAAGAACAACTGGGAGTGTCGCTACTCTCCCAAAACCAAAAGGTCTCCGCTGACTAGGGCACATCTGACAGAAGTGGAATCAAGGCTAGAAAGACTGGAACAGCTATTTCTACTGATTTTTCCTCGAGAAGACCTTGACATGATTTTGAAAATGGATTCTTTACAGGATATAAAAGCATTGTTAACAGGATTATTTGTACAAGATAATGTGAATAAAGATGCCGTCACAGATAGATTGGCTTCAGTGGAGACTGATATGCCTCTAACATTGAGACAGCATAGAATAAGTGCGACATCATCATCGGAAGAGAGTAGTAACAAAGGTCAAAGACAGTTGACTGTATAATTCACTCCTCAGGTGCAGGCTGCCTATCAGAAGGTGGTGGCTGGTGTGGCCAATGCCCTGGCTCACAAATACCACTGAGATCTTTTTCCCTCTGCCAAAAATTATGGGGACATCATGAAGCCCCTTGAGCATCTGACTTCTGGCTAATAAAGGAAATTTATTTTCATTGCAATAGTGTGTTGGAATTTTTTGTGTCTCTCACTCGGAAGGACATATGGGAGGGCAAATCATTTAAAACATCAGAATGAGTATTTGGTTTAGAGTTTGGCAACATATGCCCATATGCTGGCTGCCATGAACAAAGGTTGGCTATAAAGAGGTCATCAGTATATGAAACAGCCCCCTGCTGTCCATTCCTTATTCCATAGAAAAGCCTTGACTTGAGGTTAGATTTTTTTTATATTTTGTTTTGTGTTATTTTTTTCTTTAACATCCCTAAAATTTTCCTTACATGTTTTACTAGCCAGATTTTTCCTCCTCTCCTGACTACTCCCAGTCATAGCTGTCCCTCTTCTCTTATGGAGATC(SEQ ID NO:21)。
具有SEQ ID NO:22所述核苷酸序列的核酸编码由细胞特异性启动子hMUC1、第一microRNA为mir-199a-3p、第二microRNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白分别为Lacl、tetR-KRAB、第一和第二识别序列为4×UAS的元件组成的表达系统。
CGGCCATATGTCTTCCGCGGAGTGCTGTACTCCGCGGAGTACTGTCCTCCGCGGAGTGCCGCACTCCGAGCGGAGACTCTAGACTCCCTATCAGTGATAGAGATAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCTCCCTATCAGTGATAGAGAGAATTCGACCGCCACCATGAGACATATTATCTGCCACGGAGGTGTTATTACCGAAGAAATGGCCGCCAGTCTTTTGGACCAGCTGATCGAAGAGGTACTGGCTGATAATCTTCCACCTCCTAGCCATTTTGAACCACCTACCCTTCACGAACTGTATGATTTAGACGTGACGGCCCCCGAAGATCCCAACGAGGAGGCGGTTTCGCAGATTTTTCCCGACTCTGTAATGTTGGCGGTGCAGGAAGGGATTGACTTACTCACTTTTCCGCCGGCGCCCGGTTCTCCGGAGCCGCCTCACCTTTCCCGGCAGCCCGAGCAGCCGGAGCAGAGAGCCTTGGGTCCGGTTTCTATGCCAAACCTTGTACCGGAGGTGATCGATCTTACCTGCCACGAGGCTGGCTTTCCACCCAGTGACGACGAGGATGAAGAGGGTGAGGAGTTTGTGTTAGATTATGTGGAGCACCCCGGGCACGGTTGCAGGTCTTGTCATTATCACCGGAGGAATACGGGGGACCCAGATATTATGTGTTCGCTTTGCTATATGAGGACCTGTGGCATGTTTGTCTACAGTAAGTGAAAATTATGGGCAGTGGGTGATAGAGTGGTGGGTTTGGTGTGGTAATTTTTTTTTTAATTTTTACAGTTTTGTGGTTTAAAGAATTTTGTATTGTGATTTTTTTAAAAGGTCCTGTGTCTGAACCTGAGCCTGAGCCCGAGCCAGAACCGGAGCCTGCAAGACCTACCCGCCGTCCTAAAATGGCGCCTGCTATCCTGAGGCGCCCGACATCACCTGTGTCTAGAGAATGCAATAGTAGTACGGATAGCTGTGACTCCGGTCCTTCTAACACACCTCCTGAGATACACCCGGTGGTCCCGCTGTGCCCCATTAAACCAGTTGCCGTGAGAGTTGGTGGGCGTCGCCAGGCTGTGGAATGTATCGAGGACTTGCTTAACGAGCCTGGGCAACCTTTGGACTTGAGCTGTAAACGCCCCAGGCCACTCGAGGGGTCCGGAGCTACAAATTTTTCCCTCCTCAAACAGGCTGGAGATGTCGAAGAAAATCCTGGGCCTACCGGTCCCATGAGTGTGCCCACTCAGGTCCTGGGGTTGCTGCTGCTGTGGCTTACAGACGCTCGCTGCCAGGTGCAGCTGGTGGAGAGCGGCGGCGGCGTGGTGCAGCCCGGCAGGAGCCTGAGGCTGGACTGCAAGGCCAGCGGCATCACCTTCAGCAACAGCGGCATGCACTGGGTGAGGCAGGCCCCCGGCAAGGGCCTGGAGTGGGTGGCCGTGATCTGGTACGACGGCAGCAAGAGGTACTACGCCGACAGCGTGAAGGGCAGGTTCACCATCAGCAGGGACAACAGCAAGAACACCCTGTTCCTGCAGATGAACAGCCTGAGGGCCGAGGACACCGCCGTGTACTACTGCGCCACCAACGACGACTACTGGGGCCAGGGCACCCTGGTGACCGTGAGCAGCGGTGGAGGCGGTTCAGGCGGAGGTGGCTCTGGCGGTGGCGGATCGGAGATCGTGCTGACCCAGAGCCCCGCCACCCTGAGCCTGAGCCCCGGCGAGAGGGCCACCCTGAGCTGCAGGGCCAGCCAGAGCGTGAGCAGCTACCTGGCCTGGTACCAGCAGAAGCCCGGCCAGGCCCCCAGGCTGCTGATCTACGACGCCAGCAACAGGGCCACCGGCATCCCCGCCAGGTTCAGCGGCAGCGGCAGCGGCACCGACTTCACCCTGACCATCAGCAGCCTGGAGCCCGAGGACTTCGCCGTGTACTACTGCCAGCAGAGCAGCAACTGGCCCAGGACCTTCGGCCAGGGCACCAAGGTGGAGATCAAGAGATCTGAGGGCCGCGGCAGCCTGCTGACCTGCGGCGACGTGGAGGAAAACCCCGGCCCCGGATCCATGAAACCAGTAACGTTATACGATGTCGCAGAGTATGCCGGTGTCTCTTATCAGACCGTTTCCCGCGTGGTGAACCAGGCCAGCCACGTTTCTGCGAAAACGCGGGAAAAAGTGGAAGCGGCGATGGCGGAGCTGAATTACATTCCCAACCGCGTGGCACAACAACTGGCGGGCAAACAGTCGTTGCTGATTGGCGTTGCCACCTCCAGTCTGGCCCTGCACGCGCCGTCGCAAATTGTCGCGGCGATTAAATCTCGCGCCGATCAACTGGGTGCCAGCGTGGTGGTGTCGATGGTAGAACGAAGCGGCGTCGAAGCCTGTAAAGCGGCGGTGCACAATCTTCTCGCGCAACGCGTCAGTGGGCTGATCATTAACTATCCGCTGGATGACCAGGATGCCATTGCTGTGGAAGCTGCCTGCACTAATGTTCCGGCGTTATTTCTTGATGTCTCTGACCAGACACCCATCAACAGTATTATTTTCTCCCATGAAGACGGTACGCGACTGGGCGTGGAGCATCTGGTCGCATTGGGTCACCAGCAAATCGCGCTGTTAGCGGGCCCATTAAGTTCTGTCTCGGCGCGTCTGCGTCTGGCTGGCTGGCATAAATATCTCACTCGCAATCAAATTCAGCCGATAGCGGAACGGGAAGGCGACTGGAGTGCCATGTCCGGTTTTCAACAAACCATGCAAATGCTGAATGAGGGCATCGTTCCCACTGCGATGCTGGTTGCCAACGATCAGATGGCGCTGGGCGCAATGCGCGCCATTACCGAGTCCGGGCTGCGCGTTGGTGCGGATATCTCGGTAGTGGGATACGACGATACCGAAGACAGCTCATGTTATATCCCGCCGTTAACCACCATCAAACAGGATTTTCGCCTGCTGGGGCAAACCAGCGTGGACCGCTTGCTGCAACTCTCTCAGGGCCAGGCGGTGAAGGGCAATCAGCTGTTGCCGGTCTCACTGGTGAAAAGAAAAACCACCCTGGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGAGCAGCCTGAGGCCTCCTAAGAAGAAGAGGAAGGTTGCGGCCGCTAACCAATGTGCAGACTACTGTTAACCAATGTGCAGACTACTGTGAATTCTAACCAATGTGCAGACTACTGTTAACCAATGTGCAGACTACTGTAAGCTTCGAATTGAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGGTCGACAATACTAGTTAAGAAAGCGGTCCACTGTGTGCCGCGGGTGACAGCCCTCCGCGGCGGTCTTTCGTCCGCGGGTGACCGCCCTCCGAGCGGAGACTCTAGAAATTGTGAGCGGATAACAATTGGGTATATAATGGGGGCCGTCAGCTACTACCAGAGCCGATCGAACACTCGAGCCGAGCAGACGTGCCTACAATTGTGAGCGCTCACAATTACTAGAAGATCTTGAATTCGCCACCATGGCTAGATTAGATAAAAGTAAAGTGATTAACAGCGCATTAGAGCTGCTTAATGAGGTCGGAATCGAAGGTTTAACAACCCGTAAACTCGCCCAGAAGCTAGGTGTAGAGCAGCCTACATTGTATTGGCATGTAAAAAATAAGCGGGCTTTGCTCGACGCCTTAGCCATTGAGATGTTAGATAGGCACCATACTCACTTTTGCCCTTTAGAAGGGGAAAGCTGGCAAGATTTTTTACGTAATAACGCTAAAAGTTTTAGATGTGCTTTACTAAGTCATCGCGATGGAGCAAAAGTACATTTAGGTACACGGCCTACAGAAAAACAGTATGAAACTCTCGAAAATCAATTAGCCTTTTTATGCCAACAAGGTTTTTCACTAGAGAATGCATTATATGCACTCAGCGCTGTGGGGCATTTTACTTTAGGTTGCGTATTGGAAGATCAAGAGCATCAAGTCGCTAAAGAAGAAAGGGAAACACCTACTACTGATAGTATGCCGCCATTATTACGACAAGCTATCGAATTATTTGATCACCAAGGTGCAGAGCCAGCCTTCTTATTCGGCCTTGAATTGATCATATGCGGATTAGAAAAACAACTTAAATGTGAAAGTGGGTCGCCAAAAAAGAAGAGAAAGGTGGATGGTGGCGGAGCACTGTCTCCCCAGCATTCAGCTGTGACCCAGGGATCAATCATTAAGAACAAGGAAGGTATGGATGCTAAGTCCCTGACCGCTTGGTCAAGAACTTTGGTAACTTTTAAGGACGTGTTCGTCGATTTTACTCGGGAGGAGTGGAAACTATTGGATACCGCCCAGCAAATCGTGTATAGAAACGTTATGCTCGAGAACTATAAGAATCTTGTGTCACTGGGCTACCAACTGACAAAGCCTGACGTTATCCTGAGGCTGGAAAAGGGTGAGGAGCCTTGGTTGGTCGAGCGGGAGATCCACCAGGAAACTCACCCAGACAGTGAGACTGCTTTCGAGATCAAGTCCTCTGTGTAATACAAGGCGGCCGCAAATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTAAAGCTTCGAATTCTGATAATCAGCCATACCACATTTGTAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCAGCTCGACTAGTGTTCATCGGAGCCCAGGTTTACTCCCTTAAGTGGAAATTTCTTCCCCCACTCCCTCCTTGGCTTTCTCCAAGGAGGGAACCCAGGCTACTGGAAAGTCCGGCTGGGGCGGGGACTGTGGGTTTCAGGGTAGAACTGCGTGTGGAACGGGACAGGGAGCGGTTAGAAGGGTGGGGCTATTCCGGGAAGTGGTGGGGGGAGGGAGCCCAAAACTAGCACCTAGTCCACTCATTATCCAGCCCTCTTATTTCTCGGCCCCGCTCTGCTTCAGTGGACCCGGGGAGGGCGGGGAAGTGGAGTGGGAGACCTAGGGGTGGGCTTCCCGACCTTGCTGTACAGGACCTCGACCTAGCTGGCTTTGTTCCCCATCCCCACGTTAGTTGTTGCCCTGAGGCTAAAACTAGAGCCCAGGGGCCCCAAGTTCCAGACTGCCCCTCCCCCCTCCCCCGGAGCCAGGGAGTGGTTGGTGAAAGGGGGAGGCCAGCTGGAGAACAAACGGGTAGTCAGGGGGTTGAGCGATTAGAGCCCTTGTACCCTACCCAGGAATGGTTGGGGAGGAGGAGGAAGAGGTAGGAGGTAGGGGAGGGGGCGGGGTTTTGTCACCTGTCACCTGCTCCGGCTGTGCCTAGGGCGGGCGGGCGGGGAGTGGGGGGACCGGTATAAAGCGGTAGGCGCCTGTGCCCGCTCCACCTCTCAAGCAGCCAGCGCCTGCCTGAATCTGTTCTGCCCCCTCCCCACCCATTTCACCACCACCCTCGAGGGATCCGGCCACCATGGCCCCCCCGACCGATGTCAGCCTGGGGGACGAGCTCCACTTAGACGGCGAGGACGTGGCGATGGCGCATGCCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGGACGGGGATTCCCCGGGTCCGGGATTTACCCCCCACGACTCCGCCCCCTACGGCGCTCTGGATATGGCCGACTTCGAGTTTGAGCAGATGTTTACCGATGCCCTTGGAATTGACGAGTACGGTGGGACGCGTATGAAGCTACTGTCTTCTATCGAACAAGCATGCGATATTTGCCGACTTAAAAAGCTCAAGTGCTCCAAAGAAAAACCGAAGTGCGCCAAGTGTCTGAAGAACAACTGGGAGTGTCGCTACTCTCCCAAAACCAAAAGGTCTCCGCTGACTAGGGCACATCTGACAGAAGTGGAATCAAGGCTAGAAAGACTGGAACAGCTATTTCTACTGATTTTTCCTCGAGAAGACCTTGACATGATTTTGAAAATGGATTCTTTACAGGATATAAAAGCATTGTTAACAGGATTATTTGTACAAGATAATGTGAATAAAGATGCCGTCACAGATAGATTGGCTTCAGTGGAGACTGATATGCCTCTAACATTGAGACAGCATAGAATAAGTGCGACATCATCATCGGAAGAGAGTAGTAACAAAGGTCAAAGACAGTTGACTGTATAATTCACTCCTCAGGTGCAGGCTGCCTATCAGAAGGTGGTGGCTGGTGTGGCCAATGCCCTGGCTCACAAATACCACTGAGATCTTTTTCCCTCTGCCAAAAATTATGGGGACATCATGAAGCCCCTTGAGCATCTGACTTCTGGCTAATAAAGGAAATTTATTTTCATTGCAATAGTGTGTTGGAATTTTTTGTGTCTCTCACTCGGAAGGACATATGGGAGGGCAAATCATTTAAAACATCAGAATGAGTATTTGGTTTAGAGTTTGGCAACATATGCCCATATGCTGGCTGCCATGAACAAAGGTTGGCTATAAAGAGGTCATCAGTATATGAAACAGCCCCCTGCTGTCCATTCCTTATTCCATAGAAAAGCCTTGACTTGAGGTTAGATTTTTTTTATATTTTGTTTTGTGTTATTTTTTTCTTTAACATCCCTAAAATTTTCCTTACATGTTTTACTAGCCAGATTTTTCCTCCTCTCCTGACTACTCCCAGTCATAGCTGTCCCTCTTCTCTTATGGAGATC(SEQ IDNO:22)。
具有SEQ ID NO:23所述核苷酸序列的核酸编码由细胞特异性启动子hMUC1、第一microRNA为mir-199a-3p、第二microRNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白分别为TALE蛋白、第一和第二识别序列为4×UAS的元件组成的表达系统。
CGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTTCTGTCCTCCGAGCGGAGACTCTAGATACCCCTCTCCGCTTCTGAATTCTAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCCTCGAGTACCCCTCTCCGCTTCTTACCCCTCTCCGCTTCTTACCCCTCTCCGCTTCTGAATTCGACCGCCACCATGAGACATATTATCTGCCACGGAGGTGTTATTACCGAAGAAATGGCCGCCAGTCTTTTGGACCAGCTGATCGAAGAGGTACTGGCTGATAATCTTCCACCTCCTAGCCATTTTGAACCACCTACCCTTCACGAACTGTATGATTTAGACGTGACGGCCCCCGAAGATCCCAACGAGGAGGCGGTTTCGCAGATTTTTCCCGACTCTGTAATGTTGGCGGTGCAGGAAGGGATTGACTTACTCACTTTTCCGCCGGCGCCCGGTTCTCCGGAGCCGCCTCACCTTTCCCGGCAGCCCGAGCAGCCGGAGCAGAGAGCCTTGGGTCCGGTTTCTATGCCAAACCTTGTACCGGAGGTGATCGATCTTACCTGCCACGAGGCTGGCTTTCCACCCAGTGACGACGAGGATGAAGAGGGTGAGGAGTTTGTGTTAGATTATGTGGAGCACCCCGGGCACGGTTGCAGGTCTTGTCATTATCACCGGAGGAATACGGGGGACCCAGATATTATGTGTTCGCTTTGCTATATGAGGACCTGTGGCATGTTTGTCTACAGTAAGTGAAAATTATGGGCAGTGGGTGATAGAGTGGTGGGTTTGGTGTGGTAATTTTTTTTTTAATTTTTACAGTTTTGTGGTTTAAAGAATTTTGTATTGTGATTTTTTTAAAAGGTCCTGTGTCTGAACCTGAGCCTGAGCCCGAGCCAGAACCGGAGCCTGCAAGACCTACCCGCCGTCCTAAAATGGCGCCTGCTATCCTGAGGCGCCCGACATCACCTGTGTCTAGAGAATGCAATAGTAGTACGGATAGCTGTGACTCCGGTCCTTCTAACACACCTCCTGAGATACACCCGGTGGTCCCGCTGTGCCCCATTAAACCAGTTGCCGTGAGAGTTGGTGGGCGTCGCCAGGCTGTGGAATGTATCGAGGACTTGCTTAACGAGCCTGGGCAACCTTTGGACTTGAGCTGTAAACGCCCCAGGCCAGGGTCCGGAGCTACAAATTTTTCCCTCCTCAAACAGGCTGGAGATGTCGAAGAAAATCCTGGGCCTATGGAGGCTTGGGAGTGTTTGGAAGATTTTTCTGCTGTGCGTAACTTGCTGGAACAGAGCTCTAACAGTACCTCTTGGTTTTGGAGGTTTCTGTGGGGCTCATCCCAGGCAAAGTTAGTCTGCAGAATTAAGGAGGATTACAAGTGGGAATTTGAAGAGCTTTTGAAATCCTGTGGTGAGCTGTTTGATTCTTTGAATCTGGGTCACCAGGCGCTTTTCCAAGAGAAGGTCATCAAGACTTTGGATTTTTCCACACCGGGGCGCGCTGCGGCTGCTGTTGCTTTTTTGAGTTTTATAAAGGATAAATGGAGCGAAGAAACCCATCTGAGCGGGGGGTACCTGCTGGATTTTCTGGCCATGCATCTGTGGAGAGCGGTTGTGAGACACAAGAATCGCCTGCTACTGTTGTCTTCCGTCCGCCCGGCGATAATACCGACGGAGGAGCAGCAGCAGCAGCAGGAGGAAGCCAGGCGGCGGCGGCAGGAGCAGAGCCCATGGAACCCGAGAGCCGGCCTGGACCCTCGGGAATGAATGTTGTACAGGTGGCTGAACTGTATCCAGAACTGAGACGCATTTTGACAATTACAGAGGATGGGCAGGGGCTAAAGGGGGTAAAGAGGGAGCGGGGGGCTTGTGAGGCTACAGAGGAGGCTAGGAATCTAGCTTTTAGCTTAATGACCAGACACCGTCCTGAGTGTATTACTTTTCAACAGATCAAGGATAATTGCGCTAATGAGCTTGATCTGCTGGCGCAGAAGTATTCCATAGAGCAGCTGACCACTTACTGGCTGCAGCCAGGGGATGATTTTGAGGAGGCTATTAGGGTATATGCAAAGGTGGCACTTAGGCCAGATTGCAAGTACAAGATCAGCAAACTTGTAAATATCAGGAATTGTTGCTACATTTCTGGGAACGGGGCCGAGGTGGAGATAGATACGGAGGATAGGGTGGCCTTTAGATGTAGCATGATAAATATGTGGCCGGGGGTGCTTGGCATGGACGGGGTGGTTATTATGAATGTAAGGTTTACTGGCCCCAATTTTAGCGGTACGGTTTTCCTGGCCAATACCAACCTTATCCTACACGGTGTAAGCTTCTATGGGTTTAACAATACCTGTGTGGAAGCCTGGACCGATGTAAGGGTTCGGGGCTGTGCCTTTTACTGCTGCTGGAAGGGGGTGGTGTGTCGCCCCAAAAGCAGGGCTTCAATTAAGAAATGCCTCTTTGAAAGGTGTACCTTGGGTATCCTGTCTGAGGGTAACTCCAGGGTGCGCCACAATGTGGCCTCCGACTGTGGTTGCTTCATGCTAGTGAAAAGCGTGGCTGTGATTAAGCATAACATGGTATGTGGCAACTGCGAGGACAGGGCCTCTCAGATGCTGACCTGCTCGGACGGCAACTGTCACCTGCTGAAGACCATTCACGTAGCCAGCCACTCTCGCAAGGCCTGGCCAGTGTTTGAGCATAACATACTGACCCGCTGTTCCTTGCATTTGGGTAACAGGAGGGGGGTGTTCCTACCTTACCAATGCAATTTGAGTCACACTAAGATATTGCTTGAGCCCGAGAGCATGTCCAAGGTGAACCTGAACGGGGTGTTTGACATGACCATGAAGATCTGGAAGGTGCTGAGGTACGATGAGACCCGCACCAGGTGCAGACCCTGCGAGTGTGGCGGTAAACATATTAGGAACCAGCCTGTGATGCTGGATGTGACCGAGGAGCTGAGGCCCGATCACTTGGTGCTGGCCTGCACCCGCGCTGAGTTTGGCTCTAGCGATGAAGATACAGATCTCGAGGGGTCCGGAGCTACAAATTTTTCCCTCCTCAAACAGGCTGGAGATGTCGAAGAAAATCCTGGGCCTACCGGTATGTATAGGATGCAGTTGCTCTCTTGCATCGCCCTGTCTCTAGCGTTGGTGACTAATAGCGCCCCTACTTCTTCTTCCACGAAGAAAACCCAGCTGCAACTGGAACACCTTCTGCTGGATCTCCAGATGATCCTTAACGGGATTAACAACTATAAGAACCCGAAGCTCACGGCTATGCTGACGGCGAAGTTCGCTATGCCTAAAAAGGCTACTGAGCTGAAACACTTACAGTGTCTCGAAGAGGCACTCAAACCCCTCGAGGAGGTACTCAATCTGGCGCAAAGCAAAAACTTCCATCTCCGCCCACGGGACCTAATCTCCAACATCAACGTGATTGTGCTGGAGCTCAAGGGGTCTGAAACCACGTTTATGTGCGAGTACGCAGATGAAACTGCTACCATAGTGGAATTCCTCAATCGTTGGATCACTTTCTGCCAGTCAATCATTTCAACCCTGACCAGATCTGAGGGCCGCGGCAGCCTGCTGACCTGCGGCGACGTGGAGGAAAACCCCGGCCCCGGATCCTTGTCGACCATTCGTCCGCGCAGGCCAAGTCCTGCCCGCGAGCTTCTGCCCGGACCCCAACCGGATAGGGTTCAGCCGACTGCAGATCGTGGGGTGTCTGCGCCTGCTGGCAGCCCTCTGGATGGCTTGCCCGCTCGGCGGACGGTGTCCCGGACCCGGCTGCCATCTCCCCCTGCGCCCTCACCTGCGTTCTCGGCGGGCAGCTTCAGCGATCTGCTCCGTCCGTTCGATCCGTCGCTTCTTGATACATCGCTTCTTGATTCGATGCCTGCCGTCGGCACGCCGCATACAGCGGCTGCCCCAGCAGAGTGGGATGAGGCGCAATCGGCTCTGCGTGCAGCCGATGACCCGCCACCCACCGTGCGTGTCGCTGTCACTGCCGCGCGGCCGCCGCGCGCCAAGCCGGCCCCGCGACGGCGTGCTGCGCAACCCTCCGACGCTTCGCCGGCCGCGCAGGTGGATCTACGCACGCTCGGCTACAGTCAGCAGCAGCAAGAGAAGATCAAACCGAAGGTGCGTTCGACAGTGGCGCAGCACCACGAGGCACTGGTGGGCCATGGGTTTACACACGCGCACATCGTTGCGCTCAGCCAACACCCGGCAGCGTTAGGGACCGTCGCTGTCACGTATCAGCACATAATCACGGCGTTGCCAGAGGCGACACACGAAGACATCGTTGGCGTCGGCAAACAGTGGTCCGGCGCACGCGCCCTGGAGGCCTTGCTCACGGATGCGGGGGAGTTGAGAGGTCCGCCGTTACAGTTGGACACAGGCCAACTTGTGAAGATTGCAAAACGTGGCGGCGTGACCGCAATGGAGGCAGTGCATGCATCGCGCAATGCACTGACGGGTGCCCCCCTGAACCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACAATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACAATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAAGCATTGTGGCCCAGCTGAGCCGGCCTGATCCGGCGTTGGCCGCGTTGACCAACGACCACCTCGTCGCCTTGGCCTGCCTCGGCGGACGTCCTGCCATGGATGCAGTGAAAAAGGGATTGCCGCACGCGCCGGAATTGATCAGAAGAGTCAATCGCCGTATTGGCGAACGCACGTCCCATCGCGTTGCCGACTACGCGCAAGTGGTTCGCGTGCTGGAGTTTTTCCAGTGCCACTCCCACCCAGCGTACGCATTTGATGAGGCCATGACGCAGTTCGGGATGAGCAGGAACGGGTTGGTACAGCTCTTTCGCAGAGTGGGCGTCACCGAACTCGAAGCCCGCGGTGGAACGCTCCCCCCAGCCTCGCAGCGTTGGGACCGTATCCTCCAGGCATCAGGGATGAAAAGGGCCAAACCGTCCCCTACTTCAGCTCAAACACCGGATCAGGCGTCTTTGCATGCATTCGCCGATTCGCTGGAGCGTGACCTTGATGCGCCCAGCCCAATGCACGAGGGAGATCAGACGCGGGCAAGCAGCCGTAAACGGTCCCGATCGGATCGTGCTGTCACCGGCCCCTCCGCACAGCAGGCTGTCGAGGTGCGCGTTCCCGAACAGCGCGATGCGCTGCATTTGCCCCTCAGCTGGAGGGTAAAACGCCCGCGTACCAGGATCTGGGGCGGCCTCCCGGTTAAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGTAACCAATGTGCAGACTACTGTTAACCAATGTGCAGACTACTGTTAACCAATGTGCAGACTACTGTTAACCAATGTGCAGACTACTGTGAAAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTTCTGTCCTCCGAGCGGAGACTCTAGATATAGAACCGATCCTCCCATTGAATTCTAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCCTCGAGTATAGAACCGATCCTCCCATTTATAGAACCGATCCTCCCATTTATAGAACCGATCCTCCCATTACTAGAAGATCTTGAATTCGCCACCTTGTCGACCATTCGTCCGCGCAGGCCAAGTCCTGCCCGCGAGCTTCTGCCCGGACCCCAACCGGATAGGGTTCAGCCGACTGCAGATCGTGGGGTGTCTGCGCCTGCTGGCAGCCCTCTGGATGGCTTGCCCGCTCGGCGGACGGTGTCCCGGACCCGGCTGCCATCTCCCCCTGCGCCCTCACCTGCGTTCTCGGCGGGCAGCTTCAGCGATCTGCTCCGTCCGTTCGATCCGTCGCTTCTTGATACATCGCTTCTTGATTCGATGCCTGCCGTCGGCACGCCGCATACAGCGGCTGCCCCAGCAGAGTGGGATGAGGCGCAATCGGCTCTGCGTGCAGCCGATGACCCGCCACCCACCGTGCGTGTCGCTGTCACTGCCGCGCGGCCGCCGCGCGCCAAGCCGGCCCCGCGACGGCGTGCTGCGCAACCCTCCGACGCTTCGCCGGCCGCGCAGGTGGATCTACGCACGCTCGGCTACAGTCAGCAGCAGCAAGAGAAGATCAAACCGAAGGTGCGTTCGACAGTGGCGCAGCACCACGAGGCACTGGTGGGCCATGGGTTTACACACGCGCACATCGTTGCGCTCAGCCAACACCCGGCAGCGTTAGGGACCGTCGCTGTCACGTATCAGCACATAATCACGGCGTTGCCAGAGGCGACACACGAAGACATCGTTGGCGTCGGCAAACAGTGGTCCGGCGCACGCGCCCTGGAGGCCTTGCTCACGGATGCGGGGGAGTTGAGAGGTCCGCCGTTACAGTTGGACACAGGCCAACTTGTGAAGATTGCAAAACGTGGCGGCGTGACCGCAATGGAGGCAGTGCATGCATCGCGCAATGCACTGACGGGTGCCCCCCTGAACCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACAATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAAGCATTGTGGCCCAGCTGAGCCGGCCTGATCCGGCGTTGGCCGCGTTGACCAACGACCACCTCGTCGCCTTGGCCTGCCTCGGCGGACGTCCTGCCATGGATGCAGTGAAAAAGGGATTGCCGCACGCGCCGGAATTGATCAGAAGAGTCAATCGCCGTATTGGCGAACGCACGTCCCATCGCGTTGCCGACTACGCGCAAGTGGTTCGCGTGCTGGAGTTTTTCCAGTGCCACTCCCACCCAGCGTACGCATTTGATGAGGCCATGACGCAGTTCGGGATGAGCAGGAACGGGTTGGTACAGCTCTTTCGCAGAGTGGGCGTCACCGAACTCGAAGCCCGCGGTGGAACGCTCCCCCCAGCCTCGCAGCGTTGGGACCGTATCCTCCAGGCATCAGGGATGAAAAGGGCCAAACCGTCCCCTACTTCAGCTCAAACACCGGATCAGGCGTCTTTGCATGCATTCGCCGATTCGCTGGAGCGTGACCTTGATGCGCCCAGCCCAATGCACGAGGGAGATCAGACGCGGGCAAGCAGCCGTAAACGGTCCCGATCGGATCGTGCTGTCACCGGCCCCTCCGCACAGCAGGCTGTCGAGGTGCGCGTTCCCGAACAGCGCGATGCGCTGCATTTGCCCCTCAGCTGGAGGGTAAAACGCCCGCGTACCAGGATCTGGGGCGGCCTCCCGTAATACAAGGCGGCCGCAAATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTAAAGCTTCGAATTCTGATAATCAGCCATACCACATTTGTAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCGTTCATCGGAGCCCAGGTTTACTCCCTTAAGTGGAAATTTCTTCCCCCACTCCCTCCTTGGCTTTCTCCAAGGAGGGAACCCAGGCTACTGGAAAGTCCGGCTGGGGCGGGGACTGTGGGTTTCAGGGTAGAACTGCGTGTGGAACGGGACAGGGAGCGGTTAGAAGGGTGGGGCTATTCCGGGAAGTGGTGGGGGGAGGGAGCCCAAAACTAGCACCTAGTCCACTCATTATCCAGCCCTCTTATTTCTCGGCCCCGCTCTGCTTCAGTGGACCCGGGGAGGGCGGGGAAGTGGAGTGGGAGACCTAGGGGTGGGCTTCCCGACCTTGCTGTACAGGACCTCGACCTAGCTGGCTTTGTTCCCCATCCCCACGTTAGTTGTTGCCCTGAGGCTAAAACTAGAGCCCAGGGGCCCCAAGTTCCAGACTGCCCCTCCCCCCTCCCCCGGAGCCAGGGAGTGGTTGGTGAAAGGGGGAGGCCAGCTGGAGAACAAACGGGTAGTCAGGGGGTTGAGCGATTAGAGCCCTTGTACCCTACCCAGGAATGGTTGGGGAGGAGGAGGAAGAGGTAGGAGGTAGGGGAGGGGGCGGGGTTTTGTCACCTGTCACCTGCTCCGGCTGTGCCTAGGGCGGGCGGGCGGGGAGTGGGGGGACCGGTATAAAGCGGTAGGCGCCTGTGCCCGCTCCACCTCTCAAGCAGCCAGCGCCTGCCTGAATCTGTTCTGCCCCCTCCCCACCCATTTCACCACCACCATGGCCCCCCCGACCGATGTCAGCCTGGGGGACGAGCTCCACTTAGACGGCGAGGACGTGGCGATGGCGCATGCCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGAGCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGAGCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGAGCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGGACGGGGATTCCCCGGGTCCGGGATTTACCCCCCACGACTCCGCCCCCTACGGCGCTCTGGATATGGCCGACTTCGAGTTTGAGCAGATGTTTACCGATGCCCTTGGAATTGACGAGTACGGTGGGACGCGTATGAAGCTACTGTCTTCTATCGAACAAGCATGCGATATTTGCCGACTTAAAAAGCTCAAGTGCTCCAAAGAAAAACCGAAGTGCGCCAAGTGTCTGAAGAACAACTGGGAGTGTCGCTACTCTCCCAAAACCAAAAGGTCTCCGCTGACTAGGGCACATCTGACAGAAGTGGAATCAAGGCTAGAAAGACTGGAACAGCTATTTCTACTGATTTTTCCTCGAGAAGACCTTGACATGATTTTGAAAATGGATTCTTTACAGGATATAAAAGCATTGTTAACAGGATTATTTGTACAAGATAATGTGAATAAAGATGCCGTCACAGATAGATTGGCTTCAGTGGAGACTGATATGCCTCTAACATTGAGACAGCATAGAATAAGTGCGACATCATCATCGGAAGAGAGTAGTAACAAAGGTCAAAGACAGTTGACTGTATAAAAGCTTCACTCCTCAGGTGCAGGCTGCCTATCAGAAGGTGGTGGCTGGTGTGGCCAATGCCCTGGCTCACAAATACCACTGAGATCTTTTTCCCTCTGCCAAAAATTATGGGGACATCATGAAGCCCCTTGAGCATCTGACTTCTGGCTAATAAAGGAAATTTATTTTCATTGCAATAGTGTGTTGGAATTTTTTGTGTCTCTCACTCGGAAGGACATATGGGAGGGCAAATCATTTAAAACATCAGAATGAGTATTTGGTTTAGAGTTTGGCAACATATGCCCATATGCTGGCTGCCATGAACAAAGGTTGGCTATAAAGAGGTCATCAGTATATGAAACAGCCCCCTGCTGTCCATTCCTTATTCCATAGAAAAGCCTTGACTTGAGGTTAGATTTTTTTTATATTTTGTTTTGTGTTATTTTTTTCTTTAACATCCCTAAAATTTTCCTTACATGTTTTACTAGCCAGATTTTTCCTCCTCTCCTGACTACTCCCAGTCATAGCTGTCCCTCTTCTCTTATGGAGATC(SEQ ID NO:23)。
具有SEQ ID NO:24所述核苷酸序列的核酸编码由细胞特异性启动子hMUC1、第一microRNA为mir-199a-3p、第二microRNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白分别为TALE蛋白、第一和第二识别序列为5×UAS的元件组成的表达系统。
CGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTTCTGTCCTCCGAGCGGAGACTCTAGATACCCCTCTCCGCTTCTGAATTCTAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCCTCGAGTACCCCTCTCCGCTTCTTACCCCTCTCCGCTTCTTACCCCTCTCCGCTTCTGAATTCGACCGCCACCATGAGACATATTATCTGCCACGGAGGTGTTATTACCGAAGAAATGGCCGCCAGTCTTTTGGACCAGCTGATCGAAGAGGTACTGGCTGATAATCTTCCACCTCCTAGCCATTTTGAACCACCTACCCTTCACGAACTGTATGATTTAGACGTGACGGCCCCCGAAGATCCCAACGAGGAGGCGGTTTCGCAGATTTTTCCCGACTCTGTAATGTTGGCGGTGCAGGAAGGGATTGACTTACTCACTTTTCCGCCGGCGCCCGGTTCTCCGGAGCCGCCTCACCTTTCCCGGCAGCCCGAGCAGCCGGAGCAGAGAGCCTTGGGTCCGGTTTCTATGCCAAACCTTGTACCGGAGGTGATCGATCTTACCTGCCACGAGGCTGGCTTTCCACCCAGTGACGACGAGGATGAAGAGGGTGAGGAGTTTGTGTTAGATTATGTGGAGCACCCCGGGCACGGTTGCAGGTCTTGTCATTATCACCGGAGGAATACGGGGGACCCAGATATTATGTGTTCGCTTTGCTATATGAGGACCTGTGGCATGTTTGTCTACAGTAAGTGAAAATTATGGGCAGTGGGTGATAGAGTGGTGGGTTTGGTGTGGTAATTTTTTTTTTAATTTTTACAGTTTTGTGGTTTAAAGAATTTTGTATTGTGATTTTTTTAAAAGGTCCTGTGTCTGAACCTGAGCCTGAGCCCGAGCCAGAACCGGAGCCTGCAAGACCTACCCGCCGTCCTAAAATGGCGCCTGCTATCCTGAGGCGCCCGACATCACCTGTGTCTAGAGAATGCAATAGTAGTACGGATAGCTGTGACTCCGGTCCTTCTAACACACCTCCTGAGATACACCCGGTGGTCCCGCTGTGCCCCATTAAACCAGTTGCCGTGAGAGTTGGTGGGCGTCGCCAGGCTGTGGAATGTATCGAGGACTTGCTTAACGAGCCTGGGCAACCTTTGGACTTGAGCTGTAAACGCCCCAGGCCAGGGTCCGGAGCTACAAATTTTTCCCTCCTCAAACAGGCTGGAGATGTCGAAGAAAATCCTGGGCCTATGGAGGCTTGGGAGTGTTTGGAAGATTTTTCTGCTGTGCGTAACTTGCTGGAACAGAGCTCTAACAGTACCTCTTGGTTTTGGAGGTTTCTGTGGGGCTCATCCCAGGCAAAGTTAGTCTGCAGAATTAAGGAGGATTACAAGTGGGAATTTGAAGAGCTTTTGAAATCCTGTGGTGAGCTGTTTGATTCTTTGAATCTGGGTCACCAGGCGCTTTTCCAAGAGAAGGTCATCAAGACTTTGGATTTTTCCACACCGGGGCGCGCTGCGGCTGCTGTTGCTTTTTTGAGTTTTATAAAGGATAAATGGAGCGAAGAAACCCATCTGAGCGGGGGGTACCTGCTGGATTTTCTGGCCATGCATCTGTGGAGAGCGGTTGTGAGACACAAGAATCGCCTGCTACTGTTGTCTTCCGTCCGCCCGGCGATAATACCGACGGAGGAGCAGCAGCAGCAGCAGGAGGAAGCCAGGCGGCGGCGGCAGGAGCAGAGCCCATGGAACCCGAGAGCCGGCCTGGACCCTCGGGAATGAATGTTGTACAGGTGGCTGAACTGTATCCAGAACTGAGACGCATTTTGACAATTACAGAGGATGGGCAGGGGCTAAAGGGGGTAAAGAGGGAGCGGGGGGCTTGTGAGGCTACAGAGGAGGCTAGGAATCTAGCTTTTAGCTTAATGACCAGACACCGTCCTGAGTGTATTACTTTTCAACAGATCAAGGATAATTGCGCTAATGAGCTTGATCTGCTGGCGCAGAAGTATTCCATAGAGCAGCTGACCACTTACTGGCTGCAGCCAGGGGATGATTTTGAGGAGGCTATTAGGGTATATGCAAAGGTGGCACTTAGGCCAGATTGCAAGTACAAGATCAGCAAACTTGTAAATATCAGGAATTGTTGCTACATTTCTGGGAACGGGGCCGAGGTGGAGATAGATACGGAGGATAGGGTGGCCTTTAGATGTAGCATGATAAATATGTGGCCGGGGGTGCTTGGCATGGACGGGGTGGTTATTATGAATGTAAGGTTTACTGGCCCCAATTTTAGCGGTACGGTTTTCCTGGCCAATACCAACCTTATCCTACACGGTGTAAGCTTCTATGGGTTTAACAATACCTGTGTGGAAGCCTGGACCGATGTAAGGGTTCGGGGCTGTGCCTTTTACTGCTGCTGGAAGGGGGTGGTGTGTCGCCCCAAAAGCAGGGCTTCAATTAAGAAATGCCTCTTTGAAAGGTGTACCTTGGGTATCCTGTCTGAGGGTAACTCCAGGGTGCGCCACAATGTGGCCTCCGACTGTGGTTGCTTCATGCTAGTGAAAAGCGTGGCTGTGATTAAGCATAACATGGTATGTGGCAACTGCGAGGACAGGGCCTCTCAGATGCTGACCTGCTCGGACGGCAACTGTCACCTGCTGAAGACCATTCACGTAGCCAGCCACTCTCGCAAGGCCTGGCCAGTGTTTGAGCATAACATACTGACCCGCTGTTCCTTGCATTTGGGTAACAGGAGGGGGGTGTTCCTACCTTACCAATGCAATTTGAGTCACACTAAGATATTGCTTGAGCCCGAGAGCATGTCCAAGGTGAACCTGAACGGGGTGTTTGACATGACCATGAAGATCTGGAAGGTGCTGAGGTACGATGAGACCCGCACCAGGTGCAGACCCTGCGAGTGTGGCGGTAAACATATTAGGAACCAGCCTGTGATGCTGGATGTGACCGAGGAGCTGAGGCCCGATCACTTGGTGCTGGCCTGCACCCGCGCTGAGTTTGGCTCTAGCGATGAAGATACAGATCTCGAGGGGTCCGGAGCTACAAATTTTTCCCTCCTCAAACAGGCTGGAGATGTCGAAGAAAATCCTGGGCCTACCGGTATGTATAGGATGCAGTTGCTCTCTTGCATCGCCCTGTCTCTAGCGTTGGTGACTAATAGCGCCCCTACTTCTTCTTCCACGAAGAAAACCCAGCTGCAACTGGAACACCTTCTGCTGGATCTCCAGATGATCCTTAACGGGATTAACAACTATAAGAACCCGAAGCTCACGGCTATGCTGACGGCGAAGTTCGCTATGCCTAAAAAGGCTACTGAGCTGAAACACTTACAGTGTCTCGAAGAGGCACTCAAACCCCTCGAGGAGGTACTCAATCTGGCGCAAAGCAAAAACTTCCATCTCCGCCCACGGGACCTAATCTCCAACATCAACGTGATTGTGCTGGAGCTCAAGGGGTCTGAAACCACGTTTATGTGCGAGTACGCAGATGAAACTGCTACCATAGTGGAATTCCTCAATCGTTGGATCACTTTCTGCCAGTCAATCATTTCAACCCTGACCAGATCTGAGGGCCGCGGCAGCCTGCTGACCTGCGGCGACGTGGAGGAAAACCCCGGCCCCGGATCCTTGTCGACCATTCGTCCGCGCAGGCCAAGTCCTGCCCGCGAGCTTCTGCCCGGACCCCAACCGGATAGGGTTCAGCCGACTGCAGATCGTGGGGTGTCTGCGCCTGCTGGCAGCCCTCTGGATGGCTTGCCCGCTCGGCGGACGGTGTCCCGGACCCGGCTGCCATCTCCCCCTGCGCCCTCACCTGCGTTCTCGGCGGGCAGCTTCAGCGATCTGCTCCGTCCGTTCGATCCGTCGCTTCTTGATACATCGCTTCTTGATTCGATGCCTGCCGTCGGCACGCCGCATACAGCGGCTGCCCCAGCAGAGTGGGATGAGGCGCAATCGGCTCTGCGTGCAGCCGATGACCCGCCACCCACCGTGCGTGTCGCTGTCACTGCCGCGCGGCCGCCGCGCGCCAAGCCGGCCCCGCGACGGCGTGCTGCGCAACCCTCCGACGCTTCGCCGGCCGCGCAGGTGGATCTACGCACGCTCGGCTACAGTCAGCAGCAGCAAGAGAAGATCAAACCGAAGGTGCGTTCGACAGTGGCGCAGCACCACGAGGCACTGGTGGGCCATGGGTTTACACACGCGCACATCGTTGCGCTCAGCCAACACCCGGCAGCGTTAGGGACCGTCGCTGTCACGTATCAGCACATAATCACGGCGTTGCCAGAGGCGACACACGAAGACATCGTTGGCGTCGGCAAACAGTGGTCCGGCGCACGCGCCCTGGAGGCCTTGCTCACGGATGCGGGGGAGTTGAGAGGTCCGCCGTTACAGTTGGACACAGGCCAACTTGTGAAGATTGCAAAACGTGGCGGCGTGACCGCAATGGAGGCAGTGCATGCATCGCGCAATGCACTGACGGGTGCCCCCCTGAACCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACAATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACAATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAAGCATTGTGGCCCAGCTGAGCCGGCCTGATCCGGCGTTGGCCGCGTTGACCAACGACCACCTCGTCGCCTTGGCCTGCCTCGGCGGACGTCCTGCCATGGATGCAGTGAAAAAGGGATTGCCGCACGCGCCGGAATTGATCAGAAGAGTCAATCGCCGTATTGGCGAACGCACGTCCCATCGCGTTGCCGACTACGCGCAAGTGGTTCGCGTGCTGGAGTTTTTCCAGTGCCACTCCCACCCAGCGTACGCATTTGATGAGGCCATGACGCAGTTCGGGATGAGCAGGAACGGGTTGGTACAGCTCTTTCGCAGAGTGGGCGTCACCGAACTCGAAGCCCGCGGTGGAACGCTCCCCCCAGCCTCGCAGCGTTGGGACCGTATCCTCCAGGCATCAGGGATGAAAAGGGCCAAACCGTCCCCTACTTCAGCTCAAACACCGGATCAGGCGTCTTTGCATGCATTCGCCGATTCGCTGGAGCGTGACCTTGATGCGCCCAGCCCAATGCACGAGGGAGATCAGACGCGGGCAAGCAGCCGTAAACGGTCCCGATCGGATCGTGCTGTCACCGGCCCCTCCGCACAGCAGGCTGTCGAGGTGCGCGTTCCCGAACAGCGCGATGCGCTGCATTTGCCCCTCAGCTGGAGGGTAAAACGCCCGCGTACCAGGATCTGGGGCGGCCTCCCGGTTAAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGTAACCAATGTGCAGACTACTGTTAACCAATGTGCAGACTACTGTTAACCAATGTGCAGACTACTGTTAACCAATGTGCAGACTACTGTGAAAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTTCTGTCCTCCGAGCGGAGACTCTAGATATAGAACCGATCCTCCCATTGAATTCTAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCCTCGAGTATAGAACCGATCCTCCCATTTATAGAACCGATCCTCCCATTTATAGAACCGATCCTCCCATTACTAGAAGATCTTGAATTCGCCACCTTGTCGACCATTCGTCCGCGCAGGCCAAGTCCTGCCCGCGAGCTTCTGCCCGGACCCCAACCGGATAGGGTTCAGCCGACTGCAGATCGTGGGGTGTCTGCGCCTGCTGGCAGCCCTCTGGATGGCTTGCCCGCTCGGCGGACGGTGTCCCGGACCCGGCTGCCATCTCCCCCTGCGCCCTCACCTGCGTTCTCGGCGGGCAGCTTCAGCGATCTGCTCCGTCCGTTCGATCCGTCGCTTCTTGATACATCGCTTCTTGATTCGATGCCTGCCGTCGGCACGCCGCATACAGCGGCTGCCCCAGCAGAGTGGGATGAGGCGCAATCGGCTCTGCGTGCAGCCGATGACCCGCCACCCACCGTGCGTGTCGCTGTCACTGCCGCGCGGCCGCCGCGCGCCAAGCCGGCCCCGCGACGGCGTGCTGCGCAACCCTCCGACGCTTCGCCGGCCGCGCAGGTGGATCTACGCACGCTCGGCTACAGTCAGCAGCAGCAAGAGAAGATCAAACCGAAGGTGCGTTCGACAGTGGCGCAGCACCACGAGGCACTGGTGGGCCATGGGTTTACACACGCGCACATCGTTGCGCTCAGCCAACACCCGGCAGCGTTAGGGACCGTCGCTGTCACGTATCAGCACATAATCACGGCGTTGCCAGAGGCGACACACGAAGACATCGTTGGCGTCGGCAAACAGTGGTCCGGCGCACGCGCCCTGGAGGCCTTGCTCACGGATGCGGGGGAGTTGAGAGGTCCGCCGTTACAGTTGGACACAGGCCAACTTGTGAAGATTGCAAAACGTGGCGGCGTGACCGCAATGGAGGCAGTGCATGCATCGCGCAATGCACTGACGGGTGCCCCCCTGAACCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACAATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAAGCATTGTGGCCCAGCTGAGCCGGCCTGATCCGGCGTTGGCCGCGTTGACCAACGACCACCTCGTCGCCTTGGCCTGCCTCGGCGGACGTCCTGCCATGGATGCAGTGAAAAAGGGATTGCCGCACGCGCCGGAATTGATCAGAAGAGTCAATCGCCGTATTGGCGAACGCACGTCCCATCGCGTTGCCGACTACGCGCAAGTGGTTCGCGTGCTGGAGTTTTTCCAGTGCCACTCCCACCCAGCGTACGCATTTGATGAGGCCATGACGCAGTTCGGGATGAGCAGGAACGGGTTGGTACAGCTCTTTCGCAGAGTGGGCGTCACCGAACTCGAAGCCCGCGGTGGAACGCTCCCCCCAGCCTCGCAGCGTTGGGACCGTATCCTCCAGGCATCAGGGATGAAAAGGGCCAAACCGTCCCCTACTTCAGCTCAAACACCGGATCAGGCGTCTTTGCATGCATTCGCCGATTCGCTGGAGCGTGACCTTGATGCGCCCAGCCCAATGCACGAGGGAGATCAGACGCGGGCAAGCAGCCGTAAACGGTCCCGATCGGATCGTGCTGTCACCGGCCCCTCCGCACAGCAGGCTGTCGAGGTGCGCGTTCCCGAACAGCGCGATGCGCTGCATTTGCCCCTCAGCTGGAGGGTAAAACGCCCGCGTACCAGGATCTGGGGCGGCCTCCCGTAATACAAGGCGGCCGCAAATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTAAAGCTTCGAATTCTGATAATCAGCCATACCACATTTGTAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCGTTCATCGGAGCCCAGGTTTACTCCCTTAAGTGGAAATTTCTTCCCCCACTCCCTCCTTGGCTTTCTCCAAGGAGGGAACCCAGGCTACTGGAAAGTCCGGCTGGGGCGGGGACTGTGGGTTTCAGGGTAGAACTGCGTGTGGAACGGGACAGGGAGCGGTTAGAAGGGTGGGGCTATTCCGGGAAGTGGTGGGGGGAGGGAGCCCAAAACTAGCACCTAGTCCACTCATTATCCAGCCCTCTTATTTCTCGGCCCCGCTCTGCTTCAGTGGACCCGGGGAGGGCGGGGAAGTGGAGTGGGAGACCTAGGGGTGGGCTTCCCGACCTTGCTGTACAGGACCTCGACCTAGCTGGCTTTGTTCCCCATCCCCACGTTAGTTGTTGCCCTGAGGCTAAAACTAGAGCCCAGGGGCCCCAAGTTCCAGACTGCCCCTCCCCCCTCCCCCGGAGCCAGGGAGTGGTTGGTGAAAGGGGGAGGCCAGCTGGAGAACAAACGGGTAGTCAGGGGGTTGAGCGATTAGAGCCCTTGTACCCTACCCAGGAATGGTTGGGGAGGAGGAGGAAGAGGTAGGAGGTAGGGGAGGGGGCGGGGTTTTGTCACCTGTCACCTGCTCCGGCTGTGCCTAGGGCGGGCGGGCGGGGAGTGGGGGGACCGGTATAAAGCGGTAGGCGCCTGTGCCCGCTCCACCTCTCAAGCAGCCAGCGCCTGCCTGAATCTGTTCTGCCCCCTCCCCACCCATTTCACCACCACCATGGCCCCCCCGACCGATGTCAGCCTGGGGGACGAGCTCCACTTAGACGGCGAGGACGTGGCGATGGCGCATGCCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGAGCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGAGCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGAGCGACGCGCTAGACGATTTCGATCTGGACATGTTGGGGGACGGGGATTCCCCGGGTCCGGGATTTACCCCCCACGACTCCGCCCCCTACGGCGCTCTGGATATGGCCGACTTCGAGTTTGAGCAGATGTTTACCGATGCCCTTGGAATTGACGAGTACGGTGGGACGCGTATGAAGCTACTGTCTTCTATCGAACAAGCATGCGATATTTGCCGACTTAAAAAGCTCAAGTGCTCCAAAGAAAAACCGAAGTGCGCCAAGTGTCTGAAGAACAACTGGGAGTGTCGCTACTCTCCCAAAACCAAAAGGTCTCCGCTGACTAGGGCACATCTGACAGAAGTGGAATCAAGGCTAGAAAGACTGGAACAGCTATTTCTACTGATTTTTCCTCGAGAAGACCTTGACATGATTTTGAAAATGGATTCTTTACAGGATATAAAAGCATTGTTAACAGGATTATTTGTACAAGATAATGTGAATAAAGATGCCGTCACAGATAGATTGGCTTCAGTGGAGACTGATATGCCTCTAACATTGAGACAGCATAGAATAAGTGCGACATCATCATCGGAAGAGAGTAGTAACAAAGGTCAAAGACAGTTGACTGTATAAAAGCTTCACTCCTCAGGTGCAGGCTGCCTATCAGAAGGTGGTGGCTGGTGTGGCCAATGCCCTGGCTCACAAATACCACTGAGATCTTTTTCCCTCTGCCAAAAATTATGGGGACATCATGAAGCCCCTTGAGCATCTGACTTCTGGCTAATAAAGGAAATTTATTTTCATTGCAATAGTGTGTTGGAATTTTTTGTGTCTCTCACTCGGAAGGACATATGGGAGGGCAAATCATTTAAAACATCAGAATGAGTATTTGGTTTAGAGTTTGGCAACATATGCCCATATGCTGGCTGCCATGAACAAAGGTTGGCTATAAAGAGGTCATCAGTATATGAAACAGCCCCCTGCTGTCCATTCCTTATTCCATAGAAAAGCCTTGACTTGAGGTTAGATTTTTTTTATATTTTGTTTTGTGTTATTTTTTTCTTTAACATCCCTAAAATTTTCCTTACATGTTTTACTAGCCAGATTTTTCCTCCTCTCCTGACTACTCCCAGTCATAGCTGTCCCTCTTCTCTTATGGAGATC(SEQ ID NO:24)。
具有SEQ ID NO:25所述核苷酸序列的核酸编码由细胞特异性启动子hMUC1、第一microRNA为mir-199a-3p、第二microRNA为mir-21、转录激活因子为分割dCas9、第一调控蛋白和第二调控蛋白分别为TALE蛋白、第一和第二识别序列为5×UAS的元件组成的表达系统。
CGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTTCTGTCCTCCGAGCGGAGACTCTAGATACCCCTCTCCGCTTCTGAATTCTAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCCTCGAGTACCCCTCTCCGCTTCTTACCCCTCTCCGCTTCTTACCCCTCTCCGCTTCTGAATTCGACCGCCACCATGAGACATATTATCTGCCACGGAGGTGTTATTACCGAAGAAATGGCCGCCAGTCTTTTGGACCAGCTGATCGAAGAGGTACTGGCTGATAATCTTCCACCTCCTAGCCATTTTGAACCACCTACCCTTCACGAACTGTATGATTTAGACGTGACGGCCCCCGAAGATCCCAACGAGGAGGCGGTTTCGCAGATTTTTCCCGACTCTGTAATGTTGGCGGTGCAGGAAGGGATTGACTTACTCACTTTTCCGCCGGCGCCCGGTTCTCCGGAGCCGCCTCACCTTTCCCGGCAGCCCGAGCAGCCGGAGCAGAGAGCCTTGGGTCCGGTTTCTATGCCAAACCTTGTACCGGAGGTGATCGATCTTACCTGCCACGAGGCTGGCTTTCCACCCAGTGACGACGAGGATGAAGAGGGTGAGGAGTTTGTGTTAGATTATGTGGAGCACCCCGGGCACGGTTGCAGGTCTTGTCATTATCACCGGAGGAATACGGGGGACCCAGATATTATGTGTTCGCTTTGCTATATGAGGACCTGTGGCATGTTTGTCTACAGTAAGTGAAAATTATGGGCAGTGGGTGATAGAGTGGTGGGTTTGGTGTGGTAATTTTTTTTTTAATTTTTACAGTTTTGTGGTTTAAAGAATTTTGTATTGTGATTTTTTTAAAAGGTCCTGTGTCTGAACCTGAGCCTGAGCCCGAGCCAGAACCGGAGCCTGCAAGACCTACCCGCCGTCCTAAAATGGCGCCTGCTATCCTGAGGCGCCCGACATCACCTGTGTCTAGAGAATGCAATAGTAGTACGGATAGCTGTGACTCCGGTCCTTCTAACACACCTCCTGAGATACACCCGGTGGTCCCGCTGTGCCCCATTAAACCAGTTGCCGTGAGAGTTGGTGGGCGTCGCCAGGCTGTGGAATGTATCGAGGACTTGCTTAACGAGCCTGGGCAACCTTTGGACTTGAGCTGTAAACGCCCCAGGCCAGGGTCCGGAGCTACAAATTTTTCCCTCCTCAAACAGGCTGGAGATGTCGAAGAAAATCCTGGGCCTATGGAGGCTTGGGAGTGTTTGGAAGATTTTTCTGCTGTGCGTAACTTGCTGGAACAGAGCTCTAACAGTACCTCTTGGTTTTGGAGGTTTCTGTGGGGCTCATCCCAGGCAAAGTTAGTCTGCAGAATTAAGGAGGATTACAAGTGGGAATTTGAAGAGCTTTTGAAATCCTGTGGTGAGCTGTTTGATTCTTTGAATCTGGGTCACCAGGCGCTTTTCCAAGAGAAGGTCATCAAGACTTTGGATTTTTCCACACCGGGGCGCGCTGCGGCTGCTGTTGCTTTTTTGAGTTTTATAAAGGATAAATGGAGCGAAGAAACCCATCTGAGCGGGGGGTACCTGCTGGATTTTCTGGCCATGCATCTGTGGAGAGCGGTTGTGAGACACAAGAATCGCCTGCTACTGTTGTCTTCCGTCCGCCCGGCGATAATACCGACGGAGGAGCAGCAGCAGCAGCAGGAGGAAGCCAGGCGGCGGCGGCAGGAGCAGAGCCCATGGAACCCGAGAGCCGGCCTGGACCCTCGGGAATGAATGTTGTACAGGTGGCTGAACTGTATCCAGAACTGAGACGCATTTTGACAATTACAGAGGATGGGCAGGGGCTAAAGGGGGTAAAGAGGGAGCGGGGGGCTTGTGAGGCTACAGAGGAGGCTAGGAATCTAGCTTTTAGCTTAATGACCAGACACCGTCCTGAGTGTATTACTTTTCAACAGATCAAGGATAATTGCGCTAATGAGCTTGATCTGCTGGCGCAGAAGTATTCCATAGAGCAGCTGACCACTTACTGGCTGCAGCCAGGGGATGATTTTGAGGAGGCTATTAGGGTATATGCAAAGGTGGCACTTAGGCCAGATTGCAAGTACAAGATCAGCAAACTTGTAAATATCAGGAATTGTTGCTACATTTCTGGGAACGGGGCCGAGGTGGAGATAGATACGGAGGATAGGGTGGCCTTTAGATGTAGCATGATAAATATGTGGCCGGGGGTGCTTGGCATGGACGGGGTGGTTATTATGAATGTAAGGTTTACTGGCCCCAATTTTAGCGGTACGGTTTTCCTGGCCAATACCAACCTTATCCTACACGGTGTAAGCTTCTATGGGTTTAACAATACCTGTGTGGAAGCCTGGACCGATGTAAGGGTTCGGGGCTGTGCCTTTTACTGCTGCTGGAAGGGGGTGGTGTGTCGCCCCAAAAGCAGGGCTTCAATTAAGAAATGCCTCTTTGAAAGGTGTACCTTGGGTATCCTGTCTGAGGGTAACTCCAGGGTGCGCCACAATGTGGCCTCCGACTGTGGTTGCTTCATGCTAGTGAAAAGCGTGGCTGTGATTAAGCATAACATGGTATGTGGCAACTGCGAGGACAGGGCCTCTCAGATGCTGACCTGCTCGGACGGCAACTGTCACCTGCTGAAGACCATTCACGTAGCCAGCCACTCTCGCAAGGCCTGGCCAGTGTTTGAGCATAACATACTGACCCGCTGTTCCTTGCATTTGGGTAACAGGAGGGGGGTGTTCCTACCTTACCAATGCAATTTGAGTCACACTAAGATATTGCTTGAGCCCGAGAGCATGTCCAAGGTGAACCTGAACGGGGTGTTTGACATGACCATGAAGATCTGGAAGGTGCTGAGGTACGATGAGACCCGCACCAGGTGCAGACCCTGCGAGTGTGGCGGTAAACATATTAGGAACCAGCCTGTGATGCTGGATGTGACCGAGGAGCTGAGGCCCGATCACTTGGTGCTGGCCTGCACCCGCGCTGAGTTTGGCTCTAGCGATGAAGATACAGATCTCGAGGGGTCCGGAGCTACAAATTTTTCCCTCCTCAAACAGGCTGGAGATGTCGAAGAAAATCCTGGGCCTACCGGTATGTATAGGATGCAGTTGCTCTCTTGCATCGCCCTGTCTCTAGCGTTGGTGACTAATAGCGCCCCTACTTCTTCTTCCACGAAGAAAACCCAGCTGCAACTGGAACACCTTCTGCTGGATCTCCAGATGATCCTTAACGGGATTAACAACTATAAGAACCCGAAGCTCACGGCTATGCTGACGGCGAAGTTCGCTATGCCTAAAAAGGCTACTGAGCTGAAACACTTACAGTGTCTCGAAGAGGCACTCAAACCCCTCGAGGAGGTACTCAATCTGGCGCAAAGCAAAAACTTCCATCTCCGCCCACGGGACCTAATCTCCAACATCAACGTGATTGTGCTGGAGCTCAAGGGGTCTGAAACCACGTTTATGTGCGAGTACGCAGATGAAACTGCTACCATAGTGGAATTCCTCAATCGTTGGATCACTTTCTGCCAGTCAATCATTTCAACCCTGACCAGATCTGAGGGCCGCGGCAGCCTGCTGACCTGCGGCGACGTGGAGGAAAACCCCGGCCCCGGATCCTTGTCGACCATTCGTCCGCGCAGGCCAAGTCCTGCCCGCGAGCTTCTGCCCGGACCCCAACCGGATAGGGTTCAGCCGACTGCAGATCGTGGGGTGTCTGCGCCTGCTGGCAGCCCTCTGGATGGCTTGCCCGCTCGGCGGACGGTGTCCCGGACCCGGCTGCCATCTCCCCCTGCGCCCTCACCTGCGTTCTCGGCGGGCAGCTTCAGCGATCTGCTCCGTCCGTTCGATCCGTCGCTTCTTGATACATCGCTTCTTGATTCGATGCCTGCCGTCGGCACGCCGCATACAGCGGCTGCCCCAGCAGAGTGGGATGAGGCGCAATCGGCTCTGCGTGCAGCCGATGACCCGCCACCCACCGTGCGTGTCGCTGTCACTGCCGCGCGGCCGCCGCGCGCCAAGCCGGCCCCGCGACGGCGTGCTGCGCAACCCTCCGACGCTTCGCCGGCCGCGCAGGTGGATCTACGCACGCTCGGCTACAGTCAGCAGCAGCAAGAGAAGATCAAACCGAAGGTGCGTTCGACAGTGGCGCAGCACCACGAGGCACTGGTGGGCCATGGGTTTACACACGCGCACATCGTTGCGCTCAGCCAACACCCGGCAGCGTTAGGGACCGTCGCTGTCACGTATCAGCACATAATCACGGCGTTGCCAGAGGCGACACACGAAGACATCGTTGGCGTCGGCAAACAGTGGTCCGGCGCACGCGCCCTGGAGGCCTTGCTCACGGATGCGGGGGAGTTGAGAGGTCCGCCGTTACAGTTGGACACAGGCCAACTTGTGAAGATTGCAAAACGTGGCGGCGTGACCGCAATGGAGGCAGTGCATGCATCGCGCAATGCACTGACGGGTGCCCCCCTGAACCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACAATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACAATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAAGCATTGTGGCCCAGCTGAGCCGGCCTGATCCGGCGTTGGCCGCGTTGACCAACGACCACCTCGTCGCCTTGGCCTGCCTCGGCGGACGTCCTGCCATGGATGCAGTGAAAAAGGGATTGCCGCACGCGCCGGAATTGATCAGAAGAGTCAATCGCCGTATTGGCGAACGCACGTCCCATCGCGTTGCCGACTACGCGCAAGTGGTTCGCGTGCTGGAGTTTTTCCAGTGCCACTCCCACCCAGCGTACGCATTTGATGAGGCCATGACGCAGTTCGGGATGAGCAGGAACGGGTTGGTACAGCTCTTTCGCAGAGTGGGCGTCACCGAACTCGAAGCCCGCGGTGGAACGCTCCCCCCAGCCTCGCAGCGTTGGGACCGTATCCTCCAGGCATCAGGGATGAAAAGGGCCAAACCGTCCCCTACTTCAGCTCAAACACCGGATCAGGCGTCTTTGCATGCATTCGCCGATTCGCTGGAGCGTGACCTTGATGCGCCCAGCCCAATGCACGAGGGAGATCAGACGCGGGCAAGCAGCCGTAAACGGTCCCGATCGGATCGTGCTGTCACCGGCCCCTCCGCACAGCAGGCTGTCGAGGTGCGCGTTCCCGAACAGCGCGATGCGCTGCATTTGCCCCTCAGCTGGAGGGTAAAACGCCCGCGTACCAGGATCTGGGGCGGCCTCCCGGTTAAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGTAACCAATGTGCAGACTACTGTTAACCAATGTGCAGACTACTGTTAACCAATGTGCAGACTACTGTTAACCAATGTGCAGACTACTGTGAAAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTTCTGTCCTCCGAGCGGAGACTCTAGATATAGAACCGATCCTCCCATTGAATTCTAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCCTCGAGTATAGAACCGATCCTCCCATTTATAGAACCGATCCTCCCATTTATAGAACCGATCCTCCCATTACTAGAAGATCTTGAATTCGCCACCTTGTCGACCATTCGTCCGCGCAGGCCAAGTCCTGCCCGCGAGCTTCTGCCCGGACCCCAACCGGATAGGGTTCAGCCGACTGCAGATCGTGGGGTGTCTGCGCCTGCTGGCAGCCCTCTGGATGGCTTGCCCGCTCGGCGGACGGTGTCCCGGACCCGGCTGCCATCTCCCCCTGCGCCCTCACCTGCGTTCTCGGCGGGCAGCTTCAGCGATCTGCTCCGTCCGTTCGATCCGTCGCTTCTTGATACATCGCTTCTTGATTCGATGCCTGCCGTCGGCACGCCGCATACAGCGGCTGCCCCAGCAGAGTGGGATGAGGCGCAATCGGCTCTGCGTGCAGCCGATGACCCGCCACCCACCGTGCGTGTCGCTGTCACTGCCGCGCGGCCGCCGCGCGCCAAGCCGGCCCCGCGACGGCGTGCTGCGCAACCCTCCGACGCTTCGCCGGCCGCGCAGGTGGATCTACGCACGCTCGGCTACAGTCAGCAGCAGCAAGAGAAGATCAAACCGAAGGTGCGTTCGACAGTGGCGCAGCACCACGAGGCACTGGTGGGCCATGGGTTTACACACGCGCACATCGTTGCGCTCAGCCAACACCCGGCAGCGTTAGGGACCGTCGCTGTCACGTATCAGCACATAATCACGGCGTTGCCAGAGGCGACACACGAAGACATCGTTGGCGTCGGCAAACAGTGGTCCGGCGCACGCGCCCTGGAGGCCTTGCTCACGGATGCGGGGGAGTTGAGAGGTCCGCCGTTACAGTTGGACACAGGCCAACTTGTGAAGATTGCAAAACGTGGCGGCGTGACCGCAATGGAGGCAGTGCATGCATCGCGCAATGCACTGACGGGTGCCCCCCTGAACCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACAATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAAGCATTGTGGCCCAGCTGAGCCGGCCTGATCCGGCGTTGGCCGCGTTGACCAACGACCACCTCGTCGCCTTGGCCTGCCTCGGCGGACGTCCTGCCATGGATGCAGTGAAAAAGGGATTGCCGCACGCGCCGGAATTGATCAGAAGAGTCAATCGCCGTATTGGCGAACGCACGTCCCATCGCGTTGCCGACTACGCGCAAGTGGTTCGCGTGCTGGAGTTTTTCCAGTGCCACTCCCACCCAGCGTACGCATTTGATGAGGCCATGACGCAGTTCGGGATGAGCAGGAACGGGTTGGTACAGCTCTTTCGCAGAGTGGGCGTCACCGAACTCGAAGCCCGCGGTGGAACGCTCCCCCCAGCCTCGCAGCGTTGGGACCGTATCCTCCAGGCATCAGGGATGAAAAGGGCCAAACCGTCCCCTACTTCAGCTCAAACACCGGATCAGGCGTCTTTGCATGCATTCGCCGATTCGCTGGAGCGTGACCTTGATGCGCCCAGCCCAATGCACGAGGGAGATCAGACGCGGGCAAGCAGCCGTAAACGGTCCCGATCGGATCGTGCTGTCACCGGCCCCTCCGCACAGCAGGCTGTCGAGGTGCGCGTTCCCGAACAGCGCGATGCGCTGCATTTGCCCCTCAGCTGGAGGGTAAAACGCCCGCGTACCAGGATCTGGGGCGGCCTCCCGTAATACAAGGCGGCCGCAAATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTAAAGCTTCGAATTCTGATAATCAGCCATACCACATTTGTAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCGTTCATCGGAGCCCAGGTTTACTCCCTTAAGTGGAAATTTCTTCCCCCACTCCCTCCTTGGCTTTCTCCAAGGAGGGAACCCAGGCTACTGGAAAGTCCGGCTGGGGCGGGGACTGTGGGTTTCAGGGTAGAACTGCGTGTGGAACGGGACAGGGAGCGGTTAGAAGGGTGGGGCTATTCCGGGAAGTGGTGGGGGGAGGGAGCCCAAAACTAGCACCTAGTCCACTCATTATCCAGCCCTCTTATTTCTCGGCCCCGCTCTGCTTCAGTGGACCCGGGGAGGGCGGGGAAGTGGAGTGGGAGACCTAGGGGTGGGCTTCCCGACCTTGCTGTACAGGACCTCGACCTAGCTGGCTTTGTTCCCCATCCCCACGTTAGTTGTTGCCCTGAGGCTAAAACTAGAGCCCAGGGGCCCCAAGTTCCAGACTGCCCCTCCCCCCTCCCCCGGAGCCAGGGAGTGGTTGGTGAAAGGGGGAGGCCAGCTGGAGAACAAACGGGTAGTCAGGGGGTTGAGCGATTAGAGCCCTTGTACCCTACCCAGGAATGGTTGGGGAGGAGGAGGAAGAGGTAGGAGGTAGGGGAGGGGGCGGGGTTTTGTCACCTGTCACCTGCTCCGGCTGTGCCTAGGGCGGGCGGGCGGGGAGTGGGGGGACCGGTATAAAGCGGTAGGCGCCTGTGCCCGCTCCACCTCTCAAGCAGCCAGCGCCTGCCTGAATCTGTTCTGCCCCCTCCCCACCCATTTCACCACCACCATGCCAAAGAAGAAGCGGAAGGTCGGTGGCGGCTCACCCGGGGGTGGCGGCTCAATGGACAAGAAGTACTCCATTGGCCTCGCCATCGGAACAAATAGCGTGGGCTGGGCTGTCATCACAGATGAGTACAAGGTGCCTAGCAAGAAATTTAAGGTGCTGGGAAATACAGACAGACATAGCATCAAGAAGAACCTCATTGGCGCTCTCCTGTTTGACTCCGGCGAAACAGCCGAAGCTACCAGACTCAAGAGAACCGCTAGGAGAAGGTACACCAGAAGGAAAAACAGGATTTGCTACCTGCAGGAAATTTTTTCCAACGAGATGGCCAAGGTGGACGATTCCTTCTTCCATAGGCTGGAAGAGAGCTTCCTCGTGGAGGAAGACAAGAAACACGAGAGGCATCCTATTTTTGGCAATATTGTGGATGAGGTCGCCTACCATGAGAAGTATCCCACAATCTATCATCTGAGAAAAAAACTGGTGGATAGCACCGACAAGGCCGATCTCAGGCTCATTTATCTCGCTCTGGCTCACATGATCAAGTTTAGGGGCCACTTCCTGATCGAAGGCGACCTGAATCCCGACAACTCCGACGTGGACAAACTGTTCATCCAGCTCGTCCAGACCTACAATCAACTCTTCGAGGAGAACCCCATCAATGCTTCCGGCGTGGATGCCAAGGCCATCCTGAGCGCTAGGCTCTCCAAGTCCAGGAGGCTGGAAAATCTGATCGCCCAACTCCCTGGAGAGAAGAAGAACGGCCTGTTTGGCAATCTGATTGCCCTGAGCCTCGGACTCACCCCCAACTTCAAGAGCAACTTCGATCTCGCCGAAGACGCCAAACTCCAACTGAGCAAGGATACCTACGACGACGATCTCGATAATCTCCTCGCCCAGATCGGCGATCAATATGCCGACCTCTTTCTGGCCGCCAAAAACCTGAGCGACGCTATTCTGCTCAGCGACATTCTCAGGGTGAATACAGAAATCACAAAAGCCCCCCTGTCCGCCAGCATGATCAAAAGGTACGATGAACACCATCAGGACCTCACCCTGCTGAAGGCTCTGGTCAGGCAGCAACTCCCCGAAAAGTACAAGGAGATTTTCTTTGATCAGTCCAAGAATGGATATGCTGGCTATATTGATGGAGGCGCCTCCCAGGAGGAATTTTATAAATTCATCAAGCCCATTCTCGAAAAGATGGACGGAACCGAAGAGCTGCTGGTCAAACTCAATAGGGAGGATCTGCTGAGGAAGCAAAGGACCTTCGACAATGGCAGCATCCCCCACCAGATCCACCTCGGCGAACTCCACGCTATCCTCAGGAGGCAGGAAGACTTCTACCCTTTCCTGAAGGATAACAGGGAGAAAATCGAGAAAATCCTGACCTTCAGAATCCCCTACTACGTCGGACCTCTCGCCAGGGGCAATTCCAGATTCGCCTGGATGACAAGGAAGAGCGAGGAAACAATCACACCATGGAACTTCGAAGAAGTGGTCGATAAGGGCGCCAGCGCCCAGAGCTTCATTGAAAGGATGACCAACTTTGATAAGAACCTGCCCAATGAGAAGGTGCTGCCTAAGCACTCCCTGCTGTATGAGTATTTCACCGTGTATAATGAGCTGACCAAGGTCAAGTACGTCACCGAGGGAATGAGAAAGCCTGCTTTTCTCTCCGGCGAGCAGAAAAAAGCCATCGTGGACCTGCTGTTCAAGACCAACAGGAAGGTGACCGTCAAGCAACTCAAGGAGGACTACTTTAAGAAGATTGAGTGCTTTGATAGCGTGGAAATTAGCGGAGTCGAGGACAGGTTCAATGCCTCCCTCGGAACATATCACGACCTGCTGAAAATCATCAAAGACAAAGATTTTCTGGATAACGAGGAGAATGAAGACATTCTGGAGGACATTGTCCTCACCCTGACCCTGTTTGAGGACAGAGAGATGATTGAAGAGAGGCTGAAAACCTATGCCCACCTGTTCGACGACAAGGTGATGAAGCAGCTCAAAAGAAGGAGATATACCGGCTGGGGCAGACTGTCCAGGAAGCTGATCAACGGCATTAGGGACAAGCAGAGCGGCAAGACCATTCTCGACTTTCTCAAGTCCGACGGATTCGCCAACAGAAACTTTATGCAACTGATCCACGATGACAGCCTCACCTTTAAGGAAGATATTCAGAAGGCTCAGGTCTGCCTGGACTTGAAAACCCAAGTCCAGACCCCCCAAGGCATGAAGGAGATCAGCAATATACAAGTGGGCGACCTGGTGCTGTCCAACACCGGCTACAACGAGGTGTTGAATGTTTTCCCCAAGAGCAAGAAAAAAAGCTACAAGATCACTTTGGAGGACGGAAAGGAAATCATCTGCTCCGAGGAACACCTGTTCCCCACCCAGACCGGCGAGATGAACATCTCTGGTGGACTGAAAGAGGGCATGTGCCTGTACGTCAAAGAGACTAGTGGTGGCGGCTCAAAGCGTCCTGCTGCTACTAAGAAAGCTGGTCAAGCTAAGAAAAAGAAATCTAGATAGAAGCTTCACTCCTCAGGTGCAGGCTGCCTATCAGAAGGTGGTGGCTGGTGTGGCCAATGCCCTGGCTCACAAATACCACTGAGATCTTTTTCCCTCTGCCAAAAATTATGGGGACATCATGAAGCCCCTTGAGCATCTGACTTCTGGCTAATAAAGGAAATTTATTTTCATTGCAATAGTGTGTTGGAATTTTTTGTGTCTCTCACTCGGAAGGACATATGGGAGGGCAAATCATTTAAAACATCAGAATGAGTATTTGGTTTAGAGTTTGGCAACATATGCCCATATGCTGGCTGCCATGAACAAAGGTTGGCTATAAAGAGGTCATCAGTATATGAAACAGCCCCCTGCTGTCCATTCCTTATTCCATAGAAAAGCCTTGACTTGAGGTTAGATTTTTTTTATATTTTGTTTTGTGTTATTTTTTTCTTTAACATCCCTAAAATTTTCCTTACATGTTTTACTAGCCAGATTTTTCCTCCTCTCCTGACTACTCCCAGTCATAGCTGTCCCTCTTCTCTTATGGAGATC(SEQ ID NO:25)。
具有SEQ ID NO:26所述核苷酸序列的核酸编码由细胞特异性启动子hMUC1、第一microRNA为mir-199a-3p、第二microRNA为mir-21、转录激活因子为分割dCas9、第一调控蛋白和第二调控蛋白分别为TALE蛋白、第一和第二识别序列为5×UAS的元件组成的表达系统。
CGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTTCTGTCCTCCGAGCGGAGACTCTAGATACCCCTCTCCGCTTCTGAATTCTAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCCTCGAGTACCCCTCTCCGCTTCTTACCCCTCTCCGCTTCTTACCCCTCTCCGCTTCTGAATTCGACCGCCACCATGAGACATATTATCTGCCACGGAGGTGTTATTACCGAAGAAATGGCCGCCAGTCTTTTGGACCAGCTGATCGAAGAGGTACTGGCTGATAATCTTCCACCTCCTAGCCATTTTGAACCACCTACCCTTCACGAACTGTATGATTTAGACGTGACGGCCCCCGAAGATCCCAACGAGGAGGCGGTTTCGCAGATTTTTCCCGACTCTGTAATGTTGGCGGTGCAGGAAGGGATTGACTTACTCACTTTTCCGCCGGCGCCCGGTTCTCCGGAGCCGCCTCACCTTTCCCGGCAGCCCGAGCAGCCGGAGCAGAGAGCCTTGGGTCCGGTTTCTATGCCAAACCTTGTACCGGAGGTGATCGATCTTACCTGCCACGAGGCTGGCTTTCCACCCAGTGACGACGAGGATGAAGAGGGTGAGGAGTTTGTGTTAGATTATGTGGAGCACCCCGGGCACGGTTGCAGGTCTTGTCATTATCACCGGAGGAATACGGGGGACCCAGATATTATGTGTTCGCTTTGCTATATGAGGACCTGTGGCATGTTTGTCTACAGTAAGTGAAAATTATGGGCAGTGGGTGATAGAGTGGTGGGTTTGGTGTGGTAATTTTTTTTTTAATTTTTACAGTTTTGTGGTTTAAAGAATTTTGTATTGTGATTTTTTTAAAAGGTCCTGTGTCTGAACCTGAGCCTGAGCCCGAGCCAGAACCGGAGCCTGCAAGACCTACCCGCCGTCCTAAAATGGCGCCTGCTATCCTGAGGCGCCCGACATCACCTGTGTCTAGAGAATGCAATAGTAGTACGGATAGCTGTGACTCCGGTCCTTCTAACACACCTCCTGAGATACACCCGGTGGTCCCGCTGTGCCCCATTAAACCAGTTGCCGTGAGAGTTGGTGGGCGTCGCCAGGCTGTGGAATGTATCGAGGACTTGCTTAACGAGCCTGGGCAACCTTTGGACTTGAGCTGTAAACGCCCCAGGCCAGGGTCCGGAGCTACAAATTTTTCCCTCCTCAAACAGGCTGGAGATGTCGAAGAAAATCCTGGGCCTATGGAGGCTTGGGAGTGTTTGGAAGATTTTTCTGCTGTGCGTAACTTGCTGGAACAGAGCTCTAACAGTACCTCTTGGTTTTGGAGGTTTCTGTGGGGCTCATCCCAGGCAAAGTTAGTCTGCAGAATTAAGGAGGATTACAAGTGGGAATTTGAAGAGCTTTTGAAATCCTGTGGTGAGCTGTTTGATTCTTTGAATCTGGGTCACCAGGCGCTTTTCCAAGAGAAGGTCATCAAGACTTTGGATTTTTCCACACCGGGGCGCGCTGCGGCTGCTGTTGCTTTTTTGAGTTTTATAAAGGATAAATGGAGCGAAGAAACCCATCTGAGCGGGGGGTACCTGCTGGATTTTCTGGCCATGCATCTGTGGAGAGCGGTTGTGAGACACAAGAATCGCCTGCTACTGTTGTCTTCCGTCCGCCCGGCGATAATACCGACGGAGGAGCAGCAGCAGCAGCAGGAGGAAGCCAGGCGGCGGCGGCAGGAGCAGAGCCCATGGAACCCGAGAGCCGGCCTGGACCCTCGGGAATGAATGTTGTACAGGTGGCTGAACTGTATCCAGAACTGAGACGCATTTTGACAATTACAGAGGATGGGCAGGGGCTAAAGGGGGTAAAGAGGGAGCGGGGGGCTTGTGAGGCTACAGAGGAGGCTAGGAATCTAGCTTTTAGCTTAATGACCAGACACCGTCCTGAGTGTATTACTTTTCAACAGATCAAGGATAATTGCGCTAATGAGCTTGATCTGCTGGCGCAGAAGTATTCCATAGAGCAGCTGACCACTTACTGGCTGCAGCCAGGGGATGATTTTGAGGAGGCTATTAGGGTATATGCAAAGGTGGCACTTAGGCCAGATTGCAAGTACAAGATCAGCAAACTTGTAAATATCAGGAATTGTTGCTACATTTCTGGGAACGGGGCCGAGGTGGAGATAGATACGGAGGATAGGGTGGCCTTTAGATGTAGCATGATAAATATGTGGCCGGGGGTGCTTGGCATGGACGGGGTGGTTATTATGAATGTAAGGTTTACTGGCCCCAATTTTAGCGGTACGGTTTTCCTGGCCAATACCAACCTTATCCTACACGGTGTAAGCTTCTATGGGTTTAACAATACCTGTGTGGAAGCCTGGACCGATGTAAGGGTTCGGGGCTGTGCCTTTTACTGCTGCTGGAAGGGGGTGGTGTGTCGCCCCAAAAGCAGGGCTTCAATTAAGAAATGCCTCTTTGAAAGGTGTACCTTGGGTATCCTGTCTGAGGGTAACTCCAGGGTGCGCCACAATGTGGCCTCCGACTGTGGTTGCTTCATGCTAGTGAAAAGCGTGGCTGTGATTAAGCATAACATGGTATGTGGCAACTGCGAGGACAGGGCCTCTCAGATGCTGACCTGCTCGGACGGCAACTGTCACCTGCTGAAGACCATTCACGTAGCCAGCCACTCTCGCAAGGCCTGGCCAGTGTTTGAGCATAACATACTGACCCGCTGTTCCTTGCATTTGGGTAACAGGAGGGGGGTGTTCCTACCTTACCAATGCAATTTGAGTCACACTAAGATATTGCTTGAGCCCGAGAGCATGTCCAAGGTGAACCTGAACGGGGTGTTTGACATGACCATGAAGATCTGGAAGGTGCTGAGGTACGATGAGACCCGCACCAGGTGCAGACCCTGCGAGTGTGGCGGTAAACATATTAGGAACCAGCCTGTGATGCTGGATGTGACCGAGGAGCTGAGGCCCGATCACTTGGTGCTGGCCTGCACCCGCGCTGAGTTTGGCTCTAGCGATGAAGATACAGATCTCGAGGGGTCCGGAGCTACAAATTTTTCCCTCCTCAAACAGGCTGGAGATGTCGAAGAAAATCCTGGGCCTACCGGTATGTATAGGATGCAGTTGCTCTCTTGCATCGCCCTGTCTCTAGCGTTGGTGACTAATAGCGCCCCTACTTCTTCTTCCACGAAGAAAACCCAGCTGCAACTGGAACACCTTCTGCTGGATCTCCAGATGATCCTTAACGGGATTAACAACTATAAGAACCCGAAGCTCACGGCTATGCTGACGGCGAAGTTCGCTATGCCTAAAAAGGCTACTGAGCTGAAACACTTACAGTGTCTCGAAGAGGCACTCAAACCCCTCGAGGAGGTACTCAATCTGGCGCAAAGCAAAAACTTCCATCTCCGCCCACGGGACCTAATCTCCAACATCAACGTGATTGTGCTGGAGCTCAAGGGGTCTGAAACCACGTTTATGTGCGAGTACGCAGATGAAACTGCTACCATAGTGGAATTCCTCAATCGTTGGATCACTTTCTGCCAGTCAATCATTTCAACCCTGACCAGATCTGAGGGCCGCGGCAGCCTGCTGACCTGCGGCGACGTGGAGGAAAACCCCGGCCCCGGATCCTTGTCGACCATTCGTCCGCGCAGGCCAAGTCCTGCCCGCGAGCTTCTGCCCGGACCCCAACCGGATAGGGTTCAGCCGACTGCAGATCGTGGGGTGTCTGCGCCTGCTGGCAGCCCTCTGGATGGCTTGCCCGCTCGGCGGACGGTGTCCCGGACCCGGCTGCCATCTCCCCCTGCGCCCTCACCTGCGTTCTCGGCGGGCAGCTTCAGCGATCTGCTCCGTCCGTTCGATCCGTCGCTTCTTGATACATCGCTTCTTGATTCGATGCCTGCCGTCGGCACGCCGCATACAGCGGCTGCCCCAGCAGAGTGGGATGAGGCGCAATCGGCTCTGCGTGCAGCCGATGACCCGCCACCCACCGTGCGTGTCGCTGTCACTGCCGCGCGGCCGCCGCGCGCCAAGCCGGCCCCGCGACGGCGTGCTGCGCAACCCTCCGACGCTTCGCCGGCCGCGCAGGTGGATCTACGCACGCTCGGCTACAGTCAGCAGCAGCAAGAGAAGATCAAACCGAAGGTGCGTTCGACAGTGGCGCAGCACCACGAGGCACTGGTGGGCCATGGGTTTACACACGCGCACATCGTTGCGCTCAGCCAACACCCGGCAGCGTTAGGGACCGTCGCTGTCACGTATCAGCACATAATCACGGCGTTGCCAGAGGCGACACACGAAGACATCGTTGGCGTCGGCAAACAGTGGTCCGGCGCACGCGCCCTGGAGGCCTTGCTCACGGATGCGGGGGAGTTGAGAGGTCCGCCGTTACAGTTGGACACAGGCCAACTTGTGAAGATTGCAAAACGTGGCGGCGTGACCGCAATGGAGGCAGTGCATGCATCGCGCAATGCACTGACGGGTGCCCCCCTGAACCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACAATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACAATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAAGCATTGTGGCCCAGCTGAGCCGGCCTGATCCGGCGTTGGCCGCGTTGACCAACGACCACCTCGTCGCCTTGGCCTGCCTCGGCGGACGTCCTGCCATGGATGCAGTGAAAAAGGGATTGCCGCACGCGCCGGAATTGATCAGAAGAGTCAATCGCCGTATTGGCGAACGCACGTCCCATCGCGTTGCCGACTACGCGCAAGTGGTTCGCGTGCTGGAGTTTTTCCAGTGCCACTCCCACCCAGCGTACGCATTTGATGAGGCCATGACGCAGTTCGGGATGAGCAGGAACGGGTTGGTACAGCTCTTTCGCAGAGTGGGCGTCACCGAACTCGAAGCCCGCGGTGGAACGCTCCCCCCAGCCTCGCAGCGTTGGGACCGTATCCTCCAGGCATCAGGGATGAAAAGGGCCAAACCGTCCCCTACTTCAGCTCAAACACCGGATCAGGCGTCTTTGCATGCATTCGCCGATTCGCTGGAGCGTGACCTTGATGCGCCCAGCCCAATGCACGAGGGAGATCAGACGCGGGCAAGCAGCCGTAAACGGTCCCGATCGGATCGTGCTGTCACCGGCCCCTCCGCACAGCAGGCTGTCGAGGTGCGCGTTCCCGAACAGCGCGATGCGCTGCATTTGCCCCTCAGCTGGAGGGTAAAACGCCCGCGTACCAGGATCTGGGGCGGCCTCCCGGTTAAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGAGTAGTGCTTTCTACTTTATGTAACCAATGTGCAGACTACTGTTAACCAATGTGCAGACTACTGTTAACCAATGTGCAGACTACTGTTAACCAATGTGCAGACTACTGTGAAAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTACTGTCCTCCGAGCGGAGTTCTGTCCTCCGAGCGGAGACTCTAGATATAGAACCGATCCTCCCATTGAATTCTAGGCGTGTACGGTGGGAGGCCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCCTCGAGTATAGAACCGATCCTCCCATTTATAGAACCGATCCTCCCATTTATAGAACCGATCCTCCCATTACTAGAAGATCTTGAATTCGCCACCTTGTCGACCATTCGTCCGCGCAGGCCAAGTCCTGCCCGCGAGCTTCTGCCCGGACCCCAACCGGATAGGGTTCAGCCGACTGCAGATCGTGGGGTGTCTGCGCCTGCTGGCAGCCCTCTGGATGGCTTGCCCGCTCGGCGGACGGTGTCCCGGACCCGGCTGCCATCTCCCCCTGCGCCCTCACCTGCGTTCTCGGCGGGCAGCTTCAGCGATCTGCTCCGTCCGTTCGATCCGTCGCTTCTTGATACATCGCTTCTTGATTCGATGCCTGCCGTCGGCACGCCGCATACAGCGGCTGCCCCAGCAGAGTGGGATGAGGCGCAATCGGCTCTGCGTGCAGCCGATGACCCGCCACCCACCGTGCGTGTCGCTGTCACTGCCGCGCGGCCGCCGCGCGCCAAGCCGGCCCCGCGACGGCGTGCTGCGCAACCCTCCGACGCTTCGCCGGCCGCGCAGGTGGATCTACGCACGCTCGGCTACAGTCAGCAGCAGCAAGAGAAGATCAAACCGAAGGTGCGTTCGACAGTGGCGCAGCACCACGAGGCACTGGTGGGCCATGGGTTTACACACGCGCACATCGTTGCGCTCAGCCAACACCCGGCAGCGTTAGGGACCGTCGCTGTCACGTATCAGCACATAATCACGGCGTTGCCAGAGGCGACACACGAAGACATCGTTGGCGTCGGCAAACAGTGGTCCGGCGCACGCGCCCTGGAGGCCTTGCTCACGGATGCGGGGGAGTTGAGAGGTCCGCCGTTACAGTTGGACACAGGCCAACTTGTGAAGATTGCAAAACGTGGCGGCGTGACCGCAATGGAGGCAGTGCATGCATCGCGCAATGCACTGACGGGTGCCCCCCTGAACCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACATTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACAATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACTCCGGACCAAGTGGTGGCTATCGCCAGCCACGATGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCCTGACCCCGGACCAAGTGGTGGCTATCGCCAGCAACGGTGGCGGCAAGCAAGCGCTCGAAAGCATTGTGGCCCAGCTGAGCCGGCCTGATCCGGCGTTGGCCGCGTTGACCAACGACCACCTCGTCGCCTTGGCCTGCCTCGGCGGACGTCCTGCCATGGATGCAGTGAAAAAGGGATTGCCGCACGCGCCGGAATTGATCAGAAGAGTCAATCGCCGTATTGGCGAACGCACGTCCCATCGCGTTGCCGACTACGCGCAAGTGGTTCGCGTGCTGGAGTTTTTCCAGTGCCACTCCCACCCAGCGTACGCATTTGATGAGGCCATGACGCAGTTCGGGATGAGCAGGAACGGGTTGGTACAGCTCTTTCGCAGAGTGGGCGTCACCGAACTCGAAGCCCGCGGTGGAACGCTCCCCCCAGCCTCGCAGCGTTGGGACCGTATCCTCCAGGCATCAGGGATGAAAAGGGCCAAACCGTCCCCTACTTCAGCTCAAACACCGGATCAGGCGTCTTTGCATGCATTCGCCGATTCGCTGGAGCGTGACCTTGATGCGCCCAGCCCAATGCACGAGGGAGATCAGACGCGGGCAAGCAGCCGTAAACGGTCCCGATCGGATCGTGCTGTCACCGGCCCCTCCGCACAGCAGGCTGTCGAGGTGCGCGTTCCCGAACAGCGCGATGCGCTGCATTTGCCCCTCAGCTGGAGGGTAAAACGCCCGCGTACCAGGATCTGGGGCGGCCTCCCGTAATACAAGGCGGCCGCAAATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTATCAACATCAGTCTGATAAGCTAAAGCTTCGAATTCTGATAATCAGCCATACCACATTTGTAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCGTTCATCGGAGCCCAGGTTTACTCCCTTAAGTGGAAATTTCTTCCCCCACTCCCTCCTTGGCTTTCTCCAAGGAGGGAACCCAGGCTACTGGAAAGTCCGGCTGGGGCGGGGACTGTGGGTTTCAGGGTAGAACTGCGTGTGGAACGGGACAGGGAGCGGTTAGAAGGGTGGGGCTATTCCGGGAAGTGGTGGGGGGAGGGAGCCCAAAACTAGCACCTAGTCCACTCATTATCCAGCCCTCTTATTTCTCGGCCCCGCTCTGCTTCAGTGGACCCGGGGAGGGCGGGGAAGTGGAGTGGGAGACCTAGGGGTGGGCTTCCCGACCTTGCTGTACAGGACCTCGACCTAGCTGGCTTTGTTCCCCATCCCCACGTTAGTTGTTGCCCTGAGGCTAAAACTAGAGCCCAGGGGCCCCAAGTTCCAGACTGCCCCTCCCCCCTCCCCCGGAGCCAGGGAGTGGTTGGTGAAAGGGGGAGGCCAGCTGGAGAACAAACGGGTAGTCAGGGGGTTGAGCGATTAGAGCCCTTGTACCCTACCCAGGAATGGTTGGGGAGGAGGAGGAAGAGGTAGGAGGTAGGGGAGGGGGCGGGGTTTTGTCACCTGTCACCTGCTCCGGCTGTGCCTAGGGCGGGCGGGCGGGGAGTGGGGGGACCGGTATAAAGCGGTAGGCGCCTGTGCCCGCTCCACCTCTCAAGCAGCCAGCGCCTGCCTGAATCTGTTCTGCCCCCTCCCCACCCATTTCACCACCACCATGCTGAAGAAGATCCTGAAGATCGAGGAGCTGGACGAGAGAGAGCTGATCGACATCGAGGTGAGCGGCAACCACCTGTTCTACGCCAACGACATCCTGACCCACAACAGCAGTGGCCAAGGCGATTCCCTCCATGAGCACATCGCTAATCTGGCTGGCTCCCCTGCTATCAAAAAGGGCATCCTCCAGACAGTCAAAGTCGTCGATGAGCTGGTCAAGGTGATGGGCAGGCATAAACCCGAGAACATTGTGATTGAGATGGCTAGGGAGAACCAGACCACCCAGAAAGGCCAGAAAAACAGCAGAGAAAGAATGAAGAGGATCGAGGAGGGCATCAAAGAACTGGGCAGCCAAATCCTCAAGGAGCACCCCGTCGAAAATACACAACTCCAGAACGAAAAACTCTACCTCTACTATCTGCAGAACGGCAGAGACATGTACGTGGACCAGGAACTGGACATCAACAGGCTCTCCGATTACGATGTGGACGCCATCGTCCCTCAGTCCTTTCTGAAAGATGATAGCATCGACAACAAGGTGCTGACCAGGTCCGACAAGAATAGGGGCAAGAGCGATAATGTGCCCTCCGAGGAGGTCGTCAAAAAAATGAAAAACTACTGGAGACAACTCCTCAACGCTAAGCTCATCACCCAAAGAAAGTTCGACAATCTGACCAAAGCCGAGAGGGGCGGCCTCTCCGAACTGGACAAGGCCGGCTTCATCAAAAGGCAATTGGTGGAAACCAGGCAGATTACAAAGCATGTCGCTCAAATTCTCGATAGCAGGATGAATACCAAATATGACGAGAACGACAAGCTGATCAGAGAGGTCAAGGTCATCACACTCAAGTCCAAGCTCGTGAGCGACTTCAGAAAAGATTTCCAATTTTATAAAGTCAGGGAGATCAACAATTACCACCACGCTCACGACGCTTATCTCAACGCTGTCGTGGGAACCGCCCTGATCAAAAAATACCCCAAGCTGGAAAGCGAGTTCGTGTATGGCGATTATAAAGTGTACGACGTGAGGAAGATGATCGCTAAAAGCGAGCAGGAAATCGGCAAGGCTACAGCCAAGTACTTTTTCTACTCCAACATTATGAACTTCTTCAAGACCGAGATTACCCTCGCCAACGGCGAAATTAGGAAGAGGCCCCTGATTGAAACAAATGGAGAAACAGGCGAAATCGTCTGGGACAAGGGCAGGGACTTCGCCACAGTCAGAAAAGTGCTGTCCATGCCTCAAGTCAACATCGTCAAAAAGACCGAGGTGCAGACCGGCGGCTTTAGCAAAGAAAGCATCCTGCCCAAGAGAAACTCCGACAAGCTCATCGCTAGGAAGAAGGACTGGGACCCTAAGAAATACGGAGGATTTGACTCCCCTACCGTCGCCTATTCCGTCCTCGTCGTCGCTAAGGTGGAGAAGGGCAAGAGCAAGAAGCTCAAGAGCGTCAAGGAGCTGCTGGGAATCACCATCATGGAGAGGAGCTCCTTCGAAAAAAACCCTATTGATTTCCTGGAGGCCAAGGGCTACAAGGAGGTCAAGAAGGACCTCATCATCAAGCTGCCCAAATACAGCCTCTTCGAACTGGAAAATGGCAGGAAGAGAATGCTCGCTAGCGCCGGCGAGCTCCAGAAAGGAAATGAGCTGGCTCTGCCCAGCAAGTACGTCAACTTCCTCTATCTCGCCAGCCACTATGAAAAGCTCAAGGGCAGCCCCGAAGACAATGAGCAGAAGCAGCTCTTCGTCGAGCAGCACAAGCACTACCTCGATGAAATCATCGAGCAAATCAGCGAGTTTTCCAAAAGGGTGATTCTCGCCGACGCTAACCTCGATAAGGTCCTCTCCGCTTACAACAAGCATAGAGACAAGCCCATCAGAGAACAGGCCGAGAACATCATCCACCTGTTTACACTCACAAACCTCGGAGCCCCTGCCGCTTTTAAATACTTCGATACAACCATTGATAGGAAGAGGTACACCTCCACCAAGGAGGTGCTGGATGCTACCCTGATTCATCAATCCATCACAGGACTCTACGAAACAAGGATTGACCTGTCCCAACTGGGAGGCGACACTAGTGGTGGCGGCTCAAAGCGTCCTGCTGCTACTAAGAAAGCTGGTCAAGCTAAGAAAAAGAAATCTAGAATGGACGCCAAGAGCCTGACCGCCTGGTCAAGGACCCTGGTAACCTTCAAAGACGTTTTTGTGGACTTCACAAGGGAGGAATGGAAGTTGCTGGACACCGCACAGCAGATCGTGTATAGGAACGTGATGTTGGAGAATTACAAGAACCTGGTTAGCCTGGGCTATCAGTTGACCAAGCCCGACGTAATCCTTAGGCTGGAAAAGGGCGAGGAACCCTGGCTGGTGGAGAGGGAGATTCATCAGGAAACCCACCCCGACAGCGAAACCGCTTTCGAGATCAAGTCATCTGTGCTCGAGTAATCCTCAGGTGCAGGCTGCCTATCAGAAGGTGGTGGCTGGTGTGGCCAATGCCCTGGCTCACAAATACCACTGAGATCTTTTTCCCTCTGCCAAAAATTATGGGGACATCATGAAGCCCCTTGAGCATCTGACTTCTGGCTAATAAAGGAAATTTATTTTCATTGCAATAGTGTGTTGGAATTTTTTGTGTCTCTCACTCGGAAGGACATATGGGAGGGCAAATCATTTAAAACATCAGAATGAGTATTTGGTTTAGAGTTTGGCAACATATGCCCATATGCTGGCTGCCATGAACAAAGGTTGGCTATAAAGAGGTCATCAGTATATGAAACAGCCCCCTGCTGTCCATTCCTTATTCCATAGAAAAGCCTTGACTTGAGGTTAGATTTTTTTTATATTTTGTTTTGTGTTATTTTTTTCTTTAACATCCCTAAAATTTTCCTTACATGTTTTACTAGCCAGATTTTTCCTCCTCTCCTGACTACTCCCAGTCATAGCTGTCCCTCTTCTCTTATGGAGATC(SEQ ID NO:26)。
根据本发明的实施例，所述腺病毒是通过如下方式获得的：所述的腺病毒载体去除了与腺病毒复制包装相关的E1基因和部分E3基因，E1A基因通过逐级Golden Gate的方法构建到基因线路中，最终通过Gateway或Gibson的方式将基因线路插入到腺病毒载体中。上述获得腺病毒的方式实现了复杂、大片段溶瘤腺病毒载体的快速改造。具体构建方法可参考201780002478.X。
根据本发明的实施例，所述腺病毒载体为去除E1基因与部分E3基因的的B、C亚型腺病毒。
根据本发明的实施例，所述腺病毒载体为去除E1基因与部分E3基因的5、11、12、34或35型腺病毒。
根据本发明的实施例，SEQ ID NO:1～26任一所示的核苷酸序列在所述5型腺病毒载体的插入位点为E1基因区域、E3基因区域或E4基因区域。例如，去除E1基因与部分E3基因5型腺病毒载体的图谱可参见图13，其序列如SEQ ID NO:27所示，其插入位点可以选择在该序列的第458个碱基后第459个碱基前。
CATCATCAATAATATACCTTATTTTGGATTGAAGCCAATATGATAATGAGGGGGTGGAGTTTGTGACGTGGCGCGGGGCGTGGGAACGGGGCGGGTGACGTAGTAGTGTGGCGGAAGTGTGATGTTGCAAGTGTGGCGGAACACATGTAAGCGACGGATGTGGCAAAAGTGACGTTTTTGGTGTGCGCCGGTGTACACAGGAAGTGACAATTTTCGCGCGGTTTTAGGCGGATGTTGTAGTAAATTTGGGCGTAACCGAGTAAGATTTGGCCATTTTCGCGGGAAAACTGAATAAGAGGAAGTGAAATCTGAATAATTTTGTGTTACTCATAGCGCGTAATATTTGTCTAGGGCCGCGGGGACTTTGACCGTTTACGTGGAGACTCGCCCAGGTGTTTTTCTCAGGTGTTTTCTGCGTTCCGGGTCAAAGTTGGCGTTTTATTATTATAGTCAGTCGAACTGAAATGTGTGGGCGTGGCTTAAGGGTGGGAAAGAATATATAAGGTGGGGGTCTTATGTAGTTTTGTATCTGTTTTGCAGCAGCCGCCGCCGCCATGAGCACCAACTCGTTTGATGGAAGCATTGTGAGCTCATATTTGACAACGCGCATGCCCCCATGGGCCGGGGTGCGTCAGAATGTGATGGGCTCCAGCATTGATGGTCGCCCCGTCCTGCCCGCAAACTCTACTACCTTGACCTACGAGACCGTGTCTGGAACGCCGTTGGAGACTGCAGCCTCCGCCGCCGCTTCAGCCGCTGCAGCCACCGCCCGCGGGATTGTGACTGACTTTGCTTTCCTGAGCCCGCTTGCAAGCAGTGCAGCTTCCCGTTCATCCGCCCGCGATGACAAGTTGACGGCTCTTTTGGCACAATTGGATTCTTTGACCCGGGAACTTAATGTCGTTTCTCAGCAGCTGTTGGATCTGCGCCAGCAGGTTTCTGCCCTGAAGGCTTCCTCCCCTCCCAATGCGGTTTAAAACATAAATAAAAAACCAGACTCTGTTTGGATTTGGATCAAGCAAGTGTCTTGCTGTCTTTATTTAGGGGTTTTGCGCGCGCGGTAGGCCCGGGACCAGCGGTCTCGGTCGTTGAGGGTCCTGTGTATTTTTTCCAGGACGTGGTAAAGGTGACTCTGGATGTTCAGATACATGGGCATAAGCCCGTCTCTGGGGTGGAGGTAGCACCACTGCAGAGCTTCATGCTGCGGGGTGGTGTTGTAGATGATCCAGTCGTAGCAGGAGCGCTGGGCGTGGTGCCTAAAAATGTCTTTCAGTAGCAAGCTGATTGCCAGGGGCAGGCCCTTGGTGTAAGTGTTTACAAAGCGGTTAAGCTGGGATGGGTGCATACGTGGGGATATGAGATGCATCTTGGACTGTATTTTTAGGTTGGCTATGTTCCCAGCCATATCCCTCCGGGGATTCATGTTGTGCAGAACCACCAGCACAGTGTATCCGGTGCACTTGGGAAATTTGTCATGTAGCTTAGAAGGAAATGCGTGGAAGAACTTGGAGACGCCCTTGTGACCTCCAAGATTTTCCATGCATTCGTCCATAATGATGGCAATGGGCCCACGGGCGGCGGCCTGGGCGAAGATATTTCTGGGATCACTAACGTCATAGTTGTGTTCCAGGATGAGATCGTCATAGGCCATTTTTACAAAGCGCGGGCGGAGGGTGCCAGACTGCGGTATAATGGTTCCATCCGGCCCAGGGGCGTAGTTACCCTCACAGATTTGCATTTCCCACGCTTTGAGTTCAGATGGGGGGATCATGTCTACCTGCGGGGCGATGAAGAAAACGGTTTCCGGGGTAGGGGAGATCAGCTGGGAAGAAAGCAGGTTCCTGAGCAGCTGCGACTTACCGCAGCCGGTGGGCCCGTAAATCACACCTATTACCGGCTGCAACTGGTAGTTAAGAGAGCTGCAGCTGCCGTCATCCCTGAGCAGGGGGGCCACTTCGTTAAGCATGTCCCTGACTCGCATGTTTTCCCTGACCAAATCCGCCAGAAGGCGCTCGCCGCCCAGCGATAGCAGTTCTTGCAAGGAAGCAAAGTTTTTCAACGGTTTGAGACCGTCCGCCGTAGGCATGCTTTTGAGCGTTTGACCAAGCAGTTCCAGGCGGTCCCACAGCTCGGTCACCTGCTCTACGGCATCTCGATCCAGCATATCTCCTCGTTTCGCGGGTTGGGGCGGCTTTCGCTGTACGGCAGTAGTCGGTGCTCGTCCAGACGGGCCAGGGTCATGTCTTTCCACGGGCGCAGGGTCCTCGTCAGCGTAGTCTGGGTCACGGTGAAGGGGTGCGCTCCGGGCTGCGCGCTGGCCAGGGTGCGCTTGAGGCTGGTCCTGCTGGTGCTGAAGCGCTGCCGGTCTTCGCCCTGCGCGTCGGCCAGGTAGCATTTGACCATGGTGTCATAGTCCAGCCCCTCCGCGGCGTGGCCCTTGGCGCGCAGCTTGCCCTTGGAGGAGGCGCCGCACGAGGGGCAGTGCAGACTTTTGAGGGCGTAGAGCTTGGGCGCGAGAAATACCGATTCCGGGGAGTAGGCATCCGCGCCGCAGGCCCCGCAGACGGTCTCGCATTCCACGAGCCAGGTGAGCTCTGGCCGTTCGGGGTCAAAAACCAGGTTTCCCCCATGCTTTTTGATGCGTTTCTTACCTCTGGTTTCCATGAGCCGGTGTCCACGCTCGGTGACGAAAAGGCTGTCCGTGTCCCCGTATACAGACTTGAGAGGCCTGTCCTCGAGCGGTGTTCCGCGGTCCTCCTCGTATAGAAACTCGGACCACTCTGAGACAAAGGCTCGCGTCCAGGCCAGCACGAAGGAGGCTAAGTGGGAGGGGTAGCGGTCGTTGTCCACTAGGGGGTCCACTCGCTCCAGGGTGTGAAGACACATGTCGCCCTCTTCGGCATCAAGGAAGGTGATTGGTTTGTAGGTGTAGGCCACGTGACCGGGTGTTCCTGAAGGGGGGCTATAAAAGGGGGTGGGGGCGCGTTCGTCCTCACTCTCTTCCGCATCGCTGTCTGCGAGGGCCAGCTGTTGGGGTGAGTACTCCCTCTGAAAAGCGGGCATGACTTCTGCGCTAAGATTGTCAGTTTCCAAAAACGAGGAGGATTTGATATTCACCTGGCCCGCGGTGATGCCTTTGAGGGTGGCCGCATCCATCTGGTCAGAAAAGACAATCTTTTTGTTGTCAAGCTTGGTGGCAAACGACCCGTAGAGGGCGTTGGACAGCAACTTGGCGATGGAGCGCAGGGTTTGGTTTTTGTCGCGATCGGCGCGCTCCTTGGCCGCGATGTTTAGCTGCACGTATTCGCGCGCAACGCACCGCCATTCGGGAAAGACGGTGGTGCGCTCGTCGGGCACCAGGTGCACGCGCCAACCGCGGTTGTGCAGGGTGACAAGGTCAACGCTGGTGGCTACCTCTCCGCGTAGGCGCTCGTTGGTCCAGCAGAGGCGGCCGCCCTTGCGCGAGCAGAATGGCGGTAGGGGGTCTAGCTGCGTCTCGTCCGGGGGGTCTGCGTCCACGGTAAAGACCCCGGGCAGCAGGCGCGCGTCGAAGTAGTCTATCTTGCATCCTTGCAAGTCTAGCGCCTGCTGCCATGCGCGGGCGGCAAGCGCGCGCTCGTATGGGTTGAGTGGGGGACCCCATGGCATGGGGTGGGTGAGCGCGGAGGCGTACATGCCGCAAATGTCGTAAACGTAGAGGGGCTCTCTGAGTATTCCAAGATATGTAGGGTAGCATCTTCCACCGCGGATGCTGGCGCGCACGTAATCGTATAGTTCGTGCGAGGGAGCGAGGAGGTCGGGACCGAGGTTGCTACGGGCGGGCTGCTCTGCTCGGAAGACTATCTGCCTGAAGATGGCATGTGAGTTGGATGATATGGTTGGACGCTGGAAGACGTTGAAGCTGGCGTCTGTGAGACCTACCGCGTCACGCACGAAGGAGGCGTAGGAGTCGCGCAGCTTGTTGACCAGCTCGGCGGTGACCTGCACGTCTAGGGCGCAGTAGTCCAGGGTTTCCTTGATGATGTCATACTTATCCTGTCCCTTTTTTTTCCACAGCTCGCGGTTGAGGACAAACTCTTCGCGGTCTTTCCAGTACTCTTGGATCGGAAACCCGTCGGCCTCCGAACGGTAAGAGCCTAGCATGTAGAACTGGTTGACGGCCTGGTAGGCGCAGCATCCCTTTTCTACGGGTAGCGCGTATGCCTGCGCGGCCTTCCGGAGCGAGGTGTGGGTGAGCGCAAAGGTGTCCCTGACCATGACTTTGAGGTACTGGTATTTGAAGTCAGTGTCGTCGCATCCGCCCTGCTCCCAGAGCAAAAAGTCCGTGCGCTTTTTGGAACGCGGATTTGGCAGGGCGAAGGTGACATCGTTGAAGAGTATCTTTCCCGCGCGAGGCATAAAGTTGCGTGTGATGCGGAAGGGTCCCGGCACCTCGGAACGGTTGTTAATTACCTGGGCGGCGAGCACGATCTCGTCAAAGCCGTTGATGTTGTGGCCCACAATGTAAAGTTCCAAGAAGCGCGGGATGCCCTTGATGGAAGGCAATTTTTTAAGTTCCTCGTAGGTGAGCTCTTCAGGGGAGCTGAGCCCGTGCTCTGAAAGGGCCCAGTCTGCAAGATGAGGGTTGGAAGCGACGAATGAGCTCCACAGGTCACGGGCCATTAGCATTTGCAGGTGGTCGCGAAAGGTCCTAAACTGGCGACCTATGGCCATTTTTTCTGGGGTGATGCAGTAGAAGGTAAGCGGGTCTTGTTCCCAGCGGTCCCATCCAAGGTTCGCGGCTAGGTCTCGCGCGGCAGTCACTAGAGGCTCATCTCCGCCGAACTTCATGACCAGCATGAAGGGCACGAGCTGCTTCCCAAAGGCCCCCATCCAAGTATAGGTCTCTACATCGTAGGTGACAAAGAGACGCTCGGTGCGAGGATGCGAGCCGATCGGGAAGAACTGGATCTCCCGCCACCAATTGGAGGAGTGGCTATTGATGTGGTGAAAGTAGAAGTCCCTGCGACGGGCCGAACACTCGTGCTGGCTTTTGTAAAAACGTGCGCAGTACTGGCAGCGGTGCACGGGCTGTACATCCTGCACGAGGTTGACCTGACGACCGCGCACAAGGAAGCAGAGTGGGAATTTGAGCCCCTCGCCTGGCGGGTTTGGCTGGTGGTCTTCTACTTCGGCTGCTTGTCCTTGACCGTCTGGCTGCTCGAGGGGAGTTACGGTGGATCGGACCACCACGCCGCGCGAGCCCAAAGTCCAGATGTCCGCGCGCGGCGGTCGGAGCTTGATGACAACATCGCGCAGATGGGAGCTGTCCATGGTCTGGAGCTCCCGCGGCGTCAGGTCAGGCGGGAGCTCCTGCAGGTTTACCTCGCATAGACGGGTCAGGGCGCGGGCTAGATCCAGGTGATACCTAATTTCCAGGGGCTGGTTGGTGGCGGCGTCGATGGCTTGCAAGAGGCCGCATCCCCGCGGCGCGACTACGGTACCGCGCGGCGGGCGGTGGGCCGCGGGGGTGTCCTTGGATGATGCATCTAAAAGCGGTGACGCGGGCGAGCCCCCGGAGGTAGGGGGGGCTCCGGACCCGCCGGGAGAGGGGGCAGGGGCACGTCGGCGCCGCGCGCGGGCAGGAGCTGGTGCTGCGCGCGTAGGTTGCTGGCGAACGCGACGACGCGGCGGTTGATCTCCTGAATCTGGCGCCTCTGCGTGAAGACGACGGGCCCGGTGAGCTTGAACCTGAAAGAGAGTTCGACAGAATCAATTTCGGTGTCGTTGACGGCGGCCTGGCGCAAAATCTCCTGCACGTCTCCTGAGTTGTCTTGATAGGCGATCTCGGCCATGAACTGCTCGATCTCTTCCTCCTGGAGATCTCCGCGTCCGGCTCGCTCCACGGTGGCGGCGAGGTCGTTGGAAATGCGGGCCATGAGCTGCGAGAAGGCGTTGAGGCCTCCCTCGTTCCAGACGCGGCTGTAGACCACGCCCCCTTCGGCATCGCGGGCGCGCATGACCACCTGCGCGAGATTGAGCTCCACGTGCCGGGCGAAGACGGCGTAGTTTCGCAGGCGCTGAAAGAGGTAGTTGAGGGTGGTGGCGGTGTGTTCTGCCACGAAGAAGTACATAACCCAGCGTCGCAACGTGGATTCGTTGATATCCCCCAAGGCCTCAAGGCGCTCCATGGCCTCGTAGAAGTCCACGGCGAAGTTGAAAAACTGGGAGTTGCGCGCCGACACGGTTAACTCCTCCTCCAGAAGACGGATGAGCTCGGCGACAGTGTCGCGCACCTCGCGCTCAAAGGCTACAGGGGCCTCTTCTTCTTCTTCAATCTCCTCTTCCATAAGGGCCTCCCCTTCTTCTTCTTCTGGCGGCGGTGGGGGAGGGGGGACACGGCGGCGACGACGGCGCACCGGGAGGCGGTCGACAAAGCGCTCGATCATCTCCCCGCGGCGACGGCGCATGGTCTCGGTGACGGCGCGGCCGTTCTCGCGGGGGCGCAGTTGGAAGACGCCGCCCGTCATGTCCCGGTTATGGGTTGGCGGGGGGCTGCCATGCGGCAGGGATACGGCGCTAACGATGCATCTCAACAATTGTTGTGTAGGTACTCCGCCGCCGAGGGACCTGAGCGAGTCCGCATCGACCGGATCGGAAAACCTCTCGAGAAAGGCGTCTAACCAGTCACAGTCGCAAGGTAGGCTGAGCACCGTGGCGGGCGGCAGCGGGCGGCGGTCGGGGTTGTTTCTGGCGGAGGTGCTGCTGATGATGTAATTAAAGTAGGCGGTCTTGAGACGGCGGATGGTCGACAGAAGCACCATGTCCTTGGGTCCGGCCTGCTGAATGCGCAGGCGGTCGGCCATGCCCCAGGCTTCGTTTTGACATCGGCGCAGGTCTTTGTAGTAGTCTTGCATGAGCCTTTCTACCGGCACTTCTTCTTCTCCTTCCTCTTGTCCTGCATCTCTTGCATCTATCGCTGCGGCGGCGGCGGAGTTTGGCCGTAGGTGGCGCCCTCTTCCTCCCATGCGTGTGACCCCGAAGCCCCTCATCGGCTGAAGCAGGGCTAGGTCGGCGACAACGCGCTCGGCTAATATGGCCTGCTGCACCTGCGTGAGGGTAGACTGGAAGTCATCCATGTCCACAAAGCGGTGGTATGCGCCCGTGTTGATGGTGTAAGTGCAGTTGGCCATAACGGACCAGTTAACGGTCTGGTGACCCGGCTGCGAGAGCTCGGTGTACCTGAGACGCGAGTAAGCCCTCGAGTCAAATACGTAGTCGTTGCAAGTCCGCACCAGGTACTGGTATCCCACCAAAAAGTGCGGCGGCGGCTGGCGGTAGAGGGGCCAGCGTAGGGTGGCCGGGGCTCCGGGGGCGAGATCTTCCAACATAAGGCGATGATATCCGTAGATGTACCTGGACATCCAGGTGATGCCGGCGGCGGTGGTGGAGGCGCGCGGAAAGTCGCGGACGCGGTTCCAGATGTTGCGCAGCGGCAAAAAGTGCTCCATGGTCGGGACGCTCTGGCCGGTCAGGCGCGCGCAATCGTTGACGCTCTAGACCGTGCAAAAGGAGAGCCTGTAAGCGGGCACTCTTCCGTGGTCTGGTGGATAAATTCGCAAGGGTATCATGGCGGACGACCGGGGTTCGAGCCCCGTATCCGGCCGTCCGCCGTGATCCATGCGGTTACCGCCCGCGTGTCGAACCCAGGTGTGCGACGTCAGACAACGGGGGAGTGCTCCTTTTGGCTTCCTTCCAGGCGCGGCGGCTGCTGCGCTAGCTTTTTTGGCCACTGGCCGCGCGCAGCGTAAGCGGTTAGGCTGGAAAGCGAAAGCATTAAGTGGCTCGCTCCCTGTAGCCGGAGGGTTATTTTCCAAGGGTTGAGTCGCGGGACCCCCGGTTCGAGTCTCGGACCGGCCGGACTGCGGCGAACGGGGGTTTGCCTCCCCGTCATGCAAGACCCCGCTTGCAAATTCCTCCGGAAACAGGGACGAGCCCCTTTTTTGCTTTTCCCAGATGCATCCGGTGCTGCGGCAGATGCGCCCCCCTCCTCAGCAGCGGCAAGAGCAAGAGCAGCGGCAGACATGCAGGGCACCCTCCCCTCCTCCTACCGCGTCAGGAGGGGCGACATCCGCGGTTGACGCGGCAGCAGATGGTGATTACGAACCCCCGCGGCGCCGGGCCCGGCACTACCTGGACTTGGAGGAGGGCGAGGGCCTGGCGCGGCTAGGAGCGCCCTCTCCTGAGCGGCACCCAAGGGTGCAGCTGAAGCGTGATACGCGTGAGGCGTACGTGCCGCGGCAGAACCTGTTTCGCGACCGCGAGGGAGAGGAGCCCGAGGAGATGCGGGATCGAAAGTTCCACGCAGGGCGCGAGCTGCGGCATGGCCTGAATCGCGAGCGGTTGCTGCGCGAGGAGGACTTTGAGCCCGACGCGCGAACCGGGATTAGTCCCGCGCGCGCACACGTGGCGGCCGCCGACCTGGTAACCGCATACGAGCAGACGGTGAACCAGGAGATTAACTTTCAAAAAAGCTTTAACAACCACGTGCGTACGCTTGTGGCGCGCGAGGAGGTGGCTATAGGACTGATGCATCTGTGGGACTTTGTAAGCGCGCTGGAGCAAAACCCAAATAGCAAGCCGCTCATGGCGCAGCTGTTCCTTATAGTGCAGCACAGCAGGGACAACGAGGCATTCAGGGATGCGCTGCTAAACATAGTAGAGCCCGAGGGCCGCTGGCTGCTCGATTTGATAAACATCCTGCAGAGCATAGTGGTGCAGGAGCGCAGCTTGAGCCTGGCTGACAAGGTGGCCGCCATCAACTATTCCATGCTTAGCCTGGGCAAGTTTTACGCCCGCAAGATATACCATACCCCTTACGTTCCCATAGACAAGGAGGTAAAGATCGAGGGGTTCTACATGCGCATGGCGCTGAAGGTGCTTACCTTGAGCGACGACCTGGGCGTTTATCGCAACGAGCGCATCCACAAGGCCGTGAGCGTGAGCCGGCGGCGCGAGCTCAGCGACCGCGAGCTGATGCACAGCCTGCAAAGGGCCCTGGCTGGCACGGGCAGCGGCGATAGAGAGGCCGAGTCCTACTTTGACGCGGGCGCTGACCTGCGCTGGGCCCCAAGCCGACGCGCCCTGGAGGCAGCTGGGGCCGGACCTGGGCTGGCGGTGGCACCCGCGCGCGCTGGCAACGTCGGCGGCGTGGAGGAATATGACGAGGACGATGAGTACGAGCCAGAGGACGGCGAGTACTAAGCGGTGATGTTTCTGATCAGATGATGCAAGACGCAACGGACCCGGCGGTGCGGGCGGCGCTGCAGAGCCAGCCGTCCGGCCTTAACTCCACGGACGACTGGCGCCAGGTCATGGACCGCATCATGTCGCTGACTGCGCGCAATCCTGACGCGTTCCGGCAGCAGCCGCAGGCCAACCGGCTCTCCGCAATTCTGGAAGCGGTGGTCCCGGCGCGCGCAAACCCCACGCACGAGAAGGTGCTGGCGATCGTAAACGCGCTGGCCGAAAACAGGGCCATCCGGCCCGACGAGGCCGGCCTGGTCTACGACGCGCTGCTTCAGCGCGTGGCTCGTTACAACAGCGGCAACGTGCAGACCAACCTGGACCGGCTGGTGGGGGATGTGCGCGAGGCCGTGGCGCAGCGTGAGCGCGCGCAGCAGCAGGGCAACCTGGGCTCCATGGTTGCACTAAACGCCTTCCTGAGTACACAGCCCGCCAACGTGCCGCGGGGACAGGAGGACTACACCAACTTTGTGAGCGCACTGCGGCTAATGGTGACTGAGACACCGCAAAGTGAGGTGTACCAGTCTGGGCCAGACTATTTTTTCCAGACCAGTAGACAAGGCCTGCAGACCGTAAACCTGAGCCAGGCTTTCAAAAACTTGCAGGGGCTGTGGGGGGTGCGGGCTCCCACAGGCGACCGCGCGACCGTGTCTAGCTTGCTGACGCCCAACTCGCGCCTGTTGCTGCTGCTAATAGCGCCCTTCACGGACAGTGGCAGCGTGTCCCGGGACACATACCTAGGTCACTTGCTGACACTGTACCGCGAGGCCATAGGTCAGGCGCATGTGGACGAGCATACTTTCCAGGAGATTACAAGTGTCAGCCGCGCGCTGGGGCAGGAGGACACGGGCAGCCTGGAGGCAACCCTAAACTACCTGCTGACCAACCGGCGGCAGAAGATCCCCTCGTTGCACAGTTTAAACAGCGAGGAGGAGCGCATTTTGCGCTACGTGCAGCAGAGCGTGAGCCTTAACCTGATGCGCGACGGGGTAACGCCCAGCGTGGCGCTGGACATGACCGCGCGCAACATGGAACCGGGCATGTATGCCTCAAACCGGCCGTTTATCAACCGCCTAATGGACTACTTGCATCGCGCGGCCGCCGTGAACCCCGAGTATTTCACCAATGCCATCTTGAACCCGCACTGGCTACCGCCCCCTGGTTTCTACACCGGGGGATTCGAGGTGCCCGAGGGTAACGATGGATTCCTCTGGGACGACATAGACGACAGCGTGTTTTCCCCGCAACCGCAGACCCTGCTAGAGTTGCAACAGCGCGAGCAGGCAGAGGCGGCGCTGCGAAAGGAAAGCTTCCGCAGGCCAAGCAGCTTGTCCGATCTAGGCGCTGCGGCCCCGCGGTCAGATGCTAGTAGCCCATTTCCAAGCTTGATAGGGTCTCTTACCAGCACTCGCACCACCCGCCCGCGCCTGCTGGGCGAGGAGGAGTACCTAAACAACTCGCTGCTGCAGCCGCAGCGCGAAAAAAACCTGCCTCCGGCATTTCCCAACAACGGGATAGAGAGCCTAGTGGACAAGATGAGTAGATGGAAGACGTACGCGCAGGAGCACAGGGACGTGCCAGGCCCGCGCCCGCCCACCCGTCGTCAAAGGCACGACCGTCAGCGGGGTCTGGTGTGGGAGGACGATGACTCGGCAGACGACAGCAGCGTCCTGGATTTGGGAGGGAGTGGCAACCCGTTTGCGCACCTTCGCCCCAGGCTGGGGAGAATGTTTTAAAAAAAAAAAAAGCATGATGCAAAATAAAAAACTCACCAAGGCCATGGCACCGAGCGTTGGTTTTCTTGTATTCCCCTTAGTATGCGGCGCGCGGCGATGTATGAGGAAGGTCCTCCTCCCTCCTACGAGAGTGTGGTGAGCGCGGCGCCAGTGGCGGCGGCGCTGGGTTCTCCCTTCGATGCTCCCCTGGACCCGCCGTTTGTGCCTCCGCGGTACCTGCGGCCTACCGGGGGGAGAAACAGCATCCGTTACTCTGAGTTGGCACCCCTATTCGACACCACCCGTGTGTACCTGGTGGACAACAAGTCAACGGATGTGGCATCCCTGAACTACCAGAACGACCACAGCAACTTTCTGACCACGGTCATTCAAAACAATGACTACAGCCCGGGGGAGGCAAGCACACAGACCATCAATCTTGACGACCGGTCGCACTGGGGCGGCGACCTGAAAACCATCCTGCATACCAACATGCCAAATGTGAACGAGTTCATGTTTACCAATAAGTTTAAGGCGCGGGTGATGGTGTCGCGCTTGCCTACTAAGGACAATCAGGTGGAGCTGAAATACGAGTGGGTGGAGTTCACGCTGCCCGAGGGCAACTACTCCGAGACCATGACCATAGACCTTATGAACAACGCGATCGTGGAGCACTACTTGAAAGTGGGCAGACAGAACGGGGTTCTGGAAAGCGACATCGGGGTAAAGTTTGACACCCGCAACTTCAGACTGGGGTTTGACCCCGTCACTGGTCTTGTCATGCCTGGGGTATATACAAACGAAGCCTTCCATCCAGACATCATTTTGCTGCCAGGATGCGGGGTGGACTTCACCCACAGCCGCCTGAGCAACTTGTTGGGCATCCGCAAGCGGCAACCCTTCCAGGAGGGCTTTAGGATCACCTACGATGATCTGGAGGGTGGTAACATTCCCGCACTGTTGGATGTGGACGCCTACCAGGCGAGCTTGAAAGATGACACCGAACAGGGCGGGGGTGGCGCAGGCGGCAGCAACAGCAGTGGCAGCGGCGCGGAAGAGAACTCCAACGCGGCAGCCGCGGCAATGCAGCCGGTGGAGGACATGAACGATCATGCCATTCGCGGCGACACCTTTGCCACACGGGCTGAGGAGAAGCGCGCTGAGGCCGAAGCAGCGGCCGAAGCTGCCGCCCCCGCTGCGCAACCCGAGGTCGAGAAGCCTCAGAAGAAACCGGTGATCAAACCCCTGACAGAGGACAGCAAGAAACGCAGTTACAACCTAATAAGCAATGACAGCACCTTCACCCAGTACCGCAGCTGGTACCTTGCATACAACTACGGCGACCCTCAGACCGGAATCCGCTCATGGACCCTGCTTTGCACTCCTGACGTAACCTGCGGCTCGGAGCAGGTCTACTGGTCGTTGCCAGACATGATGCAAGACCCCGTGACCTTCCGCTCCACGCGCCAGATCAGCAACTTTCCGGTGGTGGGCGCCGAGCTGTTGCCCGTGCACTCCAAGAGCTTCTACAACGACCAGGCCGTCTACTCCCAACTCATCCGCCAGTTTACCTCTCTGACCCACGTGTTCAATCGCTTTCCCGAGAACCAGATTTTGGCGCGCCCGCCAGCCCCCACCATCACCACCGTCAGTGAAAACGTTCCTGCTCTCACAGATCACGGGACGCTACCGCTGCGCAACAGCATCGGAGGAGTCCAGCGAGTGACCATTACTGACGCCAGACGCCGCACCTGCCCCTACGTTTACAAGGCCCTGGGCATAGTCTCGCCGCGCGTCCTATCGAGCCGCACTTTTTGAGCAAGCATGTCCATCCTTATATCGCCCAGCAATAACACAGGCTGGGGCCTGCGCTTCCCAAGCAAGATGTTTGGCGGGGCCAAGAAGCGCTCCGACCAACACCCAGTGCGCGTGCGCGGGCACTACCGCGCGCCCTGGGGCGCGCACAAACGCGGCCGCACTGGGCGCACCACCGTCGATGACGCCATCGACGCGGTGGTGGAGGAGGCGCGCAACTACACGCCCACGCCGCCACCAGTGTCCACAGTGGACGCGGCCATTCAGACCGTGGTGCGCGGAGCCCGGCGCTATGCTAAAATGAAGAGACGGCGGAGGCGCGTAGCACGTCGCCACCGCCGCCGACCCGGCACTGCCGCCCAACGCGCGGCGGCGGCCCTGCTTAACCGCGCACGTCGCACCGGCCGACGGGCGGCCATGCGGGCCGCTCGAAGGCTGGCCGCGGGTATTGTCACTGTGCCCCCCAGGTCCAGGCGACGAGCGGCCGCCGCAGCAGCCGCGGCCATTAGTGCTATGACTCAGGGTCGCAGGGGCAACGTGTATTGGGTGCGCGACTCGGTTAGCGGCCTGCGCGTGCCCGTGCGCACCCGCCCCCCGCGCAACTAGATTGCAAGAAAAAACTACTTAGACTCGTACTGTTGTATGTATCCAGCGGCGGCGGCGCGCAACGAAGCTATGTCCAAGCGCAAAATCAAAGAAGAGATGCTCCAGGTCATCGCGCCGGAGATCTATGGCCCCCCGAAGAAGGAAGAGCAGGATTACAAGCCCCGAAAGCTAAAGCGGGTCAAAAAGAAAAAGAAAGATGATGATGATGAACTTGACGACGAGGTGGAACTGCTGCACGCTACCGCGCCCAGGCGACGGGTACAGTGGAAAGGTCGACGCGTAAAACGTGTTTTGCGACCCGGCACCACCGTAGTCTTTACGCCCGGTGAGCGCTCCACCCGCACCTACAAGCGCGTGTATGATGAGGTGTACGGCGACGAGGACCTGCTTGAGCAGGCCAACGAGCGCCTCGGGGAGTTTGCCTACGGAAAGCGGCATAAGGACATGCTGGCGTTGCCGCTGGACGAGGGCAACCCAACACCTAGCCTAAAGCCCGTAACACTGCAGCAGGTGCTGCCCGCGCTTGCACCGTCCGAAGAAAAGCGCGGCCTAAAGCGCGAGTCTGGTGACTTGGCACCCACCGTGCAGCTGATGGTACCCAAGCGCCAGCGACTGGAAGATGTCTTGGAAAAAATGACCGTGGAACCTGGGCTGGAGCCCGAGGTCCGCGTGCGGCCAATCAAGCAGGTGGCGCCGGGACTGGGCGTGCAGACCGTGGACGTTCAGATACCCACTACCAGTAGCACCAGTATTGCCACCGCCACAGAGGGCATGGAGACACAAACGTCCCCGGTTGCCTCAGCGGTGGCGGATGCCGCGGTGCAGGCGGTCGCTGCGGCCGCGTCCAAGACCTCTACGGAGGTGCAAACGGACCCGTGGATGTTTCGCGTTTCAGCCCCCCGGCGCCCGCGCCGTTCGAGGAAGTACGGCGCCGCCAGCGCGCTACTGCCCGAATATGCCCTACATCCTTCCATTGCGCCTACCCCCGGCTATCGTGGCTACACCTACCGCCCCAGAAGACGAGCAACTACCCGACGCCGAACCACCACTGGAACCCGCCGCCGCCGTCGCCGTCGCCAGCCCGTGCTGGCCCCGATTTCCGTGCGCAGGGTGGCTCGCGAAGGAGGCAGGACCCTGGTGCTGCCAACAGCGCGCTACCACCCCAGCATCGTTTAAAAGCCGGTCTTTGTGGTTCTTGCAGATATGGCCCTCACCTGCCGCCTCCGTTTCCCGGTGCCGGGATTCCGAGGAAGAATGCACCGTAGGAGGGGCATGGCCGGCCACGGCCTGACGGGCGGCATGCGTCGTGCGCACCACCGGCGGCGGCGCGCGTCGCACCGTCGCATGCGCGGCGGTATCCTGCCCCTCCTTATTCCACTGATCGCCGCGGCGATTGGCGCCGTGCCCGGAATTGCATCCGTGGCCTTGCAGGCGCAGAGACACTGATTAAAAACAAGTTGCATGTGGAAAAATCAAAATAAAAAGTCTGGACTCTCACGCTCGCTTGGTCCTGTAACTATTTTGTAGAATGGAAGACATCAACTTTGCGTCTCTGGCCCCGCGACACGGCTCGCGCCCGTTCATGGGAAACTGGCAAGATATCGGCACCAGCAATATGAGCGGTGGCGCCTTCAGCTGGGGCTCGCTGTGGAGCGGCATTAAAAATTTCGGTTCCACCGTTAAGAACTATGGCAGCAAGGCCTGGAACAGCAGCACAGGCCAGATGCTGAGGGATAAGTTGAAAGAGCAAAATTTCCAACAAAAGGTGGTAGATGGCCTGGCCTCTGGCATTAGCGGGGTGGTGGACCTGGCCAACCAGGCAGTGCAAAATAAGATTAACAGTAAGCTTGATCCCCGCCCTCCCGTAGAGGAGCCTCCACCGGCCGTGGAGACAGTGTCTCCAGAGGGGCGTGGCGAAAAGCGTCCGCGCCCCGACAGGGAAGAAACTCTGGTGACGCAAATAGACGAGCCTCCCTCGTACGAGGAGGCACTAAAGCAAGGCCTGCCCACCACCCGTCCCATCGCGCCCATGGCTACCGGAGTGCTGGGCCAGCACACACCCGTAACGCTGGACCTGCCTCCCCCCGCCGACACCCAGCAGAAACCTGTGCTGCCAGGCCCGACCGCCGTTGTTGTAACCCGTCCTAGCCGCGCGTCCCTGCGCCGCGCCGCCAGCGGTCCGCGATCGTTGCGGCCCGTAGCCAGTGGCAACTGGCAAAGCACACTGAACAGCATCGTGGGTCTGGGGGTGCAATCCCTGAAGCGCCGACGATGCTTCTGATAGCTAACGTGTCGTATGTGTGTCATGTATGCGTCCATGTCGCCGCCAGAGGAGCTGCTGAGCCGCCGCGCGCCCGCTTTCCAAGATGGCTACCCCTTCGATGATGCCGCAGTGGTCTTACATGCACATCTCGGGCCAGGACGCCTCGGAGTACCTGAGCCCCGGGCTGGTGCAGTTTGCCCGCGCCACCGAGACGTACTTCAGCCTGAATAACAAGTTTAGAAACCCCACGGTGGCGCCTACGCACGACGTGACCACAGACCGGTCCCAGCGTTTGACGCTGCGGTTCATCCCTGTGGACCGTGAGGATACTGCGTACTCGTACAAGGCGCGGTTCACCCTAGCTGTGGGTGATAACCGTGTGCTGGACATGGCTTCCACGTACTTTGACATCCGCGGCGTGCTGGACAGGGGCCCTACTTTTAAGCCCTACTCTGGCACTGCCTACAACGCCCTGGCTCCCAAGGGTGCCCCAAATCCTTGCGAATGGGATGAAGCTGCTACTGCTCTTGAAATAAACCTAGAAGAAGAGGACGATGACAACGAAGACGAAGTAGACGAGCAAGCTGAGCAGCAAAAAACTCACGTATTTGGGCAGGCGCCTTATTCTGGTATAAATATTACAAAGGAGGGTATTCAAATAGGTGTCGAAGGTCAAACACCTAAATATGCCGATAAAACATTTCAACCTGAACCTCAAATAGGAGAATCTCAGTGGTACGAAACAGAAATTAATCATGCAGCTGGGAGAGTCCTAAAAAAGACTACCCCAATGAAACCATGTTACGGTTCATATGCAAAACCCACAAATGAAAATGGAGGGCAAGGCATTCTTGTAAAGCAACAAAATGGAAAGCTAGAAAGTCAAGTGGAAATGCAATTTTTCTCAACTACTGAGGCAGCCGCAGGCAATGGTGATAACTTGACTCCTAAAGTGGTATTGTACAGTGAAGATGTAGATATAGAAACCCCAGACACTCATATTTCTTACATGCCCACTATTAAGGAAGGTAACTCACGAGAACTAATGGGCCAACAATCTATGCCCAACAGGCCTAATTACATTGCTTTTAGGGACAATTTTATTGGTCTAATGTATTACAACAGCACGGGTAATATGGGTGTTCTGGCGGGCCAAGCATCGCAGTTGAATGCTGTTGTAGATTTGCAAGACAGAAACACAGAGCTTTCATACCAGCTTTTGCTTGATTCCATTGGTGATAGAACCAGGTACTTTTCTATGTGGAATCAGGCTGTTGACAGCTATGATCCAGATGTTAGAATTATTGAAAATCATGGAACTGAAGATGAACTTCCAAATTACTGCTTTCCACTGGGAGGTGTGATTAATACAGAGACTCTTACCAAGGTAAAACCTAAAACAGGTCAGGAAAATGGATGGGAAAAAGATGCTACAGAATTTTCAGATAAAAATGAAATAAGAGTTGGAAATAATTTTGCCATGGAAATCAATCTAAATGCCAACCTGTGGAGAAATTTCCTGTACTCCAACATAGCGCTGTATTTGCCCGACAAGCTAAAGTACAGTCCTTCCAACGTAAAAATTTCTGATAACCCAAACACCTACGACTACATGAACAAGCGAGTGGTGGCTCCCGGGCTAGTGGACTGCTACATTAACCTTGGAGCACGCTGGTCCCTTGACTATATGGACAACGTCAACCCATTTAACCACCACCGCAATGCTGGCCTGCGCTACCGCTCAATGTTGCTGGGCAATGGTCGCTATGTGCCCTTCCACATCCAGGTGCCTCAGAAGTTCTTTGCCATTAAAAACCTCCTTCTCCTGCCGGGCTCATACACCTACGAGTGGAACTTCAGGAAGGATGTTAACATGGTTCTGCAGAGCTCCCTAGGAAATGACCTAAGGGTTGACGGAGCCAGCATTAAGTTTGATAGCATTTGCCTTTACGCCACCTTCTTCCCCATGGCCCACAACACCGCCTCCACGCTTGAGGCCATGCTTAGAAACGACACCAACGACCAGTCCTTTAACGACTATCTCTCCGCCGCCAACATGCTCTACCCTATACCCGCCAACGCTACCAACGTGCCCATATCCATCCCCTCCCGCAACTGGGCGGCTTTCCGCGGCTGGGCCTTCACGCGCCTTAAGACTAAGGAAACCCCATCACTGGGCTCGGGCTACGACCCTTATTACACCTACTCTGGCTCTATACCCTACCTAGATGGAACCTTTTACCTCAACCACACCTTTAAGAAGGTGGCCATTACCTTTGACTCTTCTGTCAGCTGGCCTGGCAATGACCGCCTGCTTACCCCCAACGAGTTTGAAATTAAGCGCTCAGTTGACGGGGAGGGTTACAACGTTGCCCAGTGTAACATGACCAAAGACTGGTTCCTGGTACAAATGCTAGCTAACTATAACATTGGCTACCAGGGCTTCTATATCCCAGAGAGCTACAAGGACCGCATGTACTCCTTCTTTAGAAACTTCCAGCCCATGAGCCGTCAGGTGGTGGATGATACTAAATACAAGGACTACCAACAGGTGGGCATCCTACACCAACACAACAACTCTGGATTTGTTGGCTACCTTGCCCCCACCATGCGCGAAGGACAGGCCTACCCTGCTAACTTCCCCTATCCGCTTATAGGCAAGACCGCAGTTGACAGCATTACCCAGAAAAAGTTTCTTTGCGATCGCACCCTTTGGCGCATCCCATTCTCCAGTAACTTTATGTCCATGGGCGCACTCACAGACCTGGGCCAAAACCTTCTCTACGCCAACTCCGCCCACGCGCTAGACATGACTTTTGAGGTGGATCCCATGGACGAGCCCACCCTTCTTTATGTTTTGTTTGAAGTCTTTGACGTGGTCCGTGTGCACCAGCCGCACCGCGGCGTCATCGAAACCGTGTACCTGCGCACGCCCTTCTCGGCCGGCAACGCCACAACATAAAGAAGCAAGCAACATCAACAACAGCTGCCGCCATGGGCTCCAGTGAGCAGGAACTGAAAGCCATTGTCAAAGATCTTGGTTGTGGGCCATATTTTTTGGGCACCTATGACAAGCGCTTTCCAGGCTTTGTTTCTCCACACAAGCTCGCCTGCGCCATAGTCAATACGGCCGGTCGCGAGACTGGGGGCGTACACTGGATGGCCTTTGCCTGGAACCCGCACTCAAAAACATGCTACCTCTTTGAGCCCTTTGGCTTTTCTGACCAGCGACTCAAGCAGGTTTACCAGTTTGAGTACGAGTCACTCCTGCGCCGTAGCGCCATTGCTTCTTCCCCCGACCGCTGTATAACGCTGGAAAAGTCCACCCAAAGCGTACAGGGGCCCAACTCGGCCGCCTGTGGACTATTCTGCTGCATGTTTCTCCACGCCTTTGCCAACTGGCCCCAAACTCCCATGGATCACAACCCCACCATGAACCTTATTACCGGGGTACCCAACTCCATGCTCAACAGTCCCCAGGTACAGCCCACCCTGCGTCGCAACCAGGAACAGCTCTACAGCTTCCTGGAGCGCCACTCGCCCTACTTCCGCAGCCACAGTGCGCAGATTAGGAGCGCCACTTCTTTTTGTCACTTGAAAAACATGTAAAAATAATGTACTAGAGACACTTTCAATAAAGGCAAATGCTTTTATTTGTACACTCTCGGGTGATTATTTACCCCCACCCTTGCCGTCTGCGCCGTTTAAAAATCAAAGGGGTTCTGCCGCGCATCGCTATGCGCCACTGGCAGGGACACGTTGCGATACTGGTGTTTAGTGCTCCACTTAAACTCAGGCACAACCATCCGCGGCAGCTCGGTGAAGTTTTCACTCCACAGGCTGCGCACCATCACCAACGCGTTTAGCAGGTCGGGCGCCGATATCTTGAAGTCGCAGTTGGGGCCTCCGCCCTGCGCGCGCGAGTTGCGATACACAGGGTTGCAGCACTGGAACACTATCAGCGCCGGGTGGTGCACGCTGGCCAGCACGCTCTTGTCGGAGATCAGATCCGCGTCCAGGTCCTCCGCGTTGCTCAGGGCGAACGGAGTCAACTTTGGTAGCTGCCTTCCCAAAAAGGGCGCGTGCCCAGGCTTTGAGTTGCACTCGCACCGTAGTGGCATCAAAAGGTGACCGTGCCCGGTCTGGGCGTTAGGATACAGCGCCTGCATAAAAGCCTTGATCTGCTTAAAAGCCACCTGAGCCTTTGCGCCTTCAGAGAAGAACATGCCGCAAGACTTGCCGGAAAACTGATTGGCCGGACAGGCCGCGTCGTGCACGCAGCACCTTGCGTCGGTGTTGGAGATCTGCACCACATTTCGGCCCCACCGGTTCTTCACGATCTTGGCCTTGCTAGACTGCTCCTTCAGCGCGCGCTGCCCGTTTTCGCTCGTCACATCCATTTCAATCACGTGCTCCTTATTTATCATAATGCTTCCGTGTAGACACTTAAGCTCGCCTTCGATCTCAGCGCAGCGGTGCAGCCACAACGCGCAGCCCGTGGGCTCGTGATGCTTGTAGGTCACCTCTGCAAACGACTGCAGGTACGCCTGCAGGAATCGCCCCATCATCGTCACAAAGGTCTTGTTGCTGGTGAAGGTCAGCTGCAACCCGCGGTGCTCCTCGTTCAGCCAGGTCTTGCATACGGCCGCCAGAGCTTCCACTTGGTCAGGCAGTAGTTTGAAGTTCGCCTTTAGATCGTTATCCACGTGGTACTTGTCCATCAGCGCGCGCGCAGCCTCCATGCCCTTCTCCCACGCAGACACGATCGGCACACTCAGCGGGTTCATCACCGTAATTTCACTTTCCGCTTCGCTGGGCTCTTCCTCTTCCTCTTGCGTCCGCATACCACGCGCCACTGGGTCGTCTTCATTCAGCCGCCGCACTGTGCGCTTACCTCCTTTGCCATGCTTGATTAGCACCGGTGGGTTGCTGAAACCCACCATTTGTAGCGCCACATCTTCTCTTTCTTCCTCGCTGTCCACGATTACCTCTGGTGATGGCGGGCGCTCGGGCTTGGGAGAAGGGCGCTTCTTTTTCTTCTTGGGCGCAATGGCCAAATCCGCCGCCGAGGTCGATGGCCGCGGGCTGGGTGTGCGCGGCACCAGCGCGTCTTGTGATGAGTCTTCCTCGTCCTCGGACTCGATACGCCGCCTCATCCGCTTTTTTGGGGGCGCCCGGGGAGGCGGCGGCGACGGGGACGGGGACGACACGTCCTCCATGGTTGGGGGACGTCGCGCCGCACCGCGTCCGCGCTCGGGGGTGGTTTCGCGCTGCTCCTCTTCCCGACTGGCCATTTCCTTCTCCTATAGGCAGAAAAAGATCATGGAGTCAGTCGAGAAGAAGGACAGCCTAACCGCCCCCTCTGAGTTCGCCACCACCGCCTCCACCGATGCCGCCAACGCGCCTACCACCTTCCCCGTCGAGGCACCCCCGCTTGAGGAGGAGGAAGTGATTATCGAGCAGGACCCAGGTTTTGTAAGCGAAGACGACGAGGACCGCTCAGTACCAACAGAGGATAAAAAGCAAGACCAGGACAACGCAGAGGCAAACGAGGAACAAGTCGGGCGGGGGGACGAAAGGCATGGCGACTACCTAGATGTGGGAGACGACGTGCTGTTGAAGCATCTGCAGCGCCAGTGCGCCATTATCTGCGACGCGTTGCAAGAGCGCAGCGATGTGCCCCTCGCCATAGCGGATGTCAGCCTTGCCTACGAACGCCACCTATTCTCACCGCGCGTACCCCCCAAACGCCAAGAAAACGGCACATGCGAGCCCAACCCGCGCCTCAACTTCTACCCCGTATTTGCCGTGCCAGAGGTGCTTGCCACCTATCACATCTTTTTCCAAAACTGCAAGATACCCCTATCCTGCCGTGCCAACCGCAGCCGAGCGGACAAGCAGCTGGCCTTGCGGCAGGGCGCTGTCATACCTGATATCGCCTCGCTCAACGAAGTGCCAAAAATCTTTGAGGGTCTTGGACGCGACGAGAAGCGCGCGGCAAACGCTCTGCAACAGGAAAACAGCGAAAATGAAAGTCACTCTGGAGTGTTGGTGGAACTCGAGGGTGACAACGCGCGCCTAGCCGTACTAAAACGCAGCATCGAGGTCACCCACTTTGCCTACCCGGCACTTAACCTACCCCCCAAGGTCATGAGCACAGTCATGAGTGAGCTGATCGTGCGCCGTGCGCAGCCCCTGGAGAGGGATGCAAATTTGCAAGAACAAACAGAGGAGGGCCTACCCGCAGTTGGCGACGAGCAGCTAGCGCGCTGGCTTCAAACGCGCGAGCCTGCCGACTTGGAGGAGCGACGCAAACTAATGATGGCCGCAGTGCTCGTTACCGTGGAGCTTGAGTGCATGCAGCGGTTCTTTGCTGACCCGGAGATGCAGCGCAAGCTAGAGGAAACATTGCACTACACCTTTCGACAGGGCTACGTACGCCAGGCCTGCAAGATCTCCAACGTGGAGCTCTGCAACCTGGTCTCCTACCTTGGAATTTTGCACGAAAACCGCCTTGGGCAAAACGTGCTTCATTCCACGCTCAAGGGCGAGGCGCGCCGCGACTACGTCCGCGACTGCGTTTACTTATTTCTATGCTACACCTGGCAGACGGCCATGGGCGTTTGGCAGCAGTGCTTGGAGGAGTGCAACCTCAAGGAGCTGCAGAAACTGCTAAAGCAAAACTTGAAGGACCTATGGACGGCCTTCAACGAGCGCTCCGTGGCCGCGCACCTGGCGGACATCATTTTCCCCGAACGCCTGCTTAAAACCCTGCAACAGGGTCTGCCAGACTTCACCAGTCAAAGCATGTTGCAGAACTTTAGGAACTTTATCCTAGAGCGCTCAGGAATCTTGCCCGCCACCTGCTGTGCACTTCCTAGCGACTTTGTGCCCATTAAGTACCGCGAATGCCCTCCGCCGCTTTGGGGCCACTGCTACCTTCTGCAGCTAGCCAACTACCTTGCCTACCACTCTGACATAATGGAAGACGTGAGCGGTGACGGTCTACTGGAGTGTCACTGTCGCTGCAACCTATGCACCCCGCACCGCTCCCTGGTTTGCAATTCGCAGCTGCTTAACGAAAGTCAAATTATCGGTACCTTTGAGCTGCAGGGTCCCTCGCCTGACGAAAAGTCCGCGGCTCCGGGGTTGAAACTCACTCCGGGGCTGTGGACGTCGGCTTACCTTCGCAAATTTGTACCTGAGGACTACCACGCCCACGAGATTAGGTTCTACGAAGACCAATCCCGCCCGCCTAATGCGGAGCTTACCGCCTGCGTCATTACCCAGGGCCACATTCTTGGCCAATTGCAAGCCATCAACAAAGCCCGCCAAGAGTTTCTGCTACGAAAGGGACGGGGGGTTTACTTGGACCCCCAGTCCGGCGAGGAGCTCAACCCAATCCCCCCGCCGCCGCAGCCCTATCAGCAGCAGCCGCGGGCCCTTGCTTCCCAGGATGGCACCCAAAAAGAAGCTGCAGCTGCCGCCGCCACCCACGGACGAGGAGGAATACTGGGACAGTCAGGCAGAGGAGGTTTTGGACGAGGAGGAGGAGGACATGATGGAAGACTGGGAGAGCCTAGACGAGGAAGCTTCCGAGGTCGAAGAGGTGTCAGACGAAACACCGTCACCCTCGGTCGCATTCCCCTCGCCGGCGCCCCAGAAATCGGCAACCGGTTCCAGCATGGCTACAACCTCCGCTCCTCAGGCGCCGCCGGCACTGCCCGTTCGCCGACCCAACCGTAGATGGGACACCACTGGAACCAGGGCCGGTAAGTCCAAGCAGCCGCCGCCGTTAGCCCAAGAGCAACAACAGCGCCAAGGCTACCGCTCATGGCGCGGGCACAAGAACGCCATAGTTGCTTGCTTGCAAGACTGTGGGGGCAACATCTCCTTCGCCCGCCGCTTTCTTCTCTACCATCACGGCGTGGCCTTCCCCCGTAACATCCTGCATTACTACCGTCATCTCTACAGCCCATACTGCACCGGCGGCAGCGGCAGCAACAGCAGCGGCCACACAGAAGCAAAGGCGACCGGATAGCAAGACTCTGACAAAGCCCAAGAAATCCACAGCGGCGGCAGCAGCAGGAGGAGGAGCGCTGCGTCTGGCGCCCAACGAACCCGTATCGACCCGCGAGCTTAGAAACAGGATTTTTCCCACTCTGTATGCTATATTTCAACAGAGCAGGGGCCAAGAACAAGAGCTGAAAATAAAAAACAGGTCTCTGCGATCCCTCACCCGCAGCTGCCTGTATCACAAAAGCGAAGATCAGCTTCGGCGCACGCTGGAAGACGCGGAGGCTCTCTTCAGTAAATACTGCGCGCTGACTCTTAAGGACTAGTTTCGCGCCCTTTCTCAAATTTAAGCGCGAAAACTACGTCATCTCCAGCGGCCACACCCGGCGCCAGCACCTGTTGTCAGCGCCATTATGAGCAAGGAAATTCCCACGCCCTACATGTGGAGTTACCAGCCACAAATGGGACTTGCGGCTGGAGCTGCCCAAGACTACTCAACCCGAATAAACTACATGAGCGCGGGACCCCACATGATATCCCGGGTCAACGGAATACGCGCCCACCGAAACCGAATTCTCCTGGAACAGGCGGCTATTACCACCACACCTCGTAATAACCTTAATCCCCGTAGTTGGCCCGCTGCCCTGGTGTACCAGGAAAGTCCCGCTCCCACCACTGTGGTACTTCCCAGAGACGCCCAGGCCGAAGTTCAGATGACTAACTCAGGGGCGCAGCTTGCGGGCGGCTTTCGTCACAGGGTGCGGTCGCCCGGGCAGGGTATAACTCACCTGACAATCAGAGGGCGAGGTATTCAGCTCAACGACGAGTCGGTGAGCTCCTCGCTTGGTCTCCGTCCGGACGGGACATTTCAGATCGGCGGCGCCGGCCGCTCTTCATTCACGCCTCGTCAGGCAATCCTAACTCTGCAGACCTCGTCCTCTGAGCCGCGCTCTGGAGGCATTGGAACTCTGCAATTTATTGAGGAGTTTGTGCCATCGGTCTACTTTAACCCCTTCTCGGGACCTCCCGGCCACTATCCGGATCAATTTATTCCTAACTTTGACGCGGTAAAGGACTCGGCGGACGGCTACGACTGAATGTTAAGTGGAGAGGCAGAGCAACTGCGCCTGAAACACCTGGTCCACTGTCGCCGCCACAAGTGCTTTGCCCGCGACTCCGGTGAGTTTTGCTACTTTGAATTGCCCGAGGATCATATCGAGGGCCCGGCGCACGGCGTCCGGCTTACCGCCCAGGGAGAGCTTGCCCGTAGCCTGATTCGGGAGTTTACCCAGCGCCCCCTGCTAGTTGAGCGGGACAGGGGACCCTGTGTTCTCACTGTGATTTGCAACTGTCCTAACCCTGGATTACATCAAGATCTTTGTTGCCATCTCTGTGCTGAGTATAATAAATACAGAAATTAAAATATACTGGGGCTCCTATCGCCATCCTGTAAACGCCACCGTCTTCACCCGCCCAAGCAAACCAAGGCGAACCTTACCTGGTACTTTTAACATCTCTCCCTCTGTGATTTACAACAGTTTCAACCCAGACGGAGTGAGTCTACGAGAGAACCTCTCCGAGCTCAGCTACTCCATCAGAAAAAACACCACCCTCCTTACCTGCCGGGAACGTACGAGTGCGTCACCGGCCGCTGCACCACACCTACCGCCTGACCGTAAACCAGACTTTTTCCGGACAGACCTCAATAACTCTGTTTACCAGAACAGGAGGTGAGCTTAGAAAACCCTTAGGGTATTAGGCCAAAGGCGCAGCTACTGTGGGGTTTATGAACAATTCAAGCAACTCTACGGGCTATTCTAATTCAGGTTTCTCTAGAAATGGACGGAATTATTACAGAGCAGCGCCTGCTAGAAAGACGCAGGGCAGCGGCCGAGCAACAGCGCATGAATCAAGAGCTCCAAGACATGGTTAACTTGCACCAGTGCAAAAGGGGTATCTTTTGTCTGGTAAAGCAGGCCAAAGTCACCTACGACAGTAATACCACCGGACACCGCCTTAGCTACAAGTTGCCAACCAAGCGTCAGAAATTGGTGGTCATGGTGGGAGAAAAGCCCATTACCATAACTCAGCACTCGGTAGAAACCGAAGGCTGCATTCACTCACCTTGTCAAGGACCTGAGGATCTCTGCACCCTTATTAAGACCCTGTGCGGTCTCAAAGATCTTATTCCCTTTAACTAATAAAAAAAAATAATAAAGCATCACTTACTTAAAATCAGTTAGCAAATTTCTGTCCAGTTTATTCAGCAGCACCTCCTTGCCCTCCTCCCAGCTCTGGTATTGCAGCTTCCTCCTGGCTGCAAACTTTCTCCACAATCTAAATGGAATGTCAGTTTCCTCCTGTTCCTGTCCATCCGCACCCACTATCTTCATGTTGTTGCAGATGAAGCGCGCAAGACCGTCTGAAGATACCTTCAACCCCGTGTATCCATATGACACGGAAACCGGTCCTCCAACTGTGCCTTTTCTTACTCCTCCCTTTGTATCCCCCAATGGGTTTCAAGAGAGTCCCCCTGGGGTACTCTCTTTGCGCCTATCCGAACCTCTAGTTACCTCCAATGGCATGCTTGCGCTCAAAATGGGCAACGGCCTCTCTCTGGACGAGGCCGGCAACCTTACCTCCCAAAATGTAACCACTGTGAGCCCACCTCTCAAAAAAACCAAGTCAAACATAAACCTGGAAATATCTGCACCCCTCACAGTTACCTCAGAAGCCCTAACTGTGGCTGCCGCCGCACCTCTAATGGTCGCGGGCAACACACTCACCATGCAATCACAGGCCCCGCTAACCGTGCACGACTCCAAACTTAGCATTGCCACCCAAGGACCCCTCACAGTGTCAGAAGGAAAGCTAGCCCTGCAAACATCAGGCCCCCTCACCACCACCGATAGCAGTACCCTTACTATCACTGCCTCACCCCCTCTAACTACTGCCACTGGTAGCTTGGGCATTGACTTGAAAGAGCCCATTTATACACAAAATGGAAAACTAGGACTAAAGTACGGGGCTCCTTTGCATGTAACAGACGACCTAAACACTTTGACCGTAGCAACTGGTCCAGGTGTGACTATTAATAATACTTCCTTGCAAACTAAAGTTACTGGAGCCTTGGGTTTTGATTCACAAGGCAATATGCAACTTAATGTAGCAGGAGGACTAAGGATTGATTCTCAAAACAGACGCCTTATACTTGATGTTAGTTATCCGTTTGATGCTCAAAACCAACTAAATCTAAGACTAGGACAGGGCCCTCTTTTTATAAACTCAGCCCACAACTTGGATATTAACTACAACAAAGGCCTTTACTTGTTTACAGCTTCAAACAATTCCAAAAAGCTTGAGGTTAACCTAAGCACTGCCAAGGGGTTGATGTTTGACGCTACAGCCATAGCCATTAATGCAGGAGATGGGCTTGAATTTGGTTCACCTAATGCACCAAACACAAATCCCCTCAAAACAAAAATTGGCCATGGCCTAGAATTTGATTCAAACAAGGCTATGGTTCCTAAACTAGGAACTGGCCTTAGTTTTGACAGCACAGGTGCCATTACAGTAGGAAACAAAAATAATGATAAGCTAACTTTGTGGACCACACCAGCTCCATCTCCTAACTGTAGACTAAATGCAGAGAAAGATGCTAAACTCACTTTGGTCTTAACAAAATGTGGCAGTCAAATACTTGCTACAGTTTCAGTTTTGGCTGTTAAAGGCAGTTTGGCTCCAATATCTGGAACAGTTCAAAGTGCTCATCTTATTATAAGATTTGACGAAAATGGAGTGCTACTAAACAATTCCTTCCTGGACCCAGAATATTGGAACTTTAGAAATGGAGATCTTACTGAAGGCACAGCCTATACAAACGCTGTTGGATTTATGCCTAACCTATCAGCTTATCCAAAATCTCACGGTAAAACTGCCAAAAGTAACATTGTCAGTCAAGTTTACTTAAACGGAGACAAAACTAAACCTGTAACACTAACCATTACACTAAACGGTACACAGGAAACAGGAGACACAACTCCAAGTGCATACTCTATGTCATTTTCATGGGACTGGTCTGGCCACAACTACATTAATGAAATATTTGCCACATCCTCTTACACTTTTTCATACATTGCCCAAGAATAAAGAATCGTTTGTGTTATGTTTCAACGTGTTTATTTTTCAATTGCAGAAAATTTCGAATCATTTTTCATTCAGTAGTATAGCCCCACCACCACATAGCTTATACAGATCACCGTACCTTAATCAAACTCACAGAACCCTAGTATTCAACCTGCCACCTCCCTCCCAACACACAGAGTACACAGTCCTTTCTCCCCGGCTGGCCTTAAAAAGCATCATATCATGGGTAACAGACATATTCTTAGGTGTTATATTCCACACGGTTTCCTGTCGAGCCAAACGCTCATCAGTGATATTAATAAACTCCCCGGGCAGCTCACTTAAGTTCATGTCGCTGTCCAGCTGCTGAGCCACAGGCTGCTGTCCAACTTGCGGTTGCTTAACGGGCGGCGAAGGAGAAGTCCACGCCTACATGGGGGTAGAGTCATAATCGTGCATCAGGATAGGGCGGTGGTGCTGCAGCAGCGCGCGAATAAACTGCTGCCGCCGCCGCTCCGTCCTGCAGGAATACAACATGGCAGTGGTCTCCTCAGCGATGATTCGCACCGCCCGCAGCATAAGGCGCCTTGTCCTCCGGGCACAGCAGCGCACCCTGATCTCACTTAAATCAGCACAGTAACTGCAGCACAGCACCACAATATTGTTCAAAATCCCACAGTGCAAGGCGCTGTATCCAAAGCTCATGGCGGGGACCACAGAACCCACGTGGCCATCATACCACAAGCGCAGGTAGATTAAGTGGCGACCCCTCATAAACACGCTGGACATAAACATTACCTCTTTTGGCATGTTGTAATTCACCACCTCCCGGTACCATATAAACCTCTGATTAAACATGGCGCCATCCACCACCATCCTAAACCAGCTGGCCAAAACCTGCCCGCCGGCTATACACTGCAGGGAACCGGGACTGGAACAATGACAGTGGAGAGCCCAGGACTCGTAACCATGGATCATCATGCTCGTCATGATATCAATGTTGGCACAACACAGGCACACGTGCATACACTTCCTCAGGATTACAAGCTCCTCCCGCGTTAGAACCATATCCCAGGGAACAACCCATTCCTGAATCAGCGTAAATCCCACACTGCAGGGAAGACCTCGCACGTAACTCACGTTGTGCATTGTCAAAGTGTTACATTCGGGCAGCAGCGGATGATCCTCCAGTATGGTAGCGCGGGTTTCTGTCTCAAAAGGAGGTAGACGATCCCTACTGTACGGAGTGCGCCGAGACAACCGAGATCGTGTTGGTCGTAGTGTCATGCCAAATGGAACGCCGGACGTAGTCATATTTCCTGAAGCAAAACCAGGTGCGGGCGTGACAAACAGATCTGCGTCTCCGGTCTCGCCGCTTAGATCGCTCTGTGTAGTAGTTGTAGTATATCCACTCTCTCAAAGCATCCAGGCGCCCCCTGGCTTCGGGTTCTATGTAAACTCCTTCATGCGCCGCTGCCCTGATAACATCCACCACCGCAGAATAAGCCACACCCAGCCAACCTACACATTCGTTCTGCGAGTCACACACGGGAGGAGCGGGAAGAGCTGGAAGAACCATGTTTTTTTTTTTATTCCAAAAGATTATCCAAAACCTCAAAATGAAGATCTATTAAGTGAACGCGCTCCCCTCCGGTGGCGTGGTCAAACTCTACAGCCAAAGAACAGATAATGGCATTTGTAAGATGTTGCACAATGGCTTCCAAAAGGCAAACGGCCCTCACGTCCAAGTGGACGTAAAGGCTAAACCCTTCAGGGTGAATCTCCTCTATAAACATTCCAGCACCTTCAACCATGCCCAAATAATTCTCATCTCGCCACCTTCTCAATATATCTCTAAGCAAATCCCGAATATTAAGTCCGGCCATTGTAAAAATCTGCTCCAGAGCGCCCTCCACCTTCAGCCTCAAGCAGCGAATCATGATTGCAAAAATTCAGGTTCCTCACAGACCTGTATAAGATTCAAAAGCGGAACATTAACAAAAATACCGCGATCCCGTAGGTCCCTTCGCAGGGCCAGCTGAACATAATCGTGCAGGTCTGCACGGACCAGCGCGGCCACTTCCCCGCCAGGAACCATGACAAAAGAACCCACACTGATTATGACACGCATACTCGGAGCTATGCTAACCAGCGTAGCCCCGATGTAAGCTTGTTGCATGGGCGGCGATATAAAATGCAAGGTGCTGCTCAAAAAATCAGGCAAAGCCTCGCGCAAAAAAGAAAGCACATCGTAGTCATGCTCATGCAGATAAAGGCAGGTAAGCTCCGGAACCACCACAGAAAAAGACACCATTTTTCTCTCAAACATGTCTGCGGGTTTCTGCATAAACACAAAATAAAATAACAAAAAAACATTTAAACATTAGAAGCCTGTCTTACAACAGGAAAAACAACCCTTATAAGCATAAGACGGACTACGGCCATGCCGGCGTGACCGTAAAAAAACTGGTCACCGTGATTAAAAAGCACCACCGACAGCTCCTCGGTCATGTCCGGAGTCATAATGTAAGACTCGGTAAACACATCAGGTTGATTCACATCGGTCAGTGCTAAAAAGCGACCGAAATAGCCCGGGGGAATACATACCCGCAGGCGTAGAGACAACATTACAGCCCCCATAGGAGGTATAACAAAATTAATAGGAGAGAAAAACACATAAACACCTGAAAAACCCTCCTGCCTAGGCAAAATAGCACCCTCCCGCTCCAGAACAACATACAGCGCTTCCACAGCGGCAGCCATAACAGTCAGCCTTACCAGTAAAAAAGAAAACCTATTAAAAAAACACCACTCGACACGGCACCAGCTCAATCAGTCACAGTGTAAAAAAGGGCCAAGTGCAGAGCGAGTATATATAGGACTAAAAAATGACGTAACGGTTAAAGTCCACAAAAAACACCCAGAAAACCGCACGCGAACCTACGCCCAGAAACGAAAGCCAAAAAACCCACAACTTCCTCAAATCGTCACTTCCGTTTTCCCACGTTACGTCACTTCCCATTTTAAGAAAACTACAATTCCCAACACATACAAGTTACTCCGCCCTAAAACCTACGTCACCCGCCCCGTTCCCACGCCCCGCGCCACGTCACAAACTCCACCCCCTCATTATCATATTGGCTTCAATCCAAAATAAGGTATATTATTGATGATG(SEQ ID NO:27)。
在本发明的第二方面，本发明提出了一种重组病毒。根据本发明的实施例，所述重组病毒包括：第一核酸分子，所述第一核酸分子含有肿瘤细胞特异性启动子；第二核酸分子，所述第二核酸分子与所述第一核酸分子可操作地连接，所述第二核酸分子编码转录激活因子，所述转录激活因子为Gal4VP16；第三核酸分子，所述第三核酸分子含有所述转录激活因子的第一识别序列；第四核酸分子，所述第四核酸分子与所述第三核酸分子可操作地连接，所述第四核酸分子含有第一启动子和第一调控元件，所述第一启动子为miniCMV，所述第一调控元件包括多个重复的tetO序列，所述多个重复的tetO序列的至少之一设置在所述第一启动子的下游；第五核酸分子，所述第五核酸分子与第四核酸分子可操作地连接，所述第五核酸分子编码第一调控蛋白，所述第一调控蛋白为LacI；所述第五核酸分子进一步包括编码所述目的蛋白的序列，并且所述目的蛋白包括病毒复制蛋白、免疫效应因子，所述免疫效应因子是以单独或融合蛋白的形式表达的，并且所述毒复制蛋白和所述效应因子之间通过可切割的连接肽连接的，所述目的蛋白与所述第一调控蛋白是以融合蛋白的形式表达的，并且所述目的蛋白与所述第一调控蛋白之间通过可切割的连接肽连接的；第六核酸分子，所述第六核酸分子含有所述转录激活因子的第二识别序列；第七核酸分子，所述第七核酸分子与所述第六核酸分子可操作地连接，所述第七核酸分子含有第二启动子和第二调控元件，所述第二启动子为miniCMV，所述第二调控元件包括多个重复的LacO序列，所述多个重复的LacO序列的至少之一设置在所述第二启动子的下游；第八核酸分子，所述第八核酸分子与第七核酸分子可操作地连接，并且所述第八核酸分子编码第二调控蛋白,所述第二调控蛋白为tetR-KRAB；第九核酸分子，所述第九核酸分子与所述第五核酸分子可操作地连接，所述第九核酸分子被配置为条件性抑制所述第一调控蛋白的表达，所述第九核酸分子含有被第一microRNA特异性识别的核酸序列，所述第一microRNA为正常细胞特异性microRNA；以及第十核酸分子，所述第十核酸分子与所述第八核酸分子可操作地连接，所述第十核酸分子被配置为条件性抑制所述第二调控蛋白的表达，所述第十核酸分子含有被第二microRNA特异性识别的核酸序列，所述第二microRNA为肿瘤细胞特异性microRNA，其中，所述第一调控元件适于通过结合所述第二调控蛋白抑制所述第一启动子的功能，所述第二调控元件适于通过结合所述第一调控蛋白抑制所述第二启动子的功能；所述肿瘤细胞特异性启动子为CEA205，所述第一microRNA为mir-135a-5p，所述第二microRNA为mir-21，所述第一识别序列与所述第二识别序列为5×UAS；或所述肿瘤细胞特异性启动子为CEA368，所述第一microRNA为mir-135a-5p，所述第二microRNA为mir-21，所述第一识别序列与所述第二识别序列为5×UAS；或所述肿瘤细胞特异性启动子为CXCR4，所述第一microRNA为mir-135a-5p，所述第二microRNA为mir-21，所述第一识别序列与所述第二识别序列为5×UAS；或所述肿瘤细胞特异性启动子为Survivin1，所述第一microRNA为mir-135a-5p，所述第二microRNA为mir-21，所述第一识别序列与所述第二识别序列为5×UAS；或所述肿瘤细胞特异性启动子为CEA368，所述第一microRNA为mir-143-3p，所述第二microRNA为mir-21，所述第一识别序列与所述第二识别序列为5×UAS；或所述肿瘤细胞特异性启动子为CEA205，所述第一microRNA为mir-135a-5p，所述第二microRNA为mir-21，所述第一识别序列与所述第二识别序列为4×UAS；或所述肿瘤细胞特异性启动子为CEA368，所述第一microRNA为mir-135a-5p，所述第二microRNA为mir-21，所述第一识别序列与所述第二识别序列为4×UAS；或所述肿瘤细胞特异性启动子为CXCR4，所述第一microRNA为mir-135a-5p，所述第二microRNA为mir-21，所述第一识别序列与所述第二识别序列为4×UAS；或所述肿瘤细胞特异性启动子为Survivin1，所述第一microRNA为mir-135a-5p，所述第二microRNA为mir-21，所述第一识别序列与所述第二识别序列为4×UAS；或所述肿瘤细胞特异性启动子为CEA368，所述第一microRNA为mir-143-3p，所述第二microRNA为mir-21，所述第一识别序列与所述第二识别序列为4×UAS；或所述肿瘤细胞特异性启动子为hMuc1，所述第一microRNA为mir-199a-3p，所述第二microRNA为mir-21，所述第一识别序列与所述第二识别序列为4×UAS。利用根据本申请实施例的重组病毒，在肿瘤细胞特异性启动子和第九核酸分子、第十核酸分子的共同调控下，第一调控蛋白LacI、目的蛋白在胃癌、胰腺癌细胞中特异性表达，第二调控蛋白tetR-KRAB在胃癌、胰腺癌细胞中特异性不表达或低表达，进而tetR-KRAB介导的第一启动子miniCMV的抑制机制解除，第一调控蛋白LacI、目的蛋白在第一启动子miniCMV的启动调控下有效表达，LacI介导的抑制机制有效抑制了第二启动子miniCMV的功能，tetR-KRAB的表达进一步受到抑制。进而，利用根据本发明实施例的重组病毒，可实现蛋白在胃癌、胰腺癌细胞中更加特异性的表达(如目的蛋白或LacI)或不表达(如tetR-KRAB)，在胃癌、胰腺癌细胞中的目的蛋白表达表达效率和特异性高。根据本发明实施例的重组病毒可实现对胃癌、胰腺癌细胞的特异、高效、安全的杀伤。
根据本发明的实施例，上述重组病毒还可以进一步包括如下附加技术特征至少之一：
根据本发明的实施例，所述重组病毒为选自逆转录病毒、腺病毒、疱疹病毒、牛痘病毒的至少之一。
根据本发明的实施例，所述重组病毒为腺病毒。如前所述，腺病毒作为基因治疗载体具有宿主范围广，对人致病性低、在增殖和非增殖细胞中感染和表达基因、滴度高、与人类基因同源、无插入致突变性、能在悬浮培养液中扩增、能同时表达多个基因的优点。
根据本发明的实施例，所述免疫效应因子包括选自拮抗PD-1基因的抑制序列、拮抗PD-L1基因的抑制序列、拮抗CTLA4基因的抑制序列、拮抗Tim-3基因的抑制序列、IL-2、IL-15、IL-12、GM-CSF至少之一。任选地，所述免疫效应因子可以以融合蛋白的形式存在。
在本发明的第三方面，本发明提出了一种重组细胞。根据本发明的实施例，所述重组细胞含有前面所述的表达系统。根据本发明实施例的重组细胞可有效激活人体的系统性的免疫反应，特异性攻击胃癌、胰腺癌细胞，安全性高，特异性强。
根据本发明的实施例，上述重组细胞还可以进一步包括如下附加技术特征至少之一：
根据本发明的实施例，所述表达系统的至少一部分整合于所述重组细胞的基因组中。表达系统随着重组细胞基因组的复制而复制，表达系统对目的蛋白的表达调控持续而有效。
在本发明的第四方面，本发明提出了前面所述的表达系统、前面所述的重组病毒、前面所述的重组细胞在制备药物中的用途，所述药物用于治疗胃癌或胰腺癌。
在本发明的第五方面，本发明提出了一种药物组合物。根据本发明的实施例，所述药物组合物包含前面所述的重组病毒或前面所述的重组细胞。根据本发明实施例的药物组合物对胃癌或胰腺癌的治疗效果显著。
根据本发明的实施例，上述药物组合物还可以进一步包括药学上可接受的辅料。
根据本发明的实施例，所述药物组合物进一步包括其他治疗胃癌的药物。
根据本发明的实施例，所述其他治疗胃癌的药物包括选自Pembrolizumab、Ogivri、Everolimus、Lanreotide、Ramucirumab、Apatinib、Trastuzumab的至少之一。
根据本发明的实施例，所述药物组合物进一步包括其他治疗胰腺癌的药物。
根据本发明的实施例，所述其他治疗胰腺癌的药物包括选自Lanreotide、Abraxane、Olaparib、Afinitor、Erlotinib、Everolimus、5-FU、Gemzar、Sunitinib、Onivyde、Gemzar的至少之一。
本发明的药物组合物可以在治疗或预防胃癌、胰腺癌时被给药。
在本文中所使用的术语“给药”指将预定量的物质通过某种适合的方式引入病人。本发明的药物组合物可以通过任何常见的途径被给药，只要它可以到达预期的组织。给药的各种方式是可以预期的，包括腹膜，静脉，肌肉，皮下，皮层，口服，局部，鼻腔，肺部和直肠，但是本发明不限于这些已举例的给药方式。然而，由于口服给药时，口服给药的组合物的活性成分应该被包被或被配制以防止其在胃部被降解。优选地，本发明的组合物可以注射制剂被给药。此外，本发明的药物组合物可以使用将活性成分传送到靶细胞的特定器械来给药。
本发明的药物组合物的给药频率和剂量可以通过多个相关因素被确定，该因素包括要被治疗的疾病类型，给药途径，病人年龄，性别，体重和疾病的严重程度以及作为活性成分的药物类型。根据本发明的一些实施例，日剂量可分为适宜形式的1剂、2剂或多剂，以在整个时间段内以1次、2次或多次给药，只要达到治疗有效量即可。
术语“治疗有效量”是指化合物足以显著改善某些与疾病或病症相关的症状的量，也即为给定病症和给药方案提供治疗效果的量。例如，在胃癌或胰腺癌的治疗中，减少、预防、延缓、抑制或阻滞疾病或病症的任何症状的药物或化合物应当是治疗有效的。治疗有效量的药物或化合物不需要治愈疾病或病症，但将为疾病或病症提供治疗，使得个体的疾病或病症的发作被延缓、阻止或预防，或者疾病或病症的症状得以缓解，或者疾病或病症的期限被改变，或者例如疾病或病症变得不严重，或者加速康复。
术语“治疗”用于指获得期望的药理学和/或生理学效果。所述效果就完全或部分预防疾病或其症状而言可以是预防性的，和/或就部分或完全治愈疾病和/或疾病导致的不良作用而言可以是治疗性的。本文使用的“治疗”涵盖哺乳动物、特别是人的疾病(主要指胃癌或胰腺癌)的治疗，包括：(a)在容易患病但是尚未确诊得病的个体中预防疾病(例如预防胃癌或胰腺癌)或病症发生；(b)抑制疾病，例如阻滞疾病发展；或(c)缓解疾病，例如减轻与疾病相关的症状。本文使用的“治疗”涵盖将药物或化合物给予个体以治疗、治愈、缓解、改善、减轻或抑制个体的疾病的任何用药，包括但不限于将含本文所述的药物给予有需要的个体。
根据本发明的实施例，本发明的药物组合物可与常规治疗方法和/或疗法相结合使用，或者可与常规治疗方法和/或疗法分开使用。当本发明的药物在采用与其它药物的联合疗法中给药时，它们可序贯地或同时地给予个体。或者，本发明的药物组合物可包含本发明的重组病毒或药学上可接受的赋形剂以及本领域已知的其它治疗药或预防药的组合。
需要说明的是，如无特别说明，本申请所述的融合蛋白是指在同一启动子调控下共转录的蛋白，包括蛋白间无连接的融合蛋白，或以其他连接肽(如GGGS或2A序列)连接的融合蛋白。
如无特别说明，本申请所述的闭合回路或表达系统的“翻转能力”是指受第一microRNA和第二microRNA输入信号分别控制时输出水平的差异，具体体现在目的蛋白(包括选自病毒复制包装蛋白、免疫效应因子的至少之一)表达水平的差异。
本申请所述的“几×序列”是指几段重复的序列顺序连接，重复序列之间没有间隔碱基，例如，“5×UAS”是指5段重复的UAS顺利连接所组成的序列，“6×tetO”是指6段重复的tetO顺序连接所组成的序列。
附图说明
图1为根据本发明实施例的靶向特定肿瘤的溶瘤腺病毒构建与测试流程示意图；
图2为根据本发明实施例的不同癌特异启动子在胃癌与胃正常细胞系中的表达测试；
图3为根据本发明实施例的不同癌特异启动子在胃癌与胃正常细胞系中翻转开关线路的测试；
图4为根据本发明实施例的不同microRNA靶位点在胃癌与胃正常细胞系中的抑制效率测试；
图5(包括图5A～5J)为根据本发明实施例的不同腺病毒在胃癌与正常细胞系中的杀伤能力测试；
图6为根据本发明实施例的不同癌特异启动子在乳腺癌与乳腺正常细胞系中的表达测试；
图7为根据本发明实施例的不同癌特异启动子在乳腺癌与乳腺正常细胞系中翻转开关线路的测试；
图8为根据本发明实施例的不同microRNA靶位点在乳腺癌与乳腺正常细胞系中的抑制效率测试。
图9(包括图9A～9D)为根据本发明实施例的不同腺病毒在乳腺癌与正常细胞系中的杀伤能力测试；
图10为不同癌特异启动子在胰腺癌与胰腺正常细胞系中的表达测试；
图11为根据本发明实施例的不同癌特异启动子在胰腺癌与胰腺正常细胞系中翻转开关线路的测试；
图12为根据本发明实施例的不同腺病毒在胰腺癌与正常细胞系中的杀伤能力测试；以及
图13为根据本发明实施例的5型腺病毒载体的图谱。
具体实施方式
下面详细描述本发明的实施例，所述实施例的示例在附图中示出。下面通过参考附图描述的实施例是示例性的，旨在用于解释本发明，而不能理解为对本发明的限制。实施例中未注明具体技术或条件的，按照本领域内的文献所描述的技术或条件或者按照产品说明书进行。所用试剂或仪器未注明生产厂商者，均为可以通过市购获得的常规产品。
在以下实施例中，涉及的构建靶向特定肿瘤的溶瘤腺病毒的方法以及测试流程可参照示意图，图1；所涉及的病毒代号如表1所示。
表1：
实施例1靶向胃癌的溶瘤腺病毒的构建与功能验证
实验一
测定癌特异性启动子CEA205、CEA368、cerbB2、COX2、CXCR4、hMUC1、Survivin1与Survivin3在胃癌细胞系AGS与胃正常细胞系GES1中的表达强度与特异性。
将CEA205-Gal4VP16的质粒、UAS-EYFP质粒、hEF1a-EBFP2质粒共转染胃癌细胞系AGS与胃正常细胞系GES1(每孔转染各质粒各100ng)，转染48小时后进行流式细胞术分析，检测EYFP与EBFP2荧光强度。归一化报告基因表达水平＝(实验组EYFP荧光强度/实验组EBFP2荧光强度)/(对照组EYFP荧光强度/对照组EBFP2荧光强度)。
用CEA368-Gal4VP16质粒代替CEA205-Gal4VP16质粒，进行上述步骤。用cerbB2-Gal4VP16质粒代替CEA205-Gal4VP16质粒，进行上述步骤。用COX2-Gal4VP16质粒代替CEA205-Gal4VP16质粒，进行上述步骤。用CXCR4-Gal4VP16质粒代替CEA205-Gal4VP16质粒，进行上述步骤。用hMUC1-Gal4VP16质粒代替CEA205-Gal4VP16质粒，进行上述步骤。用Survivin1-Gal4VP16质粒代替CEA205-Gal4VP16质粒，进行上述步骤。用Survivin3-Gal4VP16质粒代替CEA205-Gal4VP16质粒，进行上述步骤。用CAG-Gal4VP16质粒代替CEA205-Gal4VP16质粒，进行上述步骤。
归一化报告基因表达水平见图2。各癌特异启动子在胃正常细胞系GES1中表达量均很低，而在胃癌细胞系AGS中，CEA205、CEA368、CXCR4与Survivin1表达量较高。结果表明：癌特异启动子可以特异性地在胃癌细胞系中启动Gal4VP16的表达并激活UAS下游的报告基因。CEA205、CEA368、CXCR4与Survivin1有较好的表达强度与特异性。
实验二
在实验一的基础上，测定具有较好表达强度与特异性的CEA205、CEA368、CXCR4与Survivin1启动5×UAS或4×UAS开关线路的翻转能力。
将CEA205-Gal4VP16质粒、5×UAS-TetO-E1A-P2A-EBFP2-2A-LacI-FF4质粒、5×UAS-LacO-TetRKrab-FF5质粒、CAG-EYFP质粒、pDT7004质粒共转染胃癌细胞系AGS与胃正常细胞系GES1(每孔转染各质粒各100ng)，转染48小时后进行流式细胞术分析，检测EYFP与EBFP2荧光强度，表示无输入信号时开关线路的表达水平。报告基因表达水平＝EBFP2荧光强度/EYFP荧光强度。
用U6-shRNA-FF4-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤，表示一侧输入信号时开关线路的表达水平；用U6-shRNA-FF5-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤，表示另一侧输入信号时开关线路的表达水平。
用4×UAS-TetO-E1A-P2A-EBFP2-2A-LacI-FF4质粒代替5×UAS-TetO-E1A-P2A-EBFP2-2A-LacI-FF4质粒，4×UAS-LacO-TetRKrab-FF5质粒代替5×UAS-LacO-TetRKrab-FF5质粒后，进行上述步骤；用U6-shRNA-FF4-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤；用U6-shRNA-FF5-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤。
用CEA368-Gal4VP16质粒代替CEA205-Gal4VP16质粒后，进行上述步骤；用U6-shRNA-FF4-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤；用U6-shRNA-FF5-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤。
用CEA368-Gal4VP16质粒代替CEA205-Gal4VP16质粒，4×UAS-TetO-E1A-P2A-EBFP2-2A-LacI-FF4质粒代替5×UAS-TetO-E1A-P2A-EBFP2-2A-LacI-FF4质粒，4×UAS-LacO-TetRKrab-FF5质粒代替5×UAS-LacO-TetRKrab-FF5质粒后，进行上述步骤；用U6-shRNA-FF4-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤；用U6-shRNA-FF5-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤。
用CXCR4-Gal4VP16质粒代替CEA205-Gal4VP16质粒后，进行上述步骤；用U6-shRNA-FF4-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤；用U6-shRNA-FF5-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤。
用CXCR4-Gal4VP16质粒代替CEA205-Gal4VP16质粒，4×UAS-TetO-E1A-P2A-EBFP2-2A-LacI-FF4质粒代替5×UAS-TetO-E1A-P2A-EBFP2-2A-LacI-FF4质粒，4×UAS-LacO-TetRKrab-FF5质粒代替5×UAS-LacO-TetRKrab-FF5质粒后，进行上述步骤；用U6-shRNA-FF4-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤；用U6-shRNA-FF5-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤。
用Survivin1-Gal4VP16质粒代替CEA205-Gal4VP16质粒后，进行上述步骤；用U6-shRNA-FF4-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤；用U6-shRNA-FF5-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤。
用Survivin1-Gal4VP16质粒代替CEA205-Gal4VP16质粒，4×UAS-TetO-E1A-P2A-EBFP2-2A-LacI-FF4质粒代替5×UAS-TetO-E1A-P2A-EBFP2-2A-LacI-FF4质粒，4×UAS-LacO-TetRKrab-FF5质粒代替5×UAS-LacO-TetRKrab-FF5质粒后，进行上述步骤；用U6-shRNA-FF4-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤；用U6-shRNA-FF5-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤。
用CAG-Gal4VP16质粒代替CEA205-Gal4VP16质粒后，进行上述步骤；用U6-shRNA-FF4-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤；用U6-shRNA-FF5-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤。
用CAG-Gal4VP16质粒代替CEA205-Gal4VP16质粒，4×UAS-TetO-E1A-P2A-EBFP2-2A-LacI-FF4质粒代替5×UAS-TetO-E1A-P2A-EBFP2-2A-LacI-FF4质粒，4×UAS-LacO-TetRKrab-FF5质粒代替5×UAS-LacO-TetRKrab-FF5质粒后，进行上述步骤；用U6-shRNA-FF4-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤；用U6-shRNA-FF5-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤。
报告基因表达水平见图3。与实验一结果一致，被测试的所有启动子在胃正常细胞系GES1中表达量均很低，而在胃癌细胞系AGS中均可以启动开关线路翻转。在胃癌细胞系AGS中CEA368与Survivin1表达量略高于CEA205与CXCR4，而5×UAS开关线路表达强度略高于4×UAS开关线路。结果表明：被测试的所有启动子均可以特异性地在胃癌细胞系AGS中启动4×UAS或5×UAS开关线路翻转。
实验三
测定癌特异性microRNA mir-143-3p、mir-145-5p、mir-551b-3p、mir-1-3p、mir-133a-3p、mir-133b-3p、mir-139-3p、mir-139-5p、mir-145-3p、mir-204-5p、mir-195-3p、mir-935、mir-135a-5p、mir-135b-5p、mir-135b-3p、mir-196a-3p与mir-21在胃癌细胞系AGS与胃正常细胞系GES1中对各靶位点的抑制强度。
将CMV-EYFP-T143 3p x4质粒、CMV-EBFP2质粒、pDT7004质粒共转染胃癌细胞系AGS与胃正常细胞系GES1(每孔转染各质粒各100ng)，转染48小时后进行流式细胞术分析，检测EYFP与EBFP2荧光强度。设置用CMV-EYFP质粒代替CMV-EYFP-T143 3p x4质粒的对照处理。归一化报告基因表达水平＝(实验组EBFP2荧光强度/实验组EYFP荧光强度)/(对照组EBFP2荧光强度/对照组EYFP荧光强度)。
用CMV-EYFP-T145 5p x4质粒代替CMV-EYFP-T143 3p x4质粒后，进行上述步骤。用CMV-EYFP-T551 3p x4质粒代替CMV-EYFP-T143 3p x4质粒后，进行上述步骤。用CMV-EYFP-T1 3p x4质粒代替CMV-EYFP-T143 3p x4质粒后，进行上述步骤。用CMV-EYFP-T133a3p x4质粒代替CMV-EYFP-T143 3p x4质粒后，进行上述步骤。用CMV-EYFP-T133b 3p x4质粒代替CMV-EYFP-T143 3p x4质粒后，进行上述步骤。用CMV-EYFP-T139 3p x4质粒代替CMV-EYFP-T143 3p x4质粒后，进行上述步骤。用CMV-EYFP-T139 5p x4质粒代替CMV-EYFP-T143 3p x4质粒后，进行上述步骤。用CMV-EYFP-T145 3p x4质粒代替CMV-EYFP-T143 3px4质粒后，进行上述步骤。用CMV-EYFP-T204 5p x4质粒代替CMV-EYFP-T143 3p x4质粒后，进行上述步骤。用CMV-EYFP-T195 3p x4质粒代替CMV-EYFP-T143 3p x4质粒后，进行上述步骤。用CMV-EYFP-T935 x4质粒代替CMV-EYFP-T143 3p x4质粒后，进行上述步骤。用CMV-EYFP-T135a 5p x4质粒代替CMV-EYFP-T143 3p x4质粒后，进行上述步骤。用CMV-EYFP-T135b 5p x4质粒代替CMV-EYFP-T143 3p x4质粒后，进行上述步骤。用CMV-EYFP-T135b 3px4质粒代替CMV-EYFP-T143 3p x4质粒后，进行上述步骤。用CMV-EYFP-T196a 5p x4质粒代替CMV-EYFP-T143 3p x4质粒后，进行上述步骤。用CMV-EYFP-T196a 3p x4质粒代替CMV-EYFP-T143 3p x4质粒后，进行上述步骤。用CMV-EYFP-T21x4质粒代替CMV-EYFP-T143 3px4质粒后，进行上述步骤。
归一化报告基因表达水平见图4。mir-21可以通过靶位点特异性地在胃癌细胞系AGS中抑制报告基因表达，并有很高的抑制强度。mir-143-3p与mir-135a-5p可以通过靶位点特异性地在胃正常细胞系GES1中抑制报告基因表达，并有一定地抑制强度。结果表明：癌特异高表达或低表达的microRNA可以通过靶位点特异性地在胃癌细胞系AGS或胃正常细胞系GES1中抑制报告基因表达。癌特异高表达microRNA mir-21、癌特异低表达microRNAmir-143-3p与mir-135a-5p有较好的抑制强度。
实验四
实验一、二、三的基础上，构建使用CEA205/CEA368/CXCR4/Survivin1启动子、mir-21与mir-135a-5p/mir-143-3p调控的4×UAS或5×UAS开关线路控制的溶瘤腺病毒，测试对胃癌细胞系AGS、胃正常细胞系GES1、肝正常细胞系Chang与L02的杀伤能力。
在96孔板中每孔接种数量为1e4的细胞。接种24小时后分别加入感染复数为1、10、20、50、100、200、500的A1病毒，设置不加入病毒的空白对照。病毒侵染6天后使用MTS法检测细胞存活率。细胞存活率＝实验组MTS检测值/对照组MTS检测值。
使用A2病毒代替A1病毒，进行上述步骤。使用A3病毒代替A1病毒，进行上述步骤。使用A4病毒代替A1病毒，进行上述步骤。使用A5病毒代替A1病毒，进行上述步骤。使用A6病毒代替A1病毒，进行上述步骤。使用A7病毒代替A1病毒，进行上述步骤。使用A8病毒代替A1病毒，进行上述步骤。使用A9病毒代替A1病毒，进行上述步骤。使用A10病毒代替A1病毒，进行上述步骤。
细胞存活率见图5。与实验一、二、三结果一致，使用癌特异启动子CEA205/CEA368/CXCR4/Survivin1与癌特异microRNA mir-21及mir-135a-5p/mir-143-3p调控的开关线路控制的溶瘤腺病毒可以特异性地杀伤胃癌细胞系AGS，而对胃正常细胞系GES1及肝正常细胞系Chang、L02杀伤作用不明显。5×UAS开关线路控制的溶瘤腺病毒A1在感染复数约10～20间时就表现出明显的杀伤特异性，而A2、A4、A5与A1杀伤效果相近。4×UAS开关线路控制的溶瘤腺病毒A10与A1杀伤效果相近。结果表明：使用癌特异启动子CEA205/CEA368/Survivin1与癌特异microRNA mir-21及mir-135a-5p/mir-143-3p调控的开关线路控制的溶瘤腺病毒可以特异性地杀伤胃癌细胞系AGS，5×UAS开关线路相较于4×UAS开关线路效果更好。
实施例2靶向乳腺癌的溶瘤腺病毒的构建与功能验证
实验一
结合实施例1实验结果，测定癌特异性启动子CEA205、CEA368、CXCR4、hMUC1与Survivin1在乳腺癌细胞系MCF7、MDA-MB-231与乳腺正常细胞系MCF10A中的表达强度与特异性。
将CEA205-Gal4VP16的质粒、UAS-EYFP质粒、CMV-EBFP2质粒共转染乳腺癌细胞系MCF7、MDA-MB-231与乳腺正常细胞系MCF10A(每孔转染各质粒各100ng)，转染48小时后进行流式细胞术分析，检测EYFP与EBFP2荧光强度。归一化报告基因表达水平＝(实验组EYFP荧光强度/实验组EBFP2荧光强度)/(对照组EYFP荧光强度/对照组EBFP2荧光强度)。
用CEA368-Gal4VP16质粒代替CEA205-Gal4VP16质粒，进行上述步骤。用CXCR4-Gal4VP16质粒代替CEA205-Gal4VP16质粒，进行上述步骤。用hMUC1-Gal4VP16质粒代替CEA205-Gal4VP16质粒，进行上述步骤。用Survivin1-Gal4VP16质粒代替CEA205-Gal4VP16质粒，进行上述步骤。用CAG-Gal4VP16质粒代替CEA205-Gal4VP16质粒，进行上述步骤。
归一化报告基因表达水平见图6。所测试癌特异启动子中，CEA205与CXCR4在乳腺癌细胞系MCF7与MDA-MB-231中表达较高而在乳腺正常细胞系MCF10A中表达较低。结果表明：癌特异启动子可以特异性地在乳腺癌细胞系中启动Gal4VP16的表达并激活UAS下游的报告基因。CEA205与CXCR4有较好的表达强度与特异性。
实验二
测定CEA205、CEA368、CXCR4、hMUC1与Survivin1启动5×UAS或4×UAS开关线路的翻转能力。
将CEA205-Gal4VP16质粒、5×UAS-TetO-E1A-P2A-EBFP2-2A-LacI-FF4质粒、5×UAS-LacO-TetRKrab-FF5质粒、CAG-EYFP质粒、pDT7004质粒共转染乳腺癌细胞系MCF7与乳腺正常细胞系MCF10A(每孔转染各质粒各100ng)，转染48小时后进行流式细胞术分析，检测EYFP与EBFP2荧光强度，表示无输入信号时开关线路的表达水平。报告基因表达水平＝EBFP2荧光强度/EYFP荧光强度。
用U6-shRNA-FF4-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤，表示一侧输入信号时开关线路的表达水平；用U6-shRNA-FF5-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤，表示另一侧输入信号时开关线路的表达水平。
用4×UAS-TetO-E1A-P2A-EBFP2-2A-LacI-FF4质粒代替5×UAS-TetO-E1A-P2A-EBFP2-2A-LacI-FF4质粒，4×UAS-LacO-TetRKrab-FF5质粒代替5×UAS-LacO-TetRKrab-FF5质粒后，进行上述步骤；用U6-shRNA-FF4-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤；用U6-shRNA-FF5-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤。
用CEA368-Gal4VP16质粒代替CEA205-Gal4VP16质粒后，进行上述步骤；用U6-shRNA-FF4-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤；用U6-shRNA-FF5-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤。
用CEA368-Gal4VP16质粒代替CEA205-Gal4VP16质粒，4×UAS-TetO-E1A-P2A-EBFP2-2A-LacI-FF4质粒代替5×UAS-TetO-E1A-P2A-EBFP2-2A-LacI-FF4质粒，4×UAS-LacO-TetRKrab-FF5质粒代替5×UAS-LacO-TetRKrab-FF5质粒后，进行上述步骤；用U6-shRNA-FF4-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤；用U6-shRNA-FF5-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤。
用CXCR4-Gal4VP16质粒代替CEA205-Gal4VP16质粒后，进行上述步骤；用U6-shRNA-FF4-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤；用U6-shRNA-FF5-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤。
用CXCR4-Gal4VP16质粒代替CEA205-Gal4VP16质粒，4×UAS-TetO-E1A-P2A-EBFP2-2A-LacI-FF4质粒代替5×UAS-TetO-E1A-P2A-EBFP2-2A-LacI-FF4质粒，4×UAS-LacO-TetRKrab-FF5质粒代替5×UAS-LacO-TetRKrab-FF5质粒后，进行上述步骤；用U6-shRNA-FF4-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤；用U6-shRNA-FF5-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤。
用hMUC1-Gal4VP16质粒代替CEA205-Gal4VP16质粒后，进行上述步骤；用U6-shRNA-FF4-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤；用U6-shRNA-FF5-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤。
用hMUC1-Gal4VP16质粒代替CEA205-Gal4VP16质粒，4×UAS-TetO-E1A-P2A-EBFP2-2A-LacI-FF4质粒代替5×UAS-TetO-E1A-P2A-EBFP2-2A-LacI-FF4质粒，4×UAS-LacO-TetRKrab-FF5质粒代替5×UAS-LacO-TetRKrab-FF5质粒后，进行上述步骤；用U6-shRNA-FF4-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤；用U6-shRNA-FF5-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤。
用Survivin1-Gal4VP16质粒代替CEA205-Gal4VP16质粒后，进行上述步骤；用U6-shRNA-FF4-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤；用U6-shRNA-FF5-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤。
用Survivin1-Gal4VP16质粒代替CEA205-Gal4VP16质粒，4×UAS-TetO-E1A-P2A-EBFP2-2A-LacI-FF4质粒代替5×UAS-TetO-E1A-P2A-EBFP2-2A-LacI-FF4质粒，4×UAS-LacO-TetRKrab-FF5质粒代替5×UAS-LacO-TetRKrab-FF5质粒后，进行上述步骤；用U6-shRNA-FF4-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤；用U6-shRNA-FF5-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤。
用CAG-Gal4VP16质粒代替CEA205-Gal4VP16质粒后，进行上述步骤；用U6-shRNA-FF4-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤；用U6-shRNA-FF5-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤。
用CAG-Gal4VP16质粒代替CEA205-Gal4VP16质粒，4×UAS-TetO-E1A-P2A-EBFP2-2A-LacI-FF4质粒代替5×UAS-TetO-E1A-P2A-EBFP2-2A-LacI-FF4质粒，4×UAS-LacO-TetRKrab-FF5质粒代替5×UAS-LacO-TetRKrab-FF5质粒后，进行上述步骤；用U6-shRNA-FF4-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤；用U6-shRNA-FF5-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤。
报告基因表达水平见图7。在乳腺正常细胞系MCF10A中所测试的启动子翻转5×UAS开关线路表达量较低，而翻转4×UAS开关线路有一定表达。在乳腺癌细胞系MCF7中所测试的启动未能表现中对5×UAS与4×UAS开关线路较好的翻转能力。结果表明：所测试的启动子在乳腺癌细胞系MCF7中对开关线路的翻转能力不强。
实验三
测定癌特异性microRNA mir-205-5p、mir-141-3p与mir-21在乳腺癌细胞系MCF7、MDA-MB-231与乳腺正常细胞系MCF10A中对各靶位点的抑制强度。
将CMV-EYFP-T205 5p x4质粒、CMV-EBFP2质粒、pDT7004质粒共转染乳腺癌细胞系MCF7、MDA-MB-231与乳腺正常细胞系MCF10A(每孔转染各质粒各100ng)，转染48小时后进行流式细胞术分析，检测EYFP与EBFP2荧光强度。设置用CMV-EYFP质粒代替CMV-EYFP-T205 5px4质粒的对照处理。归一化报告基因表达水平＝(实验组EBFP2荧光强度/实验组EYFP荧光强度)/(对照组EBFP2荧光强度/对照组EYFP荧光强度)。
用CMV-EYFP-T141 3p x4质粒代替CMV-EYFP-T205 5p x4质粒后，进行上述步骤。用CMV-EYFP-T21x4质粒代替CMV-EYFP-T143 3p x4质粒后，进行上述步骤。
归一化报告基因表达水平见图8。mir-205-5p可以通过靶位点特异性地在乳腺正常细胞系MCF10A中抑制报告基因表达，并有很高的抑制强度，与调研结果一致。mir-141-3p虽然在乳腺癌细胞系MDA-MB-231中抑制强度较低，但在MCF7中有一定的抑制强度。mir-21虽然在乳腺癌细胞系MCF7与MDA-MB-231中抑制强度很高，但在乳腺正常细胞系MCF10A中抑制强度也很高。结果表明：癌特异高表达或低表达的microRNA可以通过靶位点在乳腺癌细胞系MCF7、MDA-MB-231与乳腺正常细胞系MCF10A中抑制报告基因表达，但特异性不够好。
实验四
实验一、二、三的基础上，构建使用CEA205/Survivin1启动子、mir-21与mir-141-3p调控的5×UAS开关线路控制的溶瘤腺病毒，测试对乳腺癌细胞系MCF7、MDA-MB-231、乳腺正常细胞系MCF10A、肝正常细胞系Chang与L02的杀伤能力。
在96孔板中每孔接种数量为1e4的细胞。接种24小时后分别加入感染复数为1、10、20、50、100、200、500的B1病毒，设置不加入病毒的空白对照。病毒侵染6天后使用MTS法检测细胞存活率。细胞存活率＝实验组MTS检测值/对照组MTS检测值。
使用B2病毒代替B1病毒，进行上述步骤。使用B3病毒代替B1病毒，进行上述步骤。
使用B4病毒代替B1病毒，进行上述步骤。
细胞存活率见图9。四种腺病毒均未能有效杀伤乳腺癌细胞系MCF7与MDA-MB-231而对乳腺正常细胞系MCF10A有一定杀伤作用。结果表明：使用癌特异启动子CEA205/Survivin1与癌特异microRNA mir-21及mir-141-3p调控的开关线路控制的溶瘤腺病毒未能特异性地杀伤乳腺癌细胞系MCF7与MDA-MB-231。
实施例3靶向胰腺癌的溶瘤腺病毒的构建与功能验证
实验一
结合实施例1实验结果，测定癌特异性启动子hMUC1在胰腺癌细胞系PANC1与胰腺正常细胞系HTERT-HPNE中的表达强度与特异性。
将hMUC1-Gal4VP16的质粒、UAS-EYFP质粒、CMV-EBFP2质粒共转染胰腺癌细胞系PANC1与胰腺正常细胞系HTERT-HPNE(每孔转染各质粒各100ng)，转染48小时后进行流式细胞术分析，检测EYFP与EBFP2荧光强度。归一化报告基因表达水平＝(实验组EYFP荧光强度/实验组EBFP2荧光强度)/(对照组EYFP荧光强度/对照组EBFP2荧光强度)。
用CAG-Gal4VP16质粒代替CEA205-Gal4VP16质粒，进行上述步骤。
归一化报告基因表达水平见图10。hMUC1在胰腺癌细胞系PANC1中表达较高而在胰腺正常细胞系HTERT-HPNE中表达较低。结果表明：癌特异启动子hMUC1可以特异性地在乳腺癌细胞系中启动Gal4VP16的表达并激活UAS下游的报告基因。
实验二
测定hMUC1启动5×UAS或4×UAS开关线路的翻转能力。
将hMUC1-Gal4VP16质粒、5×UAS-TetO-E1A-P2A-EBFP2-2A-LacI-FF4质粒、5×UAS-LacO-TetRKrab-FF5质粒、CAG-EYFP质粒、pDT7004质粒共转染胰腺癌细胞系PANC1与胰腺正常细胞系HTERT-HPNE(每孔转染各质粒各100ng)，转染48小时后进行流式细胞术分析，检测EYFP与EBFP2荧光强度，表示无输入信号时开关线路的表达水平。报告基因表达水平＝EBFP2荧光强度/EYFP荧光强度。
用U6-shRNA-FF4-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤，表示一侧输入信号时开关线路的表达水平；用U6-shRNA-FF5-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤，表示另一侧输入信号时开关线路的表达水平。
用4×UAS-TetO-E1A-P2A-EBFP2-2A-LacI-FF4质粒代替5×UAS-TetO-E1A-P2A-EBFP2-2A-LacI-FF4质粒，4×UAS-LacO-TetRKrab-FF5质粒代替5×UAS-LacO-TetRKrab-FF5质粒后，进行上述步骤；用U6-shRNA-FF4-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤；用U6-shRNA-FF5-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤。
用CAG-Gal4VP16质粒代替CEA205-Gal4VP16质粒后，进行上述步骤；用U6-shRNA-FF4-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤；用U6-shRNA-FF5-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤。
用CAG-Gal4VP16质粒代替CEA205-Gal4VP16质粒，4×UAS-TetO-E1A-P2A-EBFP2-2A-LacI-FF4质粒代替5×UAS-TetO-E1A-P2A-EBFP2-2A-LacI-FF4质粒，4×UAS-LacO-TetRKrab-FF5质粒代替5×UAS-LacO-TetRKrab-FF5质粒后，进行上述步骤；用U6-shRNA-FF4-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤；用U6-shRNA-FF5-CMV-iRFP质粒代替pDT7004质粒，进行上述步骤。
报告基因表达水平见图11。在胰腺癌细胞系PANC1中hMUC1可以翻转5×UAS与4×UAS开关线路，但在胰腺正常细胞系HTERT-HPNE中也有一定表达。结果表明：hMUC1在胰腺癌细胞系PANC1中对开关线路有一定翻转能力，但在胰腺正常细胞系HTERT-HPNE中有一定泄漏。
实验三
实验一、二基础上，构建用hMUC1启动子、mir-21与mir-199a-3p调控的5×UAS与4×UAS开关线路控制的溶瘤腺病毒，测试对胰腺癌细胞系PANC1、肝正常细胞系Chang与L02的杀伤能力。
在96孔板中每孔接种数量为1e4的细胞。接种24小时后分别加入感染复数为0.1、1、5、10、20、50、100的C1病毒，设置不加入病毒的空白对照。病毒侵染6天后使用MTS法检测细胞存活率。细胞存活率＝实验组MTS检测值/对照组MTS检测值。
使用C2病毒代替C1病毒，进行上述步骤。
细胞存活率见图12。两种腺病毒均能有效地特异性地杀伤胰腺癌细胞系PANC1。C1在感染复数约20～50间时就表现出明显的杀伤特异性，而C2杀伤效果略弱于pAD043。结果表明：使用癌特异启动子hMUC1与癌特异microRNA mir-21、mir-199a-3p调控的开关线路控制的溶瘤腺病毒可以特异性地杀伤胰腺癌细胞系PANC1
在本说明书的描述中，参考术语“一个实施例”、“一些实施例”、“示例”、“具体示例”、或“一些示例”等的描述意指结合该实施例或示例描述的具体特征、结构、材料或者特点包含于本发明的至少一个实施例或示例中。在本说明书中，对上述术语的示意性表述不必须针对的是相同的实施例或示例。而且，描述的具体特征、结构、材料或者特点可以在任一个或多个实施例或示例中以合适的方式结合。此外，在不相互矛盾的情况下，本领域的技术人员可以将本说明书中描述的不同实施例或示例以及不同实施例或示例的特征进行结合和组合。
尽管上面已经示出和描述了本发明的实施例，可以理解的是，上述实施例是示例性的，不能理解为对本发明的限制，本领域的普通技术人员在本发明的范围内可以对上述实施例进行变化、修改、替换和变型。
SEQUENCE LISTING
<110> 北京合生基因科技有限公司
<120> 特异杀伤肿瘤细胞的溶瘤病毒系统及其应用
<130> PIDC3184908A
<160> 27
<170> PatentIn version 3.3
<210> 1
<211> 6087
<212> DNA
<213> Artificial
<220>
<223> 编码由细胞特异性启动子CEA205、第一microRNA为mir-135a-5p、第二micr
oRNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白分
别为Lacl、tetR-KRAB、第一和第二识别序列为5×UAS的元件组成的表达系
统的核酸
<400> 1
cggggtgccg ccccccgcgg gtgaccgccc tccgcgggtg acagccctcc gcggagtgcc 60
gcactccgcg gagtgctgta ctccgagcgg agactctaga ctccctatca gtgatagaga 120
taggcgtgta cggtgggagg cctatataag cagagctcgt ttagtgaacc gtcagatcgc 180
tccctatcag tgatagagag aattcgaccg ccaccatgag acatattatc tgccacggag 240
gtgttattac cgaagaaatg gccgccagtc ttttggacca gctgatcgaa gaggtactgg 300
ctgataatct tccacctcct agccattttg aaccacctac ccttcacgaa ctgtatgatt 360
tagacgtgac ggcccccgaa gatcccaacg aggaggcggt ttcgcagatt tttcccgact 420
ctgtaatgtt ggcggtgcag gaagggattg acttactcac ttttccgccg gcgcccggtt 480
ctccggagcc gcctcacctt tcccggcagc ccgagcagcc ggagcagaga gccttgggtc 540
cggtttctat gccaaacctt gtaccggagg tgatcgatct tacctgccac gaggctggct 600
ttccacccag tgacgacgag gatgaagagg gtgaggagtt tgtgttagat tatgtggagc 660
accccgggca cggttgcagg tcttgtcatt atcaccggag gaatacgggg gacccagata 720
ttatgtgttc gctttgctat atgaggacct gtggcatgtt tgtctacagt aagtgaaaat 780
tatgggcagt gggtgataga gtggtgggtt tggtgtggta attttttttt taatttttac 840
agttttgtgg tttaaagaat tttgtattgt gattttttta aaaggtcctg tgtctgaacc 900
tgagcctgag cccgagccag aaccggagcc tgcaagacct acccgccgtc ctaaaatggc 960
gcctgctatc ctgaggcgcc cgacatcacc tgtgtctaga gaatgcaata gtagtacgga 1020
tagctgtgac tccggtcctt ctaacacacc tcctgagata cacccggtgg tcccgctgtg 1080
ccccattaaa ccagttgccg tgagagttgg tgggcgtcgc caggctgtgg aatgtatcga 1140
ggacttgctt aacgagcctg ggcaaccttt ggacttgagc tgtaaacgcc ccaggccact 1200
cgagggtacc gggtccggag ctacaaattt ttccctcctc aaacaggctg gagatgtcga 1260
agaaaatcct gggcctaccg gtatgtggct gcagagcctc cttctactgg gcaccgtggc 1320
ttgcagcatc tcggcccccg ccagaagtcc cagccccagc acccagccct gggaacacgt 1380
aaatgcaatc caggaggccc gccgcctcct gaacttgtca cgggacacgg cagccgagat 1440
gaatgaaact gtggaggtca tcagtgaaat gtttgacttg caggagccca catgcctgca 1500
gactcggctg gagctctaca aacagggcct gcgtggcagc ctcaccaagc tgaagggccc 1560
actgaccatg atggctagcc attacaagca gcactgcccg ccgacccctg agacatcctg 1620
tgccactcag atcattacct ttgagtcttt caaggagaac ctcaaggact ttctgctggt 1680
catcccattt gactgctggg aacccgtcca ggaaagatct gagggccgcg gcagcctgct 1740
gacctgcggc gacgtggagg aaaaccccgg ccccggatcc atgaaaccag taacgttata 1800
cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc 1860
cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta 1920
cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga ttggcgttgc 1980
cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc 2040
cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg 2100
taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca ttaactatcc 2160
gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc cggcgttatt 2220
tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg aagacggtac 2280
gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg 2340
cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat atctcactcg 2400
caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt ccggttttca 2460
acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg ttgccaacga 2520
tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga 2580
tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc cgccgttaac 2640
caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact 2700
ctctcagggc caggcggtga agggcaatca gctgttgccg gtctcactgg tgaaaagaaa 2760
aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat 2820
gcagctggca cgacaggttt cccgactgga aagcgggcag agcagcctga ggcctcctaa 2880
gaagaagagg aaggttgtta caaacaccat tgtcacactc cacaaacacc attgtcacac 2940
tccacaaaca ccattgtcac actccacaaa caccattgtc acactccatc acataggaat 3000
aaaaagccat atcacatagg aataaaaagc catatcacat aggaataaaa agccatatca 3060
cataggaata aaaagccata gaaagcggac tactgtagtc cgcggagtac tgtcctccgc 3120
ggacgactgt cgtccgcgga gcactgtgct ccgcggatga ctgtcatccg agcggagact 3180
ctagaaattg tgagcggata acaattgggt atataatggg ggccgtcagc tactaccaga 3240
gccgatcgaa cactcgagcc gagcagacgt gcctacaatt gtgagcgctc acaattacta 3300
gaagatcttg aattcgccac catggctaga ttagataaaa gtaaagtgat taacagcgca 3360
ttagagctgc ttaatgaggt cggaatcgaa ggtttaacaa cccgtaaact cgcccagaag 3420
ctaggtgtag agcagcctac attgtattgg catgtaaaaa ataagcgggc tttgctcgac 3480
gccttagcca ttgagatgtt agataggcac catactcact tttgcccttt agaaggggaa 3540
agctggcaag attttttacg taataacgct aaaagtttta gatgtgcttt actaagtcat 3600
cgcgatggag caaaagtaca tttaggtaca cggcctacag aaaaacagta tgaaactctc 3660
gaaaatcaat tagccttttt atgccaacaa ggtttttcac tagagaatgc attatatgca 3720
ctcagcgctg tggggcattt tactttaggt tgcgtattgg aagatcaaga gcatcaagtc 3780
gctaaagaag aaagggaaac acctactact gatagtatgc cgccattatt acgacaagct 3840
atcgaattat ttgatcacca aggtgcagag ccagccttct tattcggcct tgaattgatc 3900
atatgcggat tagaaaaaca acttaaatgt gaaagtgggt cgccaaaaaa gaagagaaag 3960
gtggatggtg gcggagcact gtctccccag cattcagctg tgacccaggg atcaatcatt 4020
aagaacaagg aaggtatgga tgctaagtcc ctgaccgctt ggtcaagaac tttggtaact 4080
tttaaggacg tgttcgtcga ttttactcgg gaggagtgga aactattgga taccgcccag 4140
caaatcgtgt atagaaacgt tatgctcgag aactataaga atcttgtgtc actgggctac 4200
caactgacaa agcctgacgt tatcctgagg ctggaaaagg gtgaggagcc ttggttggtc 4260
gagcgggaga tccaccagga aactcaccca gacagtgaga ctgctttcga gatcaagtcc 4320
tctgtgtaat acaaggcggc cgcaaatcaa catcagtctg ataagctatc aacatcagtc 4380
tgataagcta tcaacatcag tctgataagc tatcaacatc agtctgataa gctaaagctt 4440
cgaattctga taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc 4500
ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt 4560
attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca 4620
tttttttcac tgcagctcgc cttttgctca catgtgaaaa tagaagggaa aaaagaggag 4680
ggacaaaaga ggcagaaatg agaggggagg ggacagagga cacctgaata aagaccacac 4740
ccatgaccca cgtgatgctg agaagtactc ctgccctagg aagagactca gggcagaggg 4800
aggaaggaca gcagaccaga cagtcacagc agccttgaca aaacgttcct ggaactcaag 4860
ctcgagaccg gtatggcccc cccgaccgat gtcagcctgg gggacgagct ccacttagac 4920
ggcgaggacg tggcgatggc gcatgccgac gcgctagacg atttcgatct ggacatgttg 4980
ggggacgggg attccccggg tccgggattt accccccacg actccgcccc ctacggcgct 5040
ctggatatgg ccgacttcga gtttgagcag atgtttaccg atgcccttgg aattgacgag 5100
tacggtggga cgcgtatgaa gctactgtct tctatcgaac aagcatgcga tatttgccga 5160
cttaaaaagc tcaagtgctc caaagaaaaa ccgaagtgcg ccaagtgtct gaagaacaac 5220
tgggagtgtc gctactctcc caaaaccaaa aggtctccgc tgactagggc acatctgaca 5280
gaagtggaat caaggctaga aagactggaa cagctatttc tactgatttt tcctcgagaa 5340
gaccttgaca tgattttgaa aatggattct ttacaggata taaaagcatt gttaacagga 5400
ttatttgtac aagataatgt gaataaagat gccgtcacag atagattggc ttcagtggag 5460
actgatatgc ctctaacatt gagacagcat agaataagtg cgacatcatc atcggaagag 5520
agtagtaaca aaggtcaaag acagttgact gtataaaagc ttcactcctc aggtgcaggc 5580
tgcctatcag aaggtggtgg ctggtgtggc caatgccctg gctcacaaat accactgaga 5640
tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc 5700
tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc 5760
actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta 5820
gagtttggca acatatgccc atatgctggc tgccatgaac aaaggttggc tataaagagg 5880
tcatcagtat atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac 5940
ttgaggttag atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa 6000
attttcctta catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat 6060
agctgtccct cttctcttat ggagatc 6087
<210> 2
<211> 12709
<212> DNA
<213> Artificial
<220>
<223> 编码由细胞特异性启动子CEA205、第一microRNA为mir-143-3p、第二micro
RNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白分
别为TALE蛋白、第一和第二识别序列为5×UAS的元件组成的表达系统的核酸
<400> 2
cggagtactg tcctccgagc ggagtactgt cctccgagcg gagtactgtc ctccgagcgg 60
agtactgtcc tccgagcgga gttctgtcct ccgagcggag actctagata cccctctccg 120
cttctgaatt ctaggcgtgt acggtgggag gcctatataa gcagagctcg tttagtgaac 180
cgtcagatcg cctcgagtac ccctctccgc ttcttacccc tctccgcttc ttacccctct 240
ccgcttctga attcgaccgc caccatgaga catattatct gccacggagg tgttattacc 300
gaagaaatgg ccgccagtct tttggaccag ctgatcgaag aggtactggc tgataatctt 360
ccacctccta gccattttga accacctacc cttcacgaac tgtatgattt agacgtgacg 420
gcccccgaag atcccaacga ggaggcggtt tcgcagattt ttcccgactc tgtaatgttg 480
gcggtgcagg aagggattga cttactcact tttccgccgg cgcccggttc tccggagccg 540
cctcaccttt cccggcagcc cgagcagccg gagcagagag ccttgggtcc ggtttctatg 600
ccaaaccttg taccggaggt gatcgatctt acctgccacg aggctggctt tccacccagt 660
gacgacgagg atgaagaggg tgaggagttt gtgttagatt atgtggagca ccccgggcac 720
ggttgcaggt cttgtcatta tcaccggagg aatacggggg acccagatat tatgtgttcg 780
ctttgctata tgaggacctg tggcatgttt gtctacagta agtgaaaatt atgggcagtg 840
ggtgatagag tggtgggttt ggtgtggtaa tttttttttt aatttttaca gttttgtggt 900
ttaaagaatt ttgtattgtg atttttttaa aaggtcctgt gtctgaacct gagcctgagc 960
ccgagccaga accggagcct gcaagaccta cccgccgtcc taaaatggcg cctgctatcc 1020
tgaggcgccc gacatcacct gtgtctagag aatgcaatag tagtacggat agctgtgact 1080
ccggtccttc taacacacct cctgagatac acccggtggt cccgctgtgc cccattaaac 1140
cagttgccgt gagagttggt gggcgtcgcc aggctgtgga atgtatcgag gacttgctta 1200
acgagcctgg gcaacctttg gacttgagct gtaaacgccc caggccaggg tccggagcta 1260
caaatttttc cctcctcaaa caggctggag atgtcgaaga aaatcctggg cctatggagg 1320
cttgggagtg tttggaagat ttttctgctg tgcgtaactt gctggaacag agctctaaca 1380
gtacctcttg gttttggagg tttctgtggg gctcatccca ggcaaagtta gtctgcagaa 1440
ttaaggagga ttacaagtgg gaatttgaag agcttttgaa atcctgtggt gagctgtttg 1500
attctttgaa tctgggtcac caggcgcttt tccaagagaa ggtcatcaag actttggatt 1560
tttccacacc ggggcgcgct gcggctgctg ttgctttttt gagttttata aaggataaat 1620
ggagcgaaga aacccatctg agcggggggt acctgctgga ttttctggcc atgcatctgt 1680
ggagagcggt tgtgagacac aagaatcgcc tgctactgtt gtcttccgtc cgcccggcga 1740
taataccgac ggaggagcag cagcagcagc aggaggaagc caggcggcgg cggcaggagc 1800
agagcccatg gaacccgaga gccggcctgg accctcggga atgaatgttg tacaggtggc 1860
tgaactgtat ccagaactga gacgcatttt gacaattaca gaggatgggc aggggctaaa 1920
gggggtaaag agggagcggg gggcttgtga ggctacagag gaggctagga atctagcttt 1980
tagcttaatg accagacacc gtcctgagtg tattactttt caacagatca aggataattg 2040
cgctaatgag cttgatctgc tggcgcagaa gtattccata gagcagctga ccacttactg 2100
gctgcagcca ggggatgatt ttgaggaggc tattagggta tatgcaaagg tggcacttag 2160
gccagattgc aagtacaaga tcagcaaact tgtaaatatc aggaattgtt gctacatttc 2220
tgggaacggg gccgaggtgg agatagatac ggaggatagg gtggccttta gatgtagcat 2280
gataaatatg tggccggggg tgcttggcat ggacggggtg gttattatga atgtaaggtt 2340
tactggcccc aattttagcg gtacggtttt cctggccaat accaacctta tcctacacgg 2400
tgtaagcttc tatgggttta acaatacctg tgtggaagcc tggaccgatg taagggttcg 2460
gggctgtgcc ttttactgct gctggaaggg ggtggtgtgt cgccccaaaa gcagggcttc 2520
aattaagaaa tgcctctttg aaaggtgtac cttgggtatc ctgtctgagg gtaactccag 2580
ggtgcgccac aatgtggcct ccgactgtgg ttgcttcatg ctagtgaaaa gcgtggctgt 2640
gattaagcat aacatggtat gtggcaactg cgaggacagg gcctctcaga tgctgacctg 2700
ctcggacggc aactgtcacc tgctgaagac cattcacgta gccagccact ctcgcaaggc 2760
ctggccagtg tttgagcata acatactgac ccgctgttcc ttgcatttgg gtaacaggag 2820
gggggtgttc ctaccttacc aatgcaattt gagtcacact aagatattgc ttgagcccga 2880
gagcatgtcc aaggtgaacc tgaacggggt gtttgacatg accatgaaga tctggaaggt 2940
gctgaggtac gatgagaccc gcaccaggtg cagaccctgc gagtgtggcg gtaaacatat 3000
taggaaccag cctgtgatgc tggatgtgac cgaggagctg aggcccgatc acttggtgct 3060
ggcctgcacc cgcgctgagt ttggctctag cgatgaagat acagatctcg aggggtccgg 3120
agctacaaat ttttccctcc tcaaacaggc tggagatgtc gaagaaaatc ctgggcctac 3180
cggtatgtat aggatgcagt tgctctcttg catcgccctg tctctagcgt tggtgactaa 3240
tagcgcccct acttcttctt ccacgaagaa aacccagctg caactggaac accttctgct 3300
ggatctccag atgatcctta acgggattaa caactataag aacccgaagc tcacggctat 3360
gctgacggcg aagttcgcta tgcctaaaaa ggctactgag ctgaaacact tacagtgtct 3420
cgaagaggca ctcaaacccc tcgaggaggt actcaatctg gcgcaaagca aaaacttcca 3480
tctccgccca cgggacctaa tctccaacat caacgtgatt gtgctggagc tcaaggggtc 3540
tgaaaccacg tttatgtgcg agtacgcaga tgaaactgct accatagtgg aattcctcaa 3600
tcgttggatc actttctgcc agtcaatcat ttcaaccctg accagatctg agggccgcgg 3660
cagcctgctg acctgcggcg acgtggagga aaaccccggc cccggatcct tgtcgaccat 3720
tcgtccgcgc aggccaagtc ctgcccgcga gcttctgccc ggaccccaac cggatagggt 3780
tcagccgact gcagatcgtg gggtgtctgc gcctgctggc agccctctgg atggcttgcc 3840
cgctcggcgg acggtgtccc ggacccggct gccatctccc cctgcgccct cacctgcgtt 3900
ctcggcgggc agcttcagcg atctgctccg tccgttcgat ccgtcgcttc ttgatacatc 3960
gcttcttgat tcgatgcctg ccgtcggcac gccgcataca gcggctgccc cagcagagtg 4020
ggatgaggcg caatcggctc tgcgtgcagc cgatgacccg ccacccaccg tgcgtgtcgc 4080
tgtcactgcc gcgcggccgc cgcgcgccaa gccggccccg cgacggcgtg ctgcgcaacc 4140
ctccgacgct tcgccggccg cgcaggtgga tctacgcacg ctcggctaca gtcagcagca 4200
gcaagagaag atcaaaccga aggtgcgttc gacagtggcg cagcaccacg aggcactggt 4260
gggccatggg tttacacacg cgcacatcgt tgcgctcagc caacacccgg cagcgttagg 4320
gaccgtcgct gtcacgtatc agcacataat cacggcgttg ccagaggcga cacacgaaga 4380
catcgttggc gtcggcaaac agtggtccgg cgcacgcgcc ctggaggcct tgctcacgga 4440
tgcgggggag ttgagaggtc cgccgttaca gttggacaca ggccaacttg tgaagattgc 4500
aaaacgtggc ggcgtgaccg caatggaggc agtgcatgca tcgcgcaatg cactgacggg 4560
tgcccccctg aacctgaccc cggaccaagt ggtggctatc gccagcaaca ttggcggcaa 4620
gcaagcgctc gaaacggtgc agcggctgtt gccggtgctg tgccaggacc atggcctgac 4680
cccggaccaa gtggtggcta tcgccagcaa cggtggcggc aagcaagcgc tcgaaacggt 4740
gcagcggctg ttgccggtgc tgtgccagga ccatggcctg accccggacc aagtggtggc 4800
tatcgccagc aacattggcg gcaagcaagc gctcgaaacg gtgcagcggc tgttgccggt 4860
gctgtgccag gaccatggcc tgaccccgga ccaagtggtg gctatcgcca gcaacaatgg 4920
cggcaagcaa gcgctcgaaa cggtgcagcg gctgttgccg gtgctgtgcc aggaccatgg 4980
cctgaccccg gaccaagtgg tggctatcgc cagcaacatt ggcggcaagc aagcgctcga 5040
aacggtgcag cggctgttgc cggtgctgtg ccaggaccat ggcctgaccc cggaccaagt 5100
ggtggctatc gccagcaaca ttggcggcaa gcaagcgctc gaaacggtgc agcggctgtt 5160
gccggtgctg tgccaggacc atggcctgac tccggaccaa gtggtggcta tcgccagcca 5220
cgatggcggc aagcaagcgc tcgaaacggt gcagcggctg ttgccggtgc tgtgccagga 5280
ccatggcctg actccggacc aagtggtggc tatcgccagc cacgatggcg gcaagcaagc 5340
gctcgaaacg gtgcagcggc tgttgccggt gctgtgccag gaccatggcc tgaccccgga 5400
ccaagtggtg gctatcgcca gcaacaatgg cggcaagcaa gcgctcgaaa cggtgcagcg 5460
gctgttgccg gtgctgtgcc aggaccatgg cctgaccccg gaccaagtgg tggctatcgc 5520
cagcaacatt ggcggcaagc aagcgctcga aacggtgcag cggctgttgc cggtgctgtg 5580
ccaggaccat ggcctgaccc cggaccaagt ggtggctatc gccagcaacg gtggcggcaa 5640
gcaagcgctc gaaacggtgc agcggctgtt gccggtgctg tgccaggacc atggcctgac 5700
tccggaccaa gtggtggcta tcgccagcca cgatggcggc aagcaagcgc tcgaaacggt 5760
gcagcggctg ttgccggtgc tgtgccagga ccatggcctg actccggacc aagtggtggc 5820
tatcgccagc cacgatggcg gcaagcaagc gctcgaaacg gtgcagcggc tgttgccggt 5880
gctgtgccag gaccatggcc tgaccccgga ccaagtggtg gctatcgcca gcaacggtgg 5940
cggcaagcaa gcgctcgaaa cggtgcagcg gctgttgccg gtgctgtgcc aggaccatgg 6000
cctgactccg gaccaagtgg tggctatcgc cagccacgat ggcggcaagc aagcgctcga 6060
aacggtgcag cggctgttgc cggtgctgtg ccaggaccat ggcctgactc cggaccaagt 6120
ggtggctatc gccagccacg atggcggcaa gcaagcgctc gaaacggtgc agcggctgtt 6180
gccggtgctg tgccaggacc atggcctgac tccggaccaa gtggtggcta tcgccagcca 6240
cgatggcggc aagcaagcgc tcgaaacggt gcagcggctg ttgccggtgc tgtgccagga 6300
ccatggcctg accccggacc aagtggtggc tatcgccagc aacattggcg gcaagcaagc 6360
gctcgaaacg gtgcagcggc tgttgccggt gctgtgccag gaccatggcc tgaccccgga 6420
ccaagtggtg gctatcgcca gcaacggtgg cggcaagcaa gcgctcgaaa cggtgcagcg 6480
gctgttgccg gtgctgtgcc aggaccatgg cctgaccccg gaccaagtgg tggctatcgc 6540
cagcaacggt ggcggcaagc aagcgctcga aagcattgtg gcccagctga gccggcctga 6600
tccggcgttg gccgcgttga ccaacgacca cctcgtcgcc ttggcctgcc tcggcggacg 6660
tcctgccatg gatgcagtga aaaagggatt gccgcacgcg ccggaattga tcagaagagt 6720
caatcgccgt attggcgaac gcacgtccca tcgcgttgcc gactacgcgc aagtggttcg 6780
cgtgctggag tttttccagt gccactccca cccagcgtac gcatttgatg aggccatgac 6840
gcagttcggg atgagcagga acgggttggt acagctcttt cgcagagtgg gcgtcaccga 6900
actcgaagcc cgcggtggaa cgctcccccc agcctcgcag cgttgggacc gtatcctcca 6960
ggcatcaggg atgaaaaggg ccaaaccgtc ccctacttca gctcaaacac cggatcaggc 7020
gtctttgcat gcattcgccg attcgctgga gcgtgacctt gatgcgccca gcccaatgca 7080
cgagggagat cagacgcggg caagcagccg taaacggtcc cgatcggatc gtgctgtcac 7140
cggcccctcc gcacagcagg ctgtcgaggt gcgcgttccc gaacagcgcg atgcgctgca 7200
tttgcccctc agctggaggg taaaacgccc gcgtaccagg atctggggcg gcctcccggt 7260
taagtagtgc tttctacttt atgagtagtg ctttctactt tatgagtagt gctttctact 7320
ttatgagtag tgctttctac tttatggagc tacagtgctt catctcagag ctacagtgct 7380
tcatctcaga gctacagtgc ttcatctcag agctacagtg cttcatctca gaaagcggag 7440
tactgtcctc cgagcggagt actgtcctcc gagcggagta ctgtcctccg agcggagtac 7500
tgtcctccga gcggagttct gtcctccgag cggagactct agatatagaa ccgatcctcc 7560
cattgaattc taggcgtgta cggtgggagg cctatataag cagagctcgt ttagtgaacc 7620
gtcagatcgc ctcgagtata gaaccgatcc tcccatttat agaaccgatc ctcccattta 7680
tagaaccgat cctcccatta ctagaagatc ttgaattcgc caccttgtcg accattcgtc 7740
cgcgcaggcc aagtcctgcc cgcgagcttc tgcccggacc ccaaccggat agggttcagc 7800
cgactgcaga tcgtggggtg tctgcgcctg ctggcagccc tctggatggc ttgcccgctc 7860
ggcggacggt gtcccggacc cggctgccat ctccccctgc gccctcacct gcgttctcgg 7920
cgggcagctt cagcgatctg ctccgtccgt tcgatccgtc gcttcttgat acatcgcttc 7980
ttgattcgat gcctgccgtc ggcacgccgc atacagcggc tgccccagca gagtgggatg 8040
aggcgcaatc ggctctgcgt gcagccgatg acccgccacc caccgtgcgt gtcgctgtca 8100
ctgccgcgcg gccgccgcgc gccaagccgg ccccgcgacg gcgtgctgcg caaccctccg 8160
acgcttcgcc ggccgcgcag gtggatctac gcacgctcgg ctacagtcag cagcagcaag 8220
agaagatcaa accgaaggtg cgttcgacag tggcgcagca ccacgaggca ctggtgggcc 8280
atgggtttac acacgcgcac atcgttgcgc tcagccaaca cccggcagcg ttagggaccg 8340
tcgctgtcac gtatcagcac ataatcacgg cgttgccaga ggcgacacac gaagacatcg 8400
ttggcgtcgg caaacagtgg tccggcgcac gcgccctgga ggccttgctc acggatgcgg 8460
gggagttgag aggtccgccg ttacagttgg acacaggcca acttgtgaag attgcaaaac 8520
gtggcggcgt gaccgcaatg gaggcagtgc atgcatcgcg caatgcactg acgggtgccc 8580
ccctgaacct gaccccggac caagtggtgg ctatcgccag caacattggc ggcaagcaag 8640
cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc ctgactccgg 8700
accaagtggt ggctatcgcc agccacgatg gcggcaagca agcgctcgaa acggtgcagc 8760
ggctgttgcc ggtgctgtgc caggaccatg gcctgactcc ggaccaagtg gtggctatcg 8820
ccagccacga tggcggcaag caagcgctcg aaacggtgca gcggctgttg ccggtgctgt 8880
gccaggacca tggcctgact ccggaccaag tggtggctat cgccagccac gatggcggca 8940
agcaagcgct cgaaacggtg cagcggctgt tgccggtgct gtgccaggac catggcctga 9000
ctccggacca agtggtggct atcgccagcc acgatggcgg caagcaagcg ctcgaaacgg 9060
tgcagcggct gttgccggtg ctgtgccagg accatggcct gaccccggac caagtggtgg 9120
ctatcgccag caacggtggc ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg 9180
tgctgtgcca ggaccatggc ctgactccgg accaagtggt ggctatcgcc agccacgatg 9240
gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg 9300
gcctgacccc ggaccaagtg gtggctatcg ccagcaacgg tggcggcaag caagcgctcg 9360
aaacggtgca gcggctgttg ccggtgctgt gccaggacca tggcctgact ccggaccaag 9420
tggtggctat cgccagccac gatggcggca agcaagcgct cgaaacggtg cagcggctgt 9480
tgccggtgct gtgccaggac catggcctga ctccggacca agtggtggct atcgccagcc 9540
acgatggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg 9600
accatggcct gaccccggac caagtggtgg ctatcgccag caacaatggc ggcaagcaag 9660
cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc ctgactccgg 9720
accaagtggt ggctatcgcc agccacgatg gcggcaagca agcgctcgaa acggtgcagc 9780
ggctgttgcc ggtgctgtgc caggaccatg gcctgacccc ggaccaagtg gtggctatcg 9840
ccagcaacgg tggcggcaag caagcgctcg aaacggtgca gcggctgttg ccggtgctgt 9900
gccaggacca tggcctgacc ccggaccaag tggtggctat cgccagcaac ggtggcggca 9960
agcaagcgct cgaaacggtg cagcggctgt tgccggtgct gtgccaggac catggcctga 10020
ctccggacca agtggtggct atcgccagcc acgatggcgg caagcaagcg ctcgaaacgg 10080
tgcagcggct gttgccggtg ctgtgccagg accatggcct gaccccggac caagtggtgg 10140
ctatcgccag caacggtggc ggcaagcaag cgctcgaaag cattgtggcc cagctgagcc 10200
ggcctgatcc ggcgttggcc gcgttgacca acgaccacct cgtcgccttg gcctgcctcg 10260
gcggacgtcc tgccatggat gcagtgaaaa agggattgcc gcacgcgccg gaattgatca 10320
gaagagtcaa tcgccgtatt ggcgaacgca cgtcccatcg cgttgccgac tacgcgcaag 10380
tggttcgcgt gctggagttt ttccagtgcc actcccaccc agcgtacgca tttgatgagg 10440
ccatgacgca gttcgggatg agcaggaacg ggttggtaca gctctttcgc agagtgggcg 10500
tcaccgaact cgaagcccgc ggtggaacgc tccccccagc ctcgcagcgt tgggaccgta 10560
tcctccaggc atcagggatg aaaagggcca aaccgtcccc tacttcagct caaacaccgg 10620
atcaggcgtc tttgcatgca ttcgccgatt cgctggagcg tgaccttgat gcgcccagcc 10680
caatgcacga gggagatcag acgcgggcaa gcagccgtaa acggtcccga tcggatcgtg 10740
ctgtcaccgg cccctccgca cagcaggctg tcgaggtgcg cgttcccgaa cagcgcgatg 10800
cgctgcattt gcccctcagc tggagggtaa aacgcccgcg taccaggatc tggggcggcc 10860
tcccgtaata caaggcggcc gcaaatcaac atcagtctga taagctatca acatcagtct 10920
gataagctat caacatcagt ctgataagct atcaacatca gtctgataag ctaaagcttc 10980
gaattctgat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc 11040
cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta 11100
ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat 11160
ttttttcact gcgaaaatag aagggaaaaa agaggaggga caaaagaggc agaaatgaga 11220
ggggagggga cagaggacac ctgaataaag accacaccca tgacccacgt gatgctgaga 11280
agtactcctg ccctaggaag agactcaggg cagagggagg aaggacagca gaccagacag 11340
tcacagcagc cttgacaaaa cgttcctgga actcaagatg gcccccccga ccgatgtcag 11400
cctgggggac gagctccact tagacggcga ggacgtggcg atggcgcatg ccgacgcgct 11460
agacgatttc gatctggaca tgttggggag cgacgcgcta gacgatttcg atctggacat 11520
gttggggagc gacgcgctag acgatttcga tctggacatg ttggggagcg acgcgctaga 11580
cgatttcgat ctggacatgt tgggggacgg ggattccccg ggtccgggat ttacccccca 11640
cgactccgcc ccctacggcg ctctggatat ggccgacttc gagtttgagc agatgtttac 11700
cgatgccctt ggaattgacg agtacggtgg gacgcgtatg aagctactgt cttctatcga 11760
acaagcatgc gatatttgcc gacttaaaaa gctcaagtgc tccaaagaaa aaccgaagtg 11820
cgccaagtgt ctgaagaaca actgggagtg tcgctactct cccaaaacca aaaggtctcc 11880
gctgactagg gcacatctga cagaagtgga atcaaggcta gaaagactgg aacagctatt 11940
tctactgatt tttcctcgag aagaccttga catgattttg aaaatggatt ctttacagga 12000
tataaaagca ttgttaacag gattatttgt acaagataat gtgaataaag atgccgtcac 12060
agatagattg gcttcagtgg agactgatat gcctctaaca ttgagacagc atagaataag 12120
tgcgacatca tcatcggaag agagtagtaa caaaggtcaa agacagttga ctgtataaaa 12180
gcttcactcc tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg gccaatgccc 12240
tggctcacaa ataccactga gatctttttc cctctgccaa aaattatggg gacatcatga 12300
agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt 12360
gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac 12420
atcagaatga gtatttggtt tagagtttgg caacatatgc ccatatgctg gctgccatga 12480
acaaaggttg gctataaaga ggtcatcagt atatgaaaca gccccctgct gtccattcct 12540
tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt tttgtgttat 12600
ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga tttttcctcc 12660
tctcctgact actcccagtc atagctgtcc ctcttctctt atggagatc 12709
<210> 3
<211> 6250
<212> DNA
<213> Artificial
<220>
<223> 编码由细胞特异性启动子CEA368、第一microRNA为mir-135a-5p、第二micr
oRNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白分
别为LacI与tetR-KRAB、第一和第二识别序列为5×UAS的元件组成的表达系
统的核酸
<400> 3
cggggtgccg ccccccgcgg gtgaccgccc tccgcgggtg acagccctcc gcggagtgcc 60
gcactccgcg gagtgctgta ctccgagcgg agactctaga ctccctatca gtgatagaga 120
taggcgtgta cggtgggagg cctatataag cagagctcgt ttagtgaacc gtcagatcgc 180
tccctatcag tgatagagag aattcgaccg ccaccatgag acatattatc tgccacggag 240
gtgttattac cgaagaaatg gccgccagtc ttttggacca gctgatcgaa gaggtactgg 300
ctgataatct tccacctcct agccattttg aaccacctac ccttcacgaa ctgtatgatt 360
tagacgtgac ggcccccgaa gatcccaacg aggaggcggt ttcgcagatt tttcccgact 420
ctgtaatgtt ggcggtgcag gaagggattg acttactcac ttttccgccg gcgcccggtt 480
ctccggagcc gcctcacctt tcccggcagc ccgagcagcc ggagcagaga gccttgggtc 540
cggtttctat gccaaacctt gtaccggagg tgatcgatct tacctgccac gaggctggct 600
ttccacccag tgacgacgag gatgaagagg gtgaggagtt tgtgttagat tatgtggagc 660
accccgggca cggttgcagg tcttgtcatt atcaccggag gaatacgggg gacccagata 720
ttatgtgttc gctttgctat atgaggacct gtggcatgtt tgtctacagt aagtgaaaat 780
tatgggcagt gggtgataga gtggtgggtt tggtgtggta attttttttt taatttttac 840
agttttgtgg tttaaagaat tttgtattgt gattttttta aaaggtcctg tgtctgaacc 900
tgagcctgag cccgagccag aaccggagcc tgcaagacct acccgccgtc ctaaaatggc 960
gcctgctatc ctgaggcgcc cgacatcacc tgtgtctaga gaatgcaata gtagtacgga 1020
tagctgtgac tccggtcctt ctaacacacc tcctgagata cacccggtgg tcccgctgtg 1080
ccccattaaa ccagttgccg tgagagttgg tgggcgtcgc caggctgtgg aatgtatcga 1140
ggacttgctt aacgagcctg ggcaaccttt ggacttgagc tgtaaacgcc ccaggccact 1200
cgagggtacc gggtccggag ctacaaattt ttccctcctc aaacaggctg gagatgtcga 1260
agaaaatcct gggcctaccg gtatgtggct gcagagcctc cttctactgg gcaccgtggc 1320
ttgcagcatc tcggcccccg ccagaagtcc cagccccagc acccagccct gggaacacgt 1380
aaatgcaatc caggaggccc gccgcctcct gaacttgtca cgggacacgg cagccgagat 1440
gaatgaaact gtggaggtca tcagtgaaat gtttgacttg caggagccca catgcctgca 1500
gactcggctg gagctctaca aacagggcct gcgtggcagc ctcaccaagc tgaagggccc 1560
actgaccatg atggctagcc attacaagca gcactgcccg ccgacccctg agacatcctg 1620
tgccactcag atcattacct ttgagtcttt caaggagaac ctcaaggact ttctgctggt 1680
catcccattt gactgctggg aacccgtcca ggaaagatct gagggccgcg gcagcctgct 1740
gacctgcggc gacgtggagg aaaaccccgg ccccggatcc atgaaaccag taacgttata 1800
cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc 1860
cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta 1920
cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga ttggcgttgc 1980
cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc 2040
cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg 2100
taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca ttaactatcc 2160
gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc cggcgttatt 2220
tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg aagacggtac 2280
gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg 2340
cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat atctcactcg 2400
caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt ccggttttca 2460
acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg ttgccaacga 2520
tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga 2580
tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc cgccgttaac 2640
caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact 2700
ctctcagggc caggcggtga agggcaatca gctgttgccg gtctcactgg tgaaaagaaa 2760
aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat 2820
gcagctggca cgacaggttt cccgactgga aagcgggcag agcagcctga ggcctcctaa 2880
gaagaagagg aaggttgtta caaacaccat tgtcacactc cacaaacacc attgtcacac 2940
tccacaaaca ccattgtcac actccacaaa caccattgtc acactccatc acataggaat 3000
aaaaagccat atcacatagg aataaaaagc catatcacat aggaataaaa agccatatca 3060
cataggaata aaaagccata gaaagcggac tactgtagtc cgcggagtac tgtcctccgc 3120
ggacgactgt cgtccgcgga gcactgtgct ccgcggatga ctgtcatccg agcggagact 3180
ctagaaattg tgagcggata acaattgggt atataatggg ggccgtcagc tactaccaga 3240
gccgatcgaa cactcgagcc gagcagacgt gcctacaatt gtgagcgctc acaattacta 3300
gaagatcttg aattcgccac catggctaga ttagataaaa gtaaagtgat taacagcgca 3360
ttagagctgc ttaatgaggt cggaatcgaa ggtttaacaa cccgtaaact cgcccagaag 3420
ctaggtgtag agcagcctac attgtattgg catgtaaaaa ataagcgggc tttgctcgac 3480
gccttagcca ttgagatgtt agataggcac catactcact tttgcccttt agaaggggaa 3540
agctggcaag attttttacg taataacgct aaaagtttta gatgtgcttt actaagtcat 3600
cgcgatggag caaaagtaca tttaggtaca cggcctacag aaaaacagta tgaaactctc 3660
gaaaatcaat tagccttttt atgccaacaa ggtttttcac tagagaatgc attatatgca 3720
ctcagcgctg tggggcattt tactttaggt tgcgtattgg aagatcaaga gcatcaagtc 3780
gctaaagaag aaagggaaac acctactact gatagtatgc cgccattatt acgacaagct 3840
atcgaattat ttgatcacca aggtgcagag ccagccttct tattcggcct tgaattgatc 3900
atatgcggat tagaaaaaca acttaaatgt gaaagtgggt cgccaaaaaa gaagagaaag 3960
gtggatggtg gcggagcact gtctccccag cattcagctg tgacccaggg atcaatcatt 4020
aagaacaagg aaggtatgga tgctaagtcc ctgaccgctt ggtcaagaac tttggtaact 4080
tttaaggacg tgttcgtcga ttttactcgg gaggagtgga aactattgga taccgcccag 4140
caaatcgtgt atagaaacgt tatgctcgag aactataaga atcttgtgtc actgggctac 4200
caactgacaa agcctgacgt tatcctgagg ctggaaaagg gtgaggagcc ttggttggtc 4260
gagcgggaga tccaccagga aactcaccca gacagtgaga ctgctttcga gatcaagtcc 4320
tctgtgtaat acaaggcggc cgcaaatcaa catcagtctg ataagctatc aacatcagtc 4380
tgataagcta tcaacatcag tctgataagc tatcaacatc agtctgataa gctaaagctt 4440
cgaattctga taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc 4500
ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt 4560
attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca 4620
tttttttcac tgcagctcgc cttttgctca catgtctccc aggggatggg ggtccatcca 4680
ccttgccgaa aagatttgtc tgaggaactg aaaatagaag ggaaaaaaga ggagggacaa 4740
aagaggcaga aatgagaggg gaggggacag aggacacctg aataaagacc acacccatga 4800
cccacgtgat gctgagaagt actcctgccc taggaagaga ctcagggcag agggaggaag 4860
gacagcagac cagacagtca cagcagcctt gacaaaacgt tcctggaact caagctcttc 4920
tccacagagg aggacagagc agacagcaga gaccatggag tctccctcgg cccctcccca 4980
cagatggtgc atcccctggc agaggctcct gctcacaggt gaactcgaga ccggtatggc 5040
ccccccgacc gatgtcagcc tgggggacga gctccactta gacggcgagg acgtggcgat 5100
ggcgcatgcc gacgcgctag acgatttcga tctggacatg ttgggggacg gggattcccc 5160
gggtccggga tttacccccc acgactccgc cccctacggc gctctggata tggccgactt 5220
cgagtttgag cagatgttta ccgatgccct tggaattgac gagtacggtg ggacgcgtat 5280
gaagctactg tcttctatcg aacaagcatg cgatatttgc cgacttaaaa agctcaagtg 5340
ctccaaagaa aaaccgaagt gcgccaagtg tctgaagaac aactgggagt gtcgctactc 5400
tcccaaaacc aaaaggtctc cgctgactag ggcacatctg acagaagtgg aatcaaggct 5460
agaaagactg gaacagctat ttctactgat ttttcctcga gaagaccttg acatgatttt 5520
gaaaatggat tctttacagg atataaaagc attgttaaca ggattatttg tacaagataa 5580
tgtgaataaa gatgccgtca cagatagatt ggcttcagtg gagactgata tgcctctaac 5640
attgagacag catagaataa gtgcgacatc atcatcggaa gagagtagta acaaaggtca 5700
aagacagttg actgtataaa agcttcactc ctcaggtgca ggctgcctat cagaaggtgg 5760
tggctggtgt ggccaatgcc ctggctcaca aataccactg agatcttttt ccctctgcca 5820
aaaattatgg ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat 5880
ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg 5940
ggagggcaaa tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg 6000
cccatatgct ggctgccatg aacaaaggtt ggctataaag aggtcatcag tatatgaaac 6060
agccccctgc tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt 6120
ttatattttg ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt 6180
tactagccag atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct 6240
tatggagatc 6250
<210> 4
<211> 12906
<212> DNA
<213> Artificial
<220>
<223> 编码由细胞特异性启动子CEA368、第一microRNA为mir-143-3p、第二micro
RNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白分
别为TALE蛋白、第一和第二识别序列为5×UAS的元件组成的表达系统的核酸
<400> 4
cggagtactg tcctccgagc ggagtactgt cctccgagcg gagtactgtc ctccgagcgg 60
agtactgtcc tccgagcgga gttctgtcct ccgagcggag actctagata cccctctccg 120
cttctgaatt ctaggcgtgt acggtgggag gcctatataa gcagagctcg tttagtgaac 180
cgtcagatcg cctcgagtac ccctctccgc ttcttacccc tctccgcttc ttacccctct 240
ccgcttctga attcgaccgc caccatgaga catattatct gccacggagg tgttattacc 300
gaagaaatgg ccgccagtct tttggaccag ctgatcgaag aggtactggc tgataatctt 360
ccacctccta gccattttga accacctacc cttcacgaac tgtatgattt agacgtgacg 420
gcccccgaag atcccaacga ggaggcggtt tcgcagattt ttcccgactc tgtaatgttg 480
gcggtgcagg aagggattga cttactcact tttccgccgg cgcccggttc tccggagccg 540
cctcaccttt cccggcagcc cgagcagccg gagcagagag ccttgggtcc ggtttctatg 600
ccaaaccttg taccggaggt gatcgatctt acctgccacg aggctggctt tccacccagt 660
gacgacgagg atgaagaggg tgaggagttt gtgttagatt atgtggagca ccccgggcac 720
ggttgcaggt cttgtcatta tcaccggagg aatacggggg acccagatat tatgtgttcg 780
ctttgctata tgaggacctg tggcatgttt gtctacagta agtgaaaatt atgggcagtg 840
ggtgatagag tggtgggttt ggtgtggtaa tttttttttt aatttttaca gttttgtggt 900
ttaaagaatt ttgtattgtg atttttttaa aaggtcctgt gtctgaacct gagcctgagc 960
ccgagccaga accggagcct gcaagaccta cccgccgtcc taaaatggcg cctgctatcc 1020
tgaggcgccc gacatcacct gtgtctagag aatgcaatag tagtacggat agctgtgact 1080
ccggtccttc taacacacct cctgagatac acccggtggt cccgctgtgc cccattaaac 1140
cagttgccgt gagagttggt gggcgtcgcc aggctgtgga atgtatcgag gacttgctta 1200
acgagcctgg gcaacctttg gacttgagct gtaaacgccc caggccaggg tccggagcta 1260
caaatttttc cctcctcaaa caggctggag atgtcgaaga aaatcctggg cctatggagg 1320
cttgggagtg tttggaagat ttttctgctg tgcgtaactt gctggaacag agctctaaca 1380
gtacctcttg gttttggagg tttctgtggg gctcatccca ggcaaagtta gtctgcagaa 1440
ttaaggagga ttacaagtgg gaatttgaag agcttttgaa atcctgtggt gagctgtttg 1500
attctttgaa tctgggtcac caggcgcttt tccaagagaa ggtcatcaag actttggatt 1560
tttccacacc ggggcgcgct gcggctgctg ttgctttttt gagttttata aaggataaat 1620
ggagcgaaga aacccatctg agcggggggt acctgctgga ttttctggcc atgcatctgt 1680
ggagagcggt tgtgagacac aagaatcgcc tgctactgtt gtcttccgtc cgcccggcga 1740
taataccgac ggaggagcag cagcagcagc aggaggaagc caggcggcgg cggcaggagc 1800
agagcccatg gaacccgaga gccggcctgg accctcggga atgaatgttg tacaggtggc 1860
tgaactgtat ccagaactga gacgcatttt gacaattaca gaggatgggc aggggctaaa 1920
gggggtaaag agggagcggg gggcttgtga ggctacagag gaggctagga atctagcttt 1980
tagcttaatg accagacacc gtcctgagtg tattactttt caacagatca aggataattg 2040
cgctaatgag cttgatctgc tggcgcagaa gtattccata gagcagctga ccacttactg 2100
gctgcagcca ggggatgatt ttgaggaggc tattagggta tatgcaaagg tggcacttag 2160
gccagattgc aagtacaaga tcagcaaact tgtaaatatc aggaattgtt gctacatttc 2220
tgggaacggg gccgaggtgg agatagatac ggaggatagg gtggccttta gatgtagcat 2280
gataaatatg tggccggggg tgcttggcat ggacggggtg gttattatga atgtaaggtt 2340
tactggcccc aattttagcg gtacggtttt cctggccaat accaacctta tcctacacgg 2400
tgtaagcttc tatgggttta acaatacctg tgtggaagcc tggaccgatg taagggttcg 2460
gggctgtgcc ttttactgct gctggaaggg ggtggtgtgt cgccccaaaa gcagggcttc 2520
aattaagaaa tgcctctttg aaaggtgtac cttgggtatc ctgtctgagg gtaactccag 2580
ggtgcgccac aatgtggcct ccgactgtgg ttgcttcatg ctagtgaaaa gcgtggctgt 2640
gattaagcat aacatggtat gtggcaactg cgaggacagg gcctctcaga tgctgacctg 2700
ctcggacggc aactgtcacc tgctgaagac cattcacgta gccagccact ctcgcaaggc 2760
ctggccagtg tttgagcata acatactgac ccgctgttcc ttgcatttgg gtaacaggag 2820
gggggtgttc ctaccttacc aatgcaattt gagtcacact aagatattgc ttgagcccga 2880
gagcatgtcc aaggtgaacc tgaacggggt gtttgacatg accatgaaga tctggaaggt 2940
gctgaggtac gatgagaccc gcaccaggtg cagaccctgc gagtgtggcg gtaaacatat 3000
taggaaccag cctgtgatgc tggatgtgac cgaggagctg aggcccgatc acttggtgct 3060
ggcctgcacc cgcgctgagt ttggctctag cgatgaagat acagatctcg aggggtccgg 3120
agctacaaat ttttccctcc tcaaacaggc tggagatgtc gaagaaaatc ctgggcctac 3180
cggtatgtat aggatgcagt tgctctcttg catcgccctg tctctagcgt tggtgactaa 3240
tagcgcccct acttcttctt ccacgaagaa aacccagctg caactggaac accttctgct 3300
ggatctccag atgatcctta acgggattaa caactataag aacccgaagc tcacggctat 3360
gctgacggcg aagttcgcta tgcctaaaaa ggctactgag ctgaaacact tacagtgtct 3420
cgaagaggca ctcaaacccc tcgaggaggt actcaatctg gcgcaaagca aaaacttcca 3480
tctccgccca cgggacctaa tctccaacat caacgtgatt gtgctggagc tcaaggggtc 3540
tgaaaccacg tttatgtgcg agtacgcaga tgaaactgct accatagtgg aattcctcaa 3600
tcgttggatc actttctgcc agtcaatcat ttcaaccctg accagatctg agggccgcgg 3660
cagcctgctg acctgcggcg acgtggagga aaaccccggc cccggatcct tgtcgaccat 3720
tcgtccgcgc aggccaagtc ctgcccgcga gcttctgccc ggaccccaac cggatagggt 3780
tcagccgact gcagatcgtg gggtgtctgc gcctgctggc agccctctgg atggcttgcc 3840
cgctcggcgg acggtgtccc ggacccggct gccatctccc cctgcgccct cacctgcgtt 3900
ctcggcgggc agcttcagcg atctgctccg tccgttcgat ccgtcgcttc ttgatacatc 3960
gcttcttgat tcgatgcctg ccgtcggcac gccgcataca gcggctgccc cagcagagtg 4020
ggatgaggcg caatcggctc tgcgtgcagc cgatgacccg ccacccaccg tgcgtgtcgc 4080
tgtcactgcc gcgcggccgc cgcgcgccaa gccggccccg cgacggcgtg ctgcgcaacc 4140
ctccgacgct tcgccggccg cgcaggtgga tctacgcacg ctcggctaca gtcagcagca 4200
gcaagagaag atcaaaccga aggtgcgttc gacagtggcg cagcaccacg aggcactggt 4260
gggccatggg tttacacacg cgcacatcgt tgcgctcagc caacacccgg cagcgttagg 4320
gaccgtcgct gtcacgtatc agcacataat cacggcgttg ccagaggcga cacacgaaga 4380
catcgttggc gtcggcaaac agtggtccgg cgcacgcgcc ctggaggcct tgctcacgga 4440
tgcgggggag ttgagaggtc cgccgttaca gttggacaca ggccaacttg tgaagattgc 4500
aaaacgtggc ggcgtgaccg caatggaggc agtgcatgca tcgcgcaatg cactgacggg 4560
tgcccccctg aacctgaccc cggaccaagt ggtggctatc gccagcaaca ttggcggcaa 4620
gcaagcgctc gaaacggtgc agcggctgtt gccggtgctg tgccaggacc atggcctgac 4680
cccggaccaa gtggtggcta tcgccagcaa cggtggcggc aagcaagcgc tcgaaacggt 4740
gcagcggctg ttgccggtgc tgtgccagga ccatggcctg accccggacc aagtggtggc 4800
tatcgccagc aacattggcg gcaagcaagc gctcgaaacg gtgcagcggc tgttgccggt 4860
gctgtgccag gaccatggcc tgaccccgga ccaagtggtg gctatcgcca gcaacaatgg 4920
cggcaagcaa gcgctcgaaa cggtgcagcg gctgttgccg gtgctgtgcc aggaccatgg 4980
cctgaccccg gaccaagtgg tggctatcgc cagcaacatt ggcggcaagc aagcgctcga 5040
aacggtgcag cggctgttgc cggtgctgtg ccaggaccat ggcctgaccc cggaccaagt 5100
ggtggctatc gccagcaaca ttggcggcaa gcaagcgctc gaaacggtgc agcggctgtt 5160
gccggtgctg tgccaggacc atggcctgac tccggaccaa gtggtggcta tcgccagcca 5220
cgatggcggc aagcaagcgc tcgaaacggt gcagcggctg ttgccggtgc tgtgccagga 5280
ccatggcctg actccggacc aagtggtggc tatcgccagc cacgatggcg gcaagcaagc 5340
gctcgaaacg gtgcagcggc tgttgccggt gctgtgccag gaccatggcc tgaccccgga 5400
ccaagtggtg gctatcgcca gcaacaatgg cggcaagcaa gcgctcgaaa cggtgcagcg 5460
gctgttgccg gtgctgtgcc aggaccatgg cctgaccccg gaccaagtgg tggctatcgc 5520
cagcaacatt ggcggcaagc aagcgctcga aacggtgcag cggctgttgc cggtgctgtg 5580
ccaggaccat ggcctgaccc cggaccaagt ggtggctatc gccagcaacg gtggcggcaa 5640
gcaagcgctc gaaacggtgc agcggctgtt gccggtgctg tgccaggacc atggcctgac 5700
tccggaccaa gtggtggcta tcgccagcca cgatggcggc aagcaagcgc tcgaaacggt 5760
gcagcggctg ttgccggtgc tgtgccagga ccatggcctg actccggacc aagtggtggc 5820
tatcgccagc cacgatggcg gcaagcaagc gctcgaaacg gtgcagcggc tgttgccggt 5880
gctgtgccag gaccatggcc tgaccccgga ccaagtggtg gctatcgcca gcaacggtgg 5940
cggcaagcaa gcgctcgaaa cggtgcagcg gctgttgccg gtgctgtgcc aggaccatgg 6000
cctgactccg gaccaagtgg tggctatcgc cagccacgat ggcggcaagc aagcgctcga 6060
aacggtgcag cggctgttgc cggtgctgtg ccaggaccat ggcctgactc cggaccaagt 6120
ggtggctatc gccagccacg atggcggcaa gcaagcgctc gaaacggtgc agcggctgtt 6180
gccggtgctg tgccaggacc atggcctgac tccggaccaa gtggtggcta tcgccagcca 6240
cgatggcggc aagcaagcgc tcgaaacggt gcagcggctg ttgccggtgc tgtgccagga 6300
ccatggcctg accccggacc aagtggtggc tatcgccagc aacattggcg gcaagcaagc 6360
gctcgaaacg gtgcagcggc tgttgccggt gctgtgccag gaccatggcc tgaccccgga 6420
ccaagtggtg gctatcgcca gcaacggtgg cggcaagcaa gcgctcgaaa cggtgcagcg 6480
gctgttgccg gtgctgtgcc aggaccatgg cctgaccccg gaccaagtgg tggctatcgc 6540
cagcaacggt ggcggcaagc aagcgctcga aagcattgtg gcccagctga gccggcctga 6600
tccggcgttg gccgcgttga ccaacgacca cctcgtcgcc ttggcctgcc tcggcggacg 6660
tcctgccatg gatgcagtga aaaagggatt gccgcacgcg ccggaattga tcagaagagt 6720
caatcgccgt attggcgaac gcacgtccca tcgcgttgcc gactacgcgc aagtggttcg 6780
cgtgctggag tttttccagt gccactccca cccagcgtac gcatttgatg aggccatgac 6840
gcagttcggg atgagcagga acgggttggt acagctcttt cgcagagtgg gcgtcaccga 6900
actcgaagcc cgcggtggaa cgctcccccc agcctcgcag cgttgggacc gtatcctcca 6960
ggcatcaggg atgaaaaggg ccaaaccgtc ccctacttca gctcaaacac cggatcaggc 7020
gtctttgcat gcattcgccg attcgctgga gcgtgacctt gatgcgccca gcccaatgca 7080
cgagggagat cagacgcggg caagcagccg taaacggtcc cgatcggatc gtgctgtcac 7140
cggcccctcc gcacagcagg ctgtcgaggt gcgcgttccc gaacagcgcg atgcgctgca 7200
tttgcccctc agctggaggg taaaacgccc gcgtaccagg atctggggcg gcctcccggt 7260
taagtagtgc tttctacttt atgagtagtg ctttctactt tatgagtagt gctttctact 7320
ttatgagtag tgctttctac tttatggagc tacagtgctt catctcagag ctacagtgct 7380
tcatctcaga gctacagtgc ttcatctcag agctacagtg cttcatctca gaaagcggag 7440
tactgtcctc cgagcggagt actgtcctcc gagcggagta ctgtcctccg agcggagtac 7500
tgtcctccga gcggagttct gtcctccgag cggagactct agatatagaa ccgatcctcc 7560
cattgaattc taggcgtgta cggtgggagg cctatataag cagagctcgt ttagtgaacc 7620
gtcagatcgc ctcgagtata gaaccgatcc tcccatttat agaaccgatc ctcccattta 7680
tagaaccgat cctcccatta ctagaagatc ttgaattcgc caccttgtcg accattcgtc 7740
cgcgcaggcc aagtcctgcc cgcgagcttc tgcccggacc ccaaccggat agggttcagc 7800
cgactgcaga tcgtggggtg tctgcgcctg ctggcagccc tctggatggc ttgcccgctc 7860
ggcggacggt gtcccggacc cggctgccat ctccccctgc gccctcacct gcgttctcgg 7920
cgggcagctt cagcgatctg ctccgtccgt tcgatccgtc gcttcttgat acatcgcttc 7980
ttgattcgat gcctgccgtc ggcacgccgc atacagcggc tgccccagca gagtgggatg 8040
aggcgcaatc ggctctgcgt gcagccgatg acccgccacc caccgtgcgt gtcgctgtca 8100
ctgccgcgcg gccgccgcgc gccaagccgg ccccgcgacg gcgtgctgcg caaccctccg 8160
acgcttcgcc ggccgcgcag gtggatctac gcacgctcgg ctacagtcag cagcagcaag 8220
agaagatcaa accgaaggtg cgttcgacag tggcgcagca ccacgaggca ctggtgggcc 8280
atgggtttac acacgcgcac atcgttgcgc tcagccaaca cccggcagcg ttagggaccg 8340
tcgctgtcac gtatcagcac ataatcacgg cgttgccaga ggcgacacac gaagacatcg 8400
ttggcgtcgg caaacagtgg tccggcgcac gcgccctgga ggccttgctc acggatgcgg 8460
gggagttgag aggtccgccg ttacagttgg acacaggcca acttgtgaag attgcaaaac 8520
gtggcggcgt gaccgcaatg gaggcagtgc atgcatcgcg caatgcactg acgggtgccc 8580
ccctgaacct gaccccggac caagtggtgg ctatcgccag caacattggc ggcaagcaag 8640
cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc ctgactccgg 8700
accaagtggt ggctatcgcc agccacgatg gcggcaagca agcgctcgaa acggtgcagc 8760
ggctgttgcc ggtgctgtgc caggaccatg gcctgactcc ggaccaagtg gtggctatcg 8820
ccagccacga tggcggcaag caagcgctcg aaacggtgca gcggctgttg ccggtgctgt 8880
gccaggacca tggcctgact ccggaccaag tggtggctat cgccagccac gatggcggca 8940
agcaagcgct cgaaacggtg cagcggctgt tgccggtgct gtgccaggac catggcctga 9000
ctccggacca agtggtggct atcgccagcc acgatggcgg caagcaagcg ctcgaaacgg 9060
tgcagcggct gttgccggtg ctgtgccagg accatggcct gaccccggac caagtggtgg 9120
ctatcgccag caacggtggc ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg 9180
tgctgtgcca ggaccatggc ctgactccgg accaagtggt ggctatcgcc agccacgatg 9240
gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg 9300
gcctgacccc ggaccaagtg gtggctatcg ccagcaacgg tggcggcaag caagcgctcg 9360
aaacggtgca gcggctgttg ccggtgctgt gccaggacca tggcctgact ccggaccaag 9420
tggtggctat cgccagccac gatggcggca agcaagcgct cgaaacggtg cagcggctgt 9480
tgccggtgct gtgccaggac catggcctga ctccggacca agtggtggct atcgccagcc 9540
acgatggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg 9600
accatggcct gaccccggac caagtggtgg ctatcgccag caacaatggc ggcaagcaag 9660
cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc ctgactccgg 9720
accaagtggt ggctatcgcc agccacgatg gcggcaagca agcgctcgaa acggtgcagc 9780
ggctgttgcc ggtgctgtgc caggaccatg gcctgacccc ggaccaagtg gtggctatcg 9840
ccagcaacgg tggcggcaag caagcgctcg aaacggtgca gcggctgttg ccggtgctgt 9900
gccaggacca tggcctgacc ccggaccaag tggtggctat cgccagcaac ggtggcggca 9960
agcaagcgct cgaaacggtg cagcggctgt tgccggtgct gtgccaggac catggcctga 10020
ctccggacca agtggtggct atcgccagcc acgatggcgg caagcaagcg ctcgaaacgg 10080
tgcagcggct gttgccggtg ctgtgccagg accatggcct gaccccggac caagtggtgg 10140
ctatcgccag caacggtggc ggcaagcaag cgctcgaaag cattgtggcc cagctgagcc 10200
ggcctgatcc ggcgttggcc gcgttgacca acgaccacct cgtcgccttg gcctgcctcg 10260
gcggacgtcc tgccatggat gcagtgaaaa agggattgcc gcacgcgccg gaattgatca 10320
gaagagtcaa tcgccgtatt ggcgaacgca cgtcccatcg cgttgccgac tacgcgcaag 10380
tggttcgcgt gctggagttt ttccagtgcc actcccaccc agcgtacgca tttgatgagg 10440
ccatgacgca gttcgggatg agcaggaacg ggttggtaca gctctttcgc agagtgggcg 10500
tcaccgaact cgaagcccgc ggtggaacgc tccccccagc ctcgcagcgt tgggaccgta 10560
tcctccaggc atcagggatg aaaagggcca aaccgtcccc tacttcagct caaacaccgg 10620
atcaggcgtc tttgcatgca ttcgccgatt cgctggagcg tgaccttgat gcgcccagcc 10680
caatgcacga gggagatcag acgcgggcaa gcagccgtaa acggtcccga tcggatcgtg 10740
ctgtcaccgg cccctccgca cagcaggctg tcgaggtgcg cgttcccgaa cagcgcgatg 10800
cgctgcattt gcccctcagc tggagggtaa aacgcccgcg taccaggatc tggggcggcc 10860
tcccgtaata caaggcggcc gcaaatcaac atcagtctga taagctatca acatcagtct 10920
gataagctat caacatcagt ctgataagct atcaacatca gtctgataag ctaaagcttc 10980
gaattctgat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc 11040
cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta 11100
ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat 11160
ttttttcact gcagctcgcc ttttgctcac atgtctccca ggggatgggg gtccatccac 11220
cttgccgaaa agatttgtct gaggaactga aaatagaagg gaaaaaagag gagggacaaa 11280
agaggcagaa atgagagggg aggggacaga ggacacctga ataaagacca cacccatgac 11340
ccacgtgatg ctgagaagta ctcctgccct aggaagagac tcagggcaga gggaggaagg 11400
acagcagacc agacagtcac agcagccttg acaaaacgtt cctggaactc aagctcttct 11460
ccacagagga ggacagagca gacagcagag accatggagt ctccctcggc ccctccccac 11520
agatggtgca tcccctggca gaggctcctg ctcacaggtg aactcgagac cggtatggcc 11580
cccccgaccg atgtcagcct gggggacgag ctccacttag acggcgagga cgtggcgatg 11640
gcgcatgccg acgcgctaga cgatttcgat ctggacatgt tggggagcga cgcgctagac 11700
gatttcgatc tggacatgtt ggggagcgac gcgctagacg atttcgatct ggacatgttg 11760
gggagcgacg cgctagacga tttcgatctg gacatgttgg gggacgggga ttccccgggt 11820
ccgggattta ccccccacga ctccgccccc tacggcgctc tggatatggc cgacttcgag 11880
tttgagcaga tgtttaccga tgcccttgga attgacgagt acggtgggac gcgtatgaag 11940
ctactgtctt ctatcgaaca agcatgcgat atttgccgac ttaaaaagct caagtgctcc 12000
aaagaaaaac cgaagtgcgc caagtgtctg aagaacaact gggagtgtcg ctactctccc 12060
aaaaccaaaa ggtctccgct gactagggca catctgacag aagtggaatc aaggctagaa 12120
agactggaac agctatttct actgattttt cctcgagaag accttgacat gattttgaaa 12180
atggattctt tacaggatat aaaagcattg ttaacaggat tatttgtaca agataatgtg 12240
aataaagatg ccgtcacaga tagattggct tcagtggaga ctgatatgcc tctaacattg 12300
agacagcata gaataagtgc gacatcatca tcggaagaga gtagtaacaa aggtcaaaga 12360
cagttgactg tataaaagct tcactcctca ggtgcaggct gcctatcaga aggtggtggc 12420
tggtgtggcc aatgccctgg ctcacaaata ccactgagat ctttttccct ctgccaaaaa 12480
ttatggggac atcatgaagc cccttgagca tctgacttct ggctaataaa ggaaatttat 12540
tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag 12600
ggcaaatcat ttaaaacatc agaatgagta tttggtttag agtttggcaa catatgccca 12660
tatgctggct gccatgaaca aaggttggct ataaagaggt catcagtata tgaaacagcc 12720
ccctgctgtc cattccttat tccatagaaa agccttgact tgaggttaga ttttttttat 12780
attttgtttt gtgttatttt tttctttaac atccctaaaa ttttccttac atgttttact 12840
agccagattt ttcctcctct cctgactact cccagtcata gctgtccctc ttctcttatg 12900
gagatc 12906
<210> 5
<211> 6161
<212> DNA
<213> Artificial
<220>
<223> 编码由细胞特异性启动子CXCR4、第一microRNA为mir-135a-5p、第二micro
RNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白分
别为Lacl、tetR-KRAB、第一和第二识别序列为5×UAS的元件组成的表达系
统的核酸
<400> 5
cggggtgccg ccccccgcgg gtgaccgccc tccgcgggtg acagccctcc gcggagtgcc 60
gcactccgcg gagtgctgta ctccgagcgg agactctaga ctccctatca gtgatagaga 120
taggcgtgta cggtgggagg cctatataag cagagctcgt ttagtgaacc gtcagatcgc 180
tccctatcag tgatagagag aattcgaccg ccaccatgag acatattatc tgccacggag 240
gtgttattac cgaagaaatg gccgccagtc ttttggacca gctgatcgaa gaggtactgg 300
ctgataatct tccacctcct agccattttg aaccacctac ccttcacgaa ctgtatgatt 360
tagacgtgac ggcccccgaa gatcccaacg aggaggcggt ttcgcagatt tttcccgact 420
ctgtaatgtt ggcggtgcag gaagggattg acttactcac ttttccgccg gcgcccggtt 480
ctccggagcc gcctcacctt tcccggcagc ccgagcagcc ggagcagaga gccttgggtc 540
cggtttctat gccaaacctt gtaccggagg tgatcgatct tacctgccac gaggctggct 600
ttccacccag tgacgacgag gatgaagagg gtgaggagtt tgtgttagat tatgtggagc 660
accccgggca cggttgcagg tcttgtcatt atcaccggag gaatacgggg gacccagata 720
ttatgtgttc gctttgctat atgaggacct gtggcatgtt tgtctacagt aagtgaaaat 780
tatgggcagt gggtgataga gtggtgggtt tggtgtggta attttttttt taatttttac 840
agttttgtgg tttaaagaat tttgtattgt gattttttta aaaggtcctg tgtctgaacc 900
tgagcctgag cccgagccag aaccggagcc tgcaagacct acccgccgtc ctaaaatggc 960
gcctgctatc ctgaggcgcc cgacatcacc tgtgtctaga gaatgcaata gtagtacgga 1020
tagctgtgac tccggtcctt ctaacacacc tcctgagata cacccggtgg tcccgctgtg 1080
ccccattaaa ccagttgccg tgagagttgg tgggcgtcgc caggctgtgg aatgtatcga 1140
ggacttgctt aacgagcctg ggcaaccttt ggacttgagc tgtaaacgcc ccaggccact 1200
cgagggtacc gggtccggag ctacaaattt ttccctcctc aaacaggctg gagatgtcga 1260
agaaaatcct gggcctaccg gtatgtggct gcagagcctc cttctactgg gcaccgtggc 1320
ttgcagcatc tcggcccccg ccagaagtcc cagccccagc acccagccct gggaacacgt 1380
aaatgcaatc caggaggccc gccgcctcct gaacttgtca cgggacacgg cagccgagat 1440
gaatgaaact gtggaggtca tcagtgaaat gtttgacttg caggagccca catgcctgca 1500
gactcggctg gagctctaca aacagggcct gcgtggcagc ctcaccaagc tgaagggccc 1560
actgaccatg atggctagcc attacaagca gcactgcccg ccgacccctg agacatcctg 1620
tgccactcag atcattacct ttgagtcttt caaggagaac ctcaaggact ttctgctggt 1680
catcccattt gactgctggg aacccgtcca ggaaagatct gagggccgcg gcagcctgct 1740
gacctgcggc gacgtggagg aaaaccccgg ccccggatcc atgaaaccag taacgttata 1800
cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc 1860
cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta 1920
cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga ttggcgttgc 1980
cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc 2040
cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg 2100
taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca ttaactatcc 2160
gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc cggcgttatt 2220
tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg aagacggtac 2280
gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg 2340
cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat atctcactcg 2400
caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt ccggttttca 2460
acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg ttgccaacga 2520
tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga 2580
tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc cgccgttaac 2640
caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact 2700
ctctcagggc caggcggtga agggcaatca gctgttgccg gtctcactgg tgaaaagaaa 2760
aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat 2820
gcagctggca cgacaggttt cccgactgga aagcgggcag agcagcctga ggcctcctaa 2880
gaagaagagg aaggttgtta caaacaccat tgtcacactc cacaaacacc attgtcacac 2940
tccacaaaca ccattgtcac actccacaaa caccattgtc acactccatc acataggaat 3000
aaaaagccat atcacatagg aataaaaagc catatcacat aggaataaaa agccatatca 3060
cataggaata aaaagccata gaaagcggac tactgtagtc cgcggagtac tgtcctccgc 3120
ggacgactgt cgtccgcgga gcactgtgct ccgcggatga ctgtcatccg agcggagact 3180
ctagaaattg tgagcggata acaattgggt atataatggg ggccgtcagc tactaccaga 3240
gccgatcgaa cactcgagcc gagcagacgt gcctacaatt gtgagcgctc acaattacta 3300
gaagatcttg aattcgccac catggctaga ttagataaaa gtaaagtgat taacagcgca 3360
ttagagctgc ttaatgaggt cggaatcgaa ggtttaacaa cccgtaaact cgcccagaag 3420
ctaggtgtag agcagcctac attgtattgg catgtaaaaa ataagcgggc tttgctcgac 3480
gccttagcca ttgagatgtt agataggcac catactcact tttgcccttt agaaggggaa 3540
agctggcaag attttttacg taataacgct aaaagtttta gatgtgcttt actaagtcat 3600
cgcgatggag caaaagtaca tttaggtaca cggcctacag aaaaacagta tgaaactctc 3660
gaaaatcaat tagccttttt atgccaacaa ggtttttcac tagagaatgc attatatgca 3720
ctcagcgctg tggggcattt tactttaggt tgcgtattgg aagatcaaga gcatcaagtc 3780
gctaaagaag aaagggaaac acctactact gatagtatgc cgccattatt acgacaagct 3840
atcgaattat ttgatcacca aggtgcagag ccagccttct tattcggcct tgaattgatc 3900
atatgcggat tagaaaaaca acttaaatgt gaaagtgggt cgccaaaaaa gaagagaaag 3960
gtggatggtg gcggagcact gtctccccag cattcagctg tgacccaggg atcaatcatt 4020
aagaacaagg aaggtatgga tgctaagtcc ctgaccgctt ggtcaagaac tttggtaact 4080
tttaaggacg tgttcgtcga ttttactcgg gaggagtgga aactattgga taccgcccag 4140
caaatcgtgt atagaaacgt tatgctcgag aactataaga atcttgtgtc actgggctac 4200
caactgacaa agcctgacgt tatcctgagg ctggaaaagg gtgaggagcc ttggttggtc 4260
gagcgggaga tccaccagga aactcaccca gacagtgaga ctgctttcga gatcaagtcc 4320
tctgtgtaat acaaggcggc cgcaaatcaa catcagtctg ataagctatc aacatcagtc 4380
tgataagcta tcaacatcag tctgataagc tatcaacatc agtctgataa gctaaagctt 4440
cgaattctga taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc 4500
ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt 4560
attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca 4620
tttttttcac tgcagctcgc cttttgctca catgttaccg accacccgca aacagcaggg 4680
tcccctgggc ttcccaagcc gcgcacctct ccgccccgcc cctgcgccct ccttcctcgc 4740
gtctgcccct ctcccccacc ccgccttctc cctccccgcc ccagcggcgc atgcgccgcg 4800
ctcggagcgt gtttttataa aagtccggcc gcggccagaa acttcagttt gttggctgcg 4860
gcagcaggta gcaaagtgac gccgagggcc tgagtgctcc agtagccacc gcatctggag 4920
aaccagcggt taccctcgag accggtatgg cccccccgac cgatgtcagc ctgggggacg 4980
agctccactt agacggcgag gacgtggcga tggcgcatgc cgacgcgcta gacgatttcg 5040
atctggacat gttgggggac ggggattccc cgggtccggg atttaccccc cacgactccg 5100
ccccctacgg cgctctggat atggccgact tcgagtttga gcagatgttt accgatgccc 5160
ttggaattga cgagtacggt gggacgcgta tgaagctact gtcttctatc gaacaagcat 5220
gcgatatttg ccgacttaaa aagctcaagt gctccaaaga aaaaccgaag tgcgccaagt 5280
gtctgaagaa caactgggag tgtcgctact ctcccaaaac caaaaggtct ccgctgacta 5340
gggcacatct gacagaagtg gaatcaaggc tagaaagact ggaacagcta tttctactga 5400
tttttcctcg agaagacctt gacatgattt tgaaaatgga ttctttacag gatataaaag 5460
cattgttaac aggattattt gtacaagata atgtgaataa agatgccgtc acagatagat 5520
tggcttcagt ggagactgat atgcctctaa cattgagaca gcatagaata agtgcgacat 5580
catcatcgga agagagtagt aacaaaggtc aaagacagtt gactgtataa aagcttcact 5640
cctcaggtgc aggctgccta tcagaaggtg gtggctggtg tggccaatgc cctggctcac 5700
aaataccact gagatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt 5760
gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat 5820
tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat 5880
gagtatttgg tttagagttt ggcaacatat gcccatatgc tggctgccat gaacaaaggt 5940
tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat 6000
agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt atttttttct 6060
ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct cctctcctga 6120
ctactcccag tcatagctgt ccctcttctc ttatggagat c 6161
<210> 6
<211> 12783
<212> DNA
<213> Artificial
<220>
<223> 编码由细胞特异性启动子CXCR4、第一microRNA为mir-143-3p、第二microR
NA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白分别
为TALE蛋白、第一和第二识别序列为5×UAS的元件组成的表达系统的核酸
<400> 6
cggagtactg tcctccgagc ggagtactgt cctccgagcg gagtactgtc ctccgagcgg 60
agtactgtcc tccgagcgga gttctgtcct ccgagcggag actctagata cccctctccg 120
cttctgaatt ctaggcgtgt acggtgggag gcctatataa gcagagctcg tttagtgaac 180
cgtcagatcg cctcgagtac ccctctccgc ttcttacccc tctccgcttc ttacccctct 240
ccgcttctga attcgaccgc caccatgaga catattatct gccacggagg tgttattacc 300
gaagaaatgg ccgccagtct tttggaccag ctgatcgaag aggtactggc tgataatctt 360
ccacctccta gccattttga accacctacc cttcacgaac tgtatgattt agacgtgacg 420
gcccccgaag atcccaacga ggaggcggtt tcgcagattt ttcccgactc tgtaatgttg 480
gcggtgcagg aagggattga cttactcact tttccgccgg cgcccggttc tccggagccg 540
cctcaccttt cccggcagcc cgagcagccg gagcagagag ccttgggtcc ggtttctatg 600
ccaaaccttg taccggaggt gatcgatctt acctgccacg aggctggctt tccacccagt 660
gacgacgagg atgaagaggg tgaggagttt gtgttagatt atgtggagca ccccgggcac 720
ggttgcaggt cttgtcatta tcaccggagg aatacggggg acccagatat tatgtgttcg 780
ctttgctata tgaggacctg tggcatgttt gtctacagta agtgaaaatt atgggcagtg 840
ggtgatagag tggtgggttt ggtgtggtaa tttttttttt aatttttaca gttttgtggt 900
ttaaagaatt ttgtattgtg atttttttaa aaggtcctgt gtctgaacct gagcctgagc 960
ccgagccaga accggagcct gcaagaccta cccgccgtcc taaaatggcg cctgctatcc 1020
tgaggcgccc gacatcacct gtgtctagag aatgcaatag tagtacggat agctgtgact 1080
ccggtccttc taacacacct cctgagatac acccggtggt cccgctgtgc cccattaaac 1140
cagttgccgt gagagttggt gggcgtcgcc aggctgtgga atgtatcgag gacttgctta 1200
acgagcctgg gcaacctttg gacttgagct gtaaacgccc caggccaggg tccggagcta 1260
caaatttttc cctcctcaaa caggctggag atgtcgaaga aaatcctggg cctatggagg 1320
cttgggagtg tttggaagat ttttctgctg tgcgtaactt gctggaacag agctctaaca 1380
gtacctcttg gttttggagg tttctgtggg gctcatccca ggcaaagtta gtctgcagaa 1440
ttaaggagga ttacaagtgg gaatttgaag agcttttgaa atcctgtggt gagctgtttg 1500
attctttgaa tctgggtcac caggcgcttt tccaagagaa ggtcatcaag actttggatt 1560
tttccacacc ggggcgcgct gcggctgctg ttgctttttt gagttttata aaggataaat 1620
ggagcgaaga aacccatctg agcggggggt acctgctgga ttttctggcc atgcatctgt 1680
ggagagcggt tgtgagacac aagaatcgcc tgctactgtt gtcttccgtc cgcccggcga 1740
taataccgac ggaggagcag cagcagcagc aggaggaagc caggcggcgg cggcaggagc 1800
agagcccatg gaacccgaga gccggcctgg accctcggga atgaatgttg tacaggtggc 1860
tgaactgtat ccagaactga gacgcatttt gacaattaca gaggatgggc aggggctaaa 1920
gggggtaaag agggagcggg gggcttgtga ggctacagag gaggctagga atctagcttt 1980
tagcttaatg accagacacc gtcctgagtg tattactttt caacagatca aggataattg 2040
cgctaatgag cttgatctgc tggcgcagaa gtattccata gagcagctga ccacttactg 2100
gctgcagcca ggggatgatt ttgaggaggc tattagggta tatgcaaagg tggcacttag 2160
gccagattgc aagtacaaga tcagcaaact tgtaaatatc aggaattgtt gctacatttc 2220
tgggaacggg gccgaggtgg agatagatac ggaggatagg gtggccttta gatgtagcat 2280
gataaatatg tggccggggg tgcttggcat ggacggggtg gttattatga atgtaaggtt 2340
tactggcccc aattttagcg gtacggtttt cctggccaat accaacctta tcctacacgg 2400
tgtaagcttc tatgggttta acaatacctg tgtggaagcc tggaccgatg taagggttcg 2460
gggctgtgcc ttttactgct gctggaaggg ggtggtgtgt cgccccaaaa gcagggcttc 2520
aattaagaaa tgcctctttg aaaggtgtac cttgggtatc ctgtctgagg gtaactccag 2580
ggtgcgccac aatgtggcct ccgactgtgg ttgcttcatg ctagtgaaaa gcgtggctgt 2640
gattaagcat aacatggtat gtggcaactg cgaggacagg gcctctcaga tgctgacctg 2700
ctcggacggc aactgtcacc tgctgaagac cattcacgta gccagccact ctcgcaaggc 2760
ctggccagtg tttgagcata acatactgac ccgctgttcc ttgcatttgg gtaacaggag 2820
gggggtgttc ctaccttacc aatgcaattt gagtcacact aagatattgc ttgagcccga 2880
gagcatgtcc aaggtgaacc tgaacggggt gtttgacatg accatgaaga tctggaaggt 2940
gctgaggtac gatgagaccc gcaccaggtg cagaccctgc gagtgtggcg gtaaacatat 3000
taggaaccag cctgtgatgc tggatgtgac cgaggagctg aggcccgatc acttggtgct 3060
ggcctgcacc cgcgctgagt ttggctctag cgatgaagat acagatctcg aggggtccgg 3120
agctacaaat ttttccctcc tcaaacaggc tggagatgtc gaagaaaatc ctgggcctac 3180
cggtatgtat aggatgcagt tgctctcttg catcgccctg tctctagcgt tggtgactaa 3240
tagcgcccct acttcttctt ccacgaagaa aacccagctg caactggaac accttctgct 3300
ggatctccag atgatcctta acgggattaa caactataag aacccgaagc tcacggctat 3360
gctgacggcg aagttcgcta tgcctaaaaa ggctactgag ctgaaacact tacagtgtct 3420
cgaagaggca ctcaaacccc tcgaggaggt actcaatctg gcgcaaagca aaaacttcca 3480
tctccgccca cgggacctaa tctccaacat caacgtgatt gtgctggagc tcaaggggtc 3540
tgaaaccacg tttatgtgcg agtacgcaga tgaaactgct accatagtgg aattcctcaa 3600
tcgttggatc actttctgcc agtcaatcat ttcaaccctg accagatctg agggccgcgg 3660
cagcctgctg acctgcggcg acgtggagga aaaccccggc cccggatcct tgtcgaccat 3720
tcgtccgcgc aggccaagtc ctgcccgcga gcttctgccc ggaccccaac cggatagggt 3780
tcagccgact gcagatcgtg gggtgtctgc gcctgctggc agccctctgg atggcttgcc 3840
cgctcggcgg acggtgtccc ggacccggct gccatctccc cctgcgccct cacctgcgtt 3900
ctcggcgggc agcttcagcg atctgctccg tccgttcgat ccgtcgcttc ttgatacatc 3960
gcttcttgat tcgatgcctg ccgtcggcac gccgcataca gcggctgccc cagcagagtg 4020
ggatgaggcg caatcggctc tgcgtgcagc cgatgacccg ccacccaccg tgcgtgtcgc 4080
tgtcactgcc gcgcggccgc cgcgcgccaa gccggccccg cgacggcgtg ctgcgcaacc 4140
ctccgacgct tcgccggccg cgcaggtgga tctacgcacg ctcggctaca gtcagcagca 4200
gcaagagaag atcaaaccga aggtgcgttc gacagtggcg cagcaccacg aggcactggt 4260
gggccatggg tttacacacg cgcacatcgt tgcgctcagc caacacccgg cagcgttagg 4320
gaccgtcgct gtcacgtatc agcacataat cacggcgttg ccagaggcga cacacgaaga 4380
catcgttggc gtcggcaaac agtggtccgg cgcacgcgcc ctggaggcct tgctcacgga 4440
tgcgggggag ttgagaggtc cgccgttaca gttggacaca ggccaacttg tgaagattgc 4500
aaaacgtggc ggcgtgaccg caatggaggc agtgcatgca tcgcgcaatg cactgacggg 4560
tgcccccctg aacctgaccc cggaccaagt ggtggctatc gccagcaaca ttggcggcaa 4620
gcaagcgctc gaaacggtgc agcggctgtt gccggtgctg tgccaggacc atggcctgac 4680
cccggaccaa gtggtggcta tcgccagcaa cggtggcggc aagcaagcgc tcgaaacggt 4740
gcagcggctg ttgccggtgc tgtgccagga ccatggcctg accccggacc aagtggtggc 4800
tatcgccagc aacattggcg gcaagcaagc gctcgaaacg gtgcagcggc tgttgccggt 4860
gctgtgccag gaccatggcc tgaccccgga ccaagtggtg gctatcgcca gcaacaatgg 4920
cggcaagcaa gcgctcgaaa cggtgcagcg gctgttgccg gtgctgtgcc aggaccatgg 4980
cctgaccccg gaccaagtgg tggctatcgc cagcaacatt ggcggcaagc aagcgctcga 5040
aacggtgcag cggctgttgc cggtgctgtg ccaggaccat ggcctgaccc cggaccaagt 5100
ggtggctatc gccagcaaca ttggcggcaa gcaagcgctc gaaacggtgc agcggctgtt 5160
gccggtgctg tgccaggacc atggcctgac tccggaccaa gtggtggcta tcgccagcca 5220
cgatggcggc aagcaagcgc tcgaaacggt gcagcggctg ttgccggtgc tgtgccagga 5280
ccatggcctg actccggacc aagtggtggc tatcgccagc cacgatggcg gcaagcaagc 5340
gctcgaaacg gtgcagcggc tgttgccggt gctgtgccag gaccatggcc tgaccccgga 5400
ccaagtggtg gctatcgcca gcaacaatgg cggcaagcaa gcgctcgaaa cggtgcagcg 5460
gctgttgccg gtgctgtgcc aggaccatgg cctgaccccg gaccaagtgg tggctatcgc 5520
cagcaacatt ggcggcaagc aagcgctcga aacggtgcag cggctgttgc cggtgctgtg 5580
ccaggaccat ggcctgaccc cggaccaagt ggtggctatc gccagcaacg gtggcggcaa 5640
gcaagcgctc gaaacggtgc agcggctgtt gccggtgctg tgccaggacc atggcctgac 5700
tccggaccaa gtggtggcta tcgccagcca cgatggcggc aagcaagcgc tcgaaacggt 5760
gcagcggctg ttgccggtgc tgtgccagga ccatggcctg actccggacc aagtggtggc 5820
tatcgccagc cacgatggcg gcaagcaagc gctcgaaacg gtgcagcggc tgttgccggt 5880
gctgtgccag gaccatggcc tgaccccgga ccaagtggtg gctatcgcca gcaacggtgg 5940
cggcaagcaa gcgctcgaaa cggtgcagcg gctgttgccg gtgctgtgcc aggaccatgg 6000
cctgactccg gaccaagtgg tggctatcgc cagccacgat ggcggcaagc aagcgctcga 6060
aacggtgcag cggctgttgc cggtgctgtg ccaggaccat ggcctgactc cggaccaagt 6120
ggtggctatc gccagccacg atggcggcaa gcaagcgctc gaaacggtgc agcggctgtt 6180
gccggtgctg tgccaggacc atggcctgac tccggaccaa gtggtggcta tcgccagcca 6240
cgatggcggc aagcaagcgc tcgaaacggt gcagcggctg ttgccggtgc tgtgccagga 6300
ccatggcctg accccggacc aagtggtggc tatcgccagc aacattggcg gcaagcaagc 6360
gctcgaaacg gtgcagcggc tgttgccggt gctgtgccag gaccatggcc tgaccccgga 6420
ccaagtggtg gctatcgcca gcaacggtgg cggcaagcaa gcgctcgaaa cggtgcagcg 6480
gctgttgccg gtgctgtgcc aggaccatgg cctgaccccg gaccaagtgg tggctatcgc 6540
cagcaacggt ggcggcaagc aagcgctcga aagcattgtg gcccagctga gccggcctga 6600
tccggcgttg gccgcgttga ccaacgacca cctcgtcgcc ttggcctgcc tcggcggacg 6660
tcctgccatg gatgcagtga aaaagggatt gccgcacgcg ccggaattga tcagaagagt 6720
caatcgccgt attggcgaac gcacgtccca tcgcgttgcc gactacgcgc aagtggttcg 6780
cgtgctggag tttttccagt gccactccca cccagcgtac gcatttgatg aggccatgac 6840
gcagttcggg atgagcagga acgggttggt acagctcttt cgcagagtgg gcgtcaccga 6900
actcgaagcc cgcggtggaa cgctcccccc agcctcgcag cgttgggacc gtatcctcca 6960
ggcatcaggg atgaaaaggg ccaaaccgtc ccctacttca gctcaaacac cggatcaggc 7020
gtctttgcat gcattcgccg attcgctgga gcgtgacctt gatgcgccca gcccaatgca 7080
cgagggagat cagacgcggg caagcagccg taaacggtcc cgatcggatc gtgctgtcac 7140
cggcccctcc gcacagcagg ctgtcgaggt gcgcgttccc gaacagcgcg atgcgctgca 7200
tttgcccctc agctggaggg taaaacgccc gcgtaccagg atctggggcg gcctcccggt 7260
taagtagtgc tttctacttt atgagtagtg ctttctactt tatgagtagt gctttctact 7320
ttatgagtag tgctttctac tttatggagc tacagtgctt catctcagag ctacagtgct 7380
tcatctcaga gctacagtgc ttcatctcag agctacagtg cttcatctca gaaagcggag 7440
tactgtcctc cgagcggagt actgtcctcc gagcggagta ctgtcctccg agcggagtac 7500
tgtcctccga gcggagttct gtcctccgag cggagactct agatatagaa ccgatcctcc 7560
cattgaattc taggcgtgta cggtgggagg cctatataag cagagctcgt ttagtgaacc 7620
gtcagatcgc ctcgagtata gaaccgatcc tcccatttat agaaccgatc ctcccattta 7680
tagaaccgat cctcccatta ctagaagatc ttgaattcgc caccttgtcg accattcgtc 7740
cgcgcaggcc aagtcctgcc cgcgagcttc tgcccggacc ccaaccggat agggttcagc 7800
cgactgcaga tcgtggggtg tctgcgcctg ctggcagccc tctggatggc ttgcccgctc 7860
ggcggacggt gtcccggacc cggctgccat ctccccctgc gccctcacct gcgttctcgg 7920
cgggcagctt cagcgatctg ctccgtccgt tcgatccgtc gcttcttgat acatcgcttc 7980
ttgattcgat gcctgccgtc ggcacgccgc atacagcggc tgccccagca gagtgggatg 8040
aggcgcaatc ggctctgcgt gcagccgatg acccgccacc caccgtgcgt gtcgctgtca 8100
ctgccgcgcg gccgccgcgc gccaagccgg ccccgcgacg gcgtgctgcg caaccctccg 8160
acgcttcgcc ggccgcgcag gtggatctac gcacgctcgg ctacagtcag cagcagcaag 8220
agaagatcaa accgaaggtg cgttcgacag tggcgcagca ccacgaggca ctggtgggcc 8280
atgggtttac acacgcgcac atcgttgcgc tcagccaaca cccggcagcg ttagggaccg 8340
tcgctgtcac gtatcagcac ataatcacgg cgttgccaga ggcgacacac gaagacatcg 8400
ttggcgtcgg caaacagtgg tccggcgcac gcgccctgga ggccttgctc acggatgcgg 8460
gggagttgag aggtccgccg ttacagttgg acacaggcca acttgtgaag attgcaaaac 8520
gtggcggcgt gaccgcaatg gaggcagtgc atgcatcgcg caatgcactg acgggtgccc 8580
ccctgaacct gaccccggac caagtggtgg ctatcgccag caacattggc ggcaagcaag 8640
cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc ctgactccgg 8700
accaagtggt ggctatcgcc agccacgatg gcggcaagca agcgctcgaa acggtgcagc 8760
ggctgttgcc ggtgctgtgc caggaccatg gcctgactcc ggaccaagtg gtggctatcg 8820
ccagccacga tggcggcaag caagcgctcg aaacggtgca gcggctgttg ccggtgctgt 8880
gccaggacca tggcctgact ccggaccaag tggtggctat cgccagccac gatggcggca 8940
agcaagcgct cgaaacggtg cagcggctgt tgccggtgct gtgccaggac catggcctga 9000
ctccggacca agtggtggct atcgccagcc acgatggcgg caagcaagcg ctcgaaacgg 9060
tgcagcggct gttgccggtg ctgtgccagg accatggcct gaccccggac caagtggtgg 9120
ctatcgccag caacggtggc ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg 9180
tgctgtgcca ggaccatggc ctgactccgg accaagtggt ggctatcgcc agccacgatg 9240
gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg 9300
gcctgacccc ggaccaagtg gtggctatcg ccagcaacgg tggcggcaag caagcgctcg 9360
aaacggtgca gcggctgttg ccggtgctgt gccaggacca tggcctgact ccggaccaag 9420
tggtggctat cgccagccac gatggcggca agcaagcgct cgaaacggtg cagcggctgt 9480
tgccggtgct gtgccaggac catggcctga ctccggacca agtggtggct atcgccagcc 9540
acgatggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg 9600
accatggcct gaccccggac caagtggtgg ctatcgccag caacaatggc ggcaagcaag 9660
cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc ctgactccgg 9720
accaagtggt ggctatcgcc agccacgatg gcggcaagca agcgctcgaa acggtgcagc 9780
ggctgttgcc ggtgctgtgc caggaccatg gcctgacccc ggaccaagtg gtggctatcg 9840
ccagcaacgg tggcggcaag caagcgctcg aaacggtgca gcggctgttg ccggtgctgt 9900
gccaggacca tggcctgacc ccggaccaag tggtggctat cgccagcaac ggtggcggca 9960
agcaagcgct cgaaacggtg cagcggctgt tgccggtgct gtgccaggac catggcctga 10020
ctccggacca agtggtggct atcgccagcc acgatggcgg caagcaagcg ctcgaaacgg 10080
tgcagcggct gttgccggtg ctgtgccagg accatggcct gaccccggac caagtggtgg 10140
ctatcgccag caacggtggc ggcaagcaag cgctcgaaag cattgtggcc cagctgagcc 10200
ggcctgatcc ggcgttggcc gcgttgacca acgaccacct cgtcgccttg gcctgcctcg 10260
gcggacgtcc tgccatggat gcagtgaaaa agggattgcc gcacgcgccg gaattgatca 10320
gaagagtcaa tcgccgtatt ggcgaacgca cgtcccatcg cgttgccgac tacgcgcaag 10380
tggttcgcgt gctggagttt ttccagtgcc actcccaccc agcgtacgca tttgatgagg 10440
ccatgacgca gttcgggatg agcaggaacg ggttggtaca gctctttcgc agagtgggcg 10500
tcaccgaact cgaagcccgc ggtggaacgc tccccccagc ctcgcagcgt tgggaccgta 10560
tcctccaggc atcagggatg aaaagggcca aaccgtcccc tacttcagct caaacaccgg 10620
atcaggcgtc tttgcatgca ttcgccgatt cgctggagcg tgaccttgat gcgcccagcc 10680
caatgcacga gggagatcag acgcgggcaa gcagccgtaa acggtcccga tcggatcgtg 10740
ctgtcaccgg cccctccgca cagcaggctg tcgaggtgcg cgttcccgaa cagcgcgatg 10800
cgctgcattt gcccctcagc tggagggtaa aacgcccgcg taccaggatc tggggcggcc 10860
tcccgtaata caaggcggcc gcaaatcaac atcagtctga taagctatca acatcagtct 10920
gataagctat caacatcagt ctgataagct atcaacatca gtctgataag ctaaagcttc 10980
gaattctgat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc 11040
cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta 11100
ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat 11160
ttttttcact gctaccgacc acccgcaaac agcagggtcc cctgggcttc ccaagccgcg 11220
cacctctccg ccccgcccct gcgccctcct tcctcgcgtc tgcccctctc ccccaccccg 11280
ccttctccct ccccgcccca gcggcgcatg cgccgcgctc ggagcgtgtt tttataaaag 11340
tccggccgcg gccagaaact tcagtttgtt ggctgcggca gcaggtagca aagtgacgcc 11400
gagggcctga gtgctccagt agccaccgca tctggagaac cagcggttac catggccccc 11460
ccgaccgatg tcagcctggg ggacgagctc cacttagacg gcgaggacgt ggcgatggcg 11520
catgccgacg cgctagacga tttcgatctg gacatgttgg ggagcgacgc gctagacgat 11580
ttcgatctgg acatgttggg gagcgacgcg ctagacgatt tcgatctgga catgttgggg 11640
agcgacgcgc tagacgattt cgatctggac atgttggggg acggggattc cccgggtccg 11700
ggatttaccc cccacgactc cgccccctac ggcgctctgg atatggccga cttcgagttt 11760
gagcagatgt ttaccgatgc ccttggaatt gacgagtacg gtgggacgcg tatgaagcta 11820
ctgtcttcta tcgaacaagc atgcgatatt tgccgactta aaaagctcaa gtgctccaaa 11880
gaaaaaccga agtgcgccaa gtgtctgaag aacaactggg agtgtcgcta ctctcccaaa 11940
accaaaaggt ctccgctgac tagggcacat ctgacagaag tggaatcaag gctagaaaga 12000
ctggaacagc tatttctact gatttttcct cgagaagacc ttgacatgat tttgaaaatg 12060
gattctttac aggatataaa agcattgtta acaggattat ttgtacaaga taatgtgaat 12120
aaagatgccg tcacagatag attggcttca gtggagactg atatgcctct aacattgaga 12180
cagcatagaa taagtgcgac atcatcatcg gaagagagta gtaacaaagg tcaaagacag 12240
ttgactgtat aaaagcttca ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg 12300
tgtggccaat gccctggctc acaaatacca ctgagatctt tttccctctg ccaaaaatta 12360
tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga aatttatttt 12420
cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc 12480
aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat atgcccatat 12540
gctggctgcc atgaacaaag gttggctata aagaggtcat cagtatatga aacagccccc 12600
tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt tttttatatt 12660
ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc 12720
cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc tcttatggag 12780
atc 12783
<210> 7
<211> 6150
<212> DNA
<213> Artificial
<220>
<223> 编码由细胞特异性启动子Survivin1、第一microRNA为mir-135a-5p、第二m
icroRNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋
白分别为Lacl、tetR-KRAB、第一和第二识别序列为5×UAS的元件组成的表
达系统的核酸
<400> 7
cggggtgccg ccccccgcgg gtgaccgccc tccgcgggtg acagccctcc gcggagtgcc 60
gcactccgcg gagtgctgta ctccgagcgg agactctaga ctccctatca gtgatagaga 120
taggcgtgta cggtgggagg cctatataag cagagctcgt ttagtgaacc gtcagatcgc 180
tccctatcag tgatagagag aattcgaccg ccaccatgag acatattatc tgccacggag 240
gtgttattac cgaagaaatg gccgccagtc ttttggacca gctgatcgaa gaggtactgg 300
ctgataatct tccacctcct agccattttg aaccacctac ccttcacgaa ctgtatgatt 360
tagacgtgac ggcccccgaa gatcccaacg aggaggcggt ttcgcagatt tttcccgact 420
ctgtaatgtt ggcggtgcag gaagggattg acttactcac ttttccgccg gcgcccggtt 480
ctccggagcc gcctcacctt tcccggcagc ccgagcagcc ggagcagaga gccttgggtc 540
cggtttctat gccaaacctt gtaccggagg tgatcgatct tacctgccac gaggctggct 600
ttccacccag tgacgacgag gatgaagagg gtgaggagtt tgtgttagat tatgtggagc 660
accccgggca cggttgcagg tcttgtcatt atcaccggag gaatacgggg gacccagata 720
ttatgtgttc gctttgctat atgaggacct gtggcatgtt tgtctacagt aagtgaaaat 780
tatgggcagt gggtgataga gtggtgggtt tggtgtggta attttttttt taatttttac 840
agttttgtgg tttaaagaat tttgtattgt gattttttta aaaggtcctg tgtctgaacc 900
tgagcctgag cccgagccag aaccggagcc tgcaagacct acccgccgtc ctaaaatggc 960
gcctgctatc ctgaggcgcc cgacatcacc tgtgtctaga gaatgcaata gtagtacgga 1020
tagctgtgac tccggtcctt ctaacacacc tcctgagata cacccggtgg tcccgctgtg 1080
ccccattaaa ccagttgccg tgagagttgg tgggcgtcgc caggctgtgg aatgtatcga 1140
ggacttgctt aacgagcctg ggcaaccttt ggacttgagc tgtaaacgcc ccaggccact 1200
cgagggtacc gggtccggag ctacaaattt ttccctcctc aaacaggctg gagatgtcga 1260
agaaaatcct gggcctaccg gtatgtggct gcagagcctc cttctactgg gcaccgtggc 1320
ttgcagcatc tcggcccccg ccagaagtcc cagccccagc acccagccct gggaacacgt 1380
aaatgcaatc caggaggccc gccgcctcct gaacttgtca cgggacacgg cagccgagat 1440
gaatgaaact gtggaggtca tcagtgaaat gtttgacttg caggagccca catgcctgca 1500
gactcggctg gagctctaca aacagggcct gcgtggcagc ctcaccaagc tgaagggccc 1560
actgaccatg atggctagcc attacaagca gcactgcccg ccgacccctg agacatcctg 1620
tgccactcag atcattacct ttgagtcttt caaggagaac ctcaaggact ttctgctggt 1680
catcccattt gactgctggg aacccgtcca ggaaagatct gagggccgcg gcagcctgct 1740
gacctgcggc gacgtggagg aaaaccccgg ccccggatcc atgaaaccag taacgttata 1800
cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc 1860
cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta 1920
cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga ttggcgttgc 1980
cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc 2040
cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg 2100
taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca ttaactatcc 2160
gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc cggcgttatt 2220
tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg aagacggtac 2280
gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg 2340
cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat atctcactcg 2400
caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt ccggttttca 2460
acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg ttgccaacga 2520
tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga 2580
tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc cgccgttaac 2640
caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact 2700
ctctcagggc caggcggtga agggcaatca gctgttgccg gtctcactgg tgaaaagaaa 2760
aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat 2820
gcagctggca cgacaggttt cccgactgga aagcgggcag agcagcctga ggcctcctaa 2880
gaagaagagg aaggttgtta caaacaccat tgtcacactc cacaaacacc attgtcacac 2940
tccacaaaca ccattgtcac actccacaaa caccattgtc acactccatc acataggaat 3000
aaaaagccat atcacatagg aataaaaagc catatcacat aggaataaaa agccatatca 3060
cataggaata aaaagccata gaaagcggac tactgtagtc cgcggagtac tgtcctccgc 3120
ggacgactgt cgtccgcgga gcactgtgct ccgcggatga ctgtcatccg agcggagact 3180
ctagaaattg tgagcggata acaattgggt atataatggg ggccgtcagc tactaccaga 3240
gccgatcgaa cactcgagcc gagcagacgt gcctacaatt gtgagcgctc acaattacta 3300
gaagatcttg aattcgccac catggctaga ttagataaaa gtaaagtgat taacagcgca 3360
ttagagctgc ttaatgaggt cggaatcgaa ggtttaacaa cccgtaaact cgcccagaag 3420
ctaggtgtag agcagcctac attgtattgg catgtaaaaa ataagcgggc tttgctcgac 3480
gccttagcca ttgagatgtt agataggcac catactcact tttgcccttt agaaggggaa 3540
agctggcaag attttttacg taataacgct aaaagtttta gatgtgcttt actaagtcat 3600
cgcgatggag caaaagtaca tttaggtaca cggcctacag aaaaacagta tgaaactctc 3660
gaaaatcaat tagccttttt atgccaacaa ggtttttcac tagagaatgc attatatgca 3720
ctcagcgctg tggggcattt tactttaggt tgcgtattgg aagatcaaga gcatcaagtc 3780
gctaaagaag aaagggaaac acctactact gatagtatgc cgccattatt acgacaagct 3840
atcgaattat ttgatcacca aggtgcagag ccagccttct tattcggcct tgaattgatc 3900
atatgcggat tagaaaaaca acttaaatgt gaaagtgggt cgccaaaaaa gaagagaaag 3960
gtggatggtg gcggagcact gtctccccag cattcagctg tgacccaggg atcaatcatt 4020
aagaacaagg aaggtatgga tgctaagtcc ctgaccgctt ggtcaagaac tttggtaact 4080
tttaaggacg tgttcgtcga ttttactcgg gaggagtgga aactattgga taccgcccag 4140
caaatcgtgt atagaaacgt tatgctcgag aactataaga atcttgtgtc actgggctac 4200
caactgacaa agcctgacgt tatcctgagg ctggaaaagg gtgaggagcc ttggttggtc 4260
gagcgggaga tccaccagga aactcaccca gacagtgaga ctgctttcga gatcaagtcc 4320
tctgtgtaat acaaggcggc cgcaaatcaa catcagtctg ataagctatc aacatcagtc 4380
tgataagcta tcaacatcag tctgataagc tatcaacatc agtctgataa gctaaagctt 4440
cgaattctga taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc 4500
ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt 4560
attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca 4620
tttttttcac tgcagctcgc cttttgctca catgttaggt gtgggcaggg acgagctggc 4680
gcggcgtcgc tgggtgcacc gcgaccacgg gcagagccac gcggcgggag gactacaact 4740
cccggcacac cccgcgccgc cccgcctcta ctcccagaag gccgcggggg gtggaccgcc 4800
taagagggcg tgcgctcccg acatgccccg cggcgcgcca ttaaccgcca gatttgaatc 4860
gcgggacccg ttggcagagg tggcggcggc ggcatgggtg ccccgacgtt gccccctgcc 4920
tggctcgaga ccggtatggc ccccccgacc gatgtcagcc tgggggacga gctccactta 4980
gacggcgagg acgtggcgat ggcgcatgcc gacgcgctag acgatttcga tctggacatg 5040
ttgggggacg gggattcccc gggtccggga tttacccccc acgactccgc cccctacggc 5100
gctctggata tggccgactt cgagtttgag cagatgttta ccgatgccct tggaattgac 5160
gagtacggtg ggacgcgtat gaagctactg tcttctatcg aacaagcatg cgatatttgc 5220
cgacttaaaa agctcaagtg ctccaaagaa aaaccgaagt gcgccaagtg tctgaagaac 5280
aactgggagt gtcgctactc tcccaaaacc aaaaggtctc cgctgactag ggcacatctg 5340
acagaagtgg aatcaaggct agaaagactg gaacagctat ttctactgat ttttcctcga 5400
gaagaccttg acatgatttt gaaaatggat tctttacagg atataaaagc attgttaaca 5460
ggattatttg tacaagataa tgtgaataaa gatgccgtca cagatagatt ggcttcagtg 5520
gagactgata tgcctctaac attgagacag catagaataa gtgcgacatc atcatcggaa 5580
gagagtagta acaaaggtca aagacagttg actgtataaa agcttcactc ctcaggtgca 5640
ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca aataccactg 5700
agatcttttt ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac 5760
ttctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct 5820
ctcactcgga aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt 5880
ttagagtttg gcaacatatg cccatatgct ggctgccatg aacaaaggtt ggctataaag 5940
aggtcatcag tatatgaaac agccccctgc tgtccattcc ttattccata gaaaagcctt 6000
gacttgaggt tagatttttt ttatattttg ttttgtgtta tttttttctt taacatccct 6060
aaaattttcc ttacatgttt tactagccag atttttcctc ctctcctgac tactcccagt 6120
catagctgtc cctcttctct tatggagatc 6150
<210> 8
<211> 12787
<212> DNA
<213> Artificial
<220>
<223> 编码由细胞特异性启动子Survivin1、第一microRNA为mir-143-3p、第二mi
croRNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白
分别为TALE蛋白、第一和第二识别序列为5×UAS的元件组成的表达系统的核
酸
<400> 8
cggagtactg tcctccgagc ggagtactgt cctccgagcg gagtactgtc ctccgagcgg 60
agtactgtcc tccgagcgga gttctgtcct ccgagcggag actctagata cccctctccg 120
cttctgaatt ctaggcgtgt acggtgggag gcctatataa gcagagctcg tttagtgaac 180
cgtcagatcg cctcgagtac ccctctccgc ttcttacccc tctccgcttc ttacccctct 240
ccgcttctga attcgaccgc caccatgaga catattatct gccacggagg tgttattacc 300
gaagaaatgg ccgccagtct tttggaccag ctgatcgaag aggtactggc tgataatctt 360
ccacctccta gccattttga accacctacc cttcacgaac tgtatgattt agacgtgacg 420
gcccccgaag atcccaacga ggaggcggtt tcgcagattt ttcccgactc tgtaatgttg 480
gcggtgcagg aagggattga cttactcact tttccgccgg cgcccggttc tccggagccg 540
cctcaccttt cccggcagcc cgagcagccg gagcagagag ccttgggtcc ggtttctatg 600
ccaaaccttg taccggaggt gatcgatctt acctgccacg aggctggctt tccacccagt 660
gacgacgagg atgaagaggg tgaggagttt gtgttagatt atgtggagca ccccgggcac 720
ggttgcaggt cttgtcatta tcaccggagg aatacggggg acccagatat tatgtgttcg 780
ctttgctata tgaggacctg tggcatgttt gtctacagta agtgaaaatt atgggcagtg 840
ggtgatagag tggtgggttt ggtgtggtaa tttttttttt aatttttaca gttttgtggt 900
ttaaagaatt ttgtattgtg atttttttaa aaggtcctgt gtctgaacct gagcctgagc 960
ccgagccaga accggagcct gcaagaccta cccgccgtcc taaaatggcg cctgctatcc 1020
tgaggcgccc gacatcacct gtgtctagag aatgcaatag tagtacggat agctgtgact 1080
ccggtccttc taacacacct cctgagatac acccggtggt cccgctgtgc cccattaaac 1140
cagttgccgt gagagttggt gggcgtcgcc aggctgtgga atgtatcgag gacttgctta 1200
acgagcctgg gcaacctttg gacttgagct gtaaacgccc caggccaggg tccggagcta 1260
caaatttttc cctcctcaaa caggctggag atgtcgaaga aaatcctggg cctatggagg 1320
cttgggagtg tttggaagat ttttctgctg tgcgtaactt gctggaacag agctctaaca 1380
gtacctcttg gttttggagg tttctgtggg gctcatccca ggcaaagtta gtctgcagaa 1440
ttaaggagga ttacaagtgg gaatttgaag agcttttgaa atcctgtggt gagctgtttg 1500
attctttgaa tctgggtcac caggcgcttt tccaagagaa ggtcatcaag actttggatt 1560
tttccacacc ggggcgcgct gcggctgctg ttgctttttt gagttttata aaggataaat 1620
ggagcgaaga aacccatctg agcggggggt acctgctgga ttttctggcc atgcatctgt 1680
ggagagcggt tgtgagacac aagaatcgcc tgctactgtt gtcttccgtc cgcccggcga 1740
taataccgac ggaggagcag cagcagcagc aggaggaagc caggcggcgg cggcaggagc 1800
agagcccatg gaacccgaga gccggcctgg accctcggga atgaatgttg tacaggtggc 1860
tgaactgtat ccagaactga gacgcatttt gacaattaca gaggatgggc aggggctaaa 1920
gggggtaaag agggagcggg gggcttgtga ggctacagag gaggctagga atctagcttt 1980
tagcttaatg accagacacc gtcctgagtg tattactttt caacagatca aggataattg 2040
cgctaatgag cttgatctgc tggcgcagaa gtattccata gagcagctga ccacttactg 2100
gctgcagcca ggggatgatt ttgaggaggc tattagggta tatgcaaagg tggcacttag 2160
gccagattgc aagtacaaga tcagcaaact tgtaaatatc aggaattgtt gctacatttc 2220
tgggaacggg gccgaggtgg agatagatac ggaggatagg gtggccttta gatgtagcat 2280
gataaatatg tggccggggg tgcttggcat ggacggggtg gttattatga atgtaaggtt 2340
tactggcccc aattttagcg gtacggtttt cctggccaat accaacctta tcctacacgg 2400
tgtaagcttc tatgggttta acaatacctg tgtggaagcc tggaccgatg taagggttcg 2460
gggctgtgcc ttttactgct gctggaaggg ggtggtgtgt cgccccaaaa gcagggcttc 2520
aattaagaaa tgcctctttg aaaggtgtac cttgggtatc ctgtctgagg gtaactccag 2580
ggtgcgccac aatgtggcct ccgactgtgg ttgcttcatg ctagtgaaaa gcgtggctgt 2640
gattaagcat aacatggtat gtggcaactg cgaggacagg gcctctcaga tgctgacctg 2700
ctcggacggc aactgtcacc tgctgaagac cattcacgta gccagccact ctcgcaaggc 2760
ctggccagtg tttgagcata acatactgac ccgctgttcc ttgcatttgg gtaacaggag 2820
gggggtgttc ctaccttacc aatgcaattt gagtcacact aagatattgc ttgagcccga 2880
gagcatgtcc aaggtgaacc tgaacggggt gtttgacatg accatgaaga tctggaaggt 2940
gctgaggtac gatgagaccc gcaccaggtg cagaccctgc gagtgtggcg gtaaacatat 3000
taggaaccag cctgtgatgc tggatgtgac cgaggagctg aggcccgatc acttggtgct 3060
ggcctgcacc cgcgctgagt ttggctctag cgatgaagat acagatctcg aggggtccgg 3120
agctacaaat ttttccctcc tcaaacaggc tggagatgtc gaagaaaatc ctgggcctac 3180
cggtatgtat aggatgcagt tgctctcttg catcgccctg tctctagcgt tggtgactaa 3240
tagcgcccct acttcttctt ccacgaagaa aacccagctg caactggaac accttctgct 3300
ggatctccag atgatcctta acgggattaa caactataag aacccgaagc tcacggctat 3360
gctgacggcg aagttcgcta tgcctaaaaa ggctactgag ctgaaacact tacagtgtct 3420
cgaagaggca ctcaaacccc tcgaggaggt actcaatctg gcgcaaagca aaaacttcca 3480
tctccgccca cgggacctaa tctccaacat caacgtgatt gtgctggagc tcaaggggtc 3540
tgaaaccacg tttatgtgcg agtacgcaga tgaaactgct accatagtgg aattcctcaa 3600
tcgttggatc actttctgcc agtcaatcat ttcaaccctg accagatctg agggccgcgg 3660
cagcctgctg acctgcggcg acgtggagga aaaccccggc cccggatcct tgtcgaccat 3720
tcgtccgcgc aggccaagtc ctgcccgcga gcttctgccc ggaccccaac cggatagggt 3780
tcagccgact gcagatcgtg gggtgtctgc gcctgctggc agccctctgg atggcttgcc 3840
cgctcggcgg acggtgtccc ggacccggct gccatctccc cctgcgccct cacctgcgtt 3900
ctcggcgggc agcttcagcg atctgctccg tccgttcgat ccgtcgcttc ttgatacatc 3960
gcttcttgat tcgatgcctg ccgtcggcac gccgcataca gcggctgccc cagcagagtg 4020
ggatgaggcg caatcggctc tgcgtgcagc cgatgacccg ccacccaccg tgcgtgtcgc 4080
tgtcactgcc gcgcggccgc cgcgcgccaa gccggccccg cgacggcgtg ctgcgcaacc 4140
ctccgacgct tcgccggccg cgcaggtgga tctacgcacg ctcggctaca gtcagcagca 4200
gcaagagaag atcaaaccga aggtgcgttc gacagtggcg cagcaccacg aggcactggt 4260
gggccatggg tttacacacg cgcacatcgt tgcgctcagc caacacccgg cagcgttagg 4320
gaccgtcgct gtcacgtatc agcacataat cacggcgttg ccagaggcga cacacgaaga 4380
catcgttggc gtcggcaaac agtggtccgg cgcacgcgcc ctggaggcct tgctcacgga 4440
tgcgggggag ttgagaggtc cgccgttaca gttggacaca ggccaacttg tgaagattgc 4500
aaaacgtggc ggcgtgaccg caatggaggc agtgcatgca tcgcgcaatg cactgacggg 4560
tgcccccctg aacctgaccc cggaccaagt ggtggctatc gccagcaaca ttggcggcaa 4620
gcaagcgctc gaaacggtgc agcggctgtt gccggtgctg tgccaggacc atggcctgac 4680
cccggaccaa gtggtggcta tcgccagcaa cggtggcggc aagcaagcgc tcgaaacggt 4740
gcagcggctg ttgccggtgc tgtgccagga ccatggcctg accccggacc aagtggtggc 4800
tatcgccagc aacattggcg gcaagcaagc gctcgaaacg gtgcagcggc tgttgccggt 4860
gctgtgccag gaccatggcc tgaccccgga ccaagtggtg gctatcgcca gcaacaatgg 4920
cggcaagcaa gcgctcgaaa cggtgcagcg gctgttgccg gtgctgtgcc aggaccatgg 4980
cctgaccccg gaccaagtgg tggctatcgc cagcaacatt ggcggcaagc aagcgctcga 5040
aacggtgcag cggctgttgc cggtgctgtg ccaggaccat ggcctgaccc cggaccaagt 5100
ggtggctatc gccagcaaca ttggcggcaa gcaagcgctc gaaacggtgc agcggctgtt 5160
gccggtgctg tgccaggacc atggcctgac tccggaccaa gtggtggcta tcgccagcca 5220
cgatggcggc aagcaagcgc tcgaaacggt gcagcggctg ttgccggtgc tgtgccagga 5280
ccatggcctg actccggacc aagtggtggc tatcgccagc cacgatggcg gcaagcaagc 5340
gctcgaaacg gtgcagcggc tgttgccggt gctgtgccag gaccatggcc tgaccccgga 5400
ccaagtggtg gctatcgcca gcaacaatgg cggcaagcaa gcgctcgaaa cggtgcagcg 5460
gctgttgccg gtgctgtgcc aggaccatgg cctgaccccg gaccaagtgg tggctatcgc 5520
cagcaacatt ggcggcaagc aagcgctcga aacggtgcag cggctgttgc cggtgctgtg 5580
ccaggaccat ggcctgaccc cggaccaagt ggtggctatc gccagcaacg gtggcggcaa 5640
gcaagcgctc gaaacggtgc agcggctgtt gccggtgctg tgccaggacc atggcctgac 5700
tccggaccaa gtggtggcta tcgccagcca cgatggcggc aagcaagcgc tcgaaacggt 5760
gcagcggctg ttgccggtgc tgtgccagga ccatggcctg actccggacc aagtggtggc 5820
tatcgccagc cacgatggcg gcaagcaagc gctcgaaacg gtgcagcggc tgttgccggt 5880
gctgtgccag gaccatggcc tgaccccgga ccaagtggtg gctatcgcca gcaacggtgg 5940
cggcaagcaa gcgctcgaaa cggtgcagcg gctgttgccg gtgctgtgcc aggaccatgg 6000
cctgactccg gaccaagtgg tggctatcgc cagccacgat ggcggcaagc aagcgctcga 6060
aacggtgcag cggctgttgc cggtgctgtg ccaggaccat ggcctgactc cggaccaagt 6120
ggtggctatc gccagccacg atggcggcaa gcaagcgctc gaaacggtgc agcggctgtt 6180
gccggtgctg tgccaggacc atggcctgac tccggaccaa gtggtggcta tcgccagcca 6240
cgatggcggc aagcaagcgc tcgaaacggt gcagcggctg ttgccggtgc tgtgccagga 6300
ccatggcctg accccggacc aagtggtggc tatcgccagc aacattggcg gcaagcaagc 6360
gctcgaaacg gtgcagcggc tgttgccggt gctgtgccag gaccatggcc tgaccccgga 6420
ccaagtggtg gctatcgcca gcaacggtgg cggcaagcaa gcgctcgaaa cggtgcagcg 6480
gctgttgccg gtgctgtgcc aggaccatgg cctgaccccg gaccaagtgg tggctatcgc 6540
cagcaacggt ggcggcaagc aagcgctcga aagcattgtg gcccagctga gccggcctga 6600
tccggcgttg gccgcgttga ccaacgacca cctcgtcgcc ttggcctgcc tcggcggacg 6660
tcctgccatg gatgcagtga aaaagggatt gccgcacgcg ccggaattga tcagaagagt 6720
caatcgccgt attggcgaac gcacgtccca tcgcgttgcc gactacgcgc aagtggttcg 6780
cgtgctggag tttttccagt gccactccca cccagcgtac gcatttgatg aggccatgac 6840
gcagttcggg atgagcagga acgggttggt acagctcttt cgcagagtgg gcgtcaccga 6900
actcgaagcc cgcggtggaa cgctcccccc agcctcgcag cgttgggacc gtatcctcca 6960
ggcatcaggg atgaaaaggg ccaaaccgtc ccctacttca gctcaaacac cggatcaggc 7020
gtctttgcat gcattcgccg attcgctgga gcgtgacctt gatgcgccca gcccaatgca 7080
cgagggagat cagacgcggg caagcagccg taaacggtcc cgatcggatc gtgctgtcac 7140
cggcccctcc gcacagcagg ctgtcgaggt gcgcgttccc gaacagcgcg atgcgctgca 7200
tttgcccctc agctggaggg taaaacgccc gcgtaccagg atctggggcg gcctcccggt 7260
taagtagtgc tttctacttt atgagtagtg ctttctactt tatgagtagt gctttctact 7320
ttatgagtag tgctttctac tttatggagc tacagtgctt catctcagag ctacagtgct 7380
tcatctcaga gctacagtgc ttcatctcag agctacagtg cttcatctca gaaagcggag 7440
tactgtcctc cgagcggagt actgtcctcc gagcggagta ctgtcctccg agcggagtac 7500
tgtcctccga gcggagttct gtcctccgag cggagactct agatatagaa ccgatcctcc 7560
cattgaattc taggcgtgta cggtgggagg cctatataag cagagctcgt ttagtgaacc 7620
gtcagatcgc ctcgagtata gaaccgatcc tcccatttat agaaccgatc ctcccattta 7680
tagaaccgat cctcccatta ctagaagatc ttgaattcgc caccttgtcg accattcgtc 7740
cgcgcaggcc aagtcctgcc cgcgagcttc tgcccggacc ccaaccggat agggttcagc 7800
cgactgcaga tcgtggggtg tctgcgcctg ctggcagccc tctggatggc ttgcccgctc 7860
ggcggacggt gtcccggacc cggctgccat ctccccctgc gccctcacct gcgttctcgg 7920
cgggcagctt cagcgatctg ctccgtccgt tcgatccgtc gcttcttgat acatcgcttc 7980
ttgattcgat gcctgccgtc ggcacgccgc atacagcggc tgccccagca gagtgggatg 8040
aggcgcaatc ggctctgcgt gcagccgatg acccgccacc caccgtgcgt gtcgctgtca 8100
ctgccgcgcg gccgccgcgc gccaagccgg ccccgcgacg gcgtgctgcg caaccctccg 8160
acgcttcgcc ggccgcgcag gtggatctac gcacgctcgg ctacagtcag cagcagcaag 8220
agaagatcaa accgaaggtg cgttcgacag tggcgcagca ccacgaggca ctggtgggcc 8280
atgggtttac acacgcgcac atcgttgcgc tcagccaaca cccggcagcg ttagggaccg 8340
tcgctgtcac gtatcagcac ataatcacgg cgttgccaga ggcgacacac gaagacatcg 8400
ttggcgtcgg caaacagtgg tccggcgcac gcgccctgga ggccttgctc acggatgcgg 8460
gggagttgag aggtccgccg ttacagttgg acacaggcca acttgtgaag attgcaaaac 8520
gtggcggcgt gaccgcaatg gaggcagtgc atgcatcgcg caatgcactg acgggtgccc 8580
ccctgaacct gaccccggac caagtggtgg ctatcgccag caacattggc ggcaagcaag 8640
cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc ctgactccgg 8700
accaagtggt ggctatcgcc agccacgatg gcggcaagca agcgctcgaa acggtgcagc 8760
ggctgttgcc ggtgctgtgc caggaccatg gcctgactcc ggaccaagtg gtggctatcg 8820
ccagccacga tggcggcaag caagcgctcg aaacggtgca gcggctgttg ccggtgctgt 8880
gccaggacca tggcctgact ccggaccaag tggtggctat cgccagccac gatggcggca 8940
agcaagcgct cgaaacggtg cagcggctgt tgccggtgct gtgccaggac catggcctga 9000
ctccggacca agtggtggct atcgccagcc acgatggcgg caagcaagcg ctcgaaacgg 9060
tgcagcggct gttgccggtg ctgtgccagg accatggcct gaccccggac caagtggtgg 9120
ctatcgccag caacggtggc ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg 9180
tgctgtgcca ggaccatggc ctgactccgg accaagtggt ggctatcgcc agccacgatg 9240
gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg 9300
gcctgacccc ggaccaagtg gtggctatcg ccagcaacgg tggcggcaag caagcgctcg 9360
aaacggtgca gcggctgttg ccggtgctgt gccaggacca tggcctgact ccggaccaag 9420
tggtggctat cgccagccac gatggcggca agcaagcgct cgaaacggtg cagcggctgt 9480
tgccggtgct gtgccaggac catggcctga ctccggacca agtggtggct atcgccagcc 9540
acgatggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg 9600
accatggcct gaccccggac caagtggtgg ctatcgccag caacaatggc ggcaagcaag 9660
cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc ctgactccgg 9720
accaagtggt ggctatcgcc agccacgatg gcggcaagca agcgctcgaa acggtgcagc 9780
ggctgttgcc ggtgctgtgc caggaccatg gcctgacccc ggaccaagtg gtggctatcg 9840
ccagcaacgg tggcggcaag caagcgctcg aaacggtgca gcggctgttg ccggtgctgt 9900
gccaggacca tggcctgacc ccggaccaag tggtggctat cgccagcaac ggtggcggca 9960
agcaagcgct cgaaacggtg cagcggctgt tgccggtgct gtgccaggac catggcctga 10020
ctccggacca agtggtggct atcgccagcc acgatggcgg caagcaagcg ctcgaaacgg 10080
tgcagcggct gttgccggtg ctgtgccagg accatggcct gaccccggac caagtggtgg 10140
ctatcgccag caacggtggc ggcaagcaag cgctcgaaag cattgtggcc cagctgagcc 10200
ggcctgatcc ggcgttggcc gcgttgacca acgaccacct cgtcgccttg gcctgcctcg 10260
gcggacgtcc tgccatggat gcagtgaaaa agggattgcc gcacgcgccg gaattgatca 10320
gaagagtcaa tcgccgtatt ggcgaacgca cgtcccatcg cgttgccgac tacgcgcaag 10380
tggttcgcgt gctggagttt ttccagtgcc actcccaccc agcgtacgca tttgatgagg 10440
ccatgacgca gttcgggatg agcaggaacg ggttggtaca gctctttcgc agagtgggcg 10500
tcaccgaact cgaagcccgc ggtggaacgc tccccccagc ctcgcagcgt tgggaccgta 10560
tcctccaggc atcagggatg aaaagggcca aaccgtcccc tacttcagct caaacaccgg 10620
atcaggcgtc tttgcatgca ttcgccgatt cgctggagcg tgaccttgat gcgcccagcc 10680
caatgcacga gggagatcag acgcgggcaa gcagccgtaa acggtcccga tcggatcgtg 10740
ctgtcaccgg cccctccgca cagcaggctg tcgaggtgcg cgttcccgaa cagcgcgatg 10800
cgctgcattt gcccctcagc tggagggtaa aacgcccgcg taccaggatc tggggcggcc 10860
tcccgtaata caaggcggcc gcaaatcaac atcagtctga taagctatca acatcagtct 10920
gataagctat caacatcagt ctgataagct atcaacatca gtctgataag ctaaagcttc 10980
gaattctgat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc 11040
cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta 11100
ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat 11160
ttttttcact gccatgttag gtgtgggcag ggacgagctg gcgcggcgtc gctgggtgca 11220
ccgcgaccac gggcagagcc acgcggcggg aggactacaa ctcccggcac accccgcgcc 11280
gccccgcctc tactcccaga aggccgcggg gggtggaccg cctaagaggg cgtgcgctcc 11340
cgacatgccc cgcggcgcgc cattaaccgc cagatttgaa tcgcgggacc cgttggcaga 11400
ggtggcggcg gcggcatggg tgccccgacg ttgccccctg cctggctcga gaccgatggc 11460
ccccccgacc gatgtcagcc tgggggacga gctccactta gacggcgagg acgtggcgat 11520
ggcgcatgcc gacgcgctag acgatttcga tctggacatg ttggggagcg acgcgctaga 11580
cgatttcgat ctggacatgt tggggagcga cgcgctagac gatttcgatc tggacatgtt 11640
ggggagcgac gcgctagacg atttcgatct ggacatgttg ggggacgggg attccccggg 11700
tccgggattt accccccacg actccgcccc ctacggcgct ctggatatgg ccgacttcga 11760
gtttgagcag atgtttaccg atgcccttgg aattgacgag tacggtggga cgcgtatgaa 11820
gctactgtct tctatcgaac aagcatgcga tatttgccga cttaaaaagc tcaagtgctc 11880
caaagaaaaa ccgaagtgcg ccaagtgtct gaagaacaac tgggagtgtc gctactctcc 11940
caaaaccaaa aggtctccgc tgactagggc acatctgaca gaagtggaat caaggctaga 12000
aagactggaa cagctatttc tactgatttt tcctcgagaa gaccttgaca tgattttgaa 12060
aatggattct ttacaggata taaaagcatt gttaacagga ttatttgtac aagataatgt 12120
gaataaagat gccgtcacag atagattggc ttcagtggag actgatatgc ctctaacatt 12180
gagacagcat agaataagtg cgacatcatc atcggaagag agtagtaaca aaggtcaaag 12240
acagttgact gtataaaagc ttcactcctc aggtgcaggc tgcctatcag aaggtggtgg 12300
ctggtgtggc caatgccctg gctcacaaat accactgaga tctttttccc tctgccaaaa 12360
attatgggga catcatgaag ccccttgagc atctgacttc tggctaataa aggaaattta 12420
ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga 12480
gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca acatatgccc 12540
atatgctggc tgccatgaac aaaggttggc tataaagagg tcatcagtat atgaaacagc 12600
cccctgctgt ccattcctta ttccatagaa aagccttgac ttgaggttag atttttttta 12660
tattttgttt tgtgttattt ttttctttaa catccctaaa attttcctta catgttttac 12720
tagccagatt tttcctcctc tcctgactac tcccagtcat agctgtccct cttctcttat 12780
ggagatc 12787
<210> 9
<211> 6242
<212> DNA
<213> Artificial
<220>
<223> 编码由细胞特异性启动子CEA368、第一microRNA为mir-143-3p、第二micro
RNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白分
别为Lacl、tetR-KRAB、第一和第二识别序列为5×UAS的元件组成的表达系
统的核酸
<400> 9
cggggtgccg ccccccgcgg gtgaccgccc tccgcgggtg acagccctcc gcggagtgcc 60
gcactccgcg gagtgctgta ctccgagcgg agactctaga ctccctatca gtgatagaga 120
taggcgtgta cggtgggagg cctatataag cagagctcgt ttagtgaacc gtcagatcgc 180
tccctatcag tgatagagag aattcgaccg ccaccatgag acatattatc tgccacggag 240
gtgttattac cgaagaaatg gccgccagtc ttttggacca gctgatcgaa gaggtactgg 300
ctgataatct tccacctcct agccattttg aaccacctac ccttcacgaa ctgtatgatt 360
tagacgtgac ggcccccgaa gatcccaacg aggaggcggt ttcgcagatt tttcccgact 420
ctgtaatgtt ggcggtgcag gaagggattg acttactcac ttttccgccg gcgcccggtt 480
ctccggagcc gcctcacctt tcccggcagc ccgagcagcc ggagcagaga gccttgggtc 540
cggtttctat gccaaacctt gtaccggagg tgatcgatct tacctgccac gaggctggct 600
ttccacccag tgacgacgag gatgaagagg gtgaggagtt tgtgttagat tatgtggagc 660
accccgggca cggttgcagg tcttgtcatt atcaccggag gaatacgggg gacccagata 720
ttatgtgttc gctttgctat atgaggacct gtggcatgtt tgtctacagt aagtgaaaat 780
tatgggcagt gggtgataga gtggtgggtt tggtgtggta attttttttt taatttttac 840
agttttgtgg tttaaagaat tttgtattgt gattttttta aaaggtcctg tgtctgaacc 900
tgagcctgag cccgagccag aaccggagcc tgcaagacct acccgccgtc ctaaaatggc 960
gcctgctatc ctgaggcgcc cgacatcacc tgtgtctaga gaatgcaata gtagtacgga 1020
tagctgtgac tccggtcctt ctaacacacc tcctgagata cacccggtgg tcccgctgtg 1080
ccccattaaa ccagttgccg tgagagttgg tgggcgtcgc caggctgtgg aatgtatcga 1140
ggacttgctt aacgagcctg ggcaaccttt ggacttgagc tgtaaacgcc ccaggccact 1200
cgagggtacc gggtccggag ctacaaattt ttccctcctc aaacaggctg gagatgtcga 1260
agaaaatcct gggcctaccg gtatgtggct gcagagcctc cttctactgg gcaccgtggc 1320
ttgcagcatc tcggcccccg ccagaagtcc cagccccagc acccagccct gggaacacgt 1380
aaatgcaatc caggaggccc gccgcctcct gaacttgtca cgggacacgg cagccgagat 1440
gaatgaaact gtggaggtca tcagtgaaat gtttgacttg caggagccca catgcctgca 1500
gactcggctg gagctctaca aacagggcct gcgtggcagc ctcaccaagc tgaagggccc 1560
actgaccatg atggctagcc attacaagca gcactgcccg ccgacccctg agacatcctg 1620
tgccactcag atcattacct ttgagtcttt caaggagaac ctcaaggact ttctgctggt 1680
catcccattt gactgctggg aacccgtcca ggaaagatct gagggccgcg gcagcctgct 1740
gacctgcggc gacgtggagg aaaaccccgg ccccggatcc atgaaaccag taacgttata 1800
cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc 1860
cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta 1920
cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga ttggcgttgc 1980
cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc 2040
cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg 2100
taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca ttaactatcc 2160
gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc cggcgttatt 2220
tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg aagacggtac 2280
gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg 2340
cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat atctcactcg 2400
caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt ccggttttca 2460
acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg ttgccaacga 2520
tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga 2580
tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc cgccgttaac 2640
caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact 2700
ctctcagggc caggcggtga agggcaatca gctgttgccg gtctcactgg tgaaaagaaa 2760
aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat 2820
gcagctggca cgacaggttt cccgactgga aagcgggcag agcagcctga ggcctcctaa 2880
gaagaagagg aaggttgtta caaacaccat tgtcacactc cacaaacacc attgtcacac 2940
tccacaaaca ccattgtcac actccacaaa caccattgtc acactccaga gctacagtgc 3000
ttcatctcag agctacagtg cttcatctca gagctacagt gcttcatctc agagctacag 3060
tgcttcatct cagaaagcgg actactgtag tccgcggagt actgtcctcc gcggacgact 3120
gtcgtccgcg gagcactgtg ctccgcggat gactgtcatc cgagcggaga ctctagaaat 3180
tgtgagcgga taacaattgg gtatataatg ggggccgtca gctactacca gagccgatcg 3240
aacactcgag ccgagcagac gtgcctacaa ttgtgagcgc tcacaattac tagaagatct 3300
tgaattcgcc accatggcta gattagataa aagtaaagtg attaacagcg cattagagct 3360
gcttaatgag gtcggaatcg aaggtttaac aacccgtaaa ctcgcccaga agctaggtgt 3420
agagcagcct acattgtatt ggcatgtaaa aaataagcgg gctttgctcg acgccttagc 3480
cattgagatg ttagataggc accatactca cttttgccct ttagaagggg aaagctggca 3540
agatttttta cgtaataacg ctaaaagttt tagatgtgct ttactaagtc atcgcgatgg 3600
agcaaaagta catttaggta cacggcctac agaaaaacag tatgaaactc tcgaaaatca 3660
attagccttt ttatgccaac aaggtttttc actagagaat gcattatatg cactcagcgc 3720
tgtggggcat tttactttag gttgcgtatt ggaagatcaa gagcatcaag tcgctaaaga 3780
agaaagggaa acacctacta ctgatagtat gccgccatta ttacgacaag ctatcgaatt 3840
atttgatcac caaggtgcag agccagcctt cttattcggc cttgaattga tcatatgcgg 3900
attagaaaaa caacttaaat gtgaaagtgg gtcgccaaaa aagaagagaa aggtggatgg 3960
tggcggagca ctgtctcccc agcattcagc tgtgacccag ggatcaatca ttaagaacaa 4020
ggaaggtatg gatgctaagt ccctgaccgc ttggtcaaga actttggtaa cttttaagga 4080
cgtgttcgtc gattttactc gggaggagtg gaaactattg gataccgccc agcaaatcgt 4140
gtatagaaac gttatgctcg agaactataa gaatcttgtg tcactgggct accaactgac 4200
aaagcctgac gttatcctga ggctggaaaa gggtgaggag ccttggttgg tcgagcggga 4260
gatccaccag gaaactcacc cagacagtga gactgctttc gagatcaagt cctctgtgta 4320
atacaaggcg gccgcaaatc aacatcagtc tgataagcta tcaacatcag tctgataagc 4380
tatcaacatc agtctgataa gctatcaaca tcagtctgat aagctaaagc ttcgaattct 4440
gataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct 4500
ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc 4560
ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc 4620
actgcagctc gccttttgct cacatgtctc ccaggggatg ggggtccatc caccttgccg 4680
aaaagatttg tctgaggaac tgaaaataga agggaaaaaa gaggagggac aaaagaggca 4740
gaaatgagag gggaggggac agaggacacc tgaataaaga ccacacccat gacccacgtg 4800
atgctgagaa gtactcctgc cctaggaaga gactcagggc agagggagga aggacagcag 4860
accagacagt cacagcagcc ttgacaaaac gttcctggaa ctcaagctct tctccacaga 4920
ggaggacaga gcagacagca gagaccatgg agtctccctc ggcccctccc cacagatggt 4980
gcatcccctg gcagaggctc ctgctcacag gtgaactcga gaccggtatg gcccccccga 5040
ccgatgtcag cctgggggac gagctccact tagacggcga ggacgtggcg atggcgcatg 5100
ccgacgcgct agacgatttc gatctggaca tgttggggga cggggattcc ccgggtccgg 5160
gatttacccc ccacgactcc gccccctacg gcgctctgga tatggccgac ttcgagtttg 5220
agcagatgtt taccgatgcc cttggaattg acgagtacgg tgggacgcgt atgaagctac 5280
tgtcttctat cgaacaagca tgcgatattt gccgacttaa aaagctcaag tgctccaaag 5340
aaaaaccgaa gtgcgccaag tgtctgaaga acaactggga gtgtcgctac tctcccaaaa 5400
ccaaaaggtc tccgctgact agggcacatc tgacagaagt ggaatcaagg ctagaaagac 5460
tggaacagct atttctactg atttttcctc gagaagacct tgacatgatt ttgaaaatgg 5520
attctttaca ggatataaaa gcattgttaa caggattatt tgtacaagat aatgtgaata 5580
aagatgccgt cacagataga ttggcttcag tggagactga tatgcctcta acattgagac 5640
agcatagaat aagtgcgaca tcatcatcgg aagagagtag taacaaaggt caaagacagt 5700
tgactgtata aaagcttcac tcctcaggtg caggctgcct atcagaaggt ggtggctggt 5760
gtggccaatg ccctggctca caaataccac tgagatcttt ttccctctgc caaaaattat 5820
ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc 5880
attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca 5940
aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata tgcccatatg 6000
ctggctgcca tgaacaaagg ttggctataa agaggtcatc agtatatgaa acagccccct 6060
gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt ttttatattt 6120
tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt tttactagcc 6180
agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct cttatggaga 6240
tc 6242
<210> 10
<211> 12914
<212> DNA
<213> Artificial
<220>
<223> 编码由细胞特异性启动子CEA368、第一microRNA为mir-135a-5p、第二micr
oRNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白分
别为TALE蛋白、第一和第二识别序列为5×UAS的元件组成的表达系统的核酸
<400> 10
cggagtactg tcctccgagc ggagtactgt cctccgagcg gagtactgtc ctccgagcgg 60
agtactgtcc tccgagcgga gttctgtcct ccgagcggag actctagata cccctctccg 120
cttctgaatt ctaggcgtgt acggtgggag gcctatataa gcagagctcg tttagtgaac 180
cgtcagatcg cctcgagtac ccctctccgc ttcttacccc tctccgcttc ttacccctct 240
ccgcttctga attcgaccgc caccatgaga catattatct gccacggagg tgttattacc 300
gaagaaatgg ccgccagtct tttggaccag ctgatcgaag aggtactggc tgataatctt 360
ccacctccta gccattttga accacctacc cttcacgaac tgtatgattt agacgtgacg 420
gcccccgaag atcccaacga ggaggcggtt tcgcagattt ttcccgactc tgtaatgttg 480
gcggtgcagg aagggattga cttactcact tttccgccgg cgcccggttc tccggagccg 540
cctcaccttt cccggcagcc cgagcagccg gagcagagag ccttgggtcc ggtttctatg 600
ccaaaccttg taccggaggt gatcgatctt acctgccacg aggctggctt tccacccagt 660
gacgacgagg atgaagaggg tgaggagttt gtgttagatt atgtggagca ccccgggcac 720
ggttgcaggt cttgtcatta tcaccggagg aatacggggg acccagatat tatgtgttcg 780
ctttgctata tgaggacctg tggcatgttt gtctacagta agtgaaaatt atgggcagtg 840
ggtgatagag tggtgggttt ggtgtggtaa tttttttttt aatttttaca gttttgtggt 900
ttaaagaatt ttgtattgtg atttttttaa aaggtcctgt gtctgaacct gagcctgagc 960
ccgagccaga accggagcct gcaagaccta cccgccgtcc taaaatggcg cctgctatcc 1020
tgaggcgccc gacatcacct gtgtctagag aatgcaatag tagtacggat agctgtgact 1080
ccggtccttc taacacacct cctgagatac acccggtggt cccgctgtgc cccattaaac 1140
cagttgccgt gagagttggt gggcgtcgcc aggctgtgga atgtatcgag gacttgctta 1200
acgagcctgg gcaacctttg gacttgagct gtaaacgccc caggccaggg tccggagcta 1260
caaatttttc cctcctcaaa caggctggag atgtcgaaga aaatcctggg cctatggagg 1320
cttgggagtg tttggaagat ttttctgctg tgcgtaactt gctggaacag agctctaaca 1380
gtacctcttg gttttggagg tttctgtggg gctcatccca ggcaaagtta gtctgcagaa 1440
ttaaggagga ttacaagtgg gaatttgaag agcttttgaa atcctgtggt gagctgtttg 1500
attctttgaa tctgggtcac caggcgcttt tccaagagaa ggtcatcaag actttggatt 1560
tttccacacc ggggcgcgct gcggctgctg ttgctttttt gagttttata aaggataaat 1620
ggagcgaaga aacccatctg agcggggggt acctgctgga ttttctggcc atgcatctgt 1680
ggagagcggt tgtgagacac aagaatcgcc tgctactgtt gtcttccgtc cgcccggcga 1740
taataccgac ggaggagcag cagcagcagc aggaggaagc caggcggcgg cggcaggagc 1800
agagcccatg gaacccgaga gccggcctgg accctcggga atgaatgttg tacaggtggc 1860
tgaactgtat ccagaactga gacgcatttt gacaattaca gaggatgggc aggggctaaa 1920
gggggtaaag agggagcggg gggcttgtga ggctacagag gaggctagga atctagcttt 1980
tagcttaatg accagacacc gtcctgagtg tattactttt caacagatca aggataattg 2040
cgctaatgag cttgatctgc tggcgcagaa gtattccata gagcagctga ccacttactg 2100
gctgcagcca ggggatgatt ttgaggaggc tattagggta tatgcaaagg tggcacttag 2160
gccagattgc aagtacaaga tcagcaaact tgtaaatatc aggaattgtt gctacatttc 2220
tgggaacggg gccgaggtgg agatagatac ggaggatagg gtggccttta gatgtagcat 2280
gataaatatg tggccggggg tgcttggcat ggacggggtg gttattatga atgtaaggtt 2340
tactggcccc aattttagcg gtacggtttt cctggccaat accaacctta tcctacacgg 2400
tgtaagcttc tatgggttta acaatacctg tgtggaagcc tggaccgatg taagggttcg 2460
gggctgtgcc ttttactgct gctggaaggg ggtggtgtgt cgccccaaaa gcagggcttc 2520
aattaagaaa tgcctctttg aaaggtgtac cttgggtatc ctgtctgagg gtaactccag 2580
ggtgcgccac aatgtggcct ccgactgtgg ttgcttcatg ctagtgaaaa gcgtggctgt 2640
gattaagcat aacatggtat gtggcaactg cgaggacagg gcctctcaga tgctgacctg 2700
ctcggacggc aactgtcacc tgctgaagac cattcacgta gccagccact ctcgcaaggc 2760
ctggccagtg tttgagcata acatactgac ccgctgttcc ttgcatttgg gtaacaggag 2820
gggggtgttc ctaccttacc aatgcaattt gagtcacact aagatattgc ttgagcccga 2880
gagcatgtcc aaggtgaacc tgaacggggt gtttgacatg accatgaaga tctggaaggt 2940
gctgaggtac gatgagaccc gcaccaggtg cagaccctgc gagtgtggcg gtaaacatat 3000
taggaaccag cctgtgatgc tggatgtgac cgaggagctg aggcccgatc acttggtgct 3060
ggcctgcacc cgcgctgagt ttggctctag cgatgaagat acagatctcg aggggtccgg 3120
agctacaaat ttttccctcc tcaaacaggc tggagatgtc gaagaaaatc ctgggcctac 3180
cggtatgtat aggatgcagt tgctctcttg catcgccctg tctctagcgt tggtgactaa 3240
tagcgcccct acttcttctt ccacgaagaa aacccagctg caactggaac accttctgct 3300
ggatctccag atgatcctta acgggattaa caactataag aacccgaagc tcacggctat 3360
gctgacggcg aagttcgcta tgcctaaaaa ggctactgag ctgaaacact tacagtgtct 3420
cgaagaggca ctcaaacccc tcgaggaggt actcaatctg gcgcaaagca aaaacttcca 3480
tctccgccca cgggacctaa tctccaacat caacgtgatt gtgctggagc tcaaggggtc 3540
tgaaaccacg tttatgtgcg agtacgcaga tgaaactgct accatagtgg aattcctcaa 3600
tcgttggatc actttctgcc agtcaatcat ttcaaccctg accagatctg agggccgcgg 3660
cagcctgctg acctgcggcg acgtggagga aaaccccggc cccggatcct tgtcgaccat 3720
tcgtccgcgc aggccaagtc ctgcccgcga gcttctgccc ggaccccaac cggatagggt 3780
tcagccgact gcagatcgtg gggtgtctgc gcctgctggc agccctctgg atggcttgcc 3840
cgctcggcgg acggtgtccc ggacccggct gccatctccc cctgcgccct cacctgcgtt 3900
ctcggcgggc agcttcagcg atctgctccg tccgttcgat ccgtcgcttc ttgatacatc 3960
gcttcttgat tcgatgcctg ccgtcggcac gccgcataca gcggctgccc cagcagagtg 4020
ggatgaggcg caatcggctc tgcgtgcagc cgatgacccg ccacccaccg tgcgtgtcgc 4080
tgtcactgcc gcgcggccgc cgcgcgccaa gccggccccg cgacggcgtg ctgcgcaacc 4140
ctccgacgct tcgccggccg cgcaggtgga tctacgcacg ctcggctaca gtcagcagca 4200
gcaagagaag atcaaaccga aggtgcgttc gacagtggcg cagcaccacg aggcactggt 4260
gggccatggg tttacacacg cgcacatcgt tgcgctcagc caacacccgg cagcgttagg 4320
gaccgtcgct gtcacgtatc agcacataat cacggcgttg ccagaggcga cacacgaaga 4380
catcgttggc gtcggcaaac agtggtccgg cgcacgcgcc ctggaggcct tgctcacgga 4440
tgcgggggag ttgagaggtc cgccgttaca gttggacaca ggccaacttg tgaagattgc 4500
aaaacgtggc ggcgtgaccg caatggaggc agtgcatgca tcgcgcaatg cactgacggg 4560
tgcccccctg aacctgaccc cggaccaagt ggtggctatc gccagcaaca ttggcggcaa 4620
gcaagcgctc gaaacggtgc agcggctgtt gccggtgctg tgccaggacc atggcctgac 4680
cccggaccaa gtggtggcta tcgccagcaa cggtggcggc aagcaagcgc tcgaaacggt 4740
gcagcggctg ttgccggtgc tgtgccagga ccatggcctg accccggacc aagtggtggc 4800
tatcgccagc aacattggcg gcaagcaagc gctcgaaacg gtgcagcggc tgttgccggt 4860
gctgtgccag gaccatggcc tgaccccgga ccaagtggtg gctatcgcca gcaacaatgg 4920
cggcaagcaa gcgctcgaaa cggtgcagcg gctgttgccg gtgctgtgcc aggaccatgg 4980
cctgaccccg gaccaagtgg tggctatcgc cagcaacatt ggcggcaagc aagcgctcga 5040
aacggtgcag cggctgttgc cggtgctgtg ccaggaccat ggcctgaccc cggaccaagt 5100
ggtggctatc gccagcaaca ttggcggcaa gcaagcgctc gaaacggtgc agcggctgtt 5160
gccggtgctg tgccaggacc atggcctgac tccggaccaa gtggtggcta tcgccagcca 5220
cgatggcggc aagcaagcgc tcgaaacggt gcagcggctg ttgccggtgc tgtgccagga 5280
ccatggcctg actccggacc aagtggtggc tatcgccagc cacgatggcg gcaagcaagc 5340
gctcgaaacg gtgcagcggc tgttgccggt gctgtgccag gaccatggcc tgaccccgga 5400
ccaagtggtg gctatcgcca gcaacaatgg cggcaagcaa gcgctcgaaa cggtgcagcg 5460
gctgttgccg gtgctgtgcc aggaccatgg cctgaccccg gaccaagtgg tggctatcgc 5520
cagcaacatt ggcggcaagc aagcgctcga aacggtgcag cggctgttgc cggtgctgtg 5580
ccaggaccat ggcctgaccc cggaccaagt ggtggctatc gccagcaacg gtggcggcaa 5640
gcaagcgctc gaaacggtgc agcggctgtt gccggtgctg tgccaggacc atggcctgac 5700
tccggaccaa gtggtggcta tcgccagcca cgatggcggc aagcaagcgc tcgaaacggt 5760
gcagcggctg ttgccggtgc tgtgccagga ccatggcctg actccggacc aagtggtggc 5820
tatcgccagc cacgatggcg gcaagcaagc gctcgaaacg gtgcagcggc tgttgccggt 5880
gctgtgccag gaccatggcc tgaccccgga ccaagtggtg gctatcgcca gcaacggtgg 5940
cggcaagcaa gcgctcgaaa cggtgcagcg gctgttgccg gtgctgtgcc aggaccatgg 6000
cctgactccg gaccaagtgg tggctatcgc cagccacgat ggcggcaagc aagcgctcga 6060
aacggtgcag cggctgttgc cggtgctgtg ccaggaccat ggcctgactc cggaccaagt 6120
ggtggctatc gccagccacg atggcggcaa gcaagcgctc gaaacggtgc agcggctgtt 6180
gccggtgctg tgccaggacc atggcctgac tccggaccaa gtggtggcta tcgccagcca 6240
cgatggcggc aagcaagcgc tcgaaacggt gcagcggctg ttgccggtgc tgtgccagga 6300
ccatggcctg accccggacc aagtggtggc tatcgccagc aacattggcg gcaagcaagc 6360
gctcgaaacg gtgcagcggc tgttgccggt gctgtgccag gaccatggcc tgaccccgga 6420
ccaagtggtg gctatcgcca gcaacggtgg cggcaagcaa gcgctcgaaa cggtgcagcg 6480
gctgttgccg gtgctgtgcc aggaccatgg cctgaccccg gaccaagtgg tggctatcgc 6540
cagcaacggt ggcggcaagc aagcgctcga aagcattgtg gcccagctga gccggcctga 6600
tccggcgttg gccgcgttga ccaacgacca cctcgtcgcc ttggcctgcc tcggcggacg 6660
tcctgccatg gatgcagtga aaaagggatt gccgcacgcg ccggaattga tcagaagagt 6720
caatcgccgt attggcgaac gcacgtccca tcgcgttgcc gactacgcgc aagtggttcg 6780
cgtgctggag tttttccagt gccactccca cccagcgtac gcatttgatg aggccatgac 6840
gcagttcggg atgagcagga acgggttggt acagctcttt cgcagagtgg gcgtcaccga 6900
actcgaagcc cgcggtggaa cgctcccccc agcctcgcag cgttgggacc gtatcctcca 6960
ggcatcaggg atgaaaaggg ccaaaccgtc ccctacttca gctcaaacac cggatcaggc 7020
gtctttgcat gcattcgccg attcgctgga gcgtgacctt gatgcgccca gcccaatgca 7080
cgagggagat cagacgcggg caagcagccg taaacggtcc cgatcggatc gtgctgtcac 7140
cggcccctcc gcacagcagg ctgtcgaggt gcgcgttccc gaacagcgcg atgcgctgca 7200
tttgcccctc agctggaggg taaaacgccc gcgtaccagg atctggggcg gcctcccggt 7260
taagtagtgc tttctacttt atgagtagtg ctttctactt tatgagtagt gctttctact 7320
ttatgagtag tgctttctac tttatgtcac ataggaataa aaagccatat cacataggaa 7380
taaaaagcca tatcacatag gaataaaaag ccatatcaca taggaataaa aagccataga 7440
aagcggagta ctgtcctccg agcggagtac tgtcctccga gcggagtact gtcctccgag 7500
cggagtactg tcctccgagc ggagttctgt cctccgagcg gagactctag atatagaacc 7560
gatcctccca ttgaattcta ggcgtgtacg gtgggaggcc tatataagca gagctcgttt 7620
agtgaaccgt cagatcgcct cgagtataga accgatcctc ccatttatag aaccgatcct 7680
cccatttata gaaccgatcc tcccattact agaagatctt gaattcgcca ccttgtcgac 7740
cattcgtccg cgcaggccaa gtcctgcccg cgagcttctg cccggacccc aaccggatag 7800
ggttcagccg actgcagatc gtggggtgtc tgcgcctgct ggcagccctc tggatggctt 7860
gcccgctcgg cggacggtgt cccggacccg gctgccatct ccccctgcgc cctcacctgc 7920
gttctcggcg ggcagcttca gcgatctgct ccgtccgttc gatccgtcgc ttcttgatac 7980
atcgcttctt gattcgatgc ctgccgtcgg cacgccgcat acagcggctg ccccagcaga 8040
gtgggatgag gcgcaatcgg ctctgcgtgc agccgatgac ccgccaccca ccgtgcgtgt 8100
cgctgtcact gccgcgcggc cgccgcgcgc caagccggcc ccgcgacggc gtgctgcgca 8160
accctccgac gcttcgccgg ccgcgcaggt ggatctacgc acgctcggct acagtcagca 8220
gcagcaagag aagatcaaac cgaaggtgcg ttcgacagtg gcgcagcacc acgaggcact 8280
ggtgggccat gggtttacac acgcgcacat cgttgcgctc agccaacacc cggcagcgtt 8340
agggaccgtc gctgtcacgt atcagcacat aatcacggcg ttgccagagg cgacacacga 8400
agacatcgtt ggcgtcggca aacagtggtc cggcgcacgc gccctggagg ccttgctcac 8460
ggatgcgggg gagttgagag gtccgccgtt acagttggac acaggccaac ttgtgaagat 8520
tgcaaaacgt ggcggcgtga ccgcaatgga ggcagtgcat gcatcgcgca atgcactgac 8580
gggtgccccc ctgaacctga ccccggacca agtggtggct atcgccagca acattggcgg 8640
caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg accatggcct 8700
gactccggac caagtggtgg ctatcgccag ccacgatggc ggcaagcaag cgctcgaaac 8760
ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc ctgactccgg accaagtggt 8820
ggctatcgcc agccacgatg gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc 8880
ggtgctgtgc caggaccatg gcctgactcc ggaccaagtg gtggctatcg ccagccacga 8940
tggcggcaag caagcgctcg aaacggtgca gcggctgttg ccggtgctgt gccaggacca 9000
tggcctgact ccggaccaag tggtggctat cgccagccac gatggcggca agcaagcgct 9060
cgaaacggtg cagcggctgt tgccggtgct gtgccaggac catggcctga ccccggacca 9120
agtggtggct atcgccagca acggtggcgg caagcaagcg ctcgaaacgg tgcagcggct 9180
gttgccggtg ctgtgccagg accatggcct gactccggac caagtggtgg ctatcgccag 9240
ccacgatggc ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca 9300
ggaccatggc ctgaccccgg accaagtggt ggctatcgcc agcaacggtg gcggcaagca 9360
agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg gcctgactcc 9420
ggaccaagtg gtggctatcg ccagccacga tggcggcaag caagcgctcg aaacggtgca 9480
gcggctgttg ccggtgctgt gccaggacca tggcctgact ccggaccaag tggtggctat 9540
cgccagccac gatggcggca agcaagcgct cgaaacggtg cagcggctgt tgccggtgct 9600
gtgccaggac catggcctga ccccggacca agtggtggct atcgccagca acaatggcgg 9660
caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg accatggcct 9720
gactccggac caagtggtgg ctatcgccag ccacgatggc ggcaagcaag cgctcgaaac 9780
ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc ctgaccccgg accaagtggt 9840
ggctatcgcc agcaacggtg gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc 9900
ggtgctgtgc caggaccatg gcctgacccc ggaccaagtg gtggctatcg ccagcaacgg 9960
tggcggcaag caagcgctcg aaacggtgca gcggctgttg ccggtgctgt gccaggacca 10020
tggcctgact ccggaccaag tggtggctat cgccagccac gatggcggca agcaagcgct 10080
cgaaacggtg cagcggctgt tgccggtgct gtgccaggac catggcctga ccccggacca 10140
agtggtggct atcgccagca acggtggcgg caagcaagcg ctcgaaagca ttgtggccca 10200
gctgagccgg cctgatccgg cgttggccgc gttgaccaac gaccacctcg tcgccttggc 10260
ctgcctcggc ggacgtcctg ccatggatgc agtgaaaaag ggattgccgc acgcgccgga 10320
attgatcaga agagtcaatc gccgtattgg cgaacgcacg tcccatcgcg ttgccgacta 10380
cgcgcaagtg gttcgcgtgc tggagttttt ccagtgccac tcccacccag cgtacgcatt 10440
tgatgaggcc atgacgcagt tcgggatgag caggaacggg ttggtacagc tctttcgcag 10500
agtgggcgtc accgaactcg aagcccgcgg tggaacgctc cccccagcct cgcagcgttg 10560
ggaccgtatc ctccaggcat cagggatgaa aagggccaaa ccgtccccta cttcagctca 10620
aacaccggat caggcgtctt tgcatgcatt cgccgattcg ctggagcgtg accttgatgc 10680
gcccagccca atgcacgagg gagatcagac gcgggcaagc agccgtaaac ggtcccgatc 10740
ggatcgtgct gtcaccggcc cctccgcaca gcaggctgtc gaggtgcgcg ttcccgaaca 10800
gcgcgatgcg ctgcatttgc ccctcagctg gagggtaaaa cgcccgcgta ccaggatctg 10860
gggcggcctc ccgtaataca aggcggccgc aaatcaacat cagtctgata agctatcaac 10920
atcagtctga taagctatca acatcagtct gataagctat caacatcagt ctgataagct 10980
aaagcttcga attctgataa tcagccatac cacatttgta gaggttttac ttgctttaaa 11040
aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa 11100
cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa 11160
taaagcattt ttttcactgc agctcgcctt ttgctcacat gtctcccagg ggatgggggt 11220
ccatccacct tgccgaaaag atttgtctga ggaactgaaa atagaaggga aaaaagagga 11280
gggacaaaag aggcagaaat gagaggggag gggacagagg acacctgaat aaagaccaca 11340
cccatgaccc acgtgatgct gagaagtact cctgccctag gaagagactc agggcagagg 11400
gaggaaggac agcagaccag acagtcacag cagccttgac aaaacgttcc tggaactcaa 11460
gctcttctcc acagaggagg acagagcaga cagcagagac catggagtct ccctcggccc 11520
ctccccacag atggtgcatc ccctggcaga ggctcctgct cacaggtgaa ctcgagaccg 11580
gtatggcccc cccgaccgat gtcagcctgg gggacgagct ccacttagac ggcgaggacg 11640
tggcgatggc gcatgccgac gcgctagacg atttcgatct ggacatgttg gggagcgacg 11700
cgctagacga tttcgatctg gacatgttgg ggagcgacgc gctagacgat ttcgatctgg 11760
acatgttggg gagcgacgcg ctagacgatt tcgatctgga catgttgggg gacggggatt 11820
ccccgggtcc gggatttacc ccccacgact ccgcccccta cggcgctctg gatatggccg 11880
acttcgagtt tgagcagatg tttaccgatg cccttggaat tgacgagtac ggtgggacgc 11940
gtatgaagct actgtcttct atcgaacaag catgcgatat ttgccgactt aaaaagctca 12000
agtgctccaa agaaaaaccg aagtgcgcca agtgtctgaa gaacaactgg gagtgtcgct 12060
actctcccaa aaccaaaagg tctccgctga ctagggcaca tctgacagaa gtggaatcaa 12120
ggctagaaag actggaacag ctatttctac tgatttttcc tcgagaagac cttgacatga 12180
ttttgaaaat ggattcttta caggatataa aagcattgtt aacaggatta tttgtacaag 12240
ataatgtgaa taaagatgcc gtcacagata gattggcttc agtggagact gatatgcctc 12300
taacattgag acagcataga ataagtgcga catcatcatc ggaagagagt agtaacaaag 12360
gtcaaagaca gttgactgta taaaagcttc actcctcagg tgcaggctgc ctatcagaag 12420
gtggtggctg gtgtggccaa tgccctggct cacaaatacc actgagatct ttttccctct 12480
gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg 12540
aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca 12600
tatgggaggg caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca 12660
tatgcccata tgctggctgc catgaacaaa ggttggctat aaagaggtca tcagtatatg 12720
aaacagcccc ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt 12780
ttttttatat tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat 12840
gttttactag ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt 12900
ctcttatgga gatc 12914
<210> 11
<211> 6053
<212> DNA
<213> Artificial
<220>
<223> 编码由细胞特异性启动子CEA205、第一microRNA为mir-135a-5p、第二micr
oRNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白分
别为Lacl、tetR-KRAB、第一和第二识别序列为4×UAS的元件组成的表达系
统的核酸
<400> 11
cgggtgaccg ccctccgcgg agtactgtcc tccgcgggtg acagccctcc gcggagtgcc 60
gcactccgag cggagactct agactcccta tcagtgatag agataggcgt gtacggtggg 120
aggcctatat aagcagagct cgtttagtga accgtcagat cgctccctat cagtgataga 180
gagaattcga ccgccaccat gagacatatt atctgccacg gaggtgttat taccgaagaa 240
atggccgcca gtcttttgga ccagctgatc gaagaggtac tggctgataa tcttccacct 300
cctagccatt ttgaaccacc tacccttcac gaactgtatg atttagacgt gacggccccc 360
gaagatccca acgaggaggc ggtttcgcag atttttcccg actctgtaat gttggcggtg 420
caggaaggga ttgacttact cacttttccg ccggcgcccg gttctccgga gccgcctcac 480
ctttcccggc agcccgagca gccggagcag agagccttgg gtccggtttc tatgccaaac 540
cttgtaccgg aggtgatcga tcttacctgc cacgaggctg gctttccacc cagtgacgac 600
gaggatgaag agggtgagga gtttgtgtta gattatgtgg agcaccccgg gcacggttgc 660
aggtcttgtc attatcaccg gaggaatacg ggggacccag atattatgtg ttcgctttgc 720
tatatgagga cctgtggcat gtttgtctac agtaagtgaa aattatgggc agtgggtgat 780
agagtggtgg gtttggtgtg gtaatttttt ttttaatttt tacagttttg tggtttaaag 840
aattttgtat tgtgattttt ttaaaaggtc ctgtgtctga acctgagcct gagcccgagc 900
cagaaccgga gcctgcaaga cctacccgcc gtcctaaaat ggcgcctgct atcctgaggc 960
gcccgacatc acctgtgtct agagaatgca atagtagtac ggatagctgt gactccggtc 1020
cttctaacac acctcctgag atacacccgg tggtcccgct gtgccccatt aaaccagttg 1080
ccgtgagagt tggtgggcgt cgccaggctg tggaatgtat cgaggacttg cttaacgagc 1140
ctgggcaacc tttggacttg agctgtaaac gccccaggcc actcgagggt accgggtccg 1200
gagctacaaa tttttccctc ctcaaacagg ctggagatgt cgaagaaaat cctgggccta 1260
ccggtatgtg gctgcagagc ctccttctac tgggcaccgt ggcttgcagc atctcggccc 1320
ccgccagaag tcccagcccc agcacccagc cctgggaaca cgtaaatgca atccaggagg 1380
cccgccgcct cctgaacttg tcacgggaca cggcagccga gatgaatgaa actgtggagg 1440
tcatcagtga aatgtttgac ttgcaggagc ccacatgcct gcagactcgg ctggagctct 1500
acaaacaggg cctgcgtggc agcctcacca agctgaaggg cccactgacc atgatggcta 1560
gccattacaa gcagcactgc ccgccgaccc ctgagacatc ctgtgccact cagatcatta 1620
cctttgagtc tttcaaggag aacctcaagg actttctgct ggtcatccca tttgactgct 1680
gggaacccgt ccaggaaaga tctgagggcc gcggcagcct gctgacctgc ggcgacgtgg 1740
aggaaaaccc cggccccgga tccatgaaac cagtaacgtt atacgatgtc gcagagtatg 1800
ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac gtttctgcga 1860
aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg 1920
cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc agtctggccc 1980
tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca 2040
gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca 2100
atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg 2160
ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc 2220
agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg ggcgtggagc 2280
atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct 2340
cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga 2400
tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc 2460
tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg 2520
caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggatatctcg gtagtgggat 2580
acgacgatac cgaagacagc tcatgttata tcccgccgtt aaccaccatc aaacaggatt 2640
ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg 2700
tgaagggcaa tcagctgttg ccggtctcac tggtgaaaag aaaaaccacc ctggcgccca 2760
atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg 2820
tttcccgact ggaaagcggg cagagcagcc tgaggcctcc taagaagaag aggaaggttg 2880
ttacaaacac cattgtcaca ctccacaaac accattgtca cactccacaa acaccattgt 2940
cacactccac aaacaccatt gtcacactcc atcacatagg aataaaaagc catatcacat 3000
aggaataaaa agccatatca cataggaata aaaagccata tcacatagga ataaaaagcc 3060
atagaaagcg gggtgccgcc ccccgcggac tactgtagtc cgcggacgac tgtcgtccgc 3120
ggagcactgt gctccgagcg gagactctag aaattgtgag cggataacaa ttgggtatat 3180
aatgggggcc gtcagctact accagagccg atcgaacact cgagccgagc agacgtgcct 3240
acaattgtga gcgctcacaa ttactagaag atcttgaatt cgccaccatg gctagattag 3300
ataaaagtaa agtgattaac agcgcattag agctgcttaa tgaggtcgga atcgaaggtt 3360
taacaacccg taaactcgcc cagaagctag gtgtagagca gcctacattg tattggcatg 3420
taaaaaataa gcgggctttg ctcgacgcct tagccattga gatgttagat aggcaccata 3480
ctcacttttg ccctttagaa ggggaaagct ggcaagattt tttacgtaat aacgctaaaa 3540
gttttagatg tgctttacta agtcatcgcg atggagcaaa agtacattta ggtacacggc 3600
ctacagaaaa acagtatgaa actctcgaaa atcaattagc ctttttatgc caacaaggtt 3660
tttcactaga gaatgcatta tatgcactca gcgctgtggg gcattttact ttaggttgcg 3720
tattggaaga tcaagagcat caagtcgcta aagaagaaag ggaaacacct actactgata 3780
gtatgccgcc attattacga caagctatcg aattatttga tcaccaaggt gcagagccag 3840
ccttcttatt cggccttgaa ttgatcatat gcggattaga aaaacaactt aaatgtgaaa 3900
gtgggtcgcc aaaaaagaag agaaaggtgg atggtggcgg agcactgtct ccccagcatt 3960
cagctgtgac ccagggatca atcattaaga acaaggaagg tatggatgct aagtccctga 4020
ccgcttggtc aagaactttg gtaactttta aggacgtgtt cgtcgatttt actcgggagg 4080
agtggaaact attggatacc gcccagcaaa tcgtgtatag aaacgttatg ctcgagaact 4140
ataagaatct tgtgtcactg ggctaccaac tgacaaagcc tgacgttatc ctgaggctgg 4200
aaaagggtga ggagccttgg ttggtcgagc gggagatcca ccaggaaact cacccagaca 4260
gtgagactgc tttcgagatc aagtcctctg tgtaatacaa ggcggccgca aatcaacatc 4320
agtctgataa gctatcaaca tcagtctgat aagctatcaa catcagtctg ataagctatc 4380
aacatcagtc tgataagcta aagcttcgaa ttctgataat cagccatacc acatttgtag 4440
aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga 4500
atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata 4560
gcatcacaaa tttcacaaat aaagcatttt tttcactgca gctcgccttt tgctcacatg 4620
tgaaaataga agggaaaaaa gaggagggac aaaagaggca gaaatgagag gggaggggac 4680
agaggacacc tgaataaaga ccacacccat gacccacgtg atgctgagaa gtactcctgc 4740
cctaggaaga gactcagggc agagggagga aggacagcag accagacagt cacagcagcc 4800
ttgacaaaac gttcctggaa ctcaagctcg agaccggtat ggcccccccg accgatgtca 4860
gcctggggga cgagctccac ttagacggcg aggacgtggc gatggcgcat gccgacgcgc 4920
tagacgattt cgatctggac atgttggggg acggggattc cccgggtccg ggatttaccc 4980
cccacgactc cgccccctac ggcgctctgg atatggccga cttcgagttt gagcagatgt 5040
ttaccgatgc ccttggaatt gacgagtacg gtgggacgcg tatgaagcta ctgtcttcta 5100
tcgaacaagc atgcgatatt tgccgactta aaaagctcaa gtgctccaaa gaaaaaccga 5160
agtgcgccaa gtgtctgaag aacaactggg agtgtcgcta ctctcccaaa accaaaaggt 5220
ctccgctgac tagggcacat ctgacagaag tggaatcaag gctagaaaga ctggaacagc 5280
tatttctact gatttttcct cgagaagacc ttgacatgat tttgaaaatg gattctttac 5340
aggatataaa agcattgtta acaggattat ttgtacaaga taatgtgaat aaagatgccg 5400
tcacagatag attggcttca gtggagactg atatgcctct aacattgaga cagcatagaa 5460
taagtgcgac atcatcatcg gaagagagta gtaacaaagg tcaaagacag ttgactgtat 5520
aaaagcttca ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg tgtggccaat 5580
gccctggctc acaaatacca ctgagatctt tttccctctg ccaaaaatta tggggacatc 5640
atgaagcccc ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata 5700
gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta 5760
aaacatcaga atgagtattt ggtttagagt ttggcaacat atgcccatat gctggctgcc 5820
atgaacaaag gttggctata aagaggtcat cagtatatga aacagccccc tgctgtccat 5880
tccttattcc atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg 5940
ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc 6000
ctcctctcct gactactccc agtcatagct gtccctcttc tcttatggag atc 6053
<210> 12
<211> 12671
<212> DNA
<213> Artificial
<220>
<223> 编码由细胞特异性启动子CEA205、第一microRNA为mir-143-3p、第二micro
RNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白分
别为TALE蛋白、第一和第二识别序列为4×UAS的元件组成的表达系统的核酸
<400> 12
cggagtactg tcctccgagc ggagtactgt cctccgagcg gagtactgtc ctccgagcgg 60
agttctgtcc tccgagcgga gactctagat acccctctcc gcttctgaat tctaggcgtg 120
tacggtggga ggcctatata agcagagctc gtttagtgaa ccgtcagatc gcctcgagta 180
cccctctccg cttcttaccc ctctccgctt cttacccctc tccgcttctg aattcgaccg 240
ccaccatgag acatattatc tgccacggag gtgttattac cgaagaaatg gccgccagtc 300
ttttggacca gctgatcgaa gaggtactgg ctgataatct tccacctcct agccattttg 360
aaccacctac ccttcacgaa ctgtatgatt tagacgtgac ggcccccgaa gatcccaacg 420
aggaggcggt ttcgcagatt tttcccgact ctgtaatgtt ggcggtgcag gaagggattg 480
acttactcac ttttccgccg gcgcccggtt ctccggagcc gcctcacctt tcccggcagc 540
ccgagcagcc ggagcagaga gccttgggtc cggtttctat gccaaacctt gtaccggagg 600
tgatcgatct tacctgccac gaggctggct ttccacccag tgacgacgag gatgaagagg 660
gtgaggagtt tgtgttagat tatgtggagc accccgggca cggttgcagg tcttgtcatt 720
atcaccggag gaatacgggg gacccagata ttatgtgttc gctttgctat atgaggacct 780
gtggcatgtt tgtctacagt aagtgaaaat tatgggcagt gggtgataga gtggtgggtt 840
tggtgtggta attttttttt taatttttac agttttgtgg tttaaagaat tttgtattgt 900
gattttttta aaaggtcctg tgtctgaacc tgagcctgag cccgagccag aaccggagcc 960
tgcaagacct acccgccgtc ctaaaatggc gcctgctatc ctgaggcgcc cgacatcacc 1020
tgtgtctaga gaatgcaata gtagtacgga tagctgtgac tccggtcctt ctaacacacc 1080
tcctgagata cacccggtgg tcccgctgtg ccccattaaa ccagttgccg tgagagttgg 1140
tgggcgtcgc caggctgtgg aatgtatcga ggacttgctt aacgagcctg ggcaaccttt 1200
ggacttgagc tgtaaacgcc ccaggccagg gtccggagct acaaattttt ccctcctcaa 1260
acaggctgga gatgtcgaag aaaatcctgg gcctatggag gcttgggagt gtttggaaga 1320
tttttctgct gtgcgtaact tgctggaaca gagctctaac agtacctctt ggttttggag 1380
gtttctgtgg ggctcatccc aggcaaagtt agtctgcaga attaaggagg attacaagtg 1440
ggaatttgaa gagcttttga aatcctgtgg tgagctgttt gattctttga atctgggtca 1500
ccaggcgctt ttccaagaga aggtcatcaa gactttggat ttttccacac cggggcgcgc 1560
tgcggctgct gttgcttttt tgagttttat aaaggataaa tggagcgaag aaacccatct 1620
gagcgggggg tacctgctgg attttctggc catgcatctg tggagagcgg ttgtgagaca 1680
caagaatcgc ctgctactgt tgtcttccgt ccgcccggcg ataataccga cggaggagca 1740
gcagcagcag caggaggaag ccaggcggcg gcggcaggag cagagcccat ggaacccgag 1800
agccggcctg gaccctcggg aatgaatgtt gtacaggtgg ctgaactgta tccagaactg 1860
agacgcattt tgacaattac agaggatggg caggggctaa agggggtaaa gagggagcgg 1920
ggggcttgtg aggctacaga ggaggctagg aatctagctt ttagcttaat gaccagacac 1980
cgtcctgagt gtattacttt tcaacagatc aaggataatt gcgctaatga gcttgatctg 2040
ctggcgcaga agtattccat agagcagctg accacttact ggctgcagcc aggggatgat 2100
tttgaggagg ctattagggt atatgcaaag gtggcactta ggccagattg caagtacaag 2160
atcagcaaac ttgtaaatat caggaattgt tgctacattt ctgggaacgg ggccgaggtg 2220
gagatagata cggaggatag ggtggccttt agatgtagca tgataaatat gtggccgggg 2280
gtgcttggca tggacggggt ggttattatg aatgtaaggt ttactggccc caattttagc 2340
ggtacggttt tcctggccaa taccaacctt atcctacacg gtgtaagctt ctatgggttt 2400
aacaatacct gtgtggaagc ctggaccgat gtaagggttc ggggctgtgc cttttactgc 2460
tgctggaagg gggtggtgtg tcgccccaaa agcagggctt caattaagaa atgcctcttt 2520
gaaaggtgta ccttgggtat cctgtctgag ggtaactcca gggtgcgcca caatgtggcc 2580
tccgactgtg gttgcttcat gctagtgaaa agcgtggctg tgattaagca taacatggta 2640
tgtggcaact gcgaggacag ggcctctcag atgctgacct gctcggacgg caactgtcac 2700
ctgctgaaga ccattcacgt agccagccac tctcgcaagg cctggccagt gtttgagcat 2760
aacatactga cccgctgttc cttgcatttg ggtaacagga ggggggtgtt cctaccttac 2820
caatgcaatt tgagtcacac taagatattg cttgagcccg agagcatgtc caaggtgaac 2880
ctgaacgggg tgtttgacat gaccatgaag atctggaagg tgctgaggta cgatgagacc 2940
cgcaccaggt gcagaccctg cgagtgtggc ggtaaacata ttaggaacca gcctgtgatg 3000
ctggatgtga ccgaggagct gaggcccgat cacttggtgc tggcctgcac ccgcgctgag 3060
tttggctcta gcgatgaaga tacagatctc gaggggtccg gagctacaaa tttttccctc 3120
ctcaaacagg ctggagatgt cgaagaaaat cctgggccta ccggtatgta taggatgcag 3180
ttgctctctt gcatcgccct gtctctagcg ttggtgacta atagcgcccc tacttcttct 3240
tccacgaaga aaacccagct gcaactggaa caccttctgc tggatctcca gatgatcctt 3300
aacgggatta acaactataa gaacccgaag ctcacggcta tgctgacggc gaagttcgct 3360
atgcctaaaa aggctactga gctgaaacac ttacagtgtc tcgaagaggc actcaaaccc 3420
ctcgaggagg tactcaatct ggcgcaaagc aaaaacttcc atctccgccc acgggaccta 3480
atctccaaca tcaacgtgat tgtgctggag ctcaaggggt ctgaaaccac gtttatgtgc 3540
gagtacgcag atgaaactgc taccatagtg gaattcctca atcgttggat cactttctgc 3600
cagtcaatca tttcaaccct gaccagatct gagggccgcg gcagcctgct gacctgcggc 3660
gacgtggagg aaaaccccgg ccccggatcc ttgtcgacca ttcgtccgcg caggccaagt 3720
cctgcccgcg agcttctgcc cggaccccaa ccggataggg ttcagccgac tgcagatcgt 3780
ggggtgtctg cgcctgctgg cagccctctg gatggcttgc ccgctcggcg gacggtgtcc 3840
cggacccggc tgccatctcc ccctgcgccc tcacctgcgt tctcggcggg cagcttcagc 3900
gatctgctcc gtccgttcga tccgtcgctt cttgatacat cgcttcttga ttcgatgcct 3960
gccgtcggca cgccgcatac agcggctgcc ccagcagagt gggatgaggc gcaatcggct 4020
ctgcgtgcag ccgatgaccc gccacccacc gtgcgtgtcg ctgtcactgc cgcgcggccg 4080
ccgcgcgcca agccggcccc gcgacggcgt gctgcgcaac cctccgacgc ttcgccggcc 4140
gcgcaggtgg atctacgcac gctcggctac agtcagcagc agcaagagaa gatcaaaccg 4200
aaggtgcgtt cgacagtggc gcagcaccac gaggcactgg tgggccatgg gtttacacac 4260
gcgcacatcg ttgcgctcag ccaacacccg gcagcgttag ggaccgtcgc tgtcacgtat 4320
cagcacataa tcacggcgtt gccagaggcg acacacgaag acatcgttgg cgtcggcaaa 4380
cagtggtccg gcgcacgcgc cctggaggcc ttgctcacgg atgcggggga gttgagaggt 4440
ccgccgttac agttggacac aggccaactt gtgaagattg caaaacgtgg cggcgtgacc 4500
gcaatggagg cagtgcatgc atcgcgcaat gcactgacgg gtgcccccct gaacctgacc 4560
ccggaccaag tggtggctat cgccagcaac attggcggca agcaagcgct cgaaacggtg 4620
cagcggctgt tgccggtgct gtgccaggac catggcctga ccccggacca agtggtggct 4680
atcgccagca acggtggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg 4740
ctgtgccagg accatggcct gaccccggac caagtggtgg ctatcgccag caacattggc 4800
ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc 4860
ctgaccccgg accaagtggt ggctatcgcc agcaacaatg gcggcaagca agcgctcgaa 4920
acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg gcctgacccc ggaccaagtg 4980
gtggctatcg ccagcaacat tggcggcaag caagcgctcg aaacggtgca gcggctgttg 5040
ccggtgctgt gccaggacca tggcctgacc ccggaccaag tggtggctat cgccagcaac 5100
attggcggca agcaagcgct cgaaacggtg cagcggctgt tgccggtgct gtgccaggac 5160
catggcctga ctccggacca agtggtggct atcgccagcc acgatggcgg caagcaagcg 5220
ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg accatggcct gactccggac 5280
caagtggtgg ctatcgccag ccacgatggc ggcaagcaag cgctcgaaac ggtgcagcgg 5340
ctgttgccgg tgctgtgcca ggaccatggc ctgaccccgg accaagtggt ggctatcgcc 5400
agcaacaatg gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc 5460
caggaccatg gcctgacccc ggaccaagtg gtggctatcg ccagcaacat tggcggcaag 5520
caagcgctcg aaacggtgca gcggctgttg ccggtgctgt gccaggacca tggcctgacc 5580
ccggaccaag tggtggctat cgccagcaac ggtggcggca agcaagcgct cgaaacggtg 5640
cagcggctgt tgccggtgct gtgccaggac catggcctga ctccggacca agtggtggct 5700
atcgccagcc acgatggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg 5760
ctgtgccagg accatggcct gactccggac caagtggtgg ctatcgccag ccacgatggc 5820
ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc 5880
ctgaccccgg accaagtggt ggctatcgcc agcaacggtg gcggcaagca agcgctcgaa 5940
acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg gcctgactcc ggaccaagtg 6000
gtggctatcg ccagccacga tggcggcaag caagcgctcg aaacggtgca gcggctgttg 6060
ccggtgctgt gccaggacca tggcctgact ccggaccaag tggtggctat cgccagccac 6120
gatggcggca agcaagcgct cgaaacggtg cagcggctgt tgccggtgct gtgccaggac 6180
catggcctga ctccggacca agtggtggct atcgccagcc acgatggcgg caagcaagcg 6240
ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg accatggcct gaccccggac 6300
caagtggtgg ctatcgccag caacattggc ggcaagcaag cgctcgaaac ggtgcagcgg 6360
ctgttgccgg tgctgtgcca ggaccatggc ctgaccccgg accaagtggt ggctatcgcc 6420
agcaacggtg gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc 6480
caggaccatg gcctgacccc ggaccaagtg gtggctatcg ccagcaacgg tggcggcaag 6540
caagcgctcg aaagcattgt ggcccagctg agccggcctg atccggcgtt ggccgcgttg 6600
accaacgacc acctcgtcgc cttggcctgc ctcggcggac gtcctgccat ggatgcagtg 6660
aaaaagggat tgccgcacgc gccggaattg atcagaagag tcaatcgccg tattggcgaa 6720
cgcacgtccc atcgcgttgc cgactacgcg caagtggttc gcgtgctgga gtttttccag 6780
tgccactccc acccagcgta cgcatttgat gaggccatga cgcagttcgg gatgagcagg 6840
aacgggttgg tacagctctt tcgcagagtg ggcgtcaccg aactcgaagc ccgcggtgga 6900
acgctccccc cagcctcgca gcgttgggac cgtatcctcc aggcatcagg gatgaaaagg 6960
gccaaaccgt cccctacttc agctcaaaca ccggatcagg cgtctttgca tgcattcgcc 7020
gattcgctgg agcgtgacct tgatgcgccc agcccaatgc acgagggaga tcagacgcgg 7080
gcaagcagcc gtaaacggtc ccgatcggat cgtgctgtca ccggcccctc cgcacagcag 7140
gctgtcgagg tgcgcgttcc cgaacagcgc gatgcgctgc atttgcccct cagctggagg 7200
gtaaaacgcc cgcgtaccag gatctggggc ggcctcccgg ttaagtagtg ctttctactt 7260
tatgagtagt gctttctact ttatgagtag tgctttctac tttatgagta gtgctttcta 7320
ctttatggag ctacagtgct tcatctcaga gctacagtgc ttcatctcag agctacagtg 7380
cttcatctca gagctacagt gcttcatctc agaaagcgga gtactgtcct ccgagcggag 7440
tactgtcctc cgagcggagt actgtcctcc gagcggagtt ctgtcctccg agcggagact 7500
ctagatatag aaccgatcct cccattgaat tctaggcgtg tacggtggga ggcctatata 7560
agcagagctc gtttagtgaa ccgtcagatc gcctcgagta tagaaccgat cctcccattt 7620
atagaaccga tcctcccatt tatagaaccg atcctcccat tactagaaga tcttgaattc 7680
gccaccttgt cgaccattcg tccgcgcagg ccaagtcctg cccgcgagct tctgcccgga 7740
ccccaaccgg atagggttca gccgactgca gatcgtgggg tgtctgcgcc tgctggcagc 7800
cctctggatg gcttgcccgc tcggcggacg gtgtcccgga cccggctgcc atctccccct 7860
gcgccctcac ctgcgttctc ggcgggcagc ttcagcgatc tgctccgtcc gttcgatccg 7920
tcgcttcttg atacatcgct tcttgattcg atgcctgccg tcggcacgcc gcatacagcg 7980
gctgccccag cagagtggga tgaggcgcaa tcggctctgc gtgcagccga tgacccgcca 8040
cccaccgtgc gtgtcgctgt cactgccgcg cggccgccgc gcgccaagcc ggccccgcga 8100
cggcgtgctg cgcaaccctc cgacgcttcg ccggccgcgc aggtggatct acgcacgctc 8160
ggctacagtc agcagcagca agagaagatc aaaccgaagg tgcgttcgac agtggcgcag 8220
caccacgagg cactggtggg ccatgggttt acacacgcgc acatcgttgc gctcagccaa 8280
cacccggcag cgttagggac cgtcgctgtc acgtatcagc acataatcac ggcgttgcca 8340
gaggcgacac acgaagacat cgttggcgtc ggcaaacagt ggtccggcgc acgcgccctg 8400
gaggccttgc tcacggatgc gggggagttg agaggtccgc cgttacagtt ggacacaggc 8460
caacttgtga agattgcaaa acgtggcggc gtgaccgcaa tggaggcagt gcatgcatcg 8520
cgcaatgcac tgacgggtgc ccccctgaac ctgaccccgg accaagtggt ggctatcgcc 8580
agcaacattg gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc 8640
caggaccatg gcctgactcc ggaccaagtg gtggctatcg ccagccacga tggcggcaag 8700
caagcgctcg aaacggtgca gcggctgttg ccggtgctgt gccaggacca tggcctgact 8760
ccggaccaag tggtggctat cgccagccac gatggcggca agcaagcgct cgaaacggtg 8820
cagcggctgt tgccggtgct gtgccaggac catggcctga ctccggacca agtggtggct 8880
atcgccagcc acgatggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg 8940
ctgtgccagg accatggcct gactccggac caagtggtgg ctatcgccag ccacgatggc 9000
ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc 9060
ctgaccccgg accaagtggt ggctatcgcc agcaacggtg gcggcaagca agcgctcgaa 9120
acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg gcctgactcc ggaccaagtg 9180
gtggctatcg ccagccacga tggcggcaag caagcgctcg aaacggtgca gcggctgttg 9240
ccggtgctgt gccaggacca tggcctgacc ccggaccaag tggtggctat cgccagcaac 9300
ggtggcggca agcaagcgct cgaaacggtg cagcggctgt tgccggtgct gtgccaggac 9360
catggcctga ctccggacca agtggtggct atcgccagcc acgatggcgg caagcaagcg 9420
ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg accatggcct gactccggac 9480
caagtggtgg ctatcgccag ccacgatggc ggcaagcaag cgctcgaaac ggtgcagcgg 9540
ctgttgccgg tgctgtgcca ggaccatggc ctgaccccgg accaagtggt ggctatcgcc 9600
agcaacaatg gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc 9660
caggaccatg gcctgactcc ggaccaagtg gtggctatcg ccagccacga tggcggcaag 9720
caagcgctcg aaacggtgca gcggctgttg ccggtgctgt gccaggacca tggcctgacc 9780
ccggaccaag tggtggctat cgccagcaac ggtggcggca agcaagcgct cgaaacggtg 9840
cagcggctgt tgccggtgct gtgccaggac catggcctga ccccggacca agtggtggct 9900
atcgccagca acggtggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg 9960
ctgtgccagg accatggcct gactccggac caagtggtgg ctatcgccag ccacgatggc 10020
ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc 10080
ctgaccccgg accaagtggt ggctatcgcc agcaacggtg gcggcaagca agcgctcgaa 10140
agcattgtgg cccagctgag ccggcctgat ccggcgttgg ccgcgttgac caacgaccac 10200
ctcgtcgcct tggcctgcct cggcggacgt cctgccatgg atgcagtgaa aaagggattg 10260
ccgcacgcgc cggaattgat cagaagagtc aatcgccgta ttggcgaacg cacgtcccat 10320
cgcgttgccg actacgcgca agtggttcgc gtgctggagt ttttccagtg ccactcccac 10380
ccagcgtacg catttgatga ggccatgacg cagttcggga tgagcaggaa cgggttggta 10440
cagctctttc gcagagtggg cgtcaccgaa ctcgaagccc gcggtggaac gctcccccca 10500
gcctcgcagc gttgggaccg tatcctccag gcatcaggga tgaaaagggc caaaccgtcc 10560
cctacttcag ctcaaacacc ggatcaggcg tctttgcatg cattcgccga ttcgctggag 10620
cgtgaccttg atgcgcccag cccaatgcac gagggagatc agacgcgggc aagcagccgt 10680
aaacggtccc gatcggatcg tgctgtcacc ggcccctccg cacagcaggc tgtcgaggtg 10740
cgcgttcccg aacagcgcga tgcgctgcat ttgcccctca gctggagggt aaaacgcccg 10800
cgtaccagga tctggggcgg cctcccgtaa tacaaggcgg ccgcaaatca acatcagtct 10860
gataagctat caacatcagt ctgataagct atcaacatca gtctgataag ctatcaacat 10920
cagtctgata agctaaagct tcgaattctg ataatcagcc ataccacatt tgtagaggtt 10980
ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca 11040
attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc 11100
acaaatttca caaataaagc atttttttca ctgcgaaaat agaagggaaa aaagaggagg 11160
gacaaaagag gcagaaatga gaggggaggg gacagaggac acctgaataa agaccacacc 11220
catgacccac gtgatgctga gaagtactcc tgccctagga agagactcag ggcagaggga 11280
ggaaggacag cagaccagac agtcacagca gccttgacaa aacgttcctg gaactcaaga 11340
tggccccccc gaccgatgtc agcctggggg acgagctcca cttagacggc gaggacgtgg 11400
cgatggcgca tgccgacgcg ctagacgatt tcgatctgga catgttgggg agcgacgcgc 11460
tagacgattt cgatctggac atgttgggga gcgacgcgct agacgatttc gatctggaca 11520
tgttggggag cgacgcgcta gacgatttcg atctggacat gttgggggac ggggattccc 11580
cgggtccggg atttaccccc cacgactccg ccccctacgg cgctctggat atggccgact 11640
tcgagtttga gcagatgttt accgatgccc ttggaattga cgagtacggt gggacgcgta 11700
tgaagctact gtcttctatc gaacaagcat gcgatatttg ccgacttaaa aagctcaagt 11760
gctccaaaga aaaaccgaag tgcgccaagt gtctgaagaa caactgggag tgtcgctact 11820
ctcccaaaac caaaaggtct ccgctgacta gggcacatct gacagaagtg gaatcaaggc 11880
tagaaagact ggaacagcta tttctactga tttttcctcg agaagacctt gacatgattt 11940
tgaaaatgga ttctttacag gatataaaag cattgttaac aggattattt gtacaagata 12000
atgtgaataa agatgccgtc acagatagat tggcttcagt ggagactgat atgcctctaa 12060
cattgagaca gcatagaata agtgcgacat catcatcgga agagagtagt aacaaaggtc 12120
aaagacagtt gactgtataa aagcttcact cctcaggtgc aggctgccta tcagaaggtg 12180
gtggctggtg tggccaatgc cctggctcac aaataccact gagatctttt tccctctgcc 12240
aaaaattatg gggacatcat gaagcccctt gagcatctga cttctggcta ataaaggaaa 12300
tttattttca ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg aaggacatat 12360
gggagggcaa atcatttaaa acatcagaat gagtatttgg tttagagttt ggcaacatat 12420
gcccatatgc tggctgccat gaacaaaggt tggctataaa gaggtcatca gtatatgaaa 12480
cagccccctg ctgtccattc cttattccat agaaaagcct tgacttgagg ttagattttt 12540
tttatatttt gttttgtgtt atttttttct ttaacatccc taaaattttc cttacatgtt 12600
ttactagcca gatttttcct cctctcctga ctactcccag tcatagctgt ccctcttctc 12660
ttatggagat c 12671
<210> 13
<211> 6216
<212> DNA
<213> Artificial
<220>
<223> 编码由细胞特异性启动子CEA368、第一microRNA为mir-135a-5p、第二micr
oRNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白分
别为Lacl、tetR-KRAB、第一和第二识别序列为4×UAS的元件组成的表达系
统的核酸
<400> 13
cgggtgaccg ccctccgcgg agtactgtcc tccgcgggtg acagccctcc gcggagtgcc 60
gcactccgag cggagactct agactcccta tcagtgatag agataggcgt gtacggtggg 120
aggcctatat aagcagagct cgtttagtga accgtcagat cgctccctat cagtgataga 180
gagaattcga ccgccaccat gagacatatt atctgccacg gaggtgttat taccgaagaa 240
atggccgcca gtcttttgga ccagctgatc gaagaggtac tggctgataa tcttccacct 300
cctagccatt ttgaaccacc tacccttcac gaactgtatg atttagacgt gacggccccc 360
gaagatccca acgaggaggc ggtttcgcag atttttcccg actctgtaat gttggcggtg 420
caggaaggga ttgacttact cacttttccg ccggcgcccg gttctccgga gccgcctcac 480
ctttcccggc agcccgagca gccggagcag agagccttgg gtccggtttc tatgccaaac 540
cttgtaccgg aggtgatcga tcttacctgc cacgaggctg gctttccacc cagtgacgac 600
gaggatgaag agggtgagga gtttgtgtta gattatgtgg agcaccccgg gcacggttgc 660
aggtcttgtc attatcaccg gaggaatacg ggggacccag atattatgtg ttcgctttgc 720
tatatgagga cctgtggcat gtttgtctac agtaagtgaa aattatgggc agtgggtgat 780
agagtggtgg gtttggtgtg gtaatttttt ttttaatttt tacagttttg tggtttaaag 840
aattttgtat tgtgattttt ttaaaaggtc ctgtgtctga acctgagcct gagcccgagc 900
cagaaccgga gcctgcaaga cctacccgcc gtcctaaaat ggcgcctgct atcctgaggc 960
gcccgacatc acctgtgtct agagaatgca atagtagtac ggatagctgt gactccggtc 1020
cttctaacac acctcctgag atacacccgg tggtcccgct gtgccccatt aaaccagttg 1080
ccgtgagagt tggtgggcgt cgccaggctg tggaatgtat cgaggacttg cttaacgagc 1140
ctgggcaacc tttggacttg agctgtaaac gccccaggcc actcgagggt accgggtccg 1200
gagctacaaa tttttccctc ctcaaacagg ctggagatgt cgaagaaaat cctgggccta 1260
ccggtatgtg gctgcagagc ctccttctac tgggcaccgt ggcttgcagc atctcggccc 1320
ccgccagaag tcccagcccc agcacccagc cctgggaaca cgtaaatgca atccaggagg 1380
cccgccgcct cctgaacttg tcacgggaca cggcagccga gatgaatgaa actgtggagg 1440
tcatcagtga aatgtttgac ttgcaggagc ccacatgcct gcagactcgg ctggagctct 1500
acaaacaggg cctgcgtggc agcctcacca agctgaaggg cccactgacc atgatggcta 1560
gccattacaa gcagcactgc ccgccgaccc ctgagacatc ctgtgccact cagatcatta 1620
cctttgagtc tttcaaggag aacctcaagg actttctgct ggtcatccca tttgactgct 1680
gggaacccgt ccaggaaaga tctgagggcc gcggcagcct gctgacctgc ggcgacgtgg 1740
aggaaaaccc cggccccgga tccatgaaac cagtaacgtt atacgatgtc gcagagtatg 1800
ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac gtttctgcga 1860
aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg 1920
cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc agtctggccc 1980
tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca 2040
gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca 2100
atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg 2160
ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc 2220
agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg ggcgtggagc 2280
atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct 2340
cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga 2400
tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc 2460
tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg 2520
caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggatatctcg gtagtgggat 2580
acgacgatac cgaagacagc tcatgttata tcccgccgtt aaccaccatc aaacaggatt 2640
ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg 2700
tgaagggcaa tcagctgttg ccggtctcac tggtgaaaag aaaaaccacc ctggcgccca 2760
atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg 2820
tttcccgact ggaaagcggg cagagcagcc tgaggcctcc taagaagaag aggaaggttg 2880
ttacaaacac cattgtcaca ctccacaaac accattgtca cactccacaa acaccattgt 2940
cacactccac aaacaccatt gtcacactcc atcacatagg aataaaaagc catatcacat 3000
aggaataaaa agccatatca cataggaata aaaagccata tcacatagga ataaaaagcc 3060
atagaaagcg gggtgccgcc ccccgcggac tactgtagtc cgcggacgac tgtcgtccgc 3120
ggagcactgt gctccgagcg gagactctag aaattgtgag cggataacaa ttgggtatat 3180
aatgggggcc gtcagctact accagagccg atcgaacact cgagccgagc agacgtgcct 3240
acaattgtga gcgctcacaa ttactagaag atcttgaatt cgccaccatg gctagattag 3300
ataaaagtaa agtgattaac agcgcattag agctgcttaa tgaggtcgga atcgaaggtt 3360
taacaacccg taaactcgcc cagaagctag gtgtagagca gcctacattg tattggcatg 3420
taaaaaataa gcgggctttg ctcgacgcct tagccattga gatgttagat aggcaccata 3480
ctcacttttg ccctttagaa ggggaaagct ggcaagattt tttacgtaat aacgctaaaa 3540
gttttagatg tgctttacta agtcatcgcg atggagcaaa agtacattta ggtacacggc 3600
ctacagaaaa acagtatgaa actctcgaaa atcaattagc ctttttatgc caacaaggtt 3660
tttcactaga gaatgcatta tatgcactca gcgctgtggg gcattttact ttaggttgcg 3720
tattggaaga tcaagagcat caagtcgcta aagaagaaag ggaaacacct actactgata 3780
gtatgccgcc attattacga caagctatcg aattatttga tcaccaaggt gcagagccag 3840
ccttcttatt cggccttgaa ttgatcatat gcggattaga aaaacaactt aaatgtgaaa 3900
gtgggtcgcc aaaaaagaag agaaaggtgg atggtggcgg agcactgtct ccccagcatt 3960
cagctgtgac ccagggatca atcattaaga acaaggaagg tatggatgct aagtccctga 4020
ccgcttggtc aagaactttg gtaactttta aggacgtgtt cgtcgatttt actcgggagg 4080
agtggaaact attggatacc gcccagcaaa tcgtgtatag aaacgttatg ctcgagaact 4140
ataagaatct tgtgtcactg ggctaccaac tgacaaagcc tgacgttatc ctgaggctgg 4200
aaaagggtga ggagccttgg ttggtcgagc gggagatcca ccaggaaact cacccagaca 4260
gtgagactgc tttcgagatc aagtcctctg tgtaatacaa ggcggccgca aatcaacatc 4320
agtctgataa gctatcaaca tcagtctgat aagctatcaa catcagtctg ataagctatc 4380
aacatcagtc tgataagcta aagcttcgaa ttctgataat cagccatacc acatttgtag 4440
aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga 4500
atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata 4560
gcatcacaaa tttcacaaat aaagcatttt tttcactgca gctcgccttt tgctcacatg 4620
tctcccaggg gatgggggtc catccacctt gccgaaaaga tttgtctgag gaactgaaaa 4680
tagaagggaa aaaagaggag ggacaaaaga ggcagaaatg agaggggagg ggacagagga 4740
cacctgaata aagaccacac ccatgaccca cgtgatgctg agaagtactc ctgccctagg 4800
aagagactca gggcagaggg aggaaggaca gcagaccaga cagtcacagc agccttgaca 4860
aaacgttcct ggaactcaag ctcttctcca cagaggagga cagagcagac agcagagacc 4920
atggagtctc cctcggcccc tccccacaga tggtgcatcc cctggcagag gctcctgctc 4980
acaggtgaac tcgagaccgg tatggccccc ccgaccgatg tcagcctggg ggacgagctc 5040
cacttagacg gcgaggacgt ggcgatggcg catgccgacg cgctagacga tttcgatctg 5100
gacatgttgg gggacgggga ttccccgggt ccgggattta ccccccacga ctccgccccc 5160
tacggcgctc tggatatggc cgacttcgag tttgagcaga tgtttaccga tgcccttgga 5220
attgacgagt acggtgggac gcgtatgaag ctactgtctt ctatcgaaca agcatgcgat 5280
atttgccgac ttaaaaagct caagtgctcc aaagaaaaac cgaagtgcgc caagtgtctg 5340
aagaacaact gggagtgtcg ctactctccc aaaaccaaaa ggtctccgct gactagggca 5400
catctgacag aagtggaatc aaggctagaa agactggaac agctatttct actgattttt 5460
cctcgagaag accttgacat gattttgaaa atggattctt tacaggatat aaaagcattg 5520
ttaacaggat tatttgtaca agataatgtg aataaagatg ccgtcacaga tagattggct 5580
tcagtggaga ctgatatgcc tctaacattg agacagcata gaataagtgc gacatcatca 5640
tcggaagaga gtagtaacaa aggtcaaaga cagttgactg tataaaagct tcactcctca 5700
ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc aatgccctgg ctcacaaata 5760
ccactgagat ctttttccct ctgccaaaaa ttatggggac atcatgaagc cccttgagca 5820
tctgacttct ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt 5880
gtgtctctca ctcggaagga catatgggag ggcaaatcat ttaaaacatc agaatgagta 5940
tttggtttag agtttggcaa catatgccca tatgctggct gccatgaaca aaggttggct 6000
ataaagaggt catcagtata tgaaacagcc ccctgctgtc cattccttat tccatagaaa 6060
agccttgact tgaggttaga ttttttttat attttgtttt gtgttatttt tttctttaac 6120
atccctaaaa ttttccttac atgttttact agccagattt ttcctcctct cctgactact 6180
cccagtcata gctgtccctc ttctcttatg gagatc 6216
<210> 14
<211> 12868
<212> DNA
<213> Artificial
<220>
<223> 编码由细胞特异性启动子CEA368、第一microRNA为mir-143-3p、第二micro
RNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白分
别为TALE蛋白、第一和第二识别序列为4×UAS的元件组成的表达系统的核酸
<400> 14
cggagtactg tcctccgagc ggagtactgt cctccgagcg gagtactgtc ctccgagcgg 60
agttctgtcc tccgagcgga gactctagat acccctctcc gcttctgaat tctaggcgtg 120
tacggtggga ggcctatata agcagagctc gtttagtgaa ccgtcagatc gcctcgagta 180
cccctctccg cttcttaccc ctctccgctt cttacccctc tccgcttctg aattcgaccg 240
ccaccatgag acatattatc tgccacggag gtgttattac cgaagaaatg gccgccagtc 300
ttttggacca gctgatcgaa gaggtactgg ctgataatct tccacctcct agccattttg 360
aaccacctac ccttcacgaa ctgtatgatt tagacgtgac ggcccccgaa gatcccaacg 420
aggaggcggt ttcgcagatt tttcccgact ctgtaatgtt ggcggtgcag gaagggattg 480
acttactcac ttttccgccg gcgcccggtt ctccggagcc gcctcacctt tcccggcagc 540
ccgagcagcc ggagcagaga gccttgggtc cggtttctat gccaaacctt gtaccggagg 600
tgatcgatct tacctgccac gaggctggct ttccacccag tgacgacgag gatgaagagg 660
gtgaggagtt tgtgttagat tatgtggagc accccgggca cggttgcagg tcttgtcatt 720
atcaccggag gaatacgggg gacccagata ttatgtgttc gctttgctat atgaggacct 780
gtggcatgtt tgtctacagt aagtgaaaat tatgggcagt gggtgataga gtggtgggtt 840
tggtgtggta attttttttt taatttttac agttttgtgg tttaaagaat tttgtattgt 900
gattttttta aaaggtcctg tgtctgaacc tgagcctgag cccgagccag aaccggagcc 960
tgcaagacct acccgccgtc ctaaaatggc gcctgctatc ctgaggcgcc cgacatcacc 1020
tgtgtctaga gaatgcaata gtagtacgga tagctgtgac tccggtcctt ctaacacacc 1080
tcctgagata cacccggtgg tcccgctgtg ccccattaaa ccagttgccg tgagagttgg 1140
tgggcgtcgc caggctgtgg aatgtatcga ggacttgctt aacgagcctg ggcaaccttt 1200
ggacttgagc tgtaaacgcc ccaggccagg gtccggagct acaaattttt ccctcctcaa 1260
acaggctgga gatgtcgaag aaaatcctgg gcctatggag gcttgggagt gtttggaaga 1320
tttttctgct gtgcgtaact tgctggaaca gagctctaac agtacctctt ggttttggag 1380
gtttctgtgg ggctcatccc aggcaaagtt agtctgcaga attaaggagg attacaagtg 1440
ggaatttgaa gagcttttga aatcctgtgg tgagctgttt gattctttga atctgggtca 1500
ccaggcgctt ttccaagaga aggtcatcaa gactttggat ttttccacac cggggcgcgc 1560
tgcggctgct gttgcttttt tgagttttat aaaggataaa tggagcgaag aaacccatct 1620
gagcgggggg tacctgctgg attttctggc catgcatctg tggagagcgg ttgtgagaca 1680
caagaatcgc ctgctactgt tgtcttccgt ccgcccggcg ataataccga cggaggagca 1740
gcagcagcag caggaggaag ccaggcggcg gcggcaggag cagagcccat ggaacccgag 1800
agccggcctg gaccctcggg aatgaatgtt gtacaggtgg ctgaactgta tccagaactg 1860
agacgcattt tgacaattac agaggatggg caggggctaa agggggtaaa gagggagcgg 1920
ggggcttgtg aggctacaga ggaggctagg aatctagctt ttagcttaat gaccagacac 1980
cgtcctgagt gtattacttt tcaacagatc aaggataatt gcgctaatga gcttgatctg 2040
ctggcgcaga agtattccat agagcagctg accacttact ggctgcagcc aggggatgat 2100
tttgaggagg ctattagggt atatgcaaag gtggcactta ggccagattg caagtacaag 2160
atcagcaaac ttgtaaatat caggaattgt tgctacattt ctgggaacgg ggccgaggtg 2220
gagatagata cggaggatag ggtggccttt agatgtagca tgataaatat gtggccgggg 2280
gtgcttggca tggacggggt ggttattatg aatgtaaggt ttactggccc caattttagc 2340
ggtacggttt tcctggccaa taccaacctt atcctacacg gtgtaagctt ctatgggttt 2400
aacaatacct gtgtggaagc ctggaccgat gtaagggttc ggggctgtgc cttttactgc 2460
tgctggaagg gggtggtgtg tcgccccaaa agcagggctt caattaagaa atgcctcttt 2520
gaaaggtgta ccttgggtat cctgtctgag ggtaactcca gggtgcgcca caatgtggcc 2580
tccgactgtg gttgcttcat gctagtgaaa agcgtggctg tgattaagca taacatggta 2640
tgtggcaact gcgaggacag ggcctctcag atgctgacct gctcggacgg caactgtcac 2700
ctgctgaaga ccattcacgt agccagccac tctcgcaagg cctggccagt gtttgagcat 2760
aacatactga cccgctgttc cttgcatttg ggtaacagga ggggggtgtt cctaccttac 2820
caatgcaatt tgagtcacac taagatattg cttgagcccg agagcatgtc caaggtgaac 2880
ctgaacgggg tgtttgacat gaccatgaag atctggaagg tgctgaggta cgatgagacc 2940
cgcaccaggt gcagaccctg cgagtgtggc ggtaaacata ttaggaacca gcctgtgatg 3000
ctggatgtga ccgaggagct gaggcccgat cacttggtgc tggcctgcac ccgcgctgag 3060
tttggctcta gcgatgaaga tacagatctc gaggggtccg gagctacaaa tttttccctc 3120
ctcaaacagg ctggagatgt cgaagaaaat cctgggccta ccggtatgta taggatgcag 3180
ttgctctctt gcatcgccct gtctctagcg ttggtgacta atagcgcccc tacttcttct 3240
tccacgaaga aaacccagct gcaactggaa caccttctgc tggatctcca gatgatcctt 3300
aacgggatta acaactataa gaacccgaag ctcacggcta tgctgacggc gaagttcgct 3360
atgcctaaaa aggctactga gctgaaacac ttacagtgtc tcgaagaggc actcaaaccc 3420
ctcgaggagg tactcaatct ggcgcaaagc aaaaacttcc atctccgccc acgggaccta 3480
atctccaaca tcaacgtgat tgtgctggag ctcaaggggt ctgaaaccac gtttatgtgc 3540
gagtacgcag atgaaactgc taccatagtg gaattcctca atcgttggat cactttctgc 3600
cagtcaatca tttcaaccct gaccagatct gagggccgcg gcagcctgct gacctgcggc 3660
gacgtggagg aaaaccccgg ccccggatcc ttgtcgacca ttcgtccgcg caggccaagt 3720
cctgcccgcg agcttctgcc cggaccccaa ccggataggg ttcagccgac tgcagatcgt 3780
ggggtgtctg cgcctgctgg cagccctctg gatggcttgc ccgctcggcg gacggtgtcc 3840
cggacccggc tgccatctcc ccctgcgccc tcacctgcgt tctcggcggg cagcttcagc 3900
gatctgctcc gtccgttcga tccgtcgctt cttgatacat cgcttcttga ttcgatgcct 3960
gccgtcggca cgccgcatac agcggctgcc ccagcagagt gggatgaggc gcaatcggct 4020
ctgcgtgcag ccgatgaccc gccacccacc gtgcgtgtcg ctgtcactgc cgcgcggccg 4080
ccgcgcgcca agccggcccc gcgacggcgt gctgcgcaac cctccgacgc ttcgccggcc 4140
gcgcaggtgg atctacgcac gctcggctac agtcagcagc agcaagagaa gatcaaaccg 4200
aaggtgcgtt cgacagtggc gcagcaccac gaggcactgg tgggccatgg gtttacacac 4260
gcgcacatcg ttgcgctcag ccaacacccg gcagcgttag ggaccgtcgc tgtcacgtat 4320
cagcacataa tcacggcgtt gccagaggcg acacacgaag acatcgttgg cgtcggcaaa 4380
cagtggtccg gcgcacgcgc cctggaggcc ttgctcacgg atgcggggga gttgagaggt 4440
ccgccgttac agttggacac aggccaactt gtgaagattg caaaacgtgg cggcgtgacc 4500
gcaatggagg cagtgcatgc atcgcgcaat gcactgacgg gtgcccccct gaacctgacc 4560
ccggaccaag tggtggctat cgccagcaac attggcggca agcaagcgct cgaaacggtg 4620
cagcggctgt tgccggtgct gtgccaggac catggcctga ccccggacca agtggtggct 4680
atcgccagca acggtggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg 4740
ctgtgccagg accatggcct gaccccggac caagtggtgg ctatcgccag caacattggc 4800
ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc 4860
ctgaccccgg accaagtggt ggctatcgcc agcaacaatg gcggcaagca agcgctcgaa 4920
acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg gcctgacccc ggaccaagtg 4980
gtggctatcg ccagcaacat tggcggcaag caagcgctcg aaacggtgca gcggctgttg 5040
ccggtgctgt gccaggacca tggcctgacc ccggaccaag tggtggctat cgccagcaac 5100
attggcggca agcaagcgct cgaaacggtg cagcggctgt tgccggtgct gtgccaggac 5160
catggcctga ctccggacca agtggtggct atcgccagcc acgatggcgg caagcaagcg 5220
ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg accatggcct gactccggac 5280
caagtggtgg ctatcgccag ccacgatggc ggcaagcaag cgctcgaaac ggtgcagcgg 5340
ctgttgccgg tgctgtgcca ggaccatggc ctgaccccgg accaagtggt ggctatcgcc 5400
agcaacaatg gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc 5460
caggaccatg gcctgacccc ggaccaagtg gtggctatcg ccagcaacat tggcggcaag 5520
caagcgctcg aaacggtgca gcggctgttg ccggtgctgt gccaggacca tggcctgacc 5580
ccggaccaag tggtggctat cgccagcaac ggtggcggca agcaagcgct cgaaacggtg 5640
cagcggctgt tgccggtgct gtgccaggac catggcctga ctccggacca agtggtggct 5700
atcgccagcc acgatggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg 5760
ctgtgccagg accatggcct gactccggac caagtggtgg ctatcgccag ccacgatggc 5820
ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc 5880
ctgaccccgg accaagtggt ggctatcgcc agcaacggtg gcggcaagca agcgctcgaa 5940
acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg gcctgactcc ggaccaagtg 6000
gtggctatcg ccagccacga tggcggcaag caagcgctcg aaacggtgca gcggctgttg 6060
ccggtgctgt gccaggacca tggcctgact ccggaccaag tggtggctat cgccagccac 6120
gatggcggca agcaagcgct cgaaacggtg cagcggctgt tgccggtgct gtgccaggac 6180
catggcctga ctccggacca agtggtggct atcgccagcc acgatggcgg caagcaagcg 6240
ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg accatggcct gaccccggac 6300
caagtggtgg ctatcgccag caacattggc ggcaagcaag cgctcgaaac ggtgcagcgg 6360
ctgttgccgg tgctgtgcca ggaccatggc ctgaccccgg accaagtggt ggctatcgcc 6420
agcaacggtg gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc 6480
caggaccatg gcctgacccc ggaccaagtg gtggctatcg ccagcaacgg tggcggcaag 6540
caagcgctcg aaagcattgt ggcccagctg agccggcctg atccggcgtt ggccgcgttg 6600
accaacgacc acctcgtcgc cttggcctgc ctcggcggac gtcctgccat ggatgcagtg 6660
aaaaagggat tgccgcacgc gccggaattg atcagaagag tcaatcgccg tattggcgaa 6720
cgcacgtccc atcgcgttgc cgactacgcg caagtggttc gcgtgctgga gtttttccag 6780
tgccactccc acccagcgta cgcatttgat gaggccatga cgcagttcgg gatgagcagg 6840
aacgggttgg tacagctctt tcgcagagtg ggcgtcaccg aactcgaagc ccgcggtgga 6900
acgctccccc cagcctcgca gcgttgggac cgtatcctcc aggcatcagg gatgaaaagg 6960
gccaaaccgt cccctacttc agctcaaaca ccggatcagg cgtctttgca tgcattcgcc 7020
gattcgctgg agcgtgacct tgatgcgccc agcccaatgc acgagggaga tcagacgcgg 7080
gcaagcagcc gtaaacggtc ccgatcggat cgtgctgtca ccggcccctc cgcacagcag 7140
gctgtcgagg tgcgcgttcc cgaacagcgc gatgcgctgc atttgcccct cagctggagg 7200
gtaaaacgcc cgcgtaccag gatctggggc ggcctcccgg ttaagtagtg ctttctactt 7260
tatgagtagt gctttctact ttatgagtag tgctttctac tttatgagta gtgctttcta 7320
ctttatggag ctacagtgct tcatctcaga gctacagtgc ttcatctcag agctacagtg 7380
cttcatctca gagctacagt gcttcatctc agaaagcgga gtactgtcct ccgagcggag 7440
tactgtcctc cgagcggagt actgtcctcc gagcggagtt ctgtcctccg agcggagact 7500
ctagatatag aaccgatcct cccattgaat tctaggcgtg tacggtggga ggcctatata 7560
agcagagctc gtttagtgaa ccgtcagatc gcctcgagta tagaaccgat cctcccattt 7620
atagaaccga tcctcccatt tatagaaccg atcctcccat tactagaaga tcttgaattc 7680
gccaccttgt cgaccattcg tccgcgcagg ccaagtcctg cccgcgagct tctgcccgga 7740
ccccaaccgg atagggttca gccgactgca gatcgtgggg tgtctgcgcc tgctggcagc 7800
cctctggatg gcttgcccgc tcggcggacg gtgtcccgga cccggctgcc atctccccct 7860
gcgccctcac ctgcgttctc ggcgggcagc ttcagcgatc tgctccgtcc gttcgatccg 7920
tcgcttcttg atacatcgct tcttgattcg atgcctgccg tcggcacgcc gcatacagcg 7980
gctgccccag cagagtggga tgaggcgcaa tcggctctgc gtgcagccga tgacccgcca 8040
cccaccgtgc gtgtcgctgt cactgccgcg cggccgccgc gcgccaagcc ggccccgcga 8100
cggcgtgctg cgcaaccctc cgacgcttcg ccggccgcgc aggtggatct acgcacgctc 8160
ggctacagtc agcagcagca agagaagatc aaaccgaagg tgcgttcgac agtggcgcag 8220
caccacgagg cactggtggg ccatgggttt acacacgcgc acatcgttgc gctcagccaa 8280
cacccggcag cgttagggac cgtcgctgtc acgtatcagc acataatcac ggcgttgcca 8340
gaggcgacac acgaagacat cgttggcgtc ggcaaacagt ggtccggcgc acgcgccctg 8400
gaggccttgc tcacggatgc gggggagttg agaggtccgc cgttacagtt ggacacaggc 8460
caacttgtga agattgcaaa acgtggcggc gtgaccgcaa tggaggcagt gcatgcatcg 8520
cgcaatgcac tgacgggtgc ccccctgaac ctgaccccgg accaagtggt ggctatcgcc 8580
agcaacattg gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc 8640
caggaccatg gcctgactcc ggaccaagtg gtggctatcg ccagccacga tggcggcaag 8700
caagcgctcg aaacggtgca gcggctgttg ccggtgctgt gccaggacca tggcctgact 8760
ccggaccaag tggtggctat cgccagccac gatggcggca agcaagcgct cgaaacggtg 8820
cagcggctgt tgccggtgct gtgccaggac catggcctga ctccggacca agtggtggct 8880
atcgccagcc acgatggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg 8940
ctgtgccagg accatggcct gactccggac caagtggtgg ctatcgccag ccacgatggc 9000
ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc 9060
ctgaccccgg accaagtggt ggctatcgcc agcaacggtg gcggcaagca agcgctcgaa 9120
acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg gcctgactcc ggaccaagtg 9180
gtggctatcg ccagccacga tggcggcaag caagcgctcg aaacggtgca gcggctgttg 9240
ccggtgctgt gccaggacca tggcctgacc ccggaccaag tggtggctat cgccagcaac 9300
ggtggcggca agcaagcgct cgaaacggtg cagcggctgt tgccggtgct gtgccaggac 9360
catggcctga ctccggacca agtggtggct atcgccagcc acgatggcgg caagcaagcg 9420
ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg accatggcct gactccggac 9480
caagtggtgg ctatcgccag ccacgatggc ggcaagcaag cgctcgaaac ggtgcagcgg 9540
ctgttgccgg tgctgtgcca ggaccatggc ctgaccccgg accaagtggt ggctatcgcc 9600
agcaacaatg gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc 9660
caggaccatg gcctgactcc ggaccaagtg gtggctatcg ccagccacga tggcggcaag 9720
caagcgctcg aaacggtgca gcggctgttg ccggtgctgt gccaggacca tggcctgacc 9780
ccggaccaag tggtggctat cgccagcaac ggtggcggca agcaagcgct cgaaacggtg 9840
cagcggctgt tgccggtgct gtgccaggac catggcctga ccccggacca agtggtggct 9900
atcgccagca acggtggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg 9960
ctgtgccagg accatggcct gactccggac caagtggtgg ctatcgccag ccacgatggc 10020
ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc 10080
ctgaccccgg accaagtggt ggctatcgcc agcaacggtg gcggcaagca agcgctcgaa 10140
agcattgtgg cccagctgag ccggcctgat ccggcgttgg ccgcgttgac caacgaccac 10200
ctcgtcgcct tggcctgcct cggcggacgt cctgccatgg atgcagtgaa aaagggattg 10260
ccgcacgcgc cggaattgat cagaagagtc aatcgccgta ttggcgaacg cacgtcccat 10320
cgcgttgccg actacgcgca agtggttcgc gtgctggagt ttttccagtg ccactcccac 10380
ccagcgtacg catttgatga ggccatgacg cagttcggga tgagcaggaa cgggttggta 10440
cagctctttc gcagagtggg cgtcaccgaa ctcgaagccc gcggtggaac gctcccccca 10500
gcctcgcagc gttgggaccg tatcctccag gcatcaggga tgaaaagggc caaaccgtcc 10560
cctacttcag ctcaaacacc ggatcaggcg tctttgcatg cattcgccga ttcgctggag 10620
cgtgaccttg atgcgcccag cccaatgcac gagggagatc agacgcgggc aagcagccgt 10680
aaacggtccc gatcggatcg tgctgtcacc ggcccctccg cacagcaggc tgtcgaggtg 10740
cgcgttcccg aacagcgcga tgcgctgcat ttgcccctca gctggagggt aaaacgcccg 10800
cgtaccagga tctggggcgg cctcccgtaa tacaaggcgg ccgcaaatca acatcagtct 10860
gataagctat caacatcagt ctgataagct atcaacatca gtctgataag ctatcaacat 10920
cagtctgata agctaaagct tcgaattctg ataatcagcc ataccacatt tgtagaggtt 10980
ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca 11040
attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc 11100
acaaatttca caaataaagc atttttttca ctgcagctcg ccttttgctc acatgtctcc 11160
caggggatgg gggtccatcc accttgccga aaagatttgt ctgaggaact gaaaatagaa 11220
gggaaaaaag aggagggaca aaagaggcag aaatgagagg ggaggggaca gaggacacct 11280
gaataaagac cacacccatg acccacgtga tgctgagaag tactcctgcc ctaggaagag 11340
actcagggca gagggaggaa ggacagcaga ccagacagtc acagcagcct tgacaaaacg 11400
ttcctggaac tcaagctctt ctccacagag gaggacagag cagacagcag agaccatgga 11460
gtctccctcg gcccctcccc acagatggtg catcccctgg cagaggctcc tgctcacagg 11520
tgaactcgag accggtatgg cccccccgac cgatgtcagc ctgggggacg agctccactt 11580
agacggcgag gacgtggcga tggcgcatgc cgacgcgcta gacgatttcg atctggacat 11640
gttggggagc gacgcgctag acgatttcga tctggacatg ttggggagcg acgcgctaga 11700
cgatttcgat ctggacatgt tggggagcga cgcgctagac gatttcgatc tggacatgtt 11760
gggggacggg gattccccgg gtccgggatt taccccccac gactccgccc cctacggcgc 11820
tctggatatg gccgacttcg agtttgagca gatgtttacc gatgcccttg gaattgacga 11880
gtacggtggg acgcgtatga agctactgtc ttctatcgaa caagcatgcg atatttgccg 11940
acttaaaaag ctcaagtgct ccaaagaaaa accgaagtgc gccaagtgtc tgaagaacaa 12000
ctgggagtgt cgctactctc ccaaaaccaa aaggtctccg ctgactaggg cacatctgac 12060
agaagtggaa tcaaggctag aaagactgga acagctattt ctactgattt ttcctcgaga 12120
agaccttgac atgattttga aaatggattc tttacaggat ataaaagcat tgttaacagg 12180
attatttgta caagataatg tgaataaaga tgccgtcaca gatagattgg cttcagtgga 12240
gactgatatg cctctaacat tgagacagca tagaataagt gcgacatcat catcggaaga 12300
gagtagtaac aaaggtcaaa gacagttgac tgtataaaag cttcactcct caggtgcagg 12360
ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa taccactgag 12420
atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt 12480
ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct 12540
cactcggaag gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt 12600
agagtttggc aacatatgcc catatgctgg ctgccatgaa caaaggttgg ctataaagag 12660
gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga 12720
cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta acatccctaa 12780
aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta ctcccagtca 12840
tagctgtccc tcttctctta tggagatc 12868
<210> 15
<211> 6127
<212> DNA
<213> Artificial
<220>
<223> 编码由细胞特异性启动子CXCR4、第一microRNA为mir-135a-5p、第二micro
RNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白分
别为Lacl、tetR-KRAB、第一和第二识别序列为4×UAS的元件组成的表达系
统的核酸
<400> 15
cgggtgaccg ccctccgcgg agtactgtcc tccgcgggtg acagccctcc gcggagtgcc 60
gcactccgag cggagactct agactcccta tcagtgatag agataggcgt gtacggtggg 120
aggcctatat aagcagagct cgtttagtga accgtcagat cgctccctat cagtgataga 180
gagaattcga ccgccaccat gagacatatt atctgccacg gaggtgttat taccgaagaa 240
atggccgcca gtcttttgga ccagctgatc gaagaggtac tggctgataa tcttccacct 300
cctagccatt ttgaaccacc tacccttcac gaactgtatg atttagacgt gacggccccc 360
gaagatccca acgaggaggc ggtttcgcag atttttcccg actctgtaat gttggcggtg 420
caggaaggga ttgacttact cacttttccg ccggcgcccg gttctccgga gccgcctcac 480
ctttcccggc agcccgagca gccggagcag agagccttgg gtccggtttc tatgccaaac 540
cttgtaccgg aggtgatcga tcttacctgc cacgaggctg gctttccacc cagtgacgac 600
gaggatgaag agggtgagga gtttgtgtta gattatgtgg agcaccccgg gcacggttgc 660
aggtcttgtc attatcaccg gaggaatacg ggggacccag atattatgtg ttcgctttgc 720
tatatgagga cctgtggcat gtttgtctac agtaagtgaa aattatgggc agtgggtgat 780
agagtggtgg gtttggtgtg gtaatttttt ttttaatttt tacagttttg tggtttaaag 840
aattttgtat tgtgattttt ttaaaaggtc ctgtgtctga acctgagcct gagcccgagc 900
cagaaccgga gcctgcaaga cctacccgcc gtcctaaaat ggcgcctgct atcctgaggc 960
gcccgacatc acctgtgtct agagaatgca atagtagtac ggatagctgt gactccggtc 1020
cttctaacac acctcctgag atacacccgg tggtcccgct gtgccccatt aaaccagttg 1080
ccgtgagagt tggtgggcgt cgccaggctg tggaatgtat cgaggacttg cttaacgagc 1140
ctgggcaacc tttggacttg agctgtaaac gccccaggcc actcgagggt accgggtccg 1200
gagctacaaa tttttccctc ctcaaacagg ctggagatgt cgaagaaaat cctgggccta 1260
ccggtatgtg gctgcagagc ctccttctac tgggcaccgt ggcttgcagc atctcggccc 1320
ccgccagaag tcccagcccc agcacccagc cctgggaaca cgtaaatgca atccaggagg 1380
cccgccgcct cctgaacttg tcacgggaca cggcagccga gatgaatgaa actgtggagg 1440
tcatcagtga aatgtttgac ttgcaggagc ccacatgcct gcagactcgg ctggagctct 1500
acaaacaggg cctgcgtggc agcctcacca agctgaaggg cccactgacc atgatggcta 1560
gccattacaa gcagcactgc ccgccgaccc ctgagacatc ctgtgccact cagatcatta 1620
cctttgagtc tttcaaggag aacctcaagg actttctgct ggtcatccca tttgactgct 1680
gggaacccgt ccaggaaaga tctgagggcc gcggcagcct gctgacctgc ggcgacgtgg 1740
aggaaaaccc cggccccgga tccatgaaac cagtaacgtt atacgatgtc gcagagtatg 1800
ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac gtttctgcga 1860
aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg 1920
cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc agtctggccc 1980
tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca 2040
gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca 2100
atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg 2160
ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc 2220
agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg ggcgtggagc 2280
atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct 2340
cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga 2400
tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc 2460
tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg 2520
caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggatatctcg gtagtgggat 2580
acgacgatac cgaagacagc tcatgttata tcccgccgtt aaccaccatc aaacaggatt 2640
ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg 2700
tgaagggcaa tcagctgttg ccggtctcac tggtgaaaag aaaaaccacc ctggcgccca 2760
atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg 2820
tttcccgact ggaaagcggg cagagcagcc tgaggcctcc taagaagaag aggaaggttg 2880
ttacaaacac cattgtcaca ctccacaaac accattgtca cactccacaa acaccattgt 2940
cacactccac aaacaccatt gtcacactcc atcacatagg aataaaaagc catatcacat 3000
aggaataaaa agccatatca cataggaata aaaagccata tcacatagga ataaaaagcc 3060
atagaaagcg gggtgccgcc ccccgcggac tactgtagtc cgcggacgac tgtcgtccgc 3120
ggagcactgt gctccgagcg gagactctag aaattgtgag cggataacaa ttgggtatat 3180
aatgggggcc gtcagctact accagagccg atcgaacact cgagccgagc agacgtgcct 3240
acaattgtga gcgctcacaa ttactagaag atcttgaatt cgccaccatg gctagattag 3300
ataaaagtaa agtgattaac agcgcattag agctgcttaa tgaggtcgga atcgaaggtt 3360
taacaacccg taaactcgcc cagaagctag gtgtagagca gcctacattg tattggcatg 3420
taaaaaataa gcgggctttg ctcgacgcct tagccattga gatgttagat aggcaccata 3480
ctcacttttg ccctttagaa ggggaaagct ggcaagattt tttacgtaat aacgctaaaa 3540
gttttagatg tgctttacta agtcatcgcg atggagcaaa agtacattta ggtacacggc 3600
ctacagaaaa acagtatgaa actctcgaaa atcaattagc ctttttatgc caacaaggtt 3660
tttcactaga gaatgcatta tatgcactca gcgctgtggg gcattttact ttaggttgcg 3720
tattggaaga tcaagagcat caagtcgcta aagaagaaag ggaaacacct actactgata 3780
gtatgccgcc attattacga caagctatcg aattatttga tcaccaaggt gcagagccag 3840
ccttcttatt cggccttgaa ttgatcatat gcggattaga aaaacaactt aaatgtgaaa 3900
gtgggtcgcc aaaaaagaag agaaaggtgg atggtggcgg agcactgtct ccccagcatt 3960
cagctgtgac ccagggatca atcattaaga acaaggaagg tatggatgct aagtccctga 4020
ccgcttggtc aagaactttg gtaactttta aggacgtgtt cgtcgatttt actcgggagg 4080
agtggaaact attggatacc gcccagcaaa tcgtgtatag aaacgttatg ctcgagaact 4140
ataagaatct tgtgtcactg ggctaccaac tgacaaagcc tgacgttatc ctgaggctgg 4200
aaaagggtga ggagccttgg ttggtcgagc gggagatcca ccaggaaact cacccagaca 4260
gtgagactgc tttcgagatc aagtcctctg tgtaatacaa ggcggccgca aatcaacatc 4320
agtctgataa gctatcaaca tcagtctgat aagctatcaa catcagtctg ataagctatc 4380
aacatcagtc tgataagcta aagcttcgaa ttctgataat cagccatacc acatttgtag 4440
aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga 4500
atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata 4560
gcatcacaaa tttcacaaat aaagcatttt tttcactgca gctcgccttt tgctcacatg 4620
ttaccgacca cccgcaaaca gcagggtccc ctgggcttcc caagccgcgc acctctccgc 4680
cccgcccctg cgccctcctt cctcgcgtct gcccctctcc cccaccccgc cttctccctc 4740
cccgccccag cggcgcatgc gccgcgctcg gagcgtgttt ttataaaagt ccggccgcgg 4800
ccagaaactt cagtttgttg gctgcggcag caggtagcaa agtgacgccg agggcctgag 4860
tgctccagta gccaccgcat ctggagaacc agcggttacc ctcgagaccg gtatggcccc 4920
cccgaccgat gtcagcctgg gggacgagct ccacttagac ggcgaggacg tggcgatggc 4980
gcatgccgac gcgctagacg atttcgatct ggacatgttg ggggacgggg attccccggg 5040
tccgggattt accccccacg actccgcccc ctacggcgct ctggatatgg ccgacttcga 5100
gtttgagcag atgtttaccg atgcccttgg aattgacgag tacggtggga cgcgtatgaa 5160
gctactgtct tctatcgaac aagcatgcga tatttgccga cttaaaaagc tcaagtgctc 5220
caaagaaaaa ccgaagtgcg ccaagtgtct gaagaacaac tgggagtgtc gctactctcc 5280
caaaaccaaa aggtctccgc tgactagggc acatctgaca gaagtggaat caaggctaga 5340
aagactggaa cagctatttc tactgatttt tcctcgagaa gaccttgaca tgattttgaa 5400
aatggattct ttacaggata taaaagcatt gttaacagga ttatttgtac aagataatgt 5460
gaataaagat gccgtcacag atagattggc ttcagtggag actgatatgc ctctaacatt 5520
gagacagcat agaataagtg cgacatcatc atcggaagag agtagtaaca aaggtcaaag 5580
acagttgact gtataaaagc ttcactcctc aggtgcaggc tgcctatcag aaggtggtgg 5640
ctggtgtggc caatgccctg gctcacaaat accactgaga tctttttccc tctgccaaaa 5700
attatgggga catcatgaag ccccttgagc atctgacttc tggctaataa aggaaattta 5760
ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga 5820
gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca acatatgccc 5880
atatgctggc tgccatgaac aaaggttggc tataaagagg tcatcagtat atgaaacagc 5940
cccctgctgt ccattcctta ttccatagaa aagccttgac ttgaggttag atttttttta 6000
tattttgttt tgtgttattt ttttctttaa catccctaaa attttcctta catgttttac 6060
tagccagatt tttcctcctc tcctgactac tcccagtcat agctgtccct cttctcttat 6120
ggagatc 6127
<210> 16
<211> 12745
<212> DNA
<213> Artificial
<220>
<223> 编码由细胞特异性启动子CXCR4、第一microRNA为mir-143-3p、第二microR
NA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白分别
为TALE蛋白、第一和第二识别序列为4×UAS的元件组成的表达系统的核酸
<400> 16
cggagtactg tcctccgagc ggagtactgt cctccgagcg gagtactgtc ctccgagcgg 60
agttctgtcc tccgagcgga gactctagat acccctctcc gcttctgaat tctaggcgtg 120
tacggtggga ggcctatata agcagagctc gtttagtgaa ccgtcagatc gcctcgagta 180
cccctctccg cttcttaccc ctctccgctt cttacccctc tccgcttctg aattcgaccg 240
ccaccatgag acatattatc tgccacggag gtgttattac cgaagaaatg gccgccagtc 300
ttttggacca gctgatcgaa gaggtactgg ctgataatct tccacctcct agccattttg 360
aaccacctac ccttcacgaa ctgtatgatt tagacgtgac ggcccccgaa gatcccaacg 420
aggaggcggt ttcgcagatt tttcccgact ctgtaatgtt ggcggtgcag gaagggattg 480
acttactcac ttttccgccg gcgcccggtt ctccggagcc gcctcacctt tcccggcagc 540
ccgagcagcc ggagcagaga gccttgggtc cggtttctat gccaaacctt gtaccggagg 600
tgatcgatct tacctgccac gaggctggct ttccacccag tgacgacgag gatgaagagg 660
gtgaggagtt tgtgttagat tatgtggagc accccgggca cggttgcagg tcttgtcatt 720
atcaccggag gaatacgggg gacccagata ttatgtgttc gctttgctat atgaggacct 780
gtggcatgtt tgtctacagt aagtgaaaat tatgggcagt gggtgataga gtggtgggtt 840
tggtgtggta attttttttt taatttttac agttttgtgg tttaaagaat tttgtattgt 900
gattttttta aaaggtcctg tgtctgaacc tgagcctgag cccgagccag aaccggagcc 960
tgcaagacct acccgccgtc ctaaaatggc gcctgctatc ctgaggcgcc cgacatcacc 1020
tgtgtctaga gaatgcaata gtagtacgga tagctgtgac tccggtcctt ctaacacacc 1080
tcctgagata cacccggtgg tcccgctgtg ccccattaaa ccagttgccg tgagagttgg 1140
tgggcgtcgc caggctgtgg aatgtatcga ggacttgctt aacgagcctg ggcaaccttt 1200
ggacttgagc tgtaaacgcc ccaggccagg gtccggagct acaaattttt ccctcctcaa 1260
acaggctgga gatgtcgaag aaaatcctgg gcctatggag gcttgggagt gtttggaaga 1320
tttttctgct gtgcgtaact tgctggaaca gagctctaac agtacctctt ggttttggag 1380
gtttctgtgg ggctcatccc aggcaaagtt agtctgcaga attaaggagg attacaagtg 1440
ggaatttgaa gagcttttga aatcctgtgg tgagctgttt gattctttga atctgggtca 1500
ccaggcgctt ttccaagaga aggtcatcaa gactttggat ttttccacac cggggcgcgc 1560
tgcggctgct gttgcttttt tgagttttat aaaggataaa tggagcgaag aaacccatct 1620
gagcgggggg tacctgctgg attttctggc catgcatctg tggagagcgg ttgtgagaca 1680
caagaatcgc ctgctactgt tgtcttccgt ccgcccggcg ataataccga cggaggagca 1740
gcagcagcag caggaggaag ccaggcggcg gcggcaggag cagagcccat ggaacccgag 1800
agccggcctg gaccctcggg aatgaatgtt gtacaggtgg ctgaactgta tccagaactg 1860
agacgcattt tgacaattac agaggatggg caggggctaa agggggtaaa gagggagcgg 1920
ggggcttgtg aggctacaga ggaggctagg aatctagctt ttagcttaat gaccagacac 1980
cgtcctgagt gtattacttt tcaacagatc aaggataatt gcgctaatga gcttgatctg 2040
ctggcgcaga agtattccat agagcagctg accacttact ggctgcagcc aggggatgat 2100
tttgaggagg ctattagggt atatgcaaag gtggcactta ggccagattg caagtacaag 2160
atcagcaaac ttgtaaatat caggaattgt tgctacattt ctgggaacgg ggccgaggtg 2220
gagatagata cggaggatag ggtggccttt agatgtagca tgataaatat gtggccgggg 2280
gtgcttggca tggacggggt ggttattatg aatgtaaggt ttactggccc caattttagc 2340
ggtacggttt tcctggccaa taccaacctt atcctacacg gtgtaagctt ctatgggttt 2400
aacaatacct gtgtggaagc ctggaccgat gtaagggttc ggggctgtgc cttttactgc 2460
tgctggaagg gggtggtgtg tcgccccaaa agcagggctt caattaagaa atgcctcttt 2520
gaaaggtgta ccttgggtat cctgtctgag ggtaactcca gggtgcgcca caatgtggcc 2580
tccgactgtg gttgcttcat gctagtgaaa agcgtggctg tgattaagca taacatggta 2640
tgtggcaact gcgaggacag ggcctctcag atgctgacct gctcggacgg caactgtcac 2700
ctgctgaaga ccattcacgt agccagccac tctcgcaagg cctggccagt gtttgagcat 2760
aacatactga cccgctgttc cttgcatttg ggtaacagga ggggggtgtt cctaccttac 2820
caatgcaatt tgagtcacac taagatattg cttgagcccg agagcatgtc caaggtgaac 2880
ctgaacgggg tgtttgacat gaccatgaag atctggaagg tgctgaggta cgatgagacc 2940
cgcaccaggt gcagaccctg cgagtgtggc ggtaaacata ttaggaacca gcctgtgatg 3000
ctggatgtga ccgaggagct gaggcccgat cacttggtgc tggcctgcac ccgcgctgag 3060
tttggctcta gcgatgaaga tacagatctc gaggggtccg gagctacaaa tttttccctc 3120
ctcaaacagg ctggagatgt cgaagaaaat cctgggccta ccggtatgta taggatgcag 3180
ttgctctctt gcatcgccct gtctctagcg ttggtgacta atagcgcccc tacttcttct 3240
tccacgaaga aaacccagct gcaactggaa caccttctgc tggatctcca gatgatcctt 3300
aacgggatta acaactataa gaacccgaag ctcacggcta tgctgacggc gaagttcgct 3360
atgcctaaaa aggctactga gctgaaacac ttacagtgtc tcgaagaggc actcaaaccc 3420
ctcgaggagg tactcaatct ggcgcaaagc aaaaacttcc atctccgccc acgggaccta 3480
atctccaaca tcaacgtgat tgtgctggag ctcaaggggt ctgaaaccac gtttatgtgc 3540
gagtacgcag atgaaactgc taccatagtg gaattcctca atcgttggat cactttctgc 3600
cagtcaatca tttcaaccct gaccagatct gagggccgcg gcagcctgct gacctgcggc 3660
gacgtggagg aaaaccccgg ccccggatcc ttgtcgacca ttcgtccgcg caggccaagt 3720
cctgcccgcg agcttctgcc cggaccccaa ccggataggg ttcagccgac tgcagatcgt 3780
ggggtgtctg cgcctgctgg cagccctctg gatggcttgc ccgctcggcg gacggtgtcc 3840
cggacccggc tgccatctcc ccctgcgccc tcacctgcgt tctcggcggg cagcttcagc 3900
gatctgctcc gtccgttcga tccgtcgctt cttgatacat cgcttcttga ttcgatgcct 3960
gccgtcggca cgccgcatac agcggctgcc ccagcagagt gggatgaggc gcaatcggct 4020
ctgcgtgcag ccgatgaccc gccacccacc gtgcgtgtcg ctgtcactgc cgcgcggccg 4080
ccgcgcgcca agccggcccc gcgacggcgt gctgcgcaac cctccgacgc ttcgccggcc 4140
gcgcaggtgg atctacgcac gctcggctac agtcagcagc agcaagagaa gatcaaaccg 4200
aaggtgcgtt cgacagtggc gcagcaccac gaggcactgg tgggccatgg gtttacacac 4260
gcgcacatcg ttgcgctcag ccaacacccg gcagcgttag ggaccgtcgc tgtcacgtat 4320
cagcacataa tcacggcgtt gccagaggcg acacacgaag acatcgttgg cgtcggcaaa 4380
cagtggtccg gcgcacgcgc cctggaggcc ttgctcacgg atgcggggga gttgagaggt 4440
ccgccgttac agttggacac aggccaactt gtgaagattg caaaacgtgg cggcgtgacc 4500
gcaatggagg cagtgcatgc atcgcgcaat gcactgacgg gtgcccccct gaacctgacc 4560
ccggaccaag tggtggctat cgccagcaac attggcggca agcaagcgct cgaaacggtg 4620
cagcggctgt tgccggtgct gtgccaggac catggcctga ccccggacca agtggtggct 4680
atcgccagca acggtggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg 4740
ctgtgccagg accatggcct gaccccggac caagtggtgg ctatcgccag caacattggc 4800
ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc 4860
ctgaccccgg accaagtggt ggctatcgcc agcaacaatg gcggcaagca agcgctcgaa 4920
acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg gcctgacccc ggaccaagtg 4980
gtggctatcg ccagcaacat tggcggcaag caagcgctcg aaacggtgca gcggctgttg 5040
ccggtgctgt gccaggacca tggcctgacc ccggaccaag tggtggctat cgccagcaac 5100
attggcggca agcaagcgct cgaaacggtg cagcggctgt tgccggtgct gtgccaggac 5160
catggcctga ctccggacca agtggtggct atcgccagcc acgatggcgg caagcaagcg 5220
ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg accatggcct gactccggac 5280
caagtggtgg ctatcgccag ccacgatggc ggcaagcaag cgctcgaaac ggtgcagcgg 5340
ctgttgccgg tgctgtgcca ggaccatggc ctgaccccgg accaagtggt ggctatcgcc 5400
agcaacaatg gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc 5460
caggaccatg gcctgacccc ggaccaagtg gtggctatcg ccagcaacat tggcggcaag 5520
caagcgctcg aaacggtgca gcggctgttg ccggtgctgt gccaggacca tggcctgacc 5580
ccggaccaag tggtggctat cgccagcaac ggtggcggca agcaagcgct cgaaacggtg 5640
cagcggctgt tgccggtgct gtgccaggac catggcctga ctccggacca agtggtggct 5700
atcgccagcc acgatggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg 5760
ctgtgccagg accatggcct gactccggac caagtggtgg ctatcgccag ccacgatggc 5820
ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc 5880
ctgaccccgg accaagtggt ggctatcgcc agcaacggtg gcggcaagca agcgctcgaa 5940
acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg gcctgactcc ggaccaagtg 6000
gtggctatcg ccagccacga tggcggcaag caagcgctcg aaacggtgca gcggctgttg 6060
ccggtgctgt gccaggacca tggcctgact ccggaccaag tggtggctat cgccagccac 6120
gatggcggca agcaagcgct cgaaacggtg cagcggctgt tgccggtgct gtgccaggac 6180
catggcctga ctccggacca agtggtggct atcgccagcc acgatggcgg caagcaagcg 6240
ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg accatggcct gaccccggac 6300
caagtggtgg ctatcgccag caacattggc ggcaagcaag cgctcgaaac ggtgcagcgg 6360
ctgttgccgg tgctgtgcca ggaccatggc ctgaccccgg accaagtggt ggctatcgcc 6420
agcaacggtg gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc 6480
caggaccatg gcctgacccc ggaccaagtg gtggctatcg ccagcaacgg tggcggcaag 6540
caagcgctcg aaagcattgt ggcccagctg agccggcctg atccggcgtt ggccgcgttg 6600
accaacgacc acctcgtcgc cttggcctgc ctcggcggac gtcctgccat ggatgcagtg 6660
aaaaagggat tgccgcacgc gccggaattg atcagaagag tcaatcgccg tattggcgaa 6720
cgcacgtccc atcgcgttgc cgactacgcg caagtggttc gcgtgctgga gtttttccag 6780
tgccactccc acccagcgta cgcatttgat gaggccatga cgcagttcgg gatgagcagg 6840
aacgggttgg tacagctctt tcgcagagtg ggcgtcaccg aactcgaagc ccgcggtgga 6900
acgctccccc cagcctcgca gcgttgggac cgtatcctcc aggcatcagg gatgaaaagg 6960
gccaaaccgt cccctacttc agctcaaaca ccggatcagg cgtctttgca tgcattcgcc 7020
gattcgctgg agcgtgacct tgatgcgccc agcccaatgc acgagggaga tcagacgcgg 7080
gcaagcagcc gtaaacggtc ccgatcggat cgtgctgtca ccggcccctc cgcacagcag 7140
gctgtcgagg tgcgcgttcc cgaacagcgc gatgcgctgc atttgcccct cagctggagg 7200
gtaaaacgcc cgcgtaccag gatctggggc ggcctcccgg ttaagtagtg ctttctactt 7260
tatgagtagt gctttctact ttatgagtag tgctttctac tttatgagta gtgctttcta 7320
ctttatggag ctacagtgct tcatctcaga gctacagtgc ttcatctcag agctacagtg 7380
cttcatctca gagctacagt gcttcatctc agaaagcgga gtactgtcct ccgagcggag 7440
tactgtcctc cgagcggagt actgtcctcc gagcggagtt ctgtcctccg agcggagact 7500
ctagatatag aaccgatcct cccattgaat tctaggcgtg tacggtggga ggcctatata 7560
agcagagctc gtttagtgaa ccgtcagatc gcctcgagta tagaaccgat cctcccattt 7620
atagaaccga tcctcccatt tatagaaccg atcctcccat tactagaaga tcttgaattc 7680
gccaccttgt cgaccattcg tccgcgcagg ccaagtcctg cccgcgagct tctgcccgga 7740
ccccaaccgg atagggttca gccgactgca gatcgtgggg tgtctgcgcc tgctggcagc 7800
cctctggatg gcttgcccgc tcggcggacg gtgtcccgga cccggctgcc atctccccct 7860
gcgccctcac ctgcgttctc ggcgggcagc ttcagcgatc tgctccgtcc gttcgatccg 7920
tcgcttcttg atacatcgct tcttgattcg atgcctgccg tcggcacgcc gcatacagcg 7980
gctgccccag cagagtggga tgaggcgcaa tcggctctgc gtgcagccga tgacccgcca 8040
cccaccgtgc gtgtcgctgt cactgccgcg cggccgccgc gcgccaagcc ggccccgcga 8100
cggcgtgctg cgcaaccctc cgacgcttcg ccggccgcgc aggtggatct acgcacgctc 8160
ggctacagtc agcagcagca agagaagatc aaaccgaagg tgcgttcgac agtggcgcag 8220
caccacgagg cactggtggg ccatgggttt acacacgcgc acatcgttgc gctcagccaa 8280
cacccggcag cgttagggac cgtcgctgtc acgtatcagc acataatcac ggcgttgcca 8340
gaggcgacac acgaagacat cgttggcgtc ggcaaacagt ggtccggcgc acgcgccctg 8400
gaggccttgc tcacggatgc gggggagttg agaggtccgc cgttacagtt ggacacaggc 8460
caacttgtga agattgcaaa acgtggcggc gtgaccgcaa tggaggcagt gcatgcatcg 8520
cgcaatgcac tgacgggtgc ccccctgaac ctgaccccgg accaagtggt ggctatcgcc 8580
agcaacattg gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc 8640
caggaccatg gcctgactcc ggaccaagtg gtggctatcg ccagccacga tggcggcaag 8700
caagcgctcg aaacggtgca gcggctgttg ccggtgctgt gccaggacca tggcctgact 8760
ccggaccaag tggtggctat cgccagccac gatggcggca agcaagcgct cgaaacggtg 8820
cagcggctgt tgccggtgct gtgccaggac catggcctga ctccggacca agtggtggct 8880
atcgccagcc acgatggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg 8940
ctgtgccagg accatggcct gactccggac caagtggtgg ctatcgccag ccacgatggc 9000
ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc 9060
ctgaccccgg accaagtggt ggctatcgcc agcaacggtg gcggcaagca agcgctcgaa 9120
acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg gcctgactcc ggaccaagtg 9180
gtggctatcg ccagccacga tggcggcaag caagcgctcg aaacggtgca gcggctgttg 9240
ccggtgctgt gccaggacca tggcctgacc ccggaccaag tggtggctat cgccagcaac 9300
ggtggcggca agcaagcgct cgaaacggtg cagcggctgt tgccggtgct gtgccaggac 9360
catggcctga ctccggacca agtggtggct atcgccagcc acgatggcgg caagcaagcg 9420
ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg accatggcct gactccggac 9480
caagtggtgg ctatcgccag ccacgatggc ggcaagcaag cgctcgaaac ggtgcagcgg 9540
ctgttgccgg tgctgtgcca ggaccatggc ctgaccccgg accaagtggt ggctatcgcc 9600
agcaacaatg gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc 9660
caggaccatg gcctgactcc ggaccaagtg gtggctatcg ccagccacga tggcggcaag 9720
caagcgctcg aaacggtgca gcggctgttg ccggtgctgt gccaggacca tggcctgacc 9780
ccggaccaag tggtggctat cgccagcaac ggtggcggca agcaagcgct cgaaacggtg 9840
cagcggctgt tgccggtgct gtgccaggac catggcctga ccccggacca agtggtggct 9900
atcgccagca acggtggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg 9960
ctgtgccagg accatggcct gactccggac caagtggtgg ctatcgccag ccacgatggc 10020
ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc 10080
ctgaccccgg accaagtggt ggctatcgcc agcaacggtg gcggcaagca agcgctcgaa 10140
agcattgtgg cccagctgag ccggcctgat ccggcgttgg ccgcgttgac caacgaccac 10200
ctcgtcgcct tggcctgcct cggcggacgt cctgccatgg atgcagtgaa aaagggattg 10260
ccgcacgcgc cggaattgat cagaagagtc aatcgccgta ttggcgaacg cacgtcccat 10320
cgcgttgccg actacgcgca agtggttcgc gtgctggagt ttttccagtg ccactcccac 10380
ccagcgtacg catttgatga ggccatgacg cagttcggga tgagcaggaa cgggttggta 10440
cagctctttc gcagagtggg cgtcaccgaa ctcgaagccc gcggtggaac gctcccccca 10500
gcctcgcagc gttgggaccg tatcctccag gcatcaggga tgaaaagggc caaaccgtcc 10560
cctacttcag ctcaaacacc ggatcaggcg tctttgcatg cattcgccga ttcgctggag 10620
cgtgaccttg atgcgcccag cccaatgcac gagggagatc agacgcgggc aagcagccgt 10680
aaacggtccc gatcggatcg tgctgtcacc ggcccctccg cacagcaggc tgtcgaggtg 10740
cgcgttcccg aacagcgcga tgcgctgcat ttgcccctca gctggagggt aaaacgcccg 10800
cgtaccagga tctggggcgg cctcccgtaa tacaaggcgg ccgcaaatca acatcagtct 10860
gataagctat caacatcagt ctgataagct atcaacatca gtctgataag ctatcaacat 10920
cagtctgata agctaaagct tcgaattctg ataatcagcc ataccacatt tgtagaggtt 10980
ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca 11040
attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc 11100
acaaatttca caaataaagc atttttttca ctgctaccga ccacccgcaa acagcagggt 11160
cccctgggct tcccaagccg cgcacctctc cgccccgccc ctgcgccctc cttcctcgcg 11220
tctgcccctc tcccccaccc cgccttctcc ctccccgccc cagcggcgca tgcgccgcgc 11280
tcggagcgtg tttttataaa agtccggccg cggccagaaa cttcagtttg ttggctgcgg 11340
cagcaggtag caaagtgacg ccgagggcct gagtgctcca gtagccaccg catctggaga 11400
accagcggtt accatggccc ccccgaccga tgtcagcctg ggggacgagc tccacttaga 11460
cggcgaggac gtggcgatgg cgcatgccga cgcgctagac gatttcgatc tggacatgtt 11520
ggggagcgac gcgctagacg atttcgatct ggacatgttg gggagcgacg cgctagacga 11580
tttcgatctg gacatgttgg ggagcgacgc gctagacgat ttcgatctgg acatgttggg 11640
ggacggggat tccccgggtc cgggatttac cccccacgac tccgccccct acggcgctct 11700
ggatatggcc gacttcgagt ttgagcagat gtttaccgat gcccttggaa ttgacgagta 11760
cggtgggacg cgtatgaagc tactgtcttc tatcgaacaa gcatgcgata tttgccgact 11820
taaaaagctc aagtgctcca aagaaaaacc gaagtgcgcc aagtgtctga agaacaactg 11880
ggagtgtcgc tactctccca aaaccaaaag gtctccgctg actagggcac atctgacaga 11940
agtggaatca aggctagaaa gactggaaca gctatttcta ctgatttttc ctcgagaaga 12000
ccttgacatg attttgaaaa tggattcttt acaggatata aaagcattgt taacaggatt 12060
atttgtacaa gataatgtga ataaagatgc cgtcacagat agattggctt cagtggagac 12120
tgatatgcct ctaacattga gacagcatag aataagtgcg acatcatcat cggaagagag 12180
tagtaacaaa ggtcaaagac agttgactgt ataaaagctt cactcctcag gtgcaggctg 12240
cctatcagaa ggtggtggct ggtgtggcca atgccctggc tcacaaatac cactgagatc 12300
tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg 12360
gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac 12420
tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat ttggtttaga 12480
gtttggcaac atatgcccat atgctggctg ccatgaacaa aggttggcta taaagaggtc 12540
atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa gccttgactt 12600
gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca tccctaaaat 12660
tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc ccagtcatag 12720
ctgtccctct tctcttatgg agatc 12745
<210> 17
<211> 6116
<212> DNA
<213> Artificial
<220>
<223> 编码由细胞特异性启动子Survivin1、第一microRNA为mir-135a-5p、第二m
icroRNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋
白分别为Lacl、tetR-KRAB、第一和第二识别序列为4×UAS的元件组成的表
达系统的核酸
<400> 17
cgggtgaccg ccctccgcgg agtactgtcc tccgcgggtg acagccctcc gcggagtgcc 60
gcactccgag cggagactct agactcccta tcagtgatag agataggcgt gtacggtggg 120
aggcctatat aagcagagct cgtttagtga accgtcagat cgctccctat cagtgataga 180
gagaattcga ccgccaccat gagacatatt atctgccacg gaggtgttat taccgaagaa 240
atggccgcca gtcttttgga ccagctgatc gaagaggtac tggctgataa tcttccacct 300
cctagccatt ttgaaccacc tacccttcac gaactgtatg atttagacgt gacggccccc 360
gaagatccca acgaggaggc ggtttcgcag atttttcccg actctgtaat gttggcggtg 420
caggaaggga ttgacttact cacttttccg ccggcgcccg gttctccgga gccgcctcac 480
ctttcccggc agcccgagca gccggagcag agagccttgg gtccggtttc tatgccaaac 540
cttgtaccgg aggtgatcga tcttacctgc cacgaggctg gctttccacc cagtgacgac 600
gaggatgaag agggtgagga gtttgtgtta gattatgtgg agcaccccgg gcacggttgc 660
aggtcttgtc attatcaccg gaggaatacg ggggacccag atattatgtg ttcgctttgc 720
tatatgagga cctgtggcat gtttgtctac agtaagtgaa aattatgggc agtgggtgat 780
agagtggtgg gtttggtgtg gtaatttttt ttttaatttt tacagttttg tggtttaaag 840
aattttgtat tgtgattttt ttaaaaggtc ctgtgtctga acctgagcct gagcccgagc 900
cagaaccgga gcctgcaaga cctacccgcc gtcctaaaat ggcgcctgct atcctgaggc 960
gcccgacatc acctgtgtct agagaatgca atagtagtac ggatagctgt gactccggtc 1020
cttctaacac acctcctgag atacacccgg tggtcccgct gtgccccatt aaaccagttg 1080
ccgtgagagt tggtgggcgt cgccaggctg tggaatgtat cgaggacttg cttaacgagc 1140
ctgggcaacc tttggacttg agctgtaaac gccccaggcc actcgagggt accgggtccg 1200
gagctacaaa tttttccctc ctcaaacagg ctggagatgt cgaagaaaat cctgggccta 1260
ccggtatgtg gctgcagagc ctccttctac tgggcaccgt ggcttgcagc atctcggccc 1320
ccgccagaag tcccagcccc agcacccagc cctgggaaca cgtaaatgca atccaggagg 1380
cccgccgcct cctgaacttg tcacgggaca cggcagccga gatgaatgaa actgtggagg 1440
tcatcagtga aatgtttgac ttgcaggagc ccacatgcct gcagactcgg ctggagctct 1500
acaaacaggg cctgcgtggc agcctcacca agctgaaggg cccactgacc atgatggcta 1560
gccattacaa gcagcactgc ccgccgaccc ctgagacatc ctgtgccact cagatcatta 1620
cctttgagtc tttcaaggag aacctcaagg actttctgct ggtcatccca tttgactgct 1680
gggaacccgt ccaggaaaga tctgagggcc gcggcagcct gctgacctgc ggcgacgtgg 1740
aggaaaaccc cggccccgga tccatgaaac cagtaacgtt atacgatgtc gcagagtatg 1800
ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac gtttctgcga 1860
aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg 1920
cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc agtctggccc 1980
tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca 2040
gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca 2100
atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg 2160
ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc 2220
agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg ggcgtggagc 2280
atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct 2340
cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga 2400
tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc 2460
tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg 2520
caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggatatctcg gtagtgggat 2580
acgacgatac cgaagacagc tcatgttata tcccgccgtt aaccaccatc aaacaggatt 2640
ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg 2700
tgaagggcaa tcagctgttg ccggtctcac tggtgaaaag aaaaaccacc ctggcgccca 2760
atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg 2820
tttcccgact ggaaagcggg cagagcagcc tgaggcctcc taagaagaag aggaaggttg 2880
ttacaaacac cattgtcaca ctccacaaac accattgtca cactccacaa acaccattgt 2940
cacactccac aaacaccatt gtcacactcc atcacatagg aataaaaagc catatcacat 3000
aggaataaaa agccatatca cataggaata aaaagccata tcacatagga ataaaaagcc 3060
atagaaagcg gggtgccgcc ccccgcggac tactgtagtc cgcggacgac tgtcgtccgc 3120
ggagcactgt gctccgagcg gagactctag aaattgtgag cggataacaa ttgggtatat 3180
aatgggggcc gtcagctact accagagccg atcgaacact cgagccgagc agacgtgcct 3240
acaattgtga gcgctcacaa ttactagaag atcttgaatt cgccaccatg gctagattag 3300
ataaaagtaa agtgattaac agcgcattag agctgcttaa tgaggtcgga atcgaaggtt 3360
taacaacccg taaactcgcc cagaagctag gtgtagagca gcctacattg tattggcatg 3420
taaaaaataa gcgggctttg ctcgacgcct tagccattga gatgttagat aggcaccata 3480
ctcacttttg ccctttagaa ggggaaagct ggcaagattt tttacgtaat aacgctaaaa 3540
gttttagatg tgctttacta agtcatcgcg atggagcaaa agtacattta ggtacacggc 3600
ctacagaaaa acagtatgaa actctcgaaa atcaattagc ctttttatgc caacaaggtt 3660
tttcactaga gaatgcatta tatgcactca gcgctgtggg gcattttact ttaggttgcg 3720
tattggaaga tcaagagcat caagtcgcta aagaagaaag ggaaacacct actactgata 3780
gtatgccgcc attattacga caagctatcg aattatttga tcaccaaggt gcagagccag 3840
ccttcttatt cggccttgaa ttgatcatat gcggattaga aaaacaactt aaatgtgaaa 3900
gtgggtcgcc aaaaaagaag agaaaggtgg atggtggcgg agcactgtct ccccagcatt 3960
cagctgtgac ccagggatca atcattaaga acaaggaagg tatggatgct aagtccctga 4020
ccgcttggtc aagaactttg gtaactttta aggacgtgtt cgtcgatttt actcgggagg 4080
agtggaaact attggatacc gcccagcaaa tcgtgtatag aaacgttatg ctcgagaact 4140
ataagaatct tgtgtcactg ggctaccaac tgacaaagcc tgacgttatc ctgaggctgg 4200
aaaagggtga ggagccttgg ttggtcgagc gggagatcca ccaggaaact cacccagaca 4260
gtgagactgc tttcgagatc aagtcctctg tgtaatacaa ggcggccgca aatcaacatc 4320
agtctgataa gctatcaaca tcagtctgat aagctatcaa catcagtctg ataagctatc 4380
aacatcagtc tgataagcta aagcttcgaa ttctgataat cagccatacc acatttgtag 4440
aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga 4500
atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata 4560
gcatcacaaa tttcacaaat aaagcatttt tttcactgca gctcgccttt tgctcacatg 4620
ttaggtgtgg gcagggacga gctggcgcgg cgtcgctggg tgcaccgcga ccacgggcag 4680
agccacgcgg cgggaggact acaactcccg gcacaccccg cgccgccccg cctctactcc 4740
cagaaggccg cggggggtgg accgcctaag agggcgtgcg ctcccgacat gccccgcggc 4800
gcgccattaa ccgccagatt tgaatcgcgg gacccgttgg cagaggtggc ggcggcggca 4860
tgggtgcccc gacgttgccc cctgcctggc tcgagaccgg tatggccccc ccgaccgatg 4920
tcagcctggg ggacgagctc cacttagacg gcgaggacgt ggcgatggcg catgccgacg 4980
cgctagacga tttcgatctg gacatgttgg gggacgggga ttccccgggt ccgggattta 5040
ccccccacga ctccgccccc tacggcgctc tggatatggc cgacttcgag tttgagcaga 5100
tgtttaccga tgcccttgga attgacgagt acggtgggac gcgtatgaag ctactgtctt 5160
ctatcgaaca agcatgcgat atttgccgac ttaaaaagct caagtgctcc aaagaaaaac 5220
cgaagtgcgc caagtgtctg aagaacaact gggagtgtcg ctactctccc aaaaccaaaa 5280
ggtctccgct gactagggca catctgacag aagtggaatc aaggctagaa agactggaac 5340
agctatttct actgattttt cctcgagaag accttgacat gattttgaaa atggattctt 5400
tacaggatat aaaagcattg ttaacaggat tatttgtaca agataatgtg aataaagatg 5460
ccgtcacaga tagattggct tcagtggaga ctgatatgcc tctaacattg agacagcata 5520
gaataagtgc gacatcatca tcggaagaga gtagtaacaa aggtcaaaga cagttgactg 5580
tataaaagct tcactcctca ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc 5640
aatgccctgg ctcacaaata ccactgagat ctttttccct ctgccaaaaa ttatggggac 5700
atcatgaagc cccttgagca tctgacttct ggctaataaa ggaaatttat tttcattgca 5760
atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag ggcaaatcat 5820
ttaaaacatc agaatgagta tttggtttag agtttggcaa catatgccca tatgctggct 5880
gccatgaaca aaggttggct ataaagaggt catcagtata tgaaacagcc ccctgctgtc 5940
cattccttat tccatagaaa agccttgact tgaggttaga ttttttttat attttgtttt 6000
gtgttatttt tttctttaac atccctaaaa ttttccttac atgttttact agccagattt 6060
ttcctcctct cctgactact cccagtcata gctgtccctc ttctcttatg gagatc 6116
<210> 18
<211> 12749
<212> DNA
<213> Artificial
<220>
<223> 编码由细胞特异性启动子Survivin1、第一microRNA为mir-143-3p、第二mi
croRNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白
分别为TALE蛋白、第一和第二识别序列为4×UAS的元件组成的表达系统的核
酸
<400> 18
cggagtactg tcctccgagc ggagtactgt cctccgagcg gagtactgtc ctccgagcgg 60
agttctgtcc tccgagcgga gactctagat acccctctcc gcttctgaat tctaggcgtg 120
tacggtggga ggcctatata agcagagctc gtttagtgaa ccgtcagatc gcctcgagta 180
cccctctccg cttcttaccc ctctccgctt cttacccctc tccgcttctg aattcgaccg 240
ccaccatgag acatattatc tgccacggag gtgttattac cgaagaaatg gccgccagtc 300
ttttggacca gctgatcgaa gaggtactgg ctgataatct tccacctcct agccattttg 360
aaccacctac ccttcacgaa ctgtatgatt tagacgtgac ggcccccgaa gatcccaacg 420
aggaggcggt ttcgcagatt tttcccgact ctgtaatgtt ggcggtgcag gaagggattg 480
acttactcac ttttccgccg gcgcccggtt ctccggagcc gcctcacctt tcccggcagc 540
ccgagcagcc ggagcagaga gccttgggtc cggtttctat gccaaacctt gtaccggagg 600
tgatcgatct tacctgccac gaggctggct ttccacccag tgacgacgag gatgaagagg 660
gtgaggagtt tgtgttagat tatgtggagc accccgggca cggttgcagg tcttgtcatt 720
atcaccggag gaatacgggg gacccagata ttatgtgttc gctttgctat atgaggacct 780
gtggcatgtt tgtctacagt aagtgaaaat tatgggcagt gggtgataga gtggtgggtt 840
tggtgtggta attttttttt taatttttac agttttgtgg tttaaagaat tttgtattgt 900
gattttttta aaaggtcctg tgtctgaacc tgagcctgag cccgagccag aaccggagcc 960
tgcaagacct acccgccgtc ctaaaatggc gcctgctatc ctgaggcgcc cgacatcacc 1020
tgtgtctaga gaatgcaata gtagtacgga tagctgtgac tccggtcctt ctaacacacc 1080
tcctgagata cacccggtgg tcccgctgtg ccccattaaa ccagttgccg tgagagttgg 1140
tgggcgtcgc caggctgtgg aatgtatcga ggacttgctt aacgagcctg ggcaaccttt 1200
ggacttgagc tgtaaacgcc ccaggccagg gtccggagct acaaattttt ccctcctcaa 1260
acaggctgga gatgtcgaag aaaatcctgg gcctatggag gcttgggagt gtttggaaga 1320
tttttctgct gtgcgtaact tgctggaaca gagctctaac agtacctctt ggttttggag 1380
gtttctgtgg ggctcatccc aggcaaagtt agtctgcaga attaaggagg attacaagtg 1440
ggaatttgaa gagcttttga aatcctgtgg tgagctgttt gattctttga atctgggtca 1500
ccaggcgctt ttccaagaga aggtcatcaa gactttggat ttttccacac cggggcgcgc 1560
tgcggctgct gttgcttttt tgagttttat aaaggataaa tggagcgaag aaacccatct 1620
gagcgggggg tacctgctgg attttctggc catgcatctg tggagagcgg ttgtgagaca 1680
caagaatcgc ctgctactgt tgtcttccgt ccgcccggcg ataataccga cggaggagca 1740
gcagcagcag caggaggaag ccaggcggcg gcggcaggag cagagcccat ggaacccgag 1800
agccggcctg gaccctcggg aatgaatgtt gtacaggtgg ctgaactgta tccagaactg 1860
agacgcattt tgacaattac agaggatggg caggggctaa agggggtaaa gagggagcgg 1920
ggggcttgtg aggctacaga ggaggctagg aatctagctt ttagcttaat gaccagacac 1980
cgtcctgagt gtattacttt tcaacagatc aaggataatt gcgctaatga gcttgatctg 2040
ctggcgcaga agtattccat agagcagctg accacttact ggctgcagcc aggggatgat 2100
tttgaggagg ctattagggt atatgcaaag gtggcactta ggccagattg caagtacaag 2160
atcagcaaac ttgtaaatat caggaattgt tgctacattt ctgggaacgg ggccgaggtg 2220
gagatagata cggaggatag ggtggccttt agatgtagca tgataaatat gtggccgggg 2280
gtgcttggca tggacggggt ggttattatg aatgtaaggt ttactggccc caattttagc 2340
ggtacggttt tcctggccaa taccaacctt atcctacacg gtgtaagctt ctatgggttt 2400
aacaatacct gtgtggaagc ctggaccgat gtaagggttc ggggctgtgc cttttactgc 2460
tgctggaagg gggtggtgtg tcgccccaaa agcagggctt caattaagaa atgcctcttt 2520
gaaaggtgta ccttgggtat cctgtctgag ggtaactcca gggtgcgcca caatgtggcc 2580
tccgactgtg gttgcttcat gctagtgaaa agcgtggctg tgattaagca taacatggta 2640
tgtggcaact gcgaggacag ggcctctcag atgctgacct gctcggacgg caactgtcac 2700
ctgctgaaga ccattcacgt agccagccac tctcgcaagg cctggccagt gtttgagcat 2760
aacatactga cccgctgttc cttgcatttg ggtaacagga ggggggtgtt cctaccttac 2820
caatgcaatt tgagtcacac taagatattg cttgagcccg agagcatgtc caaggtgaac 2880
ctgaacgggg tgtttgacat gaccatgaag atctggaagg tgctgaggta cgatgagacc 2940
cgcaccaggt gcagaccctg cgagtgtggc ggtaaacata ttaggaacca gcctgtgatg 3000
ctggatgtga ccgaggagct gaggcccgat cacttggtgc tggcctgcac ccgcgctgag 3060
tttggctcta gcgatgaaga tacagatctc gaggggtccg gagctacaaa tttttccctc 3120
ctcaaacagg ctggagatgt cgaagaaaat cctgggccta ccggtatgta taggatgcag 3180
ttgctctctt gcatcgccct gtctctagcg ttggtgacta atagcgcccc tacttcttct 3240
tccacgaaga aaacccagct gcaactggaa caccttctgc tggatctcca gatgatcctt 3300
aacgggatta acaactataa gaacccgaag ctcacggcta tgctgacggc gaagttcgct 3360
atgcctaaaa aggctactga gctgaaacac ttacagtgtc tcgaagaggc actcaaaccc 3420
ctcgaggagg tactcaatct ggcgcaaagc aaaaacttcc atctccgccc acgggaccta 3480
atctccaaca tcaacgtgat tgtgctggag ctcaaggggt ctgaaaccac gtttatgtgc 3540
gagtacgcag atgaaactgc taccatagtg gaattcctca atcgttggat cactttctgc 3600
cagtcaatca tttcaaccct gaccagatct gagggccgcg gcagcctgct gacctgcggc 3660
gacgtggagg aaaaccccgg ccccggatcc ttgtcgacca ttcgtccgcg caggccaagt 3720
cctgcccgcg agcttctgcc cggaccccaa ccggataggg ttcagccgac tgcagatcgt 3780
ggggtgtctg cgcctgctgg cagccctctg gatggcttgc ccgctcggcg gacggtgtcc 3840
cggacccggc tgccatctcc ccctgcgccc tcacctgcgt tctcggcggg cagcttcagc 3900
gatctgctcc gtccgttcga tccgtcgctt cttgatacat cgcttcttga ttcgatgcct 3960
gccgtcggca cgccgcatac agcggctgcc ccagcagagt gggatgaggc gcaatcggct 4020
ctgcgtgcag ccgatgaccc gccacccacc gtgcgtgtcg ctgtcactgc cgcgcggccg 4080
ccgcgcgcca agccggcccc gcgacggcgt gctgcgcaac cctccgacgc ttcgccggcc 4140
gcgcaggtgg atctacgcac gctcggctac agtcagcagc agcaagagaa gatcaaaccg 4200
aaggtgcgtt cgacagtggc gcagcaccac gaggcactgg tgggccatgg gtttacacac 4260
gcgcacatcg ttgcgctcag ccaacacccg gcagcgttag ggaccgtcgc tgtcacgtat 4320
cagcacataa tcacggcgtt gccagaggcg acacacgaag acatcgttgg cgtcggcaaa 4380
cagtggtccg gcgcacgcgc cctggaggcc ttgctcacgg atgcggggga gttgagaggt 4440
ccgccgttac agttggacac aggccaactt gtgaagattg caaaacgtgg cggcgtgacc 4500
gcaatggagg cagtgcatgc atcgcgcaat gcactgacgg gtgcccccct gaacctgacc 4560
ccggaccaag tggtggctat cgccagcaac attggcggca agcaagcgct cgaaacggtg 4620
cagcggctgt tgccggtgct gtgccaggac catggcctga ccccggacca agtggtggct 4680
atcgccagca acggtggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg 4740
ctgtgccagg accatggcct gaccccggac caagtggtgg ctatcgccag caacattggc 4800
ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc 4860
ctgaccccgg accaagtggt ggctatcgcc agcaacaatg gcggcaagca agcgctcgaa 4920
acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg gcctgacccc ggaccaagtg 4980
gtggctatcg ccagcaacat tggcggcaag caagcgctcg aaacggtgca gcggctgttg 5040
ccggtgctgt gccaggacca tggcctgacc ccggaccaag tggtggctat cgccagcaac 5100
attggcggca agcaagcgct cgaaacggtg cagcggctgt tgccggtgct gtgccaggac 5160
catggcctga ctccggacca agtggtggct atcgccagcc acgatggcgg caagcaagcg 5220
ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg accatggcct gactccggac 5280
caagtggtgg ctatcgccag ccacgatggc ggcaagcaag cgctcgaaac ggtgcagcgg 5340
ctgttgccgg tgctgtgcca ggaccatggc ctgaccccgg accaagtggt ggctatcgcc 5400
agcaacaatg gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc 5460
caggaccatg gcctgacccc ggaccaagtg gtggctatcg ccagcaacat tggcggcaag 5520
caagcgctcg aaacggtgca gcggctgttg ccggtgctgt gccaggacca tggcctgacc 5580
ccggaccaag tggtggctat cgccagcaac ggtggcggca agcaagcgct cgaaacggtg 5640
cagcggctgt tgccggtgct gtgccaggac catggcctga ctccggacca agtggtggct 5700
atcgccagcc acgatggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg 5760
ctgtgccagg accatggcct gactccggac caagtggtgg ctatcgccag ccacgatggc 5820
ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc 5880
ctgaccccgg accaagtggt ggctatcgcc agcaacggtg gcggcaagca agcgctcgaa 5940
acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg gcctgactcc ggaccaagtg 6000
gtggctatcg ccagccacga tggcggcaag caagcgctcg aaacggtgca gcggctgttg 6060
ccggtgctgt gccaggacca tggcctgact ccggaccaag tggtggctat cgccagccac 6120
gatggcggca agcaagcgct cgaaacggtg cagcggctgt tgccggtgct gtgccaggac 6180
catggcctga ctccggacca agtggtggct atcgccagcc acgatggcgg caagcaagcg 6240
ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg accatggcct gaccccggac 6300
caagtggtgg ctatcgccag caacattggc ggcaagcaag cgctcgaaac ggtgcagcgg 6360
ctgttgccgg tgctgtgcca ggaccatggc ctgaccccgg accaagtggt ggctatcgcc 6420
agcaacggtg gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc 6480
caggaccatg gcctgacccc ggaccaagtg gtggctatcg ccagcaacgg tggcggcaag 6540
caagcgctcg aaagcattgt ggcccagctg agccggcctg atccggcgtt ggccgcgttg 6600
accaacgacc acctcgtcgc cttggcctgc ctcggcggac gtcctgccat ggatgcagtg 6660
aaaaagggat tgccgcacgc gccggaattg atcagaagag tcaatcgccg tattggcgaa 6720
cgcacgtccc atcgcgttgc cgactacgcg caagtggttc gcgtgctgga gtttttccag 6780
tgccactccc acccagcgta cgcatttgat gaggccatga cgcagttcgg gatgagcagg 6840
aacgggttgg tacagctctt tcgcagagtg ggcgtcaccg aactcgaagc ccgcggtgga 6900
acgctccccc cagcctcgca gcgttgggac cgtatcctcc aggcatcagg gatgaaaagg 6960
gccaaaccgt cccctacttc agctcaaaca ccggatcagg cgtctttgca tgcattcgcc 7020
gattcgctgg agcgtgacct tgatgcgccc agcccaatgc acgagggaga tcagacgcgg 7080
gcaagcagcc gtaaacggtc ccgatcggat cgtgctgtca ccggcccctc cgcacagcag 7140
gctgtcgagg tgcgcgttcc cgaacagcgc gatgcgctgc atttgcccct cagctggagg 7200
gtaaaacgcc cgcgtaccag gatctggggc ggcctcccgg ttaagtagtg ctttctactt 7260
tatgagtagt gctttctact ttatgagtag tgctttctac tttatgagta gtgctttcta 7320
ctttatggag ctacagtgct tcatctcaga gctacagtgc ttcatctcag agctacagtg 7380
cttcatctca gagctacagt gcttcatctc agaaagcgga gtactgtcct ccgagcggag 7440
tactgtcctc cgagcggagt actgtcctcc gagcggagtt ctgtcctccg agcggagact 7500
ctagatatag aaccgatcct cccattgaat tctaggcgtg tacggtggga ggcctatata 7560
agcagagctc gtttagtgaa ccgtcagatc gcctcgagta tagaaccgat cctcccattt 7620
atagaaccga tcctcccatt tatagaaccg atcctcccat tactagaaga tcttgaattc 7680
gccaccttgt cgaccattcg tccgcgcagg ccaagtcctg cccgcgagct tctgcccgga 7740
ccccaaccgg atagggttca gccgactgca gatcgtgggg tgtctgcgcc tgctggcagc 7800
cctctggatg gcttgcccgc tcggcggacg gtgtcccgga cccggctgcc atctccccct 7860
gcgccctcac ctgcgttctc ggcgggcagc ttcagcgatc tgctccgtcc gttcgatccg 7920
tcgcttcttg atacatcgct tcttgattcg atgcctgccg tcggcacgcc gcatacagcg 7980
gctgccccag cagagtggga tgaggcgcaa tcggctctgc gtgcagccga tgacccgcca 8040
cccaccgtgc gtgtcgctgt cactgccgcg cggccgccgc gcgccaagcc ggccccgcga 8100
cggcgtgctg cgcaaccctc cgacgcttcg ccggccgcgc aggtggatct acgcacgctc 8160
ggctacagtc agcagcagca agagaagatc aaaccgaagg tgcgttcgac agtggcgcag 8220
caccacgagg cactggtggg ccatgggttt acacacgcgc acatcgttgc gctcagccaa 8280
cacccggcag cgttagggac cgtcgctgtc acgtatcagc acataatcac ggcgttgcca 8340
gaggcgacac acgaagacat cgttggcgtc ggcaaacagt ggtccggcgc acgcgccctg 8400
gaggccttgc tcacggatgc gggggagttg agaggtccgc cgttacagtt ggacacaggc 8460
caacttgtga agattgcaaa acgtggcggc gtgaccgcaa tggaggcagt gcatgcatcg 8520
cgcaatgcac tgacgggtgc ccccctgaac ctgaccccgg accaagtggt ggctatcgcc 8580
agcaacattg gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc 8640
caggaccatg gcctgactcc ggaccaagtg gtggctatcg ccagccacga tggcggcaag 8700
caagcgctcg aaacggtgca gcggctgttg ccggtgctgt gccaggacca tggcctgact 8760
ccggaccaag tggtggctat cgccagccac gatggcggca agcaagcgct cgaaacggtg 8820
cagcggctgt tgccggtgct gtgccaggac catggcctga ctccggacca agtggtggct 8880
atcgccagcc acgatggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg 8940
ctgtgccagg accatggcct gactccggac caagtggtgg ctatcgccag ccacgatggc 9000
ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc 9060
ctgaccccgg accaagtggt ggctatcgcc agcaacggtg gcggcaagca agcgctcgaa 9120
acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg gcctgactcc ggaccaagtg 9180
gtggctatcg ccagccacga tggcggcaag caagcgctcg aaacggtgca gcggctgttg 9240
ccggtgctgt gccaggacca tggcctgacc ccggaccaag tggtggctat cgccagcaac 9300
ggtggcggca agcaagcgct cgaaacggtg cagcggctgt tgccggtgct gtgccaggac 9360
catggcctga ctccggacca agtggtggct atcgccagcc acgatggcgg caagcaagcg 9420
ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg accatggcct gactccggac 9480
caagtggtgg ctatcgccag ccacgatggc ggcaagcaag cgctcgaaac ggtgcagcgg 9540
ctgttgccgg tgctgtgcca ggaccatggc ctgaccccgg accaagtggt ggctatcgcc 9600
agcaacaatg gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc 9660
caggaccatg gcctgactcc ggaccaagtg gtggctatcg ccagccacga tggcggcaag 9720
caagcgctcg aaacggtgca gcggctgttg ccggtgctgt gccaggacca tggcctgacc 9780
ccggaccaag tggtggctat cgccagcaac ggtggcggca agcaagcgct cgaaacggtg 9840
cagcggctgt tgccggtgct gtgccaggac catggcctga ccccggacca agtggtggct 9900
atcgccagca acggtggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg 9960
ctgtgccagg accatggcct gactccggac caagtggtgg ctatcgccag ccacgatggc 10020
ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc 10080
ctgaccccgg accaagtggt ggctatcgcc agcaacggtg gcggcaagca agcgctcgaa 10140
agcattgtgg cccagctgag ccggcctgat ccggcgttgg ccgcgttgac caacgaccac 10200
ctcgtcgcct tggcctgcct cggcggacgt cctgccatgg atgcagtgaa aaagggattg 10260
ccgcacgcgc cggaattgat cagaagagtc aatcgccgta ttggcgaacg cacgtcccat 10320
cgcgttgccg actacgcgca agtggttcgc gtgctggagt ttttccagtg ccactcccac 10380
ccagcgtacg catttgatga ggccatgacg cagttcggga tgagcaggaa cgggttggta 10440
cagctctttc gcagagtggg cgtcaccgaa ctcgaagccc gcggtggaac gctcccccca 10500
gcctcgcagc gttgggaccg tatcctccag gcatcaggga tgaaaagggc caaaccgtcc 10560
cctacttcag ctcaaacacc ggatcaggcg tctttgcatg cattcgccga ttcgctggag 10620
cgtgaccttg atgcgcccag cccaatgcac gagggagatc agacgcgggc aagcagccgt 10680
aaacggtccc gatcggatcg tgctgtcacc ggcccctccg cacagcaggc tgtcgaggtg 10740
cgcgttcccg aacagcgcga tgcgctgcat ttgcccctca gctggagggt aaaacgcccg 10800
cgtaccagga tctggggcgg cctcccgtaa tacaaggcgg ccgcaaatca acatcagtct 10860
gataagctat caacatcagt ctgataagct atcaacatca gtctgataag ctatcaacat 10920
cagtctgata agctaaagct tcgaattctg ataatcagcc ataccacatt tgtagaggtt 10980
ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca 11040
attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc 11100
acaaatttca caaataaagc atttttttca ctgccatgtt aggtgtgggc agggacgagc 11160
tggcgcggcg tcgctgggtg caccgcgacc acgggcagag ccacgcggcg ggaggactac 11220
aactcccggc acaccccgcg ccgccccgcc tctactccca gaaggccgcg gggggtggac 11280
cgcctaagag ggcgtgcgct cccgacatgc cccgcggcgc gccattaacc gccagatttg 11340
aatcgcggga cccgttggca gaggtggcgg cggcggcatg ggtgccccga cgttgccccc 11400
tgcctggctc gagaccgatg gcccccccga ccgatgtcag cctgggggac gagctccact 11460
tagacggcga ggacgtggcg atggcgcatg ccgacgcgct agacgatttc gatctggaca 11520
tgttggggag cgacgcgcta gacgatttcg atctggacat gttggggagc gacgcgctag 11580
acgatttcga tctggacatg ttggggagcg acgcgctaga cgatttcgat ctggacatgt 11640
tgggggacgg ggattccccg ggtccgggat ttacccccca cgactccgcc ccctacggcg 11700
ctctggatat ggccgacttc gagtttgagc agatgtttac cgatgccctt ggaattgacg 11760
agtacggtgg gacgcgtatg aagctactgt cttctatcga acaagcatgc gatatttgcc 11820
gacttaaaaa gctcaagtgc tccaaagaaa aaccgaagtg cgccaagtgt ctgaagaaca 11880
actgggagtg tcgctactct cccaaaacca aaaggtctcc gctgactagg gcacatctga 11940
cagaagtgga atcaaggcta gaaagactgg aacagctatt tctactgatt tttcctcgag 12000
aagaccttga catgattttg aaaatggatt ctttacagga tataaaagca ttgttaacag 12060
gattatttgt acaagataat gtgaataaag atgccgtcac agatagattg gcttcagtgg 12120
agactgatat gcctctaaca ttgagacagc atagaataag tgcgacatca tcatcggaag 12180
agagtagtaa caaaggtcaa agacagttga ctgtataaaa gcttcactcc tcaggtgcag 12240
gctgcctatc agaaggtggt ggctggtgtg gccaatgccc tggctcacaa ataccactga 12300
gatctttttc cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact 12360
tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc 12420
tcactcggaa ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt 12480
tagagtttgg caacatatgc ccatatgctg gctgccatga acaaaggttg gctataaaga 12540
ggtcatcagt atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg 12600
acttgaggtt agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta 12660
aaattttcct tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc 12720
atagctgtcc ctcttctctt atggagatc 12749
<210> 19
<211> 6208
<212> DNA
<213> Artificial
<220>
<223> 编码由细胞特异性启动子CEA368、第一microRNA为mir-143-3p、第二micro
RNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白分
别为Lacl、tetR-KRAB、第一和第二识别序列为4×UAS的元件组成的表达系
统的核酸
<400> 19
cgggtgaccg ccctccgcgg agtactgtcc tccgcgggtg acagccctcc gcggagtgcc 60
gcactccgag cggagactct agactcccta tcagtgatag agataggcgt gtacggtggg 120
aggcctatat aagcagagct cgtttagtga accgtcagat cgctccctat cagtgataga 180
gagaattcga ccgccaccat gagacatatt atctgccacg gaggtgttat taccgaagaa 240
atggccgcca gtcttttgga ccagctgatc gaagaggtac tggctgataa tcttccacct 300
cctagccatt ttgaaccacc tacccttcac gaactgtatg atttagacgt gacggccccc 360
gaagatccca acgaggaggc ggtttcgcag atttttcccg actctgtaat gttggcggtg 420
caggaaggga ttgacttact cacttttccg ccggcgcccg gttctccgga gccgcctcac 480
ctttcccggc agcccgagca gccggagcag agagccttgg gtccggtttc tatgccaaac 540
cttgtaccgg aggtgatcga tcttacctgc cacgaggctg gctttccacc cagtgacgac 600
gaggatgaag agggtgagga gtttgtgtta gattatgtgg agcaccccgg gcacggttgc 660
aggtcttgtc attatcaccg gaggaatacg ggggacccag atattatgtg ttcgctttgc 720
tatatgagga cctgtggcat gtttgtctac agtaagtgaa aattatgggc agtgggtgat 780
agagtggtgg gtttggtgtg gtaatttttt ttttaatttt tacagttttg tggtttaaag 840
aattttgtat tgtgattttt ttaaaaggtc ctgtgtctga acctgagcct gagcccgagc 900
cagaaccgga gcctgcaaga cctacccgcc gtcctaaaat ggcgcctgct atcctgaggc 960
gcccgacatc acctgtgtct agagaatgca atagtagtac ggatagctgt gactccggtc 1020
cttctaacac acctcctgag atacacccgg tggtcccgct gtgccccatt aaaccagttg 1080
ccgtgagagt tggtgggcgt cgccaggctg tggaatgtat cgaggacttg cttaacgagc 1140
ctgggcaacc tttggacttg agctgtaaac gccccaggcc actcgagggt accgggtccg 1200
gagctacaaa tttttccctc ctcaaacagg ctggagatgt cgaagaaaat cctgggccta 1260
ccggtatgtg gctgcagagc ctccttctac tgggcaccgt ggcttgcagc atctcggccc 1320
ccgccagaag tcccagcccc agcacccagc cctgggaaca cgtaaatgca atccaggagg 1380
cccgccgcct cctgaacttg tcacgggaca cggcagccga gatgaatgaa actgtggagg 1440
tcatcagtga aatgtttgac ttgcaggagc ccacatgcct gcagactcgg ctggagctct 1500
acaaacaggg cctgcgtggc agcctcacca agctgaaggg cccactgacc atgatggcta 1560
gccattacaa gcagcactgc ccgccgaccc ctgagacatc ctgtgccact cagatcatta 1620
cctttgagtc tttcaaggag aacctcaagg actttctgct ggtcatccca tttgactgct 1680
gggaacccgt ccaggaaaga tctgagggcc gcggcagcct gctgacctgc ggcgacgtgg 1740
aggaaaaccc cggccccgga tccatgaaac cagtaacgtt atacgatgtc gcagagtatg 1800
ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac gtttctgcga 1860
aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg 1920
cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc agtctggccc 1980
tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca 2040
gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca 2100
atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg 2160
ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc 2220
agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg ggcgtggagc 2280
atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct 2340
cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga 2400
tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc 2460
tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg 2520
caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggatatctcg gtagtgggat 2580
acgacgatac cgaagacagc tcatgttata tcccgccgtt aaccaccatc aaacaggatt 2640
ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg 2700
tgaagggcaa tcagctgttg ccggtctcac tggtgaaaag aaaaaccacc ctggcgccca 2760
atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg 2820
tttcccgact ggaaagcggg cagagcagcc tgaggcctcc taagaagaag aggaaggttg 2880
ttacaaacac cattgtcaca ctccacaaac accattgtca cactccacaa acaccattgt 2940
cacactccac aaacaccatt gtcacactcc agagctacag tgcttcatct cagagctaca 3000
gtgcttcatc tcagagctac agtgcttcat ctcagagcta cagtgcttca tctcagaaag 3060
cggggtgccg ccccccgcgg actactgtag tccgcggacg actgtcgtcc gcggagcact 3120
gtgctccgag cggagactct agaaattgtg agcggataac aattgggtat ataatggggg 3180
ccgtcagcta ctaccagagc cgatcgaaca ctcgagccga gcagacgtgc ctacaattgt 3240
gagcgctcac aattactaga agatcttgaa ttcgccacca tggctagatt agataaaagt 3300
aaagtgatta acagcgcatt agagctgctt aatgaggtcg gaatcgaagg tttaacaacc 3360
cgtaaactcg cccagaagct aggtgtagag cagcctacat tgtattggca tgtaaaaaat 3420
aagcgggctt tgctcgacgc cttagccatt gagatgttag ataggcacca tactcacttt 3480
tgccctttag aaggggaaag ctggcaagat tttttacgta ataacgctaa aagttttaga 3540
tgtgctttac taagtcatcg cgatggagca aaagtacatt taggtacacg gcctacagaa 3600
aaacagtatg aaactctcga aaatcaatta gcctttttat gccaacaagg tttttcacta 3660
gagaatgcat tatatgcact cagcgctgtg gggcatttta ctttaggttg cgtattggaa 3720
gatcaagagc atcaagtcgc taaagaagaa agggaaacac ctactactga tagtatgccg 3780
ccattattac gacaagctat cgaattattt gatcaccaag gtgcagagcc agccttctta 3840
ttcggccttg aattgatcat atgcggatta gaaaaacaac ttaaatgtga aagtgggtcg 3900
ccaaaaaaga agagaaaggt ggatggtggc ggagcactgt ctccccagca ttcagctgtg 3960
acccagggat caatcattaa gaacaaggaa ggtatggatg ctaagtccct gaccgcttgg 4020
tcaagaactt tggtaacttt taaggacgtg ttcgtcgatt ttactcggga ggagtggaaa 4080
ctattggata ccgcccagca aatcgtgtat agaaacgtta tgctcgagaa ctataagaat 4140
cttgtgtcac tgggctacca actgacaaag cctgacgtta tcctgaggct ggaaaagggt 4200
gaggagcctt ggttggtcga gcgggagatc caccaggaaa ctcacccaga cagtgagact 4260
gctttcgaga tcaagtcctc tgtgtaatac aaggcggccg caaatcaaca tcagtctgat 4320
aagctatcaa catcagtctg ataagctatc aacatcagtc tgataagcta tcaacatcag 4380
tctgataagc taaagcttcg aattctgata atcagccata ccacatttgt agaggtttta 4440
cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt 4500
gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca 4560
aatttcacaa ataaagcatt tttttcactg cagctcgcct tttgctcaca tgtctcccag 4620
gggatggggg tccatccacc ttgccgaaaa gatttgtctg aggaactgaa aatagaaggg 4680
aaaaaagagg agggacaaaa gaggcagaaa tgagagggga ggggacagag gacacctgaa 4740
taaagaccac acccatgacc cacgtgatgc tgagaagtac tcctgcccta ggaagagact 4800
cagggcagag ggaggaagga cagcagacca gacagtcaca gcagccttga caaaacgttc 4860
ctggaactca agctcttctc cacagaggag gacagagcag acagcagaga ccatggagtc 4920
tccctcggcc cctccccaca gatggtgcat cccctggcag aggctcctgc tcacaggtga 4980
actcgagacc ggtatggccc ccccgaccga tgtcagcctg ggggacgagc tccacttaga 5040
cggcgaggac gtggcgatgg cgcatgccga cgcgctagac gatttcgatc tggacatgtt 5100
gggggacggg gattccccgg gtccgggatt taccccccac gactccgccc cctacggcgc 5160
tctggatatg gccgacttcg agtttgagca gatgtttacc gatgcccttg gaattgacga 5220
gtacggtggg acgcgtatga agctactgtc ttctatcgaa caagcatgcg atatttgccg 5280
acttaaaaag ctcaagtgct ccaaagaaaa accgaagtgc gccaagtgtc tgaagaacaa 5340
ctgggagtgt cgctactctc ccaaaaccaa aaggtctccg ctgactaggg cacatctgac 5400
agaagtggaa tcaaggctag aaagactgga acagctattt ctactgattt ttcctcgaga 5460
agaccttgac atgattttga aaatggattc tttacaggat ataaaagcat tgttaacagg 5520
attatttgta caagataatg tgaataaaga tgccgtcaca gatagattgg cttcagtgga 5580
gactgatatg cctctaacat tgagacagca tagaataagt gcgacatcat catcggaaga 5640
gagtagtaac aaaggtcaaa gacagttgac tgtataaaag cttcactcct caggtgcagg 5700
ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa taccactgag 5760
atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt 5820
ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct 5880
cactcggaag gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt 5940
agagtttggc aacatatgcc catatgctgg ctgccatgaa caaaggttgg ctataaagag 6000
gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga 6060
cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta acatccctaa 6120
aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta ctcccagtca 6180
tagctgtccc tcttctctta tggagatc 6208
<210> 20
<211> 12876
<212> DNA
<213> Artificial
<220>
<223> 编码由细胞特异性启动子CEA368、第一microRNA为mir-135a-5p、第二micr
oRNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白分
别为TALE蛋白、第一和第二识别序列为4×UAS的元件组成的表达系统的核酸
<400> 20
cggagtactg tcctccgagc ggagtactgt cctccgagcg gagtactgtc ctccgagcgg 60
agttctgtcc tccgagcgga gactctagat acccctctcc gcttctgaat tctaggcgtg 120
tacggtggga ggcctatata agcagagctc gtttagtgaa ccgtcagatc gcctcgagta 180
cccctctccg cttcttaccc ctctccgctt cttacccctc tccgcttctg aattcgaccg 240
ccaccatgag acatattatc tgccacggag gtgttattac cgaagaaatg gccgccagtc 300
ttttggacca gctgatcgaa gaggtactgg ctgataatct tccacctcct agccattttg 360
aaccacctac ccttcacgaa ctgtatgatt tagacgtgac ggcccccgaa gatcccaacg 420
aggaggcggt ttcgcagatt tttcccgact ctgtaatgtt ggcggtgcag gaagggattg 480
acttactcac ttttccgccg gcgcccggtt ctccggagcc gcctcacctt tcccggcagc 540
ccgagcagcc ggagcagaga gccttgggtc cggtttctat gccaaacctt gtaccggagg 600
tgatcgatct tacctgccac gaggctggct ttccacccag tgacgacgag gatgaagagg 660
gtgaggagtt tgtgttagat tatgtggagc accccgggca cggttgcagg tcttgtcatt 720
atcaccggag gaatacgggg gacccagata ttatgtgttc gctttgctat atgaggacct 780
gtggcatgtt tgtctacagt aagtgaaaat tatgggcagt gggtgataga gtggtgggtt 840
tggtgtggta attttttttt taatttttac agttttgtgg tttaaagaat tttgtattgt 900
gattttttta aaaggtcctg tgtctgaacc tgagcctgag cccgagccag aaccggagcc 960
tgcaagacct acccgccgtc ctaaaatggc gcctgctatc ctgaggcgcc cgacatcacc 1020
tgtgtctaga gaatgcaata gtagtacgga tagctgtgac tccggtcctt ctaacacacc 1080
tcctgagata cacccggtgg tcccgctgtg ccccattaaa ccagttgccg tgagagttgg 1140
tgggcgtcgc caggctgtgg aatgtatcga ggacttgctt aacgagcctg ggcaaccttt 1200
ggacttgagc tgtaaacgcc ccaggccagg gtccggagct acaaattttt ccctcctcaa 1260
acaggctgga gatgtcgaag aaaatcctgg gcctatggag gcttgggagt gtttggaaga 1320
tttttctgct gtgcgtaact tgctggaaca gagctctaac agtacctctt ggttttggag 1380
gtttctgtgg ggctcatccc aggcaaagtt agtctgcaga attaaggagg attacaagtg 1440
ggaatttgaa gagcttttga aatcctgtgg tgagctgttt gattctttga atctgggtca 1500
ccaggcgctt ttccaagaga aggtcatcaa gactttggat ttttccacac cggggcgcgc 1560
tgcggctgct gttgcttttt tgagttttat aaaggataaa tggagcgaag aaacccatct 1620
gagcgggggg tacctgctgg attttctggc catgcatctg tggagagcgg ttgtgagaca 1680
caagaatcgc ctgctactgt tgtcttccgt ccgcccggcg ataataccga cggaggagca 1740
gcagcagcag caggaggaag ccaggcggcg gcggcaggag cagagcccat ggaacccgag 1800
agccggcctg gaccctcggg aatgaatgtt gtacaggtgg ctgaactgta tccagaactg 1860
agacgcattt tgacaattac agaggatggg caggggctaa agggggtaaa gagggagcgg 1920
ggggcttgtg aggctacaga ggaggctagg aatctagctt ttagcttaat gaccagacac 1980
cgtcctgagt gtattacttt tcaacagatc aaggataatt gcgctaatga gcttgatctg 2040
ctggcgcaga agtattccat agagcagctg accacttact ggctgcagcc aggggatgat 2100
tttgaggagg ctattagggt atatgcaaag gtggcactta ggccagattg caagtacaag 2160
atcagcaaac ttgtaaatat caggaattgt tgctacattt ctgggaacgg ggccgaggtg 2220
gagatagata cggaggatag ggtggccttt agatgtagca tgataaatat gtggccgggg 2280
gtgcttggca tggacggggt ggttattatg aatgtaaggt ttactggccc caattttagc 2340
ggtacggttt tcctggccaa taccaacctt atcctacacg gtgtaagctt ctatgggttt 2400
aacaatacct gtgtggaagc ctggaccgat gtaagggttc ggggctgtgc cttttactgc 2460
tgctggaagg gggtggtgtg tcgccccaaa agcagggctt caattaagaa atgcctcttt 2520
gaaaggtgta ccttgggtat cctgtctgag ggtaactcca gggtgcgcca caatgtggcc 2580
tccgactgtg gttgcttcat gctagtgaaa agcgtggctg tgattaagca taacatggta 2640
tgtggcaact gcgaggacag ggcctctcag atgctgacct gctcggacgg caactgtcac 2700
ctgctgaaga ccattcacgt agccagccac tctcgcaagg cctggccagt gtttgagcat 2760
aacatactga cccgctgttc cttgcatttg ggtaacagga ggggggtgtt cctaccttac 2820
caatgcaatt tgagtcacac taagatattg cttgagcccg agagcatgtc caaggtgaac 2880
ctgaacgggg tgtttgacat gaccatgaag atctggaagg tgctgaggta cgatgagacc 2940
cgcaccaggt gcagaccctg cgagtgtggc ggtaaacata ttaggaacca gcctgtgatg 3000
ctggatgtga ccgaggagct gaggcccgat cacttggtgc tggcctgcac ccgcgctgag 3060
tttggctcta gcgatgaaga tacagatctc gaggggtccg gagctacaaa tttttccctc 3120
ctcaaacagg ctggagatgt cgaagaaaat cctgggccta ccggtatgta taggatgcag 3180
ttgctctctt gcatcgccct gtctctagcg ttggtgacta atagcgcccc tacttcttct 3240
tccacgaaga aaacccagct gcaactggaa caccttctgc tggatctcca gatgatcctt 3300
aacgggatta acaactataa gaacccgaag ctcacggcta tgctgacggc gaagttcgct 3360
atgcctaaaa aggctactga gctgaaacac ttacagtgtc tcgaagaggc actcaaaccc 3420
ctcgaggagg tactcaatct ggcgcaaagc aaaaacttcc atctccgccc acgggaccta 3480
atctccaaca tcaacgtgat tgtgctggag ctcaaggggt ctgaaaccac gtttatgtgc 3540
gagtacgcag atgaaactgc taccatagtg gaattcctca atcgttggat cactttctgc 3600
cagtcaatca tttcaaccct gaccagatct gagggccgcg gcagcctgct gacctgcggc 3660
gacgtggagg aaaaccccgg ccccggatcc ttgtcgacca ttcgtccgcg caggccaagt 3720
cctgcccgcg agcttctgcc cggaccccaa ccggataggg ttcagccgac tgcagatcgt 3780
ggggtgtctg cgcctgctgg cagccctctg gatggcttgc ccgctcggcg gacggtgtcc 3840
cggacccggc tgccatctcc ccctgcgccc tcacctgcgt tctcggcggg cagcttcagc 3900
gatctgctcc gtccgttcga tccgtcgctt cttgatacat cgcttcttga ttcgatgcct 3960
gccgtcggca cgccgcatac agcggctgcc ccagcagagt gggatgaggc gcaatcggct 4020
ctgcgtgcag ccgatgaccc gccacccacc gtgcgtgtcg ctgtcactgc cgcgcggccg 4080
ccgcgcgcca agccggcccc gcgacggcgt gctgcgcaac cctccgacgc ttcgccggcc 4140
gcgcaggtgg atctacgcac gctcggctac agtcagcagc agcaagagaa gatcaaaccg 4200
aaggtgcgtt cgacagtggc gcagcaccac gaggcactgg tgggccatgg gtttacacac 4260
gcgcacatcg ttgcgctcag ccaacacccg gcagcgttag ggaccgtcgc tgtcacgtat 4320
cagcacataa tcacggcgtt gccagaggcg acacacgaag acatcgttgg cgtcggcaaa 4380
cagtggtccg gcgcacgcgc cctggaggcc ttgctcacgg atgcggggga gttgagaggt 4440
ccgccgttac agttggacac aggccaactt gtgaagattg caaaacgtgg cggcgtgacc 4500
gcaatggagg cagtgcatgc atcgcgcaat gcactgacgg gtgcccccct gaacctgacc 4560
ccggaccaag tggtggctat cgccagcaac attggcggca agcaagcgct cgaaacggtg 4620
cagcggctgt tgccggtgct gtgccaggac catggcctga ccccggacca agtggtggct 4680
atcgccagca acggtggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg 4740
ctgtgccagg accatggcct gaccccggac caagtggtgg ctatcgccag caacattggc 4800
ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc 4860
ctgaccccgg accaagtggt ggctatcgcc agcaacaatg gcggcaagca agcgctcgaa 4920
acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg gcctgacccc ggaccaagtg 4980
gtggctatcg ccagcaacat tggcggcaag caagcgctcg aaacggtgca gcggctgttg 5040
ccggtgctgt gccaggacca tggcctgacc ccggaccaag tggtggctat cgccagcaac 5100
attggcggca agcaagcgct cgaaacggtg cagcggctgt tgccggtgct gtgccaggac 5160
catggcctga ctccggacca agtggtggct atcgccagcc acgatggcgg caagcaagcg 5220
ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg accatggcct gactccggac 5280
caagtggtgg ctatcgccag ccacgatggc ggcaagcaag cgctcgaaac ggtgcagcgg 5340
ctgttgccgg tgctgtgcca ggaccatggc ctgaccccgg accaagtggt ggctatcgcc 5400
agcaacaatg gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc 5460
caggaccatg gcctgacccc ggaccaagtg gtggctatcg ccagcaacat tggcggcaag 5520
caagcgctcg aaacggtgca gcggctgttg ccggtgctgt gccaggacca tggcctgacc 5580
ccggaccaag tggtggctat cgccagcaac ggtggcggca agcaagcgct cgaaacggtg 5640
cagcggctgt tgccggtgct gtgccaggac catggcctga ctccggacca agtggtggct 5700
atcgccagcc acgatggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg 5760
ctgtgccagg accatggcct gactccggac caagtggtgg ctatcgccag ccacgatggc 5820
ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc 5880
ctgaccccgg accaagtggt ggctatcgcc agcaacggtg gcggcaagca agcgctcgaa 5940
acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg gcctgactcc ggaccaagtg 6000
gtggctatcg ccagccacga tggcggcaag caagcgctcg aaacggtgca gcggctgttg 6060
ccggtgctgt gccaggacca tggcctgact ccggaccaag tggtggctat cgccagccac 6120
gatggcggca agcaagcgct cgaaacggtg cagcggctgt tgccggtgct gtgccaggac 6180
catggcctga ctccggacca agtggtggct atcgccagcc acgatggcgg caagcaagcg 6240
ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg accatggcct gaccccggac 6300
caagtggtgg ctatcgccag caacattggc ggcaagcaag cgctcgaaac ggtgcagcgg 6360
ctgttgccgg tgctgtgcca ggaccatggc ctgaccccgg accaagtggt ggctatcgcc 6420
agcaacggtg gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc 6480
caggaccatg gcctgacccc ggaccaagtg gtggctatcg ccagcaacgg tggcggcaag 6540
caagcgctcg aaagcattgt ggcccagctg agccggcctg atccggcgtt ggccgcgttg 6600
accaacgacc acctcgtcgc cttggcctgc ctcggcggac gtcctgccat ggatgcagtg 6660
aaaaagggat tgccgcacgc gccggaattg atcagaagag tcaatcgccg tattggcgaa 6720
cgcacgtccc atcgcgttgc cgactacgcg caagtggttc gcgtgctgga gtttttccag 6780
tgccactccc acccagcgta cgcatttgat gaggccatga cgcagttcgg gatgagcagg 6840
aacgggttgg tacagctctt tcgcagagtg ggcgtcaccg aactcgaagc ccgcggtgga 6900
acgctccccc cagcctcgca gcgttgggac cgtatcctcc aggcatcagg gatgaaaagg 6960
gccaaaccgt cccctacttc agctcaaaca ccggatcagg cgtctttgca tgcattcgcc 7020
gattcgctgg agcgtgacct tgatgcgccc agcccaatgc acgagggaga tcagacgcgg 7080
gcaagcagcc gtaaacggtc ccgatcggat cgtgctgtca ccggcccctc cgcacagcag 7140
gctgtcgagg tgcgcgttcc cgaacagcgc gatgcgctgc atttgcccct cagctggagg 7200
gtaaaacgcc cgcgtaccag gatctggggc ggcctcccgg ttaagtagtg ctttctactt 7260
tatgagtagt gctttctact ttatgagtag tgctttctac tttatgagta gtgctttcta 7320
ctttatgtca cataggaata aaaagccata tcacatagga ataaaaagcc atatcacata 7380
ggaataaaaa gccatatcac ataggaataa aaagccatag aaagcggagt actgtcctcc 7440
gagcggagta ctgtcctccg agcggagtac tgtcctccga gcggagttct gtcctccgag 7500
cggagactct agatatagaa ccgatcctcc cattgaattc taggcgtgta cggtgggagg 7560
cctatataag cagagctcgt ttagtgaacc gtcagatcgc ctcgagtata gaaccgatcc 7620
tcccatttat agaaccgatc ctcccattta tagaaccgat cctcccatta ctagaagatc 7680
ttgaattcgc caccttgtcg accattcgtc cgcgcaggcc aagtcctgcc cgcgagcttc 7740
tgcccggacc ccaaccggat agggttcagc cgactgcaga tcgtggggtg tctgcgcctg 7800
ctggcagccc tctggatggc ttgcccgctc ggcggacggt gtcccggacc cggctgccat 7860
ctccccctgc gccctcacct gcgttctcgg cgggcagctt cagcgatctg ctccgtccgt 7920
tcgatccgtc gcttcttgat acatcgcttc ttgattcgat gcctgccgtc ggcacgccgc 7980
atacagcggc tgccccagca gagtgggatg aggcgcaatc ggctctgcgt gcagccgatg 8040
acccgccacc caccgtgcgt gtcgctgtca ctgccgcgcg gccgccgcgc gccaagccgg 8100
ccccgcgacg gcgtgctgcg caaccctccg acgcttcgcc ggccgcgcag gtggatctac 8160
gcacgctcgg ctacagtcag cagcagcaag agaagatcaa accgaaggtg cgttcgacag 8220
tggcgcagca ccacgaggca ctggtgggcc atgggtttac acacgcgcac atcgttgcgc 8280
tcagccaaca cccggcagcg ttagggaccg tcgctgtcac gtatcagcac ataatcacgg 8340
cgttgccaga ggcgacacac gaagacatcg ttggcgtcgg caaacagtgg tccggcgcac 8400
gcgccctgga ggccttgctc acggatgcgg gggagttgag aggtccgccg ttacagttgg 8460
acacaggcca acttgtgaag attgcaaaac gtggcggcgt gaccgcaatg gaggcagtgc 8520
atgcatcgcg caatgcactg acgggtgccc ccctgaacct gaccccggac caagtggtgg 8580
ctatcgccag caacattggc ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg 8640
tgctgtgcca ggaccatggc ctgactccgg accaagtggt ggctatcgcc agccacgatg 8700
gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg 8760
gcctgactcc ggaccaagtg gtggctatcg ccagccacga tggcggcaag caagcgctcg 8820
aaacggtgca gcggctgttg ccggtgctgt gccaggacca tggcctgact ccggaccaag 8880
tggtggctat cgccagccac gatggcggca agcaagcgct cgaaacggtg cagcggctgt 8940
tgccggtgct gtgccaggac catggcctga ctccggacca agtggtggct atcgccagcc 9000
acgatggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg 9060
accatggcct gaccccggac caagtggtgg ctatcgccag caacggtggc ggcaagcaag 9120
cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc ctgactccgg 9180
accaagtggt ggctatcgcc agccacgatg gcggcaagca agcgctcgaa acggtgcagc 9240
ggctgttgcc ggtgctgtgc caggaccatg gcctgacccc ggaccaagtg gtggctatcg 9300
ccagcaacgg tggcggcaag caagcgctcg aaacggtgca gcggctgttg ccggtgctgt 9360
gccaggacca tggcctgact ccggaccaag tggtggctat cgccagccac gatggcggca 9420
agcaagcgct cgaaacggtg cagcggctgt tgccggtgct gtgccaggac catggcctga 9480
ctccggacca agtggtggct atcgccagcc acgatggcgg caagcaagcg ctcgaaacgg 9540
tgcagcggct gttgccggtg ctgtgccagg accatggcct gaccccggac caagtggtgg 9600
ctatcgccag caacaatggc ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg 9660
tgctgtgcca ggaccatggc ctgactccgg accaagtggt ggctatcgcc agccacgatg 9720
gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg 9780
gcctgacccc ggaccaagtg gtggctatcg ccagcaacgg tggcggcaag caagcgctcg 9840
aaacggtgca gcggctgttg ccggtgctgt gccaggacca tggcctgacc ccggaccaag 9900
tggtggctat cgccagcaac ggtggcggca agcaagcgct cgaaacggtg cagcggctgt 9960
tgccggtgct gtgccaggac catggcctga ctccggacca agtggtggct atcgccagcc 10020
acgatggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg 10080
accatggcct gaccccggac caagtggtgg ctatcgccag caacggtggc ggcaagcaag 10140
cgctcgaaag cattgtggcc cagctgagcc ggcctgatcc ggcgttggcc gcgttgacca 10200
acgaccacct cgtcgccttg gcctgcctcg gcggacgtcc tgccatggat gcagtgaaaa 10260
agggattgcc gcacgcgccg gaattgatca gaagagtcaa tcgccgtatt ggcgaacgca 10320
cgtcccatcg cgttgccgac tacgcgcaag tggttcgcgt gctggagttt ttccagtgcc 10380
actcccaccc agcgtacgca tttgatgagg ccatgacgca gttcgggatg agcaggaacg 10440
ggttggtaca gctctttcgc agagtgggcg tcaccgaact cgaagcccgc ggtggaacgc 10500
tccccccagc ctcgcagcgt tgggaccgta tcctccaggc atcagggatg aaaagggcca 10560
aaccgtcccc tacttcagct caaacaccgg atcaggcgtc tttgcatgca ttcgccgatt 10620
cgctggagcg tgaccttgat gcgcccagcc caatgcacga gggagatcag acgcgggcaa 10680
gcagccgtaa acggtcccga tcggatcgtg ctgtcaccgg cccctccgca cagcaggctg 10740
tcgaggtgcg cgttcccgaa cagcgcgatg cgctgcattt gcccctcagc tggagggtaa 10800
aacgcccgcg taccaggatc tggggcggcc tcccgtaata caaggcggcc gcaaatcaac 10860
atcagtctga taagctatca acatcagtct gataagctat caacatcagt ctgataagct 10920
atcaacatca gtctgataag ctaaagcttc gaattctgat aatcagccat accacatttg 10980
tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa 11040
tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca 11100
atagcatcac aaatttcaca aataaagcat ttttttcact gcagctcgcc ttttgctcac 11160
atgtctccca ggggatgggg gtccatccac cttgccgaaa agatttgtct gaggaactga 11220
aaatagaagg gaaaaaagag gagggacaaa agaggcagaa atgagagggg aggggacaga 11280
ggacacctga ataaagacca cacccatgac ccacgtgatg ctgagaagta ctcctgccct 11340
aggaagagac tcagggcaga gggaggaagg acagcagacc agacagtcac agcagccttg 11400
acaaaacgtt cctggaactc aagctcttct ccacagagga ggacagagca gacagcagag 11460
accatggagt ctccctcggc ccctccccac agatggtgca tcccctggca gaggctcctg 11520
ctcacaggtg aactcgagac cggtatggcc cccccgaccg atgtcagcct gggggacgag 11580
ctccacttag acggcgagga cgtggcgatg gcgcatgccg acgcgctaga cgatttcgat 11640
ctggacatgt tggggagcga cgcgctagac gatttcgatc tggacatgtt ggggagcgac 11700
gcgctagacg atttcgatct ggacatgttg gggagcgacg cgctagacga tttcgatctg 11760
gacatgttgg gggacgggga ttccccgggt ccgggattta ccccccacga ctccgccccc 11820
tacggcgctc tggatatggc cgacttcgag tttgagcaga tgtttaccga tgcccttgga 11880
attgacgagt acggtgggac gcgtatgaag ctactgtctt ctatcgaaca agcatgcgat 11940
atttgccgac ttaaaaagct caagtgctcc aaagaaaaac cgaagtgcgc caagtgtctg 12000
aagaacaact gggagtgtcg ctactctccc aaaaccaaaa ggtctccgct gactagggca 12060
catctgacag aagtggaatc aaggctagaa agactggaac agctatttct actgattttt 12120
cctcgagaag accttgacat gattttgaaa atggattctt tacaggatat aaaagcattg 12180
ttaacaggat tatttgtaca agataatgtg aataaagatg ccgtcacaga tagattggct 12240
tcagtggaga ctgatatgcc tctaacattg agacagcata gaataagtgc gacatcatca 12300
tcggaagaga gtagtaacaa aggtcaaaga cagttgactg tataaaagct tcactcctca 12360
ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc aatgccctgg ctcacaaata 12420
ccactgagat ctttttccct ctgccaaaaa ttatggggac atcatgaagc cccttgagca 12480
tctgacttct ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt 12540
gtgtctctca ctcggaagga catatgggag ggcaaatcat ttaaaacatc agaatgagta 12600
tttggtttag agtttggcaa catatgccca tatgctggct gccatgaaca aaggttggct 12660
ataaagaggt catcagtata tgaaacagcc ccctgctgtc cattccttat tccatagaaa 12720
agccttgact tgaggttaga ttttttttat attttgtttt gtgttatttt tttctttaac 12780
atccctaaaa ttttccttac atgttttact agccagattt ttcctcctct cctgactact 12840
cccagtcata gctgtccctc ttctcttatg gagatc 12876
<210> 21
<211> 6647
<212> DNA
<213> Artificial
<220>
<223> 编码由细胞特异性启动子hMUC1、第一microRNA为mir-199a-3p、第二micro
RNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白分
别为Lacl、tetR-KRAB、第一和第二识别序列为4×UAS的元件组成的表达系
统的核酸
<400> 21
cggccatatg tcttccgcgg agtgctgtac tccgcggagt actgtcctcc gcggagtgcc 60
gcactccgag cggagactct agactcccta tcagtgatag agataggcgt gtacggtggg 120
aggcctatat aagcagagct cgtttagtga accgtcagat cgctccctat cagtgataga 180
gagaattcga ccgccaccat gagacatatt atctgccacg gaggtgttat taccgaagaa 240
atggccgcca gtcttttgga ccagctgatc gaagaggtac tggctgataa tcttccacct 300
cctagccatt ttgaaccacc tacccttcac gaactgtatg atttagacgt gacggccccc 360
gaagatccca acgaggaggc ggtttcgcag atttttcccg actctgtaat gttggcggtg 420
caggaaggga ttgacttact cacttttccg ccggcgcccg gttctccgga gccgcctcac 480
ctttcccggc agcccgagca gccggagcag agagccttgg gtccggtttc tatgccaaac 540
cttgtaccgg aggtgatcga tcttacctgc cacgaggctg gctttccacc cagtgacgac 600
gaggatgaag agggtgagga gtttgtgtta gattatgtgg agcaccccgg gcacggttgc 660
aggtcttgtc attatcaccg gaggaatacg ggggacccag atattatgtg ttcgctttgc 720
tatatgagga cctgtggcat gtttgtctac agtaagtgaa aattatgggc agtgggtgat 780
agagtggtgg gtttggtgtg gtaatttttt ttttaatttt tacagttttg tggtttaaag 840
aattttgtat tgtgattttt ttaaaaggtc ctgtgtctga acctgagcct gagcccgagc 900
cagaaccgga gcctgcaaga cctacccgcc gtcctaaaat ggcgcctgct atcctgaggc 960
gcccgacatc acctgtgtct agagaatgca atagtagtac ggatagctgt gactccggtc 1020
cttctaacac acctcctgag atacacccgg tggtcccgct gtgccccatt aaaccagttg 1080
ccgtgagagt tggtgggcgt cgccaggctg tggaatgtat cgaggacttg cttaacgagc 1140
ctgggcaacc tttggacttg agctgtaaac gccccaggcc actcgagggt accgggtccg 1200
gagctacaaa tttttccctc ctcaaacagg ctggagatgt cgaagaaaat cctgggccta 1260
ccggtatgtg gctgcagagc ctccttctac tgggcaccgt ggcttgcagc atctcggccc 1320
ccgccagaag tcccagcccc agcacccagc cctgggaaca cgtaaatgca atccaggagg 1380
cccgccgcct cctgaacttg tcacgggaca cggcagccga gatgaatgaa actgtggagg 1440
tcatcagtga aatgtttgac ttgcaggagc ccacatgcct gcagactcgg ctggagctct 1500
acaaacaggg cctgcgtggc agcctcacca agctgaaggg cccactgacc atgatggcta 1560
gccattacaa gcagcactgc ccgccgaccc ctgagacatc ctgtgccact cagatcatta 1620
cctttgagtc tttcaaggag aacctcaagg actttctgct ggtcatccca tttgactgct 1680
gggaacccgt ccaggaaaga tctgagggcc gcggcagcct gctgacctgc ggcgacgtgg 1740
aggaaaaccc cggccccgga tccatgaaac cagtaacgtt atacgatgtc gcagagtatg 1800
ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac gtttctgcga 1860
aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg 1920
cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc agtctggccc 1980
tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca 2040
gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca 2100
atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg 2160
ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc 2220
agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg ggcgtggagc 2280
atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct 2340
cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga 2400
tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc 2460
tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg 2520
caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggatatctcg gtagtgggat 2580
acgacgatac cgaagacagc tcatgttata tcccgccgtt aaccaccatc aaacaggatt 2640
ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg 2700
tgaagggcaa tcagctgttg ccggtctcac tggtgaaaag aaaaaccacc ctggcgccca 2760
atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg 2820
tttcccgact ggaaagcggg cagagcagcc tgaggcctcc taagaagaag aggaaggttg 2880
cggccgctaa ccaatgtgca gactactgtt aaccaatgtg cagactactg tgaattctaa 2940
ccaatgtgca gactactgtt aaccaatgtg cagactactg taagcttcga attgagtagt 3000
gctttctact ttatgagtag tgctttctac tttatgagta gtgctttcta ctttatgagt 3060
agtgctttct actttatgag tagtgctttc tactttatgg tcgacaatac tagttaagaa 3120
agcggtccac tgtgtgccgc gggtgacagc cctccgcggc ggtctttcgt ccgcgggtga 3180
ccgccctccg agcggagact ctagaaattg tgagcggata acaattgggt atataatggg 3240
ggccgtcagc tactaccaga gccgatcgaa cactcgagcc gagcagacgt gcctacaatt 3300
gtgagcgctc acaattacta gaagatcttg aattcgccac catggctaga ttagataaaa 3360
gtaaagtgat taacagcgca ttagagctgc ttaatgaggt cggaatcgaa ggtttaacaa 3420
cccgtaaact cgcccagaag ctaggtgtag agcagcctac attgtattgg catgtaaaaa 3480
ataagcgggc tttgctcgac gccttagcca ttgagatgtt agataggcac catactcact 3540
tttgcccttt agaaggggaa agctggcaag attttttacg taataacgct aaaagtttta 3600
gatgtgcttt actaagtcat cgcgatggag caaaagtaca tttaggtaca cggcctacag 3660
aaaaacagta tgaaactctc gaaaatcaat tagccttttt atgccaacaa ggtttttcac 3720
tagagaatgc attatatgca ctcagcgctg tggggcattt tactttaggt tgcgtattgg 3780
aagatcaaga gcatcaagtc gctaaagaag aaagggaaac acctactact gatagtatgc 3840
cgccattatt acgacaagct atcgaattat ttgatcacca aggtgcagag ccagccttct 3900
tattcggcct tgaattgatc atatgcggat tagaaaaaca acttaaatgt gaaagtgggt 3960
cgccaaaaaa gaagagaaag gtggatggtg gcggagcact gtctccccag cattcagctg 4020
tgacccaggg atcaatcatt aagaacaagg aaggtatgga tgctaagtcc ctgaccgctt 4080
ggtcaagaac tttggtaact tttaaggacg tgttcgtcga ttttactcgg gaggagtgga 4140
aactattgga taccgcccag caaatcgtgt atagaaacgt tatgctcgag aactataaga 4200
atcttgtgtc actgggctac caactgacaa agcctgacgt tatcctgagg ctggaaaagg 4260
gtgaggagcc ttggttggtc gagcgggaga tccaccagga aactcaccca gacagtgaga 4320
ctgctttcga gatcaagtcc tctgtgtaat acaaggcggc cgcaaatcaa catcagtctg 4380
ataagctatc aacatcagtc tgataagcta tcaacatcag tctgataagc tatcaacatc 4440
agtctgataa gctaaagctt cgaattctga taatcagcca taccacattt gtagaggttt 4500
tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa 4560
ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca 4620
caaatttcac aaataaagca tttttttcac tgcagctcga ctagtgttca tcggagccca 4680
ggtttactcc cttaagtgga aatttcttcc cccactccct ccttggcttt ctccaaggag 4740
ggaacccagg ctactggaaa gtccggctgg ggcggggact gtgggtttca gggtagaact 4800
gcgtgtggaa cgggacaggg agcggttaga agggtggggc tattccggga agtggtgggg 4860
ggagggagcc caaaactagc acctagtcca ctcattatcc agccctctta tttctcggcc 4920
ccgctctgct tcagtggacc cggggagggc ggggaagtgg agtgggagac ctaggggtgg 4980
gcttcccgac cttgctgtac aggacctcga cctagctggc tttgttcccc atccccacgt 5040
tagttgttgc cctgaggcta aaactagagc ccaggggccc caagttccag actgcccctc 5100
ccccctcccc cggagccagg gagtggttgg tgaaaggggg aggccagctg gagaacaaac 5160
gggtagtcag ggggttgagc gattagagcc cttgtaccct acccaggaat ggttggggag 5220
gaggaggaag aggtaggagg taggggaggg ggcggggttt tgtcacctgt cacctgctcc 5280
ggctgtgcct agggcgggcg ggcggggagt ggggggaccg gtataaagcg gtaggcgcct 5340
gtgcccgctc cacctctcaa gcagccagcg cctgcctgaa tctgttctgc cccctcccca 5400
cccatttcac caccaccctc gagggatccg gccaccatgg cccccccgac cgatgtcagc 5460
ctgggggacg agctccactt agacggcgag gacgtggcga tggcgcatgc cgacgcgcta 5520
gacgatttcg atctggacat gttgggggac ggggattccc cgggtccggg atttaccccc 5580
cacgactccg ccccctacgg cgctctggat atggccgact tcgagtttga gcagatgttt 5640
accgatgccc ttggaattga cgagtacggt gggacgcgta tgaagctact gtcttctatc 5700
gaacaagcat gcgatatttg ccgacttaaa aagctcaagt gctccaaaga aaaaccgaag 5760
tgcgccaagt gtctgaagaa caactgggag tgtcgctact ctcccaaaac caaaaggtct 5820
ccgctgacta gggcacatct gacagaagtg gaatcaaggc tagaaagact ggaacagcta 5880
tttctactga tttttcctcg agaagacctt gacatgattt tgaaaatgga ttctttacag 5940
gatataaaag cattgttaac aggattattt gtacaagata atgtgaataa agatgccgtc 6000
acagatagat tggcttcagt ggagactgat atgcctctaa cattgagaca gcatagaata 6060
agtgcgacat catcatcgga agagagtagt aacaaaggtc aaagacagtt gactgtataa 6120
ttcactcctc aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc caatgccctg 6180
gctcacaaat accactgaga tctttttccc tctgccaaaa attatgggga catcatgaag 6240
ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt 6300
tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat 6360
cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc tgccatgaac 6420
aaaggttggc tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta 6480
ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt 6540
ttttctttaa catccctaaa attttcctta catgttttac tagccagatt tttcctcctc 6600
tcctgactac tcccagtcat agctgtccct cttctcttat ggagatc 6647
<210> 22
<211> 6977
<212> DNA
<213> Artificial
<220>
<223> 编码由细胞特异性启动子hMUC1、第一microRNA为mir-199a-3p、第二micro
RNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白分
别为Lacl、tetR-KRAB、第一和第二识别序列为4×UAS的元件组成的表达系
统的核酸
<400> 22
cggccatatg tcttccgcgg agtgctgtac tccgcggagt actgtcctcc gcggagtgcc 60
gcactccgag cggagactct agactcccta tcagtgatag agataggcgt gtacggtggg 120
aggcctatat aagcagagct cgtttagtga accgtcagat cgctccctat cagtgataga 180
gagaattcga ccgccaccat gagacatatt atctgccacg gaggtgttat taccgaagaa 240
atggccgcca gtcttttgga ccagctgatc gaagaggtac tggctgataa tcttccacct 300
cctagccatt ttgaaccacc tacccttcac gaactgtatg atttagacgt gacggccccc 360
gaagatccca acgaggaggc ggtttcgcag atttttcccg actctgtaat gttggcggtg 420
caggaaggga ttgacttact cacttttccg ccggcgcccg gttctccgga gccgcctcac 480
ctttcccggc agcccgagca gccggagcag agagccttgg gtccggtttc tatgccaaac 540
cttgtaccgg aggtgatcga tcttacctgc cacgaggctg gctttccacc cagtgacgac 600
gaggatgaag agggtgagga gtttgtgtta gattatgtgg agcaccccgg gcacggttgc 660
aggtcttgtc attatcaccg gaggaatacg ggggacccag atattatgtg ttcgctttgc 720
tatatgagga cctgtggcat gtttgtctac agtaagtgaa aattatgggc agtgggtgat 780
agagtggtgg gtttggtgtg gtaatttttt ttttaatttt tacagttttg tggtttaaag 840
aattttgtat tgtgattttt ttaaaaggtc ctgtgtctga acctgagcct gagcccgagc 900
cagaaccgga gcctgcaaga cctacccgcc gtcctaaaat ggcgcctgct atcctgaggc 960
gcccgacatc acctgtgtct agagaatgca atagtagtac ggatagctgt gactccggtc 1020
cttctaacac acctcctgag atacacccgg tggtcccgct gtgccccatt aaaccagttg 1080
ccgtgagagt tggtgggcgt cgccaggctg tggaatgtat cgaggacttg cttaacgagc 1140
ctgggcaacc tttggacttg agctgtaaac gccccaggcc actcgagggg tccggagcta 1200
caaatttttc cctcctcaaa caggctggag atgtcgaaga aaatcctggg cctaccggtc 1260
ccatgagtgt gcccactcag gtcctggggt tgctgctgct gtggcttaca gacgctcgct 1320
gccaggtgca gctggtggag agcggcggcg gcgtggtgca gcccggcagg agcctgaggc 1380
tggactgcaa ggccagcggc atcaccttca gcaacagcgg catgcactgg gtgaggcagg 1440
cccccggcaa gggcctggag tgggtggccg tgatctggta cgacggcagc aagaggtact 1500
acgccgacag cgtgaagggc aggttcacca tcagcaggga caacagcaag aacaccctgt 1560
tcctgcagat gaacagcctg agggccgagg acaccgccgt gtactactgc gccaccaacg 1620
acgactactg gggccagggc accctggtga ccgtgagcag cggtggaggc ggttcaggcg 1680
gaggtggctc tggcggtggc ggatcggaga tcgtgctgac ccagagcccc gccaccctga 1740
gcctgagccc cggcgagagg gccaccctga gctgcagggc cagccagagc gtgagcagct 1800
acctggcctg gtaccagcag aagcccggcc aggcccccag gctgctgatc tacgacgcca 1860
gcaacagggc caccggcatc cccgccaggt tcagcggcag cggcagcggc accgacttca 1920
ccctgaccat cagcagcctg gagcccgagg acttcgccgt gtactactgc cagcagagca 1980
gcaactggcc caggaccttc ggccagggca ccaaggtgga gatcaagaga tctgagggcc 2040
gcggcagcct gctgacctgc ggcgacgtgg aggaaaaccc cggccccgga tccatgaaac 2100
cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg 2160
tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg 2220
cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc 2280
tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga 2340
ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg 2400
gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga 2460
tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg 2520
ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt attttctccc 2580
atgaagacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg 2640
cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata 2700
aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca 2760
tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc 2820
tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc 2880
gcgttggtgc ggatatctcg gtagtgggat acgacgatac cgaagacagc tcatgttata 2940
tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc 3000
gcttgctgca actctctcag ggccaggcgg tgaagggcaa tcagctgttg ccggtctcac 3060
tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg 3120
ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagagcagcc 3180
tgaggcctcc taagaagaag aggaaggttg cggccgctaa ccaatgtgca gactactgtt 3240
aaccaatgtg cagactactg tgaattctaa ccaatgtgca gactactgtt aaccaatgtg 3300
cagactactg taagcttcga attgagtagt gctttctact ttatgagtag tgctttctac 3360
tttatgagta gtgctttcta ctttatgagt agtgctttct actttatgag tagtgctttc 3420
tactttatgg tcgacaatac tagttaagaa agcggtccac tgtgtgccgc gggtgacagc 3480
cctccgcggc ggtctttcgt ccgcgggtga ccgccctccg agcggagact ctagaaattg 3540
tgagcggata acaattgggt atataatggg ggccgtcagc tactaccaga gccgatcgaa 3600
cactcgagcc gagcagacgt gcctacaatt gtgagcgctc acaattacta gaagatcttg 3660
aattcgccac catggctaga ttagataaaa gtaaagtgat taacagcgca ttagagctgc 3720
ttaatgaggt cggaatcgaa ggtttaacaa cccgtaaact cgcccagaag ctaggtgtag 3780
agcagcctac attgtattgg catgtaaaaa ataagcgggc tttgctcgac gccttagcca 3840
ttgagatgtt agataggcac catactcact tttgcccttt agaaggggaa agctggcaag 3900
attttttacg taataacgct aaaagtttta gatgtgcttt actaagtcat cgcgatggag 3960
caaaagtaca tttaggtaca cggcctacag aaaaacagta tgaaactctc gaaaatcaat 4020
tagccttttt atgccaacaa ggtttttcac tagagaatgc attatatgca ctcagcgctg 4080
tggggcattt tactttaggt tgcgtattgg aagatcaaga gcatcaagtc gctaaagaag 4140
aaagggaaac acctactact gatagtatgc cgccattatt acgacaagct atcgaattat 4200
ttgatcacca aggtgcagag ccagccttct tattcggcct tgaattgatc atatgcggat 4260
tagaaaaaca acttaaatgt gaaagtgggt cgccaaaaaa gaagagaaag gtggatggtg 4320
gcggagcact gtctccccag cattcagctg tgacccaggg atcaatcatt aagaacaagg 4380
aaggtatgga tgctaagtcc ctgaccgctt ggtcaagaac tttggtaact tttaaggacg 4440
tgttcgtcga ttttactcgg gaggagtgga aactattgga taccgcccag caaatcgtgt 4500
atagaaacgt tatgctcgag aactataaga atcttgtgtc actgggctac caactgacaa 4560
agcctgacgt tatcctgagg ctggaaaagg gtgaggagcc ttggttggtc gagcgggaga 4620
tccaccagga aactcaccca gacagtgaga ctgctttcga gatcaagtcc tctgtgtaat 4680
acaaggcggc cgcaaatcaa catcagtctg ataagctatc aacatcagtc tgataagcta 4740
tcaacatcag tctgataagc tatcaacatc agtctgataa gctaaagctt cgaattctga 4800
taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc 4860
ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt 4920
ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac 4980
tgcagctcga ctagtgttca tcggagccca ggtttactcc cttaagtgga aatttcttcc 5040
cccactccct ccttggcttt ctccaaggag ggaacccagg ctactggaaa gtccggctgg 5100
ggcggggact gtgggtttca gggtagaact gcgtgtggaa cgggacaggg agcggttaga 5160
agggtggggc tattccggga agtggtgggg ggagggagcc caaaactagc acctagtcca 5220
ctcattatcc agccctctta tttctcggcc ccgctctgct tcagtggacc cggggagggc 5280
ggggaagtgg agtgggagac ctaggggtgg gcttcccgac cttgctgtac aggacctcga 5340
cctagctggc tttgttcccc atccccacgt tagttgttgc cctgaggcta aaactagagc 5400
ccaggggccc caagttccag actgcccctc ccccctcccc cggagccagg gagtggttgg 5460
tgaaaggggg aggccagctg gagaacaaac gggtagtcag ggggttgagc gattagagcc 5520
cttgtaccct acccaggaat ggttggggag gaggaggaag aggtaggagg taggggaggg 5580
ggcggggttt tgtcacctgt cacctgctcc ggctgtgcct agggcgggcg ggcggggagt 5640
ggggggaccg gtataaagcg gtaggcgcct gtgcccgctc cacctctcaa gcagccagcg 5700
cctgcctgaa tctgttctgc cccctcccca cccatttcac caccaccctc gagggatccg 5760
gccaccatgg cccccccgac cgatgtcagc ctgggggacg agctccactt agacggcgag 5820
gacgtggcga tggcgcatgc cgacgcgcta gacgatttcg atctggacat gttgggggac 5880
ggggattccc cgggtccggg atttaccccc cacgactccg ccccctacgg cgctctggat 5940
atggccgact tcgagtttga gcagatgttt accgatgccc ttggaattga cgagtacggt 6000
gggacgcgta tgaagctact gtcttctatc gaacaagcat gcgatatttg ccgacttaaa 6060
aagctcaagt gctccaaaga aaaaccgaag tgcgccaagt gtctgaagaa caactgggag 6120
tgtcgctact ctcccaaaac caaaaggtct ccgctgacta gggcacatct gacagaagtg 6180
gaatcaaggc tagaaagact ggaacagcta tttctactga tttttcctcg agaagacctt 6240
gacatgattt tgaaaatgga ttctttacag gatataaaag cattgttaac aggattattt 6300
gtacaagata atgtgaataa agatgccgtc acagatagat tggcttcagt ggagactgat 6360
atgcctctaa cattgagaca gcatagaata agtgcgacat catcatcgga agagagtagt 6420
aacaaaggtc aaagacagtt gactgtataa ttcactcctc aggtgcaggc tgcctatcag 6480
aaggtggtgg ctggtgtggc caatgccctg gctcacaaat accactgaga tctttttccc 6540
tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc tggctaataa 6600
aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg 6660
acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca 6720
acatatgccc atatgctggc tgccatgaac aaaggttggc tataaagagg tcatcagtat 6780
atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac ttgaggttag 6840
atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa attttcctta 6900
catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat agctgtccct 6960
cttctcttat ggagatc 6977
<210> 23
<211> 13222
<212> DNA
<213> Artificial
<220>
<223> 编码由细胞特异性启动子hMUC1、第一microRNA为mir-199a-3p、第二micro
RNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白分
别为TALE蛋白、第一和第二识别序列为4×UAS的元件组成的表达系统的核酸
<400> 23
cggagtactg tcctccgagc ggagtactgt cctccgagcg gagtactgtc ctccgagcgg 60
agttctgtcc tccgagcgga gactctagat acccctctcc gcttctgaat tctaggcgtg 120
tacggtggga ggcctatata agcagagctc gtttagtgaa ccgtcagatc gcctcgagta 180
cccctctccg cttcttaccc ctctccgctt cttacccctc tccgcttctg aattcgaccg 240
ccaccatgag acatattatc tgccacggag gtgttattac cgaagaaatg gccgccagtc 300
ttttggacca gctgatcgaa gaggtactgg ctgataatct tccacctcct agccattttg 360
aaccacctac ccttcacgaa ctgtatgatt tagacgtgac ggcccccgaa gatcccaacg 420
aggaggcggt ttcgcagatt tttcccgact ctgtaatgtt ggcggtgcag gaagggattg 480
acttactcac ttttccgccg gcgcccggtt ctccggagcc gcctcacctt tcccggcagc 540
ccgagcagcc ggagcagaga gccttgggtc cggtttctat gccaaacctt gtaccggagg 600
tgatcgatct tacctgccac gaggctggct ttccacccag tgacgacgag gatgaagagg 660
gtgaggagtt tgtgttagat tatgtggagc accccgggca cggttgcagg tcttgtcatt 720
atcaccggag gaatacgggg gacccagata ttatgtgttc gctttgctat atgaggacct 780
gtggcatgtt tgtctacagt aagtgaaaat tatgggcagt gggtgataga gtggtgggtt 840
tggtgtggta attttttttt taatttttac agttttgtgg tttaaagaat tttgtattgt 900
gattttttta aaaggtcctg tgtctgaacc tgagcctgag cccgagccag aaccggagcc 960
tgcaagacct acccgccgtc ctaaaatggc gcctgctatc ctgaggcgcc cgacatcacc 1020
tgtgtctaga gaatgcaata gtagtacgga tagctgtgac tccggtcctt ctaacacacc 1080
tcctgagata cacccggtgg tcccgctgtg ccccattaaa ccagttgccg tgagagttgg 1140
tgggcgtcgc caggctgtgg aatgtatcga ggacttgctt aacgagcctg ggcaaccttt 1200
ggacttgagc tgtaaacgcc ccaggccagg gtccggagct acaaattttt ccctcctcaa 1260
acaggctgga gatgtcgaag aaaatcctgg gcctatggag gcttgggagt gtttggaaga 1320
tttttctgct gtgcgtaact tgctggaaca gagctctaac agtacctctt ggttttggag 1380
gtttctgtgg ggctcatccc aggcaaagtt agtctgcaga attaaggagg attacaagtg 1440
ggaatttgaa gagcttttga aatcctgtgg tgagctgttt gattctttga atctgggtca 1500
ccaggcgctt ttccaagaga aggtcatcaa gactttggat ttttccacac cggggcgcgc 1560
tgcggctgct gttgcttttt tgagttttat aaaggataaa tggagcgaag aaacccatct 1620
gagcgggggg tacctgctgg attttctggc catgcatctg tggagagcgg ttgtgagaca 1680
caagaatcgc ctgctactgt tgtcttccgt ccgcccggcg ataataccga cggaggagca 1740
gcagcagcag caggaggaag ccaggcggcg gcggcaggag cagagcccat ggaacccgag 1800
agccggcctg gaccctcggg aatgaatgtt gtacaggtgg ctgaactgta tccagaactg 1860
agacgcattt tgacaattac agaggatggg caggggctaa agggggtaaa gagggagcgg 1920
ggggcttgtg aggctacaga ggaggctagg aatctagctt ttagcttaat gaccagacac 1980
cgtcctgagt gtattacttt tcaacagatc aaggataatt gcgctaatga gcttgatctg 2040
ctggcgcaga agtattccat agagcagctg accacttact ggctgcagcc aggggatgat 2100
tttgaggagg ctattagggt atatgcaaag gtggcactta ggccagattg caagtacaag 2160
atcagcaaac ttgtaaatat caggaattgt tgctacattt ctgggaacgg ggccgaggtg 2220
gagatagata cggaggatag ggtggccttt agatgtagca tgataaatat gtggccgggg 2280
gtgcttggca tggacggggt ggttattatg aatgtaaggt ttactggccc caattttagc 2340
ggtacggttt tcctggccaa taccaacctt atcctacacg gtgtaagctt ctatgggttt 2400
aacaatacct gtgtggaagc ctggaccgat gtaagggttc ggggctgtgc cttttactgc 2460
tgctggaagg gggtggtgtg tcgccccaaa agcagggctt caattaagaa atgcctcttt 2520
gaaaggtgta ccttgggtat cctgtctgag ggtaactcca gggtgcgcca caatgtggcc 2580
tccgactgtg gttgcttcat gctagtgaaa agcgtggctg tgattaagca taacatggta 2640
tgtggcaact gcgaggacag ggcctctcag atgctgacct gctcggacgg caactgtcac 2700
ctgctgaaga ccattcacgt agccagccac tctcgcaagg cctggccagt gtttgagcat 2760
aacatactga cccgctgttc cttgcatttg ggtaacagga ggggggtgtt cctaccttac 2820
caatgcaatt tgagtcacac taagatattg cttgagcccg agagcatgtc caaggtgaac 2880
ctgaacgggg tgtttgacat gaccatgaag atctggaagg tgctgaggta cgatgagacc 2940
cgcaccaggt gcagaccctg cgagtgtggc ggtaaacata ttaggaacca gcctgtgatg 3000
ctggatgtga ccgaggagct gaggcccgat cacttggtgc tggcctgcac ccgcgctgag 3060
tttggctcta gcgatgaaga tacagatctc gaggggtccg gagctacaaa tttttccctc 3120
ctcaaacagg ctggagatgt cgaagaaaat cctgggccta ccggtatgta taggatgcag 3180
ttgctctctt gcatcgccct gtctctagcg ttggtgacta atagcgcccc tacttcttct 3240
tccacgaaga aaacccagct gcaactggaa caccttctgc tggatctcca gatgatcctt 3300
aacgggatta acaactataa gaacccgaag ctcacggcta tgctgacggc gaagttcgct 3360
atgcctaaaa aggctactga gctgaaacac ttacagtgtc tcgaagaggc actcaaaccc 3420
ctcgaggagg tactcaatct ggcgcaaagc aaaaacttcc atctccgccc acgggaccta 3480
atctccaaca tcaacgtgat tgtgctggag ctcaaggggt ctgaaaccac gtttatgtgc 3540
gagtacgcag atgaaactgc taccatagtg gaattcctca atcgttggat cactttctgc 3600
cagtcaatca tttcaaccct gaccagatct gagggccgcg gcagcctgct gacctgcggc 3660
gacgtggagg aaaaccccgg ccccggatcc ttgtcgacca ttcgtccgcg caggccaagt 3720
cctgcccgcg agcttctgcc cggaccccaa ccggataggg ttcagccgac tgcagatcgt 3780
ggggtgtctg cgcctgctgg cagccctctg gatggcttgc ccgctcggcg gacggtgtcc 3840
cggacccggc tgccatctcc ccctgcgccc tcacctgcgt tctcggcggg cagcttcagc 3900
gatctgctcc gtccgttcga tccgtcgctt cttgatacat cgcttcttga ttcgatgcct 3960
gccgtcggca cgccgcatac agcggctgcc ccagcagagt gggatgaggc gcaatcggct 4020
ctgcgtgcag ccgatgaccc gccacccacc gtgcgtgtcg ctgtcactgc cgcgcggccg 4080
ccgcgcgcca agccggcccc gcgacggcgt gctgcgcaac cctccgacgc ttcgccggcc 4140
gcgcaggtgg atctacgcac gctcggctac agtcagcagc agcaagagaa gatcaaaccg 4200
aaggtgcgtt cgacagtggc gcagcaccac gaggcactgg tgggccatgg gtttacacac 4260
gcgcacatcg ttgcgctcag ccaacacccg gcagcgttag ggaccgtcgc tgtcacgtat 4320
cagcacataa tcacggcgtt gccagaggcg acacacgaag acatcgttgg cgtcggcaaa 4380
cagtggtccg gcgcacgcgc cctggaggcc ttgctcacgg atgcggggga gttgagaggt 4440
ccgccgttac agttggacac aggccaactt gtgaagattg caaaacgtgg cggcgtgacc 4500
gcaatggagg cagtgcatgc atcgcgcaat gcactgacgg gtgcccccct gaacctgacc 4560
ccggaccaag tggtggctat cgccagcaac attggcggca agcaagcgct cgaaacggtg 4620
cagcggctgt tgccggtgct gtgccaggac catggcctga ccccggacca agtggtggct 4680
atcgccagca acggtggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg 4740
ctgtgccagg accatggcct gaccccggac caagtggtgg ctatcgccag caacattggc 4800
ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc 4860
ctgaccccgg accaagtggt ggctatcgcc agcaacaatg gcggcaagca agcgctcgaa 4920
acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg gcctgacccc ggaccaagtg 4980
gtggctatcg ccagcaacat tggcggcaag caagcgctcg aaacggtgca gcggctgttg 5040
ccggtgctgt gccaggacca tggcctgacc ccggaccaag tggtggctat cgccagcaac 5100
attggcggca agcaagcgct cgaaacggtg cagcggctgt tgccggtgct gtgccaggac 5160
catggcctga ctccggacca agtggtggct atcgccagcc acgatggcgg caagcaagcg 5220
ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg accatggcct gactccggac 5280
caagtggtgg ctatcgccag ccacgatggc ggcaagcaag cgctcgaaac ggtgcagcgg 5340
ctgttgccgg tgctgtgcca ggaccatggc ctgaccccgg accaagtggt ggctatcgcc 5400
agcaacaatg gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc 5460
caggaccatg gcctgacccc ggaccaagtg gtggctatcg ccagcaacat tggcggcaag 5520
caagcgctcg aaacggtgca gcggctgttg ccggtgctgt gccaggacca tggcctgacc 5580
ccggaccaag tggtggctat cgccagcaac ggtggcggca agcaagcgct cgaaacggtg 5640
cagcggctgt tgccggtgct gtgccaggac catggcctga ctccggacca agtggtggct 5700
atcgccagcc acgatggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg 5760
ctgtgccagg accatggcct gactccggac caagtggtgg ctatcgccag ccacgatggc 5820
ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc 5880
ctgaccccgg accaagtggt ggctatcgcc agcaacggtg gcggcaagca agcgctcgaa 5940
acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg gcctgactcc ggaccaagtg 6000
gtggctatcg ccagccacga tggcggcaag caagcgctcg aaacggtgca gcggctgttg 6060
ccggtgctgt gccaggacca tggcctgact ccggaccaag tggtggctat cgccagccac 6120
gatggcggca agcaagcgct cgaaacggtg cagcggctgt tgccggtgct gtgccaggac 6180
catggcctga ctccggacca agtggtggct atcgccagcc acgatggcgg caagcaagcg 6240
ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg accatggcct gaccccggac 6300
caagtggtgg ctatcgccag caacattggc ggcaagcaag cgctcgaaac ggtgcagcgg 6360
ctgttgccgg tgctgtgcca ggaccatggc ctgaccccgg accaagtggt ggctatcgcc 6420
agcaacggtg gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc 6480
caggaccatg gcctgacccc ggaccaagtg gtggctatcg ccagcaacgg tggcggcaag 6540
caagcgctcg aaagcattgt ggcccagctg agccggcctg atccggcgtt ggccgcgttg 6600
accaacgacc acctcgtcgc cttggcctgc ctcggcggac gtcctgccat ggatgcagtg 6660
aaaaagggat tgccgcacgc gccggaattg atcagaagag tcaatcgccg tattggcgaa 6720
cgcacgtccc atcgcgttgc cgactacgcg caagtggttc gcgtgctgga gtttttccag 6780
tgccactccc acccagcgta cgcatttgat gaggccatga cgcagttcgg gatgagcagg 6840
aacgggttgg tacagctctt tcgcagagtg ggcgtcaccg aactcgaagc ccgcggtgga 6900
acgctccccc cagcctcgca gcgttgggac cgtatcctcc aggcatcagg gatgaaaagg 6960
gccaaaccgt cccctacttc agctcaaaca ccggatcagg cgtctttgca tgcattcgcc 7020
gattcgctgg agcgtgacct tgatgcgccc agcccaatgc acgagggaga tcagacgcgg 7080
gcaagcagcc gtaaacggtc ccgatcggat cgtgctgtca ccggcccctc cgcacagcag 7140
gctgtcgagg tgcgcgttcc cgaacagcgc gatgcgctgc atttgcccct cagctggagg 7200
gtaaaacgcc cgcgtaccag gatctggggc ggcctcccgg ttaagtagtg ctttctactt 7260
tatgagtagt gctttctact ttatgagtag tgctttctac tttatgagta gtgctttcta 7320
ctttatgtaa ccaatgtgca gactactgtt aaccaatgtg cagactactg ttaaccaatg 7380
tgcagactac tgttaaccaa tgtgcagact actgtgaaag cggagtactg tcctccgagc 7440
ggagtactgt cctccgagcg gagtactgtc ctccgagcgg agttctgtcc tccgagcgga 7500
gactctagat atagaaccga tcctcccatt gaattctagg cgtgtacggt gggaggccta 7560
tataagcaga gctcgtttag tgaaccgtca gatcgcctcg agtatagaac cgatcctccc 7620
atttatagaa ccgatcctcc catttataga accgatcctc ccattactag aagatcttga 7680
attcgccacc ttgtcgacca ttcgtccgcg caggccaagt cctgcccgcg agcttctgcc 7740
cggaccccaa ccggataggg ttcagccgac tgcagatcgt ggggtgtctg cgcctgctgg 7800
cagccctctg gatggcttgc ccgctcggcg gacggtgtcc cggacccggc tgccatctcc 7860
ccctgcgccc tcacctgcgt tctcggcggg cagcttcagc gatctgctcc gtccgttcga 7920
tccgtcgctt cttgatacat cgcttcttga ttcgatgcct gccgtcggca cgccgcatac 7980
agcggctgcc ccagcagagt gggatgaggc gcaatcggct ctgcgtgcag ccgatgaccc 8040
gccacccacc gtgcgtgtcg ctgtcactgc cgcgcggccg ccgcgcgcca agccggcccc 8100
gcgacggcgt gctgcgcaac cctccgacgc ttcgccggcc gcgcaggtgg atctacgcac 8160
gctcggctac agtcagcagc agcaagagaa gatcaaaccg aaggtgcgtt cgacagtggc 8220
gcagcaccac gaggcactgg tgggccatgg gtttacacac gcgcacatcg ttgcgctcag 8280
ccaacacccg gcagcgttag ggaccgtcgc tgtcacgtat cagcacataa tcacggcgtt 8340
gccagaggcg acacacgaag acatcgttgg cgtcggcaaa cagtggtccg gcgcacgcgc 8400
cctggaggcc ttgctcacgg atgcggggga gttgagaggt ccgccgttac agttggacac 8460
aggccaactt gtgaagattg caaaacgtgg cggcgtgacc gcaatggagg cagtgcatgc 8520
atcgcgcaat gcactgacgg gtgcccccct gaacctgacc ccggaccaag tggtggctat 8580
cgccagcaac attggcggca agcaagcgct cgaaacggtg cagcggctgt tgccggtgct 8640
gtgccaggac catggcctga ctccggacca agtggtggct atcgccagcc acgatggcgg 8700
caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg accatggcct 8760
gactccggac caagtggtgg ctatcgccag ccacgatggc ggcaagcaag cgctcgaaac 8820
ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc ctgactccgg accaagtggt 8880
ggctatcgcc agccacgatg gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc 8940
ggtgctgtgc caggaccatg gcctgactcc ggaccaagtg gtggctatcg ccagccacga 9000
tggcggcaag caagcgctcg aaacggtgca gcggctgttg ccggtgctgt gccaggacca 9060
tggcctgacc ccggaccaag tggtggctat cgccagcaac ggtggcggca agcaagcgct 9120
cgaaacggtg cagcggctgt tgccggtgct gtgccaggac catggcctga ctccggacca 9180
agtggtggct atcgccagcc acgatggcgg caagcaagcg ctcgaaacgg tgcagcggct 9240
gttgccggtg ctgtgccagg accatggcct gaccccggac caagtggtgg ctatcgccag 9300
caacggtggc ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca 9360
ggaccatggc ctgactccgg accaagtggt ggctatcgcc agccacgatg gcggcaagca 9420
agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg gcctgactcc 9480
ggaccaagtg gtggctatcg ccagccacga tggcggcaag caagcgctcg aaacggtgca 9540
gcggctgttg ccggtgctgt gccaggacca tggcctgacc ccggaccaag tggtggctat 9600
cgccagcaac aatggcggca agcaagcgct cgaaacggtg cagcggctgt tgccggtgct 9660
gtgccaggac catggcctga ctccggacca agtggtggct atcgccagcc acgatggcgg 9720
caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg accatggcct 9780
gaccccggac caagtggtgg ctatcgccag caacggtggc ggcaagcaag cgctcgaaac 9840
ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc ctgaccccgg accaagtggt 9900
ggctatcgcc agcaacggtg gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc 9960
ggtgctgtgc caggaccatg gcctgactcc ggaccaagtg gtggctatcg ccagccacga 10020
tggcggcaag caagcgctcg aaacggtgca gcggctgttg ccggtgctgt gccaggacca 10080
tggcctgacc ccggaccaag tggtggctat cgccagcaac ggtggcggca agcaagcgct 10140
cgaaagcatt gtggcccagc tgagccggcc tgatccggcg ttggccgcgt tgaccaacga 10200
ccacctcgtc gccttggcct gcctcggcgg acgtcctgcc atggatgcag tgaaaaaggg 10260
attgccgcac gcgccggaat tgatcagaag agtcaatcgc cgtattggcg aacgcacgtc 10320
ccatcgcgtt gccgactacg cgcaagtggt tcgcgtgctg gagtttttcc agtgccactc 10380
ccacccagcg tacgcatttg atgaggccat gacgcagttc gggatgagca ggaacgggtt 10440
ggtacagctc tttcgcagag tgggcgtcac cgaactcgaa gcccgcggtg gaacgctccc 10500
cccagcctcg cagcgttggg accgtatcct ccaggcatca gggatgaaaa gggccaaacc 10560
gtcccctact tcagctcaaa caccggatca ggcgtctttg catgcattcg ccgattcgct 10620
ggagcgtgac cttgatgcgc ccagcccaat gcacgaggga gatcagacgc gggcaagcag 10680
ccgtaaacgg tcccgatcgg atcgtgctgt caccggcccc tccgcacagc aggctgtcga 10740
ggtgcgcgtt cccgaacagc gcgatgcgct gcatttgccc ctcagctgga gggtaaaacg 10800
cccgcgtacc aggatctggg gcggcctccc gtaatacaag gcggccgcaa atcaacatca 10860
gtctgataag ctatcaacat cagtctgata agctatcaac atcagtctga taagctatca 10920
acatcagtct gataagctaa agcttcgaat tctgataatc agccatacca catttgtaga 10980
ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa 11040
tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag 11100
catcacaaat ttcacaaata aagcattttt ttcactgcgt tcatcggagc ccaggtttac 11160
tcccttaagt ggaaatttct tcccccactc cctccttggc tttctccaag gagggaaccc 11220
aggctactgg aaagtccggc tggggcgggg actgtgggtt tcagggtaga actgcgtgtg 11280
gaacgggaca gggagcggtt agaagggtgg ggctattccg ggaagtggtg gggggaggga 11340
gcccaaaact agcacctagt ccactcatta tccagccctc ttatttctcg gccccgctct 11400
gcttcagtgg acccggggag ggcggggaag tggagtggga gacctagggg tgggcttccc 11460
gaccttgctg tacaggacct cgacctagct ggctttgttc cccatcccca cgttagttgt 11520
tgccctgagg ctaaaactag agcccagggg ccccaagttc cagactgccc ctcccccctc 11580
ccccggagcc agggagtggt tggtgaaagg gggaggccag ctggagaaca aacgggtagt 11640
cagggggttg agcgattaga gcccttgtac cctacccagg aatggttggg gaggaggagg 11700
aagaggtagg aggtagggga gggggcgggg ttttgtcacc tgtcacctgc tccggctgtg 11760
cctagggcgg gcgggcgggg agtgggggga ccggtataaa gcggtaggcg cctgtgcccg 11820
ctccacctct caagcagcca gcgcctgcct gaatctgttc tgccccctcc ccacccattt 11880
caccaccacc atggcccccc cgaccgatgt cagcctgggg gacgagctcc acttagacgg 11940
cgaggacgtg gcgatggcgc atgccgacgc gctagacgat ttcgatctgg acatgttggg 12000
gagcgacgcg ctagacgatt tcgatctgga catgttgggg agcgacgcgc tagacgattt 12060
cgatctggac atgttgggga gcgacgcgct agacgatttc gatctggaca tgttggggga 12120
cggggattcc ccgggtccgg gatttacccc ccacgactcc gccccctacg gcgctctgga 12180
tatggccgac ttcgagtttg agcagatgtt taccgatgcc cttggaattg acgagtacgg 12240
tgggacgcgt atgaagctac tgtcttctat cgaacaagca tgcgatattt gccgacttaa 12300
aaagctcaag tgctccaaag aaaaaccgaa gtgcgccaag tgtctgaaga acaactggga 12360
gtgtcgctac tctcccaaaa ccaaaaggtc tccgctgact agggcacatc tgacagaagt 12420
ggaatcaagg ctagaaagac tggaacagct atttctactg atttttcctc gagaagacct 12480
tgacatgatt ttgaaaatgg attctttaca ggatataaaa gcattgttaa caggattatt 12540
tgtacaagat aatgtgaata aagatgccgt cacagataga ttggcttcag tggagactga 12600
tatgcctcta acattgagac agcatagaat aagtgcgaca tcatcatcgg aagagagtag 12660
taacaaaggt caaagacagt tgactgtata aaagcttcac tcctcaggtg caggctgcct 12720
atcagaaggt ggtggctggt gtggccaatg ccctggctca caaataccac tgagatcttt 12780
ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct 12840
aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg 12900
gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt 12960
tggcaacata tgcccatatg ctggctgcca tgaacaaagg ttggctataa agaggtcatc 13020
agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag 13080
gttagatttt ttttatattt tgttttgtgt tatttttttc tttaacatcc ctaaaatttt 13140
ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg 13200
tccctcttct cttatggaga tc 13222
<210> 24
<211> 13260
<212> DNA
<213> Artificial
<220>
<223> 编码由细胞特异性启动子hMUC1、第一microRNA为mir-199a-3p、第二micro
RNA为mir-21、转录激活因子为Gal4VP16、第一调控蛋白和第二调控蛋白分
别为TALE蛋白、第一和第二识别序列为5×UAS的元件组成的表达系统的核酸
<400> 24
cggagtactg tcctccgagc ggagtactgt cctccgagcg gagtactgtc ctccgagcgg 60
agtactgtcc tccgagcgga gttctgtcct ccgagcggag actctagata cccctctccg 120
cttctgaatt ctaggcgtgt acggtgggag gcctatataa gcagagctcg tttagtgaac 180
cgtcagatcg cctcgagtac ccctctccgc ttcttacccc tctccgcttc ttacccctct 240
ccgcttctga attcgaccgc caccatgaga catattatct gccacggagg tgttattacc 300
gaagaaatgg ccgccagtct tttggaccag ctgatcgaag aggtactggc tgataatctt 360
ccacctccta gccattttga accacctacc cttcacgaac tgtatgattt agacgtgacg 420
gcccccgaag atcccaacga ggaggcggtt tcgcagattt ttcccgactc tgtaatgttg 480
gcggtgcagg aagggattga cttactcact tttccgccgg cgcccggttc tccggagccg 540
cctcaccttt cccggcagcc cgagcagccg gagcagagag ccttgggtcc ggtttctatg 600
ccaaaccttg taccggaggt gatcgatctt acctgccacg aggctggctt tccacccagt 660
gacgacgagg atgaagaggg tgaggagttt gtgttagatt atgtggagca ccccgggcac 720
ggttgcaggt cttgtcatta tcaccggagg aatacggggg acccagatat tatgtgttcg 780
ctttgctata tgaggacctg tggcatgttt gtctacagta agtgaaaatt atgggcagtg 840
ggtgatagag tggtgggttt ggtgtggtaa tttttttttt aatttttaca gttttgtggt 900
ttaaagaatt ttgtattgtg atttttttaa aaggtcctgt gtctgaacct gagcctgagc 960
ccgagccaga accggagcct gcaagaccta cccgccgtcc taaaatggcg cctgctatcc 1020
tgaggcgccc gacatcacct gtgtctagag aatgcaatag tagtacggat agctgtgact 1080
ccggtccttc taacacacct cctgagatac acccggtggt cccgctgtgc cccattaaac 1140
cagttgccgt gagagttggt gggcgtcgcc aggctgtgga atgtatcgag gacttgctta 1200
acgagcctgg gcaacctttg gacttgagct gtaaacgccc caggccaggg tccggagcta 1260
caaatttttc cctcctcaaa caggctggag atgtcgaaga aaatcctggg cctatggagg 1320
cttgggagtg tttggaagat ttttctgctg tgcgtaactt gctggaacag agctctaaca 1380
gtacctcttg gttttggagg tttctgtggg gctcatccca ggcaaagtta gtctgcagaa 1440
ttaaggagga ttacaagtgg gaatttgaag agcttttgaa atcctgtggt gagctgtttg 1500
attctttgaa tctgggtcac caggcgcttt tccaagagaa ggtcatcaag actttggatt 1560
tttccacacc ggggcgcgct gcggctgctg ttgctttttt gagttttata aaggataaat 1620
ggagcgaaga aacccatctg agcggggggt acctgctgga ttttctggcc atgcatctgt 1680
ggagagcggt tgtgagacac aagaatcgcc tgctactgtt gtcttccgtc cgcccggcga 1740
taataccgac ggaggagcag cagcagcagc aggaggaagc caggcggcgg cggcaggagc 1800
agagcccatg gaacccgaga gccggcctgg accctcggga atgaatgttg tacaggtggc 1860
tgaactgtat ccagaactga gacgcatttt gacaattaca gaggatgggc aggggctaaa 1920
gggggtaaag agggagcggg gggcttgtga ggctacagag gaggctagga atctagcttt 1980
tagcttaatg accagacacc gtcctgagtg tattactttt caacagatca aggataattg 2040
cgctaatgag cttgatctgc tggcgcagaa gtattccata gagcagctga ccacttactg 2100
gctgcagcca ggggatgatt ttgaggaggc tattagggta tatgcaaagg tggcacttag 2160
gccagattgc aagtacaaga tcagcaaact tgtaaatatc aggaattgtt gctacatttc 2220
tgggaacggg gccgaggtgg agatagatac ggaggatagg gtggccttta gatgtagcat 2280
gataaatatg tggccggggg tgcttggcat ggacggggtg gttattatga atgtaaggtt 2340
tactggcccc aattttagcg gtacggtttt cctggccaat accaacctta tcctacacgg 2400
tgtaagcttc tatgggttta acaatacctg tgtggaagcc tggaccgatg taagggttcg 2460
gggctgtgcc ttttactgct gctggaaggg ggtggtgtgt cgccccaaaa gcagggcttc 2520
aattaagaaa tgcctctttg aaaggtgtac cttgggtatc ctgtctgagg gtaactccag 2580
ggtgcgccac aatgtggcct ccgactgtgg ttgcttcatg ctagtgaaaa gcgtggctgt 2640
gattaagcat aacatggtat gtggcaactg cgaggacagg gcctctcaga tgctgacctg 2700
ctcggacggc aactgtcacc tgctgaagac cattcacgta gccagccact ctcgcaaggc 2760
ctggccagtg tttgagcata acatactgac ccgctgttcc ttgcatttgg gtaacaggag 2820
gggggtgttc ctaccttacc aatgcaattt gagtcacact aagatattgc ttgagcccga 2880
gagcatgtcc aaggtgaacc tgaacggggt gtttgacatg accatgaaga tctggaaggt 2940
gctgaggtac gatgagaccc gcaccaggtg cagaccctgc gagtgtggcg gtaaacatat 3000
taggaaccag cctgtgatgc tggatgtgac cgaggagctg aggcccgatc acttggtgct 3060
ggcctgcacc cgcgctgagt ttggctctag cgatgaagat acagatctcg aggggtccgg 3120
agctacaaat ttttccctcc tcaaacaggc tggagatgtc gaagaaaatc ctgggcctac 3180
cggtatgtat aggatgcagt tgctctcttg catcgccctg tctctagcgt tggtgactaa 3240
tagcgcccct acttcttctt ccacgaagaa aacccagctg caactggaac accttctgct 3300
ggatctccag atgatcctta acgggattaa caactataag aacccgaagc tcacggctat 3360
gctgacggcg aagttcgcta tgcctaaaaa ggctactgag ctgaaacact tacagtgtct 3420
cgaagaggca ctcaaacccc tcgaggaggt actcaatctg gcgcaaagca aaaacttcca 3480
tctccgccca cgggacctaa tctccaacat caacgtgatt gtgctggagc tcaaggggtc 3540
tgaaaccacg tttatgtgcg agtacgcaga tgaaactgct accatagtgg aattcctcaa 3600
tcgttggatc actttctgcc agtcaatcat ttcaaccctg accagatctg agggccgcgg 3660
cagcctgctg acctgcggcg acgtggagga aaaccccggc cccggatcct tgtcgaccat 3720
tcgtccgcgc aggccaagtc ctgcccgcga gcttctgccc ggaccccaac cggatagggt 3780
tcagccgact gcagatcgtg gggtgtctgc gcctgctggc agccctctgg atggcttgcc 3840
cgctcggcgg acggtgtccc ggacccggct gccatctccc cctgcgccct cacctgcgtt 3900
ctcggcgggc agcttcagcg atctgctccg tccgttcgat ccgtcgcttc ttgatacatc 3960
gcttcttgat tcgatgcctg ccgtcggcac gccgcataca gcggctgccc cagcagagtg 4020
ggatgaggcg caatcggctc tgcgtgcagc cgatgacccg ccacccaccg tgcgtgtcgc 4080
tgtcactgcc gcgcggccgc cgcgcgccaa gccggccccg cgacggcgtg ctgcgcaacc 4140
ctccgacgct tcgccggccg cgcaggtgga tctacgcacg ctcggctaca gtcagcagca 4200
gcaagagaag atcaaaccga aggtgcgttc gacagtggcg cagcaccacg aggcactggt 4260
gggccatggg tttacacacg cgcacatcgt tgcgctcagc caacacccgg cagcgttagg 4320
gaccgtcgct gtcacgtatc agcacataat cacggcgttg ccagaggcga cacacgaaga 4380
catcgttggc gtcggcaaac agtggtccgg cgcacgcgcc ctggaggcct tgctcacgga 4440
tgcgggggag ttgagaggtc cgccgttaca gttggacaca ggccaacttg tgaagattgc 4500
aaaacgtggc ggcgtgaccg caatggaggc agtgcatgca tcgcgcaatg cactgacggg 4560
tgcccccctg aacctgaccc cggaccaagt ggtggctatc gccagcaaca ttggcggcaa 4620
gcaagcgctc gaaacggtgc agcggctgtt gccggtgctg tgccaggacc atggcctgac 4680
cccggaccaa gtggtggcta tcgccagcaa cggtggcggc aagcaagcgc tcgaaacggt 4740
gcagcggctg ttgccggtgc tgtgccagga ccatggcctg accccggacc aagtggtggc 4800
tatcgccagc aacattggcg gcaagcaagc gctcgaaacg gtgcagcggc tgttgccggt 4860
gctgtgccag gaccatggcc tgaccccgga ccaagtggtg gctatcgcca gcaacaatgg 4920
cggcaagcaa gcgctcgaaa cggtgcagcg gctgttgccg gtgctgtgcc aggaccatgg 4980
cctgaccccg gaccaagtgg tggctatcgc cagcaacatt ggcggcaagc aagcgctcga 5040
aacggtgcag cggctgttgc cggtgctgtg ccaggaccat ggcctgaccc cggaccaagt 5100
ggtggctatc gccagcaaca ttggcggcaa gcaagcgctc gaaacggtgc agcggctgtt 5160
gccggtgctg tgccaggacc atggcctgac tccggaccaa gtggtggcta tcgccagcca 5220
cgatggcggc aagcaagcgc tcgaaacggt gcagcggctg ttgccggtgc tgtgccagga 5280
ccatggcctg actccggacc aagtggtggc tatcgccagc cacgatggcg gcaagcaagc 5340
gctcgaaacg gtgcagcggc tgttgccggt gctgtgccag gaccatggcc tgaccccgga 5400
ccaagtggtg gctatcgcca gcaacaatgg cggcaagcaa gcgctcgaaa cggtgcagcg 5460
gctgttgccg gtgctgtgcc aggaccatgg cctgaccccg gaccaagtgg tggctatcgc 5520
cagcaacatt ggcggcaagc aagcgctcga aacggtgcag cggctgttgc cggtgctgtg 5580
ccaggaccat ggcctgaccc cggaccaagt ggtggctatc gccagcaacg gtggcggcaa 5640
gcaagcgctc gaaacggtgc agcggctgtt gccggtgctg tgccaggacc atggcctgac 5700
tccggaccaa gtggtggcta tcgccagcca cgatggcggc aagcaagcgc tcgaaacggt 5760
gcagcggctg ttgccggtgc tgtgccagga ccatggcctg actccggacc aagtggtggc 5820
tatcgccagc cacgatggcg gcaagcaagc gctcgaaacg gtgcagcggc tgttgccggt 5880
gctgtgccag gaccatggcc tgaccccgga ccaagtggtg gctatcgcca gcaacggtgg 5940
cggcaagcaa gcgctcgaaa cggtgcagcg gctgttgccg gtgctgtgcc aggaccatgg 6000
cctgactccg gaccaagtgg tggctatcgc cagccacgat ggcggcaagc aagcgctcga 6060
aacggtgcag cggctgttgc cggtgctgtg ccaggaccat ggcctgactc cggaccaagt 6120
ggtggctatc gccagccacg atggcggcaa gcaagcgctc gaaacggtgc agcggctgtt 6180
gccggtgctg tgccaggacc atggcctgac tccggaccaa gtggtggcta tcgccagcca 6240
cgatggcggc aagcaagcgc tcgaaacggt gcagcggctg ttgccggtgc tgtgccagga 6300
ccatggcctg accccggacc aagtggtggc tatcgccagc aacattggcg gcaagcaagc 6360
gctcgaaacg gtgcagcggc tgttgccggt gctgtgccag gaccatggcc tgaccccgga 6420
ccaagtggtg gctatcgcca gcaacggtgg cggcaagcaa gcgctcgaaa cggtgcagcg 6480
gctgttgccg gtgctgtgcc aggaccatgg cctgaccccg gaccaagtgg tggctatcgc 6540
cagcaacggt ggcggcaagc aagcgctcga aagcattgtg gcccagctga gccggcctga 6600
tccggcgttg gccgcgttga ccaacgacca cctcgtcgcc ttggcctgcc tcggcggacg 6660
tcctgccatg gatgcagtga aaaagggatt gccgcacgcg ccggaattga tcagaagagt 6720
caatcgccgt attggcgaac gcacgtccca tcgcgttgcc gactacgcgc aagtggttcg 6780
cgtgctggag tttttccagt gccactccca cccagcgtac gcatttgatg aggccatgac 6840
gcagttcggg atgagcagga acgggttggt acagctcttt cgcagagtgg gcgtcaccga 6900
actcgaagcc cgcggtggaa cgctcccccc agcctcgcag cgttgggacc gtatcctcca 6960
ggcatcaggg atgaaaaggg ccaaaccgtc ccctacttca gctcaaacac cggatcaggc 7020
gtctttgcat gcattcgccg attcgctgga gcgtgacctt gatgcgccca gcccaatgca 7080
cgagggagat cagacgcggg caagcagccg taaacggtcc cgatcggatc gtgctgtcac 7140
cggcccctcc gcacagcagg ctgtcgaggt gcgcgttccc gaacagcgcg atgcgctgca 7200
tttgcccctc agctggaggg taaaacgccc gcgtaccagg atctggggcg gcctcccggt 7260
taagtagtgc tttctacttt atgagtagtg ctttctactt tatgagtagt gctttctact 7320
ttatgagtag tgctttctac tttatgtaac caatgtgcag actactgtta accaatgtgc 7380
agactactgt taaccaatgt gcagactact gttaaccaat gtgcagacta ctgtgaaagc 7440
ggagtactgt cctccgagcg gagtactgtc ctccgagcgg agtactgtcc tccgagcgga 7500
gtactgtcct ccgagcggag ttctgtcctc cgagcggaga ctctagatat agaaccgatc 7560
ctcccattga attctaggcg tgtacggtgg gaggcctata taagcagagc tcgtttagtg 7620
aaccgtcaga tcgcctcgag tatagaaccg atcctcccat ttatagaacc gatcctccca 7680
tttatagaac cgatcctccc attactagaa gatcttgaat tcgccacctt gtcgaccatt 7740
cgtccgcgca ggccaagtcc tgcccgcgag cttctgcccg gaccccaacc ggatagggtt 7800
cagccgactg cagatcgtgg ggtgtctgcg cctgctggca gccctctgga tggcttgccc 7860
gctcggcgga cggtgtcccg gacccggctg ccatctcccc ctgcgccctc acctgcgttc 7920
tcggcgggca gcttcagcga tctgctccgt ccgttcgatc cgtcgcttct tgatacatcg 7980
cttcttgatt cgatgcctgc cgtcggcacg ccgcatacag cggctgcccc agcagagtgg 8040
gatgaggcgc aatcggctct gcgtgcagcc gatgacccgc cacccaccgt gcgtgtcgct 8100
gtcactgccg cgcggccgcc gcgcgccaag ccggccccgc gacggcgtgc tgcgcaaccc 8160
tccgacgctt cgccggccgc gcaggtggat ctacgcacgc tcggctacag tcagcagcag 8220
caagagaaga tcaaaccgaa ggtgcgttcg acagtggcgc agcaccacga ggcactggtg 8280
ggccatgggt ttacacacgc gcacatcgtt gcgctcagcc aacacccggc agcgttaggg 8340
accgtcgctg tcacgtatca gcacataatc acggcgttgc cagaggcgac acacgaagac 8400
atcgttggcg tcggcaaaca gtggtccggc gcacgcgccc tggaggcctt gctcacggat 8460
gcgggggagt tgagaggtcc gccgttacag ttggacacag gccaacttgt gaagattgca 8520
aaacgtggcg gcgtgaccgc aatggaggca gtgcatgcat cgcgcaatgc actgacgggt 8580
gcccccctga acctgacccc ggaccaagtg gtggctatcg ccagcaacat tggcggcaag 8640
caagcgctcg aaacggtgca gcggctgttg ccggtgctgt gccaggacca tggcctgact 8700
ccggaccaag tggtggctat cgccagccac gatggcggca agcaagcgct cgaaacggtg 8760
cagcggctgt tgccggtgct gtgccaggac catggcctga ctccggacca agtggtggct 8820
atcgccagcc acgatggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg 8880
ctgtgccagg accatggcct gactccggac caagtggtgg ctatcgccag ccacgatggc 8940
ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc 9000
ctgactccgg accaagtggt ggctatcgcc agccacgatg gcggcaagca agcgctcgaa 9060
acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg gcctgacccc ggaccaagtg 9120
gtggctatcg ccagcaacgg tggcggcaag caagcgctcg aaacggtgca gcggctgttg 9180
ccggtgctgt gccaggacca tggcctgact ccggaccaag tggtggctat cgccagccac 9240
gatggcggca agcaagcgct cgaaacggtg cagcggctgt tgccggtgct gtgccaggac 9300
catggcctga ccccggacca agtggtggct atcgccagca acggtggcgg caagcaagcg 9360
ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg accatggcct gactccggac 9420
caagtggtgg ctatcgccag ccacgatggc ggcaagcaag cgctcgaaac ggtgcagcgg 9480
ctgttgccgg tgctgtgcca ggaccatggc ctgactccgg accaagtggt ggctatcgcc 9540
agccacgatg gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc 9600
caggaccatg gcctgacccc ggaccaagtg gtggctatcg ccagcaacaa tggcggcaag 9660
caagcgctcg aaacggtgca gcggctgttg ccggtgctgt gccaggacca tggcctgact 9720
ccggaccaag tggtggctat cgccagccac gatggcggca agcaagcgct cgaaacggtg 9780
cagcggctgt tgccggtgct gtgccaggac catggcctga ccccggacca agtggtggct 9840
atcgccagca acggtggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg 9900
ctgtgccagg accatggcct gaccccggac caagtggtgg ctatcgccag caacggtggc 9960
ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc 10020
ctgactccgg accaagtggt ggctatcgcc agccacgatg gcggcaagca agcgctcgaa 10080
acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg gcctgacccc ggaccaagtg 10140
gtggctatcg ccagcaacgg tggcggcaag caagcgctcg aaagcattgt ggcccagctg 10200
agccggcctg atccggcgtt ggccgcgttg accaacgacc acctcgtcgc cttggcctgc 10260
ctcggcggac gtcctgccat ggatgcagtg aaaaagggat tgccgcacgc gccggaattg 10320
atcagaagag tcaatcgccg tattggcgaa cgcacgtccc atcgcgttgc cgactacgcg 10380
caagtggttc gcgtgctgga gtttttccag tgccactccc acccagcgta cgcatttgat 10440
gaggccatga cgcagttcgg gatgagcagg aacgggttgg tacagctctt tcgcagagtg 10500
ggcgtcaccg aactcgaagc ccgcggtgga acgctccccc cagcctcgca gcgttgggac 10560
cgtatcctcc aggcatcagg gatgaaaagg gccaaaccgt cccctacttc agctcaaaca 10620
ccggatcagg cgtctttgca tgcattcgcc gattcgctgg agcgtgacct tgatgcgccc 10680
agcccaatgc acgagggaga tcagacgcgg gcaagcagcc gtaaacggtc ccgatcggat 10740
cgtgctgtca ccggcccctc cgcacagcag gctgtcgagg tgcgcgttcc cgaacagcgc 10800
gatgcgctgc atttgcccct cagctggagg gtaaaacgcc cgcgtaccag gatctggggc 10860
ggcctcccgt aatacaaggc ggccgcaaat caacatcagt ctgataagct atcaacatca 10920
gtctgataag ctatcaacat cagtctgata agctatcaac atcagtctga taagctaaag 10980
cttcgaattc tgataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac 11040
ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg 11100
tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa 11160
gcattttttt cactgcgttc atcggagccc aggtttactc ccttaagtgg aaatttcttc 11220
ccccactccc tccttggctt tctccaagga gggaacccag gctactggaa agtccggctg 11280
gggcggggac tgtgggtttc agggtagaac tgcgtgtgga acgggacagg gagcggttag 11340
aagggtgggg ctattccggg aagtggtggg gggagggagc ccaaaactag cacctagtcc 11400
actcattatc cagccctctt atttctcggc cccgctctgc ttcagtggac ccggggaggg 11460
cggggaagtg gagtgggaga cctaggggtg ggcttcccga ccttgctgta caggacctcg 11520
acctagctgg ctttgttccc catccccacg ttagttgttg ccctgaggct aaaactagag 11580
cccaggggcc ccaagttcca gactgcccct cccccctccc ccggagccag ggagtggttg 11640
gtgaaagggg gaggccagct ggagaacaaa cgggtagtca gggggttgag cgattagagc 11700
ccttgtaccc tacccaggaa tggttgggga ggaggaggaa gaggtaggag gtaggggagg 11760
gggcggggtt ttgtcacctg tcacctgctc cggctgtgcc tagggcgggc gggcggggag 11820
tggggggacc ggtataaagc ggtaggcgcc tgtgcccgct ccacctctca agcagccagc 11880
gcctgcctga atctgttctg ccccctcccc acccatttca ccaccaccat ggcccccccg 11940
accgatgtca gcctggggga cgagctccac ttagacggcg aggacgtggc gatggcgcat 12000
gccgacgcgc tagacgattt cgatctggac atgttgggga gcgacgcgct agacgatttc 12060
gatctggaca tgttggggag cgacgcgcta gacgatttcg atctggacat gttggggagc 12120
gacgcgctag acgatttcga tctggacatg ttgggggacg gggattcccc gggtccggga 12180
tttacccccc acgactccgc cccctacggc gctctggata tggccgactt cgagtttgag 12240
cagatgttta ccgatgccct tggaattgac gagtacggtg ggacgcgtat gaagctactg 12300
tcttctatcg aacaagcatg cgatatttgc cgacttaaaa agctcaagtg ctccaaagaa 12360
aaaccgaagt gcgccaagtg tctgaagaac aactgggagt gtcgctactc tcccaaaacc 12420
aaaaggtctc cgctgactag ggcacatctg acagaagtgg aatcaaggct agaaagactg 12480
gaacagctat ttctactgat ttttcctcga gaagaccttg acatgatttt gaaaatggat 12540
tctttacagg atataaaagc attgttaaca ggattatttg tacaagataa tgtgaataaa 12600
gatgccgtca cagatagatt ggcttcagtg gagactgata tgcctctaac attgagacag 12660
catagaataa gtgcgacatc atcatcggaa gagagtagta acaaaggtca aagacagttg 12720
actgtataaa agcttcactc ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt 12780
ggccaatgcc ctggctcaca aataccactg agatcttttt ccctctgcca aaaattatgg 12840
ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat 12900
tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa 12960
tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg cccatatgct 13020
ggctgccatg aacaaaggtt ggctataaag aggtcatcag tatatgaaac agccccctgc 13080
tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt ttatattttg 13140
ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt tactagccag 13200
atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct tatggagatc 13260
<210> 25
<211> 14991
<212> DNA
<213> Artificial
<220>
<223> 编码由细胞特异性启动子hMUC1、第一microRNA为mir-199a-3p、第二micro
RNA为mir-21、转录激活因子为分割dCas9、第一调控蛋白和第二调控蛋白分
别为TALE蛋白、第一和第二识别序列为5×UAS的元件组成的表达系统的核酸
<400> 25
cggagtactg tcctccgagc ggagtactgt cctccgagcg gagtactgtc ctccgagcgg 60
agtactgtcc tccgagcgga gttctgtcct ccgagcggag actctagata cccctctccg 120
cttctgaatt ctaggcgtgt acggtgggag gcctatataa gcagagctcg tttagtgaac 180
cgtcagatcg cctcgagtac ccctctccgc ttcttacccc tctccgcttc ttacccctct 240
ccgcttctga attcgaccgc caccatgaga catattatct gccacggagg tgttattacc 300
gaagaaatgg ccgccagtct tttggaccag ctgatcgaag aggtactggc tgataatctt 360
ccacctccta gccattttga accacctacc cttcacgaac tgtatgattt agacgtgacg 420
gcccccgaag atcccaacga ggaggcggtt tcgcagattt ttcccgactc tgtaatgttg 480
gcggtgcagg aagggattga cttactcact tttccgccgg cgcccggttc tccggagccg 540
cctcaccttt cccggcagcc cgagcagccg gagcagagag ccttgggtcc ggtttctatg 600
ccaaaccttg taccggaggt gatcgatctt acctgccacg aggctggctt tccacccagt 660
gacgacgagg atgaagaggg tgaggagttt gtgttagatt atgtggagca ccccgggcac 720
ggttgcaggt cttgtcatta tcaccggagg aatacggggg acccagatat tatgtgttcg 780
ctttgctata tgaggacctg tggcatgttt gtctacagta agtgaaaatt atgggcagtg 840
ggtgatagag tggtgggttt ggtgtggtaa tttttttttt aatttttaca gttttgtggt 900
ttaaagaatt ttgtattgtg atttttttaa aaggtcctgt gtctgaacct gagcctgagc 960
ccgagccaga accggagcct gcaagaccta cccgccgtcc taaaatggcg cctgctatcc 1020
tgaggcgccc gacatcacct gtgtctagag aatgcaatag tagtacggat agctgtgact 1080
ccggtccttc taacacacct cctgagatac acccggtggt cccgctgtgc cccattaaac 1140
cagttgccgt gagagttggt gggcgtcgcc aggctgtgga atgtatcgag gacttgctta 1200
acgagcctgg gcaacctttg gacttgagct gtaaacgccc caggccaggg tccggagcta 1260
caaatttttc cctcctcaaa caggctggag atgtcgaaga aaatcctggg cctatggagg 1320
cttgggagtg tttggaagat ttttctgctg tgcgtaactt gctggaacag agctctaaca 1380
gtacctcttg gttttggagg tttctgtggg gctcatccca ggcaaagtta gtctgcagaa 1440
ttaaggagga ttacaagtgg gaatttgaag agcttttgaa atcctgtggt gagctgtttg 1500
attctttgaa tctgggtcac caggcgcttt tccaagagaa ggtcatcaag actttggatt 1560
tttccacacc ggggcgcgct gcggctgctg ttgctttttt gagttttata aaggataaat 1620
ggagcgaaga aacccatctg agcggggggt acctgctgga ttttctggcc atgcatctgt 1680
ggagagcggt tgtgagacac aagaatcgcc tgctactgtt gtcttccgtc cgcccggcga 1740
taataccgac ggaggagcag cagcagcagc aggaggaagc caggcggcgg cggcaggagc 1800
agagcccatg gaacccgaga gccggcctgg accctcggga atgaatgttg tacaggtggc 1860
tgaactgtat ccagaactga gacgcatttt gacaattaca gaggatgggc aggggctaaa 1920
gggggtaaag agggagcggg gggcttgtga ggctacagag gaggctagga atctagcttt 1980
tagcttaatg accagacacc gtcctgagtg tattactttt caacagatca aggataattg 2040
cgctaatgag cttgatctgc tggcgcagaa gtattccata gagcagctga ccacttactg 2100
gctgcagcca ggggatgatt ttgaggaggc tattagggta tatgcaaagg tggcacttag 2160
gccagattgc aagtacaaga tcagcaaact tgtaaatatc aggaattgtt gctacatttc 2220
tgggaacggg gccgaggtgg agatagatac ggaggatagg gtggccttta gatgtagcat 2280
gataaatatg tggccggggg tgcttggcat ggacggggtg gttattatga atgtaaggtt 2340
tactggcccc aattttagcg gtacggtttt cctggccaat accaacctta tcctacacgg 2400
tgtaagcttc tatgggttta acaatacctg tgtggaagcc tggaccgatg taagggttcg 2460
gggctgtgcc ttttactgct gctggaaggg ggtggtgtgt cgccccaaaa gcagggcttc 2520
aattaagaaa tgcctctttg aaaggtgtac cttgggtatc ctgtctgagg gtaactccag 2580
ggtgcgccac aatgtggcct ccgactgtgg ttgcttcatg ctagtgaaaa gcgtggctgt 2640
gattaagcat aacatggtat gtggcaactg cgaggacagg gcctctcaga tgctgacctg 2700
ctcggacggc aactgtcacc tgctgaagac cattcacgta gccagccact ctcgcaaggc 2760
ctggccagtg tttgagcata acatactgac ccgctgttcc ttgcatttgg gtaacaggag 2820
gggggtgttc ctaccttacc aatgcaattt gagtcacact aagatattgc ttgagcccga 2880
gagcatgtcc aaggtgaacc tgaacggggt gtttgacatg accatgaaga tctggaaggt 2940
gctgaggtac gatgagaccc gcaccaggtg cagaccctgc gagtgtggcg gtaaacatat 3000
taggaaccag cctgtgatgc tggatgtgac cgaggagctg aggcccgatc acttggtgct 3060
ggcctgcacc cgcgctgagt ttggctctag cgatgaagat acagatctcg aggggtccgg 3120
agctacaaat ttttccctcc tcaaacaggc tggagatgtc gaagaaaatc ctgggcctac 3180
cggtatgtat aggatgcagt tgctctcttg catcgccctg tctctagcgt tggtgactaa 3240
tagcgcccct acttcttctt ccacgaagaa aacccagctg caactggaac accttctgct 3300
ggatctccag atgatcctta acgggattaa caactataag aacccgaagc tcacggctat 3360
gctgacggcg aagttcgcta tgcctaaaaa ggctactgag ctgaaacact tacagtgtct 3420
cgaagaggca ctcaaacccc tcgaggaggt actcaatctg gcgcaaagca aaaacttcca 3480
tctccgccca cgggacctaa tctccaacat caacgtgatt gtgctggagc tcaaggggtc 3540
tgaaaccacg tttatgtgcg agtacgcaga tgaaactgct accatagtgg aattcctcaa 3600
tcgttggatc actttctgcc agtcaatcat ttcaaccctg accagatctg agggccgcgg 3660
cagcctgctg acctgcggcg acgtggagga aaaccccggc cccggatcct tgtcgaccat 3720
tcgtccgcgc aggccaagtc ctgcccgcga gcttctgccc ggaccccaac cggatagggt 3780
tcagccgact gcagatcgtg gggtgtctgc gcctgctggc agccctctgg atggcttgcc 3840
cgctcggcgg acggtgtccc ggacccggct gccatctccc cctgcgccct cacctgcgtt 3900
ctcggcgggc agcttcagcg atctgctccg tccgttcgat ccgtcgcttc ttgatacatc 3960
gcttcttgat tcgatgcctg ccgtcggcac gccgcataca gcggctgccc cagcagagtg 4020
ggatgaggcg caatcggctc tgcgtgcagc cgatgacccg ccacccaccg tgcgtgtcgc 4080
tgtcactgcc gcgcggccgc cgcgcgccaa gccggccccg cgacggcgtg ctgcgcaacc 4140
ctccgacgct tcgccggccg cgcaggtgga tctacgcacg ctcggctaca gtcagcagca 4200
gcaagagaag atcaaaccga aggtgcgttc gacagtggcg cagcaccacg aggcactggt 4260
gggccatggg tttacacacg cgcacatcgt tgcgctcagc caacacccgg cagcgttagg 4320
gaccgtcgct gtcacgtatc agcacataat cacggcgttg ccagaggcga cacacgaaga 4380
catcgttggc gtcggcaaac agtggtccgg cgcacgcgcc ctggaggcct tgctcacgga 4440
tgcgggggag ttgagaggtc cgccgttaca gttggacaca ggccaacttg tgaagattgc 4500
aaaacgtggc ggcgtgaccg caatggaggc agtgcatgca tcgcgcaatg cactgacggg 4560
tgcccccctg aacctgaccc cggaccaagt ggtggctatc gccagcaaca ttggcggcaa 4620
gcaagcgctc gaaacggtgc agcggctgtt gccggtgctg tgccaggacc atggcctgac 4680
cccggaccaa gtggtggcta tcgccagcaa cggtggcggc aagcaagcgc tcgaaacggt 4740
gcagcggctg ttgccggtgc tgtgccagga ccatggcctg accccggacc aagtggtggc 4800
tatcgccagc aacattggcg gcaagcaagc gctcgaaacg gtgcagcggc tgttgccggt 4860
gctgtgccag gaccatggcc tgaccccgga ccaagtggtg gctatcgcca gcaacaatgg 4920
cggcaagcaa gcgctcgaaa cggtgcagcg gctgttgccg gtgctgtgcc aggaccatgg 4980
cctgaccccg gaccaagtgg tggctatcgc cagcaacatt ggcggcaagc aagcgctcga 5040
aacggtgcag cggctgttgc cggtgctgtg ccaggaccat ggcctgaccc cggaccaagt 5100
ggtggctatc gccagcaaca ttggcggcaa gcaagcgctc gaaacggtgc agcggctgtt 5160
gccggtgctg tgccaggacc atggcctgac tccggaccaa gtggtggcta tcgccagcca 5220
cgatggcggc aagcaagcgc tcgaaacggt gcagcggctg ttgccggtgc tgtgccagga 5280
ccatggcctg actccggacc aagtggtggc tatcgccagc cacgatggcg gcaagcaagc 5340
gctcgaaacg gtgcagcggc tgttgccggt gctgtgccag gaccatggcc tgaccccgga 5400
ccaagtggtg gctatcgcca gcaacaatgg cggcaagcaa gcgctcgaaa cggtgcagcg 5460
gctgttgccg gtgctgtgcc aggaccatgg cctgaccccg gaccaagtgg tggctatcgc 5520
cagcaacatt ggcggcaagc aagcgctcga aacggtgcag cggctgttgc cggtgctgtg 5580
ccaggaccat ggcctgaccc cggaccaagt ggtggctatc gccagcaacg gtggcggcaa 5640
gcaagcgctc gaaacggtgc agcggctgtt gccggtgctg tgccaggacc atggcctgac 5700
tccggaccaa gtggtggcta tcgccagcca cgatggcggc aagcaagcgc tcgaaacggt 5760
gcagcggctg ttgccggtgc tgtgccagga ccatggcctg actccggacc aagtggtggc 5820
tatcgccagc cacgatggcg gcaagcaagc gctcgaaacg gtgcagcggc tgttgccggt 5880
gctgtgccag gaccatggcc tgaccccgga ccaagtggtg gctatcgcca gcaacggtgg 5940
cggcaagcaa gcgctcgaaa cggtgcagcg gctgttgccg gtgctgtgcc aggaccatgg 6000
cctgactccg gaccaagtgg tggctatcgc cagccacgat ggcggcaagc aagcgctcga 6060
aacggtgcag cggctgttgc cggtgctgtg ccaggaccat ggcctgactc cggaccaagt 6120
ggtggctatc gccagccacg atggcggcaa gcaagcgctc gaaacggtgc agcggctgtt 6180
gccggtgctg tgccaggacc atggcctgac tccggaccaa gtggtggcta tcgccagcca 6240
cgatggcggc aagcaagcgc tcgaaacggt gcagcggctg ttgccggtgc tgtgccagga 6300
ccatggcctg accccggacc aagtggtggc tatcgccagc aacattggcg gcaagcaagc 6360
gctcgaaacg gtgcagcggc tgttgccggt gctgtgccag gaccatggcc tgaccccgga 6420
ccaagtggtg gctatcgcca gcaacggtgg cggcaagcaa gcgctcgaaa cggtgcagcg 6480
gctgttgccg gtgctgtgcc aggaccatgg cctgaccccg gaccaagtgg tggctatcgc 6540
cagcaacggt ggcggcaagc aagcgctcga aagcattgtg gcccagctga gccggcctga 6600
tccggcgttg gccgcgttga ccaacgacca cctcgtcgcc ttggcctgcc tcggcggacg 6660
tcctgccatg gatgcagtga aaaagggatt gccgcacgcg ccggaattga tcagaagagt 6720
caatcgccgt attggcgaac gcacgtccca tcgcgttgcc gactacgcgc aagtggttcg 6780
cgtgctggag tttttccagt gccactccca cccagcgtac gcatttgatg aggccatgac 6840
gcagttcggg atgagcagga acgggttggt acagctcttt cgcagagtgg gcgtcaccga 6900
actcgaagcc cgcggtggaa cgctcccccc agcctcgcag cgttgggacc gtatcctcca 6960
ggcatcaggg atgaaaaggg ccaaaccgtc ccctacttca gctcaaacac cggatcaggc 7020
gtctttgcat gcattcgccg attcgctgga gcgtgacctt gatgcgccca gcccaatgca 7080
cgagggagat cagacgcggg caagcagccg taaacggtcc cgatcggatc gtgctgtcac 7140
cggcccctcc gcacagcagg ctgtcgaggt gcgcgttccc gaacagcgcg atgcgctgca 7200
tttgcccctc agctggaggg taaaacgccc gcgtaccagg atctggggcg gcctcccggt 7260
taagtagtgc tttctacttt atgagtagtg ctttctactt tatgagtagt gctttctact 7320
ttatgagtag tgctttctac tttatgtaac caatgtgcag actactgtta accaatgtgc 7380
agactactgt taaccaatgt gcagactact gttaaccaat gtgcagacta ctgtgaaagc 7440
ggagtactgt cctccgagcg gagtactgtc ctccgagcgg agtactgtcc tccgagcgga 7500
gtactgtcct ccgagcggag ttctgtcctc cgagcggaga ctctagatat agaaccgatc 7560
ctcccattga attctaggcg tgtacggtgg gaggcctata taagcagagc tcgtttagtg 7620
aaccgtcaga tcgcctcgag tatagaaccg atcctcccat ttatagaacc gatcctccca 7680
tttatagaac cgatcctccc attactagaa gatcttgaat tcgccacctt gtcgaccatt 7740
cgtccgcgca ggccaagtcc tgcccgcgag cttctgcccg gaccccaacc ggatagggtt 7800
cagccgactg cagatcgtgg ggtgtctgcg cctgctggca gccctctgga tggcttgccc 7860
gctcggcgga cggtgtcccg gacccggctg ccatctcccc ctgcgccctc acctgcgttc 7920
tcggcgggca gcttcagcga tctgctccgt ccgttcgatc cgtcgcttct tgatacatcg 7980
cttcttgatt cgatgcctgc cgtcggcacg ccgcatacag cggctgcccc agcagagtgg 8040
gatgaggcgc aatcggctct gcgtgcagcc gatgacccgc cacccaccgt gcgtgtcgct 8100
gtcactgccg cgcggccgcc gcgcgccaag ccggccccgc gacggcgtgc tgcgcaaccc 8160
tccgacgctt cgccggccgc gcaggtggat ctacgcacgc tcggctacag tcagcagcag 8220
caagagaaga tcaaaccgaa ggtgcgttcg acagtggcgc agcaccacga ggcactggtg 8280
ggccatgggt ttacacacgc gcacatcgtt gcgctcagcc aacacccggc agcgttaggg 8340
accgtcgctg tcacgtatca gcacataatc acggcgttgc cagaggcgac acacgaagac 8400
atcgttggcg tcggcaaaca gtggtccggc gcacgcgccc tggaggcctt gctcacggat 8460
gcgggggagt tgagaggtcc gccgttacag ttggacacag gccaacttgt gaagattgca 8520
aaacgtggcg gcgtgaccgc aatggaggca gtgcatgcat cgcgcaatgc actgacgggt 8580
gcccccctga acctgacccc ggaccaagtg gtggctatcg ccagcaacat tggcggcaag 8640
caagcgctcg aaacggtgca gcggctgttg ccggtgctgt gccaggacca tggcctgact 8700
ccggaccaag tggtggctat cgccagccac gatggcggca agcaagcgct cgaaacggtg 8760
cagcggctgt tgccggtgct gtgccaggac catggcctga ctccggacca agtggtggct 8820
atcgccagcc acgatggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg 8880
ctgtgccagg accatggcct gactccggac caagtggtgg ctatcgccag ccacgatggc 8940
ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc 9000
ctgactccgg accaagtggt ggctatcgcc agccacgatg gcggcaagca agcgctcgaa 9060
acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg gcctgacccc ggaccaagtg 9120
gtggctatcg ccagcaacgg tggcggcaag caagcgctcg aaacggtgca gcggctgttg 9180
ccggtgctgt gccaggacca tggcctgact ccggaccaag tggtggctat cgccagccac 9240
gatggcggca agcaagcgct cgaaacggtg cagcggctgt tgccggtgct gtgccaggac 9300
catggcctga ccccggacca agtggtggct atcgccagca acggtggcgg caagcaagcg 9360
ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg accatggcct gactccggac 9420
caagtggtgg ctatcgccag ccacgatggc ggcaagcaag cgctcgaaac ggtgcagcgg 9480
ctgttgccgg tgctgtgcca ggaccatggc ctgactccgg accaagtggt ggctatcgcc 9540
agccacgatg gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc 9600
caggaccatg gcctgacccc ggaccaagtg gtggctatcg ccagcaacaa tggcggcaag 9660
caagcgctcg aaacggtgca gcggctgttg ccggtgctgt gccaggacca tggcctgact 9720
ccggaccaag tggtggctat cgccagccac gatggcggca agcaagcgct cgaaacggtg 9780
cagcggctgt tgccggtgct gtgccaggac catggcctga ccccggacca agtggtggct 9840
atcgccagca acggtggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg 9900
ctgtgccagg accatggcct gaccccggac caagtggtgg ctatcgccag caacggtggc 9960
ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc 10020
ctgactccgg accaagtggt ggctatcgcc agccacgatg gcggcaagca agcgctcgaa 10080
acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg gcctgacccc ggaccaagtg 10140
gtggctatcg ccagcaacgg tggcggcaag caagcgctcg aaagcattgt ggcccagctg 10200
agccggcctg atccggcgtt ggccgcgttg accaacgacc acctcgtcgc cttggcctgc 10260
ctcggcggac gtcctgccat ggatgcagtg aaaaagggat tgccgcacgc gccggaattg 10320
atcagaagag tcaatcgccg tattggcgaa cgcacgtccc atcgcgttgc cgactacgcg 10380
caagtggttc gcgtgctgga gtttttccag tgccactccc acccagcgta cgcatttgat 10440
gaggccatga cgcagttcgg gatgagcagg aacgggttgg tacagctctt tcgcagagtg 10500
ggcgtcaccg aactcgaagc ccgcggtgga acgctccccc cagcctcgca gcgttgggac 10560
cgtatcctcc aggcatcagg gatgaaaagg gccaaaccgt cccctacttc agctcaaaca 10620
ccggatcagg cgtctttgca tgcattcgcc gattcgctgg agcgtgacct tgatgcgccc 10680
agcccaatgc acgagggaga tcagacgcgg gcaagcagcc gtaaacggtc ccgatcggat 10740
cgtgctgtca ccggcccctc cgcacagcag gctgtcgagg tgcgcgttcc cgaacagcgc 10800
gatgcgctgc atttgcccct cagctggagg gtaaaacgcc cgcgtaccag gatctggggc 10860
ggcctcccgt aatacaaggc ggccgcaaat caacatcagt ctgataagct atcaacatca 10920
gtctgataag ctatcaacat cagtctgata agctatcaac atcagtctga taagctaaag 10980
cttcgaattc tgataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac 11040
ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg 11100
tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa 11160
gcattttttt cactgcgttc atcggagccc aggtttactc ccttaagtgg aaatttcttc 11220
ccccactccc tccttggctt tctccaagga gggaacccag gctactggaa agtccggctg 11280
gggcggggac tgtgggtttc agggtagaac tgcgtgtgga acgggacagg gagcggttag 11340
aagggtgggg ctattccggg aagtggtggg gggagggagc ccaaaactag cacctagtcc 11400
actcattatc cagccctctt atttctcggc cccgctctgc ttcagtggac ccggggaggg 11460
cggggaagtg gagtgggaga cctaggggtg ggcttcccga ccttgctgta caggacctcg 11520
acctagctgg ctttgttccc catccccacg ttagttgttg ccctgaggct aaaactagag 11580
cccaggggcc ccaagttcca gactgcccct cccccctccc ccggagccag ggagtggttg 11640
gtgaaagggg gaggccagct ggagaacaaa cgggtagtca gggggttgag cgattagagc 11700
ccttgtaccc tacccaggaa tggttgggga ggaggaggaa gaggtaggag gtaggggagg 11760
gggcggggtt ttgtcacctg tcacctgctc cggctgtgcc tagggcgggc gggcggggag 11820
tggggggacc ggtataaagc ggtaggcgcc tgtgcccgct ccacctctca agcagccagc 11880
gcctgcctga atctgttctg ccccctcccc acccatttca ccaccaccat gccaaagaag 11940
aagcggaagg tcggtggcgg ctcacccggg ggtggcggct caatggacaa gaagtactcc 12000
attggcctcg ccatcggaac aaatagcgtg ggctgggctg tcatcacaga tgagtacaag 12060
gtgcctagca agaaatttaa ggtgctggga aatacagaca gacatagcat caagaagaac 12120
ctcattggcg ctctcctgtt tgactccggc gaaacagccg aagctaccag actcaagaga 12180
accgctagga gaaggtacac cagaaggaaa aacaggattt gctacctgca ggaaattttt 12240
tccaacgaga tggccaaggt ggacgattcc ttcttccata ggctggaaga gagcttcctc 12300
gtggaggaag acaagaaaca cgagaggcat cctatttttg gcaatattgt ggatgaggtc 12360
gcctaccatg agaagtatcc cacaatctat catctgagaa aaaaactggt ggatagcacc 12420
gacaaggccg atctcaggct catttatctc gctctggctc acatgatcaa gtttaggggc 12480
cacttcctga tcgaaggcga cctgaatccc gacaactccg acgtggacaa actgttcatc 12540
cagctcgtcc agacctacaa tcaactcttc gaggagaacc ccatcaatgc ttccggcgtg 12600
gatgccaagg ccatcctgag cgctaggctc tccaagtcca ggaggctgga aaatctgatc 12660
gcccaactcc ctggagagaa gaagaacggc ctgtttggca atctgattgc cctgagcctc 12720
ggactcaccc ccaacttcaa gagcaacttc gatctcgccg aagacgccaa actccaactg 12780
agcaaggata cctacgacga cgatctcgat aatctcctcg cccagatcgg cgatcaatat 12840
gccgacctct ttctggccgc caaaaacctg agcgacgcta ttctgctcag cgacattctc 12900
agggtgaata cagaaatcac aaaagccccc ctgtccgcca gcatgatcaa aaggtacgat 12960
gaacaccatc aggacctcac cctgctgaag gctctggtca ggcagcaact ccccgaaaag 13020
tacaaggaga ttttctttga tcagtccaag aatggatatg ctggctatat tgatggaggc 13080
gcctcccagg aggaatttta taaattcatc aagcccattc tcgaaaagat ggacggaacc 13140
gaagagctgc tggtcaaact caatagggag gatctgctga ggaagcaaag gaccttcgac 13200
aatggcagca tcccccacca gatccacctc ggcgaactcc acgctatcct caggaggcag 13260
gaagacttct accctttcct gaaggataac agggagaaaa tcgagaaaat cctgaccttc 13320
agaatcccct actacgtcgg acctctcgcc aggggcaatt ccagattcgc ctggatgaca 13380
aggaagagcg aggaaacaat cacaccatgg aacttcgaag aagtggtcga taagggcgcc 13440
agcgcccaga gcttcattga aaggatgacc aactttgata agaacctgcc caatgagaag 13500
gtgctgccta agcactccct gctgtatgag tatttcaccg tgtataatga gctgaccaag 13560
gtcaagtacg tcaccgaggg aatgagaaag cctgcttttc tctccggcga gcagaaaaaa 13620
gccatcgtgg acctgctgtt caagaccaac aggaaggtga ccgtcaagca actcaaggag 13680
gactacttta agaagattga gtgctttgat agcgtggaaa ttagcggagt cgaggacagg 13740
ttcaatgcct ccctcggaac atatcacgac ctgctgaaaa tcatcaaaga caaagatttt 13800
ctggataacg aggagaatga agacattctg gaggacattg tcctcaccct gaccctgttt 13860
gaggacagag agatgattga agagaggctg aaaacctatg cccacctgtt cgacgacaag 13920
gtgatgaagc agctcaaaag aaggagatat accggctggg gcagactgtc caggaagctg 13980
atcaacggca ttagggacaa gcagagcggc aagaccattc tcgactttct caagtccgac 14040
ggattcgcca acagaaactt tatgcaactg atccacgatg acagcctcac ctttaaggaa 14100
gatattcaga aggctcaggt ctgcctggac ttgaaaaccc aagtccagac cccccaaggc 14160
atgaaggaga tcagcaatat acaagtgggc gacctggtgc tgtccaacac cggctacaac 14220
gaggtgttga atgttttccc caagagcaag aaaaaaagct acaagatcac tttggaggac 14280
ggaaaggaaa tcatctgctc cgaggaacac ctgttcccca cccagaccgg cgagatgaac 14340
atctctggtg gactgaaaga gggcatgtgc ctgtacgtca aagagactag tggtggcggc 14400
tcaaagcgtc ctgctgctac taagaaagct ggtcaagcta agaaaaagaa atctagatag 14460
aagcttcact cctcaggtgc aggctgccta tcagaaggtg gtggctggtg tggccaatgc 14520
cctggctcac aaataccact gagatctttt tccctctgcc aaaaattatg gggacatcat 14580
gaagcccctt gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt 14640
gtgttggaat tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa 14700
acatcagaat gagtatttgg tttagagttt ggcaacatat gcccatatgc tggctgccat 14760
gaacaaaggt tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc 14820
cttattccat agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt 14880
atttttttct ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct 14940
cctctcctga ctactcccag tcatagctgt ccctcttctc ttatggagat c 14991
<210> 26
<211> 14898
<212> DNA
<213> Artificial
<220>
<223> 编码由细胞特异性启动子hMUC1、第一microRNA为mir-199a-3p、第二micro
RNA为mir-21、转录激活因子为分割dCas9、第一调控蛋白和第二调控蛋白分
别为TALE蛋白、第一和第二识别序列为5×UAS的元件组成的表达系统的核酸
<400> 26
cggagtactg tcctccgagc ggagtactgt cctccgagcg gagtactgtc ctccgagcgg 60
agtactgtcc tccgagcgga gttctgtcct ccgagcggag actctagata cccctctccg 120
cttctgaatt ctaggcgtgt acggtgggag gcctatataa gcagagctcg tttagtgaac 180
cgtcagatcg cctcgagtac ccctctccgc ttcttacccc tctccgcttc ttacccctct 240
ccgcttctga attcgaccgc caccatgaga catattatct gccacggagg tgttattacc 300
gaagaaatgg ccgccagtct tttggaccag ctgatcgaag aggtactggc tgataatctt 360
ccacctccta gccattttga accacctacc cttcacgaac tgtatgattt agacgtgacg 420
gcccccgaag atcccaacga ggaggcggtt tcgcagattt ttcccgactc tgtaatgttg 480
gcggtgcagg aagggattga cttactcact tttccgccgg cgcccggttc tccggagccg 540
cctcaccttt cccggcagcc cgagcagccg gagcagagag ccttgggtcc ggtttctatg 600
ccaaaccttg taccggaggt gatcgatctt acctgccacg aggctggctt tccacccagt 660
gacgacgagg atgaagaggg tgaggagttt gtgttagatt atgtggagca ccccgggcac 720
ggttgcaggt cttgtcatta tcaccggagg aatacggggg acccagatat tatgtgttcg 780
ctttgctata tgaggacctg tggcatgttt gtctacagta agtgaaaatt atgggcagtg 840
ggtgatagag tggtgggttt ggtgtggtaa tttttttttt aatttttaca gttttgtggt 900
ttaaagaatt ttgtattgtg atttttttaa aaggtcctgt gtctgaacct gagcctgagc 960
ccgagccaga accggagcct gcaagaccta cccgccgtcc taaaatggcg cctgctatcc 1020
tgaggcgccc gacatcacct gtgtctagag aatgcaatag tagtacggat agctgtgact 1080
ccggtccttc taacacacct cctgagatac acccggtggt cccgctgtgc cccattaaac 1140
cagttgccgt gagagttggt gggcgtcgcc aggctgtgga atgtatcgag gacttgctta 1200
acgagcctgg gcaacctttg gacttgagct gtaaacgccc caggccaggg tccggagcta 1260
caaatttttc cctcctcaaa caggctggag atgtcgaaga aaatcctggg cctatggagg 1320
cttgggagtg tttggaagat ttttctgctg tgcgtaactt gctggaacag agctctaaca 1380
gtacctcttg gttttggagg tttctgtggg gctcatccca ggcaaagtta gtctgcagaa 1440
ttaaggagga ttacaagtgg gaatttgaag agcttttgaa atcctgtggt gagctgtttg 1500
attctttgaa tctgggtcac caggcgcttt tccaagagaa ggtcatcaag actttggatt 1560
tttccacacc ggggcgcgct gcggctgctg ttgctttttt gagttttata aaggataaat 1620
ggagcgaaga aacccatctg agcggggggt acctgctgga ttttctggcc atgcatctgt 1680
ggagagcggt tgtgagacac aagaatcgcc tgctactgtt gtcttccgtc cgcccggcga 1740
taataccgac ggaggagcag cagcagcagc aggaggaagc caggcggcgg cggcaggagc 1800
agagcccatg gaacccgaga gccggcctgg accctcggga atgaatgttg tacaggtggc 1860
tgaactgtat ccagaactga gacgcatttt gacaattaca gaggatgggc aggggctaaa 1920
gggggtaaag agggagcggg gggcttgtga ggctacagag gaggctagga atctagcttt 1980
tagcttaatg accagacacc gtcctgagtg tattactttt caacagatca aggataattg 2040
cgctaatgag cttgatctgc tggcgcagaa gtattccata gagcagctga ccacttactg 2100
gctgcagcca ggggatgatt ttgaggaggc tattagggta tatgcaaagg tggcacttag 2160
gccagattgc aagtacaaga tcagcaaact tgtaaatatc aggaattgtt gctacatttc 2220
tgggaacggg gccgaggtgg agatagatac ggaggatagg gtggccttta gatgtagcat 2280
gataaatatg tggccggggg tgcttggcat ggacggggtg gttattatga atgtaaggtt 2340
tactggcccc aattttagcg gtacggtttt cctggccaat accaacctta tcctacacgg 2400
tgtaagcttc tatgggttta acaatacctg tgtggaagcc tggaccgatg taagggttcg 2460
gggctgtgcc ttttactgct gctggaaggg ggtggtgtgt cgccccaaaa gcagggcttc 2520
aattaagaaa tgcctctttg aaaggtgtac cttgggtatc ctgtctgagg gtaactccag 2580
ggtgcgccac aatgtggcct ccgactgtgg ttgcttcatg ctagtgaaaa gcgtggctgt 2640
gattaagcat aacatggtat gtggcaactg cgaggacagg gcctctcaga tgctgacctg 2700
ctcggacggc aactgtcacc tgctgaagac cattcacgta gccagccact ctcgcaaggc 2760
ctggccagtg tttgagcata acatactgac ccgctgttcc ttgcatttgg gtaacaggag 2820
gggggtgttc ctaccttacc aatgcaattt gagtcacact aagatattgc ttgagcccga 2880
gagcatgtcc aaggtgaacc tgaacggggt gtttgacatg accatgaaga tctggaaggt 2940
gctgaggtac gatgagaccc gcaccaggtg cagaccctgc gagtgtggcg gtaaacatat 3000
taggaaccag cctgtgatgc tggatgtgac cgaggagctg aggcccgatc acttggtgct 3060
ggcctgcacc cgcgctgagt ttggctctag cgatgaagat acagatctcg aggggtccgg 3120
agctacaaat ttttccctcc tcaaacaggc tggagatgtc gaagaaaatc ctgggcctac 3180
cggtatgtat aggatgcagt tgctctcttg catcgccctg tctctagcgt tggtgactaa 3240
tagcgcccct acttcttctt ccacgaagaa aacccagctg caactggaac accttctgct 3300
ggatctccag atgatcctta acgggattaa caactataag aacccgaagc tcacggctat 3360
gctgacggcg aagttcgcta tgcctaaaaa ggctactgag ctgaaacact tacagtgtct 3420
cgaagaggca ctcaaacccc tcgaggaggt actcaatctg gcgcaaagca aaaacttcca 3480
tctccgccca cgggacctaa tctccaacat caacgtgatt gtgctggagc tcaaggggtc 3540
tgaaaccacg tttatgtgcg agtacgcaga tgaaactgct accatagtgg aattcctcaa 3600
tcgttggatc actttctgcc agtcaatcat ttcaaccctg accagatctg agggccgcgg 3660
cagcctgctg acctgcggcg acgtggagga aaaccccggc cccggatcct tgtcgaccat 3720
tcgtccgcgc aggccaagtc ctgcccgcga gcttctgccc ggaccccaac cggatagggt 3780
tcagccgact gcagatcgtg gggtgtctgc gcctgctggc agccctctgg atggcttgcc 3840
cgctcggcgg acggtgtccc ggacccggct gccatctccc cctgcgccct cacctgcgtt 3900
ctcggcgggc agcttcagcg atctgctccg tccgttcgat ccgtcgcttc ttgatacatc 3960
gcttcttgat tcgatgcctg ccgtcggcac gccgcataca gcggctgccc cagcagagtg 4020
ggatgaggcg caatcggctc tgcgtgcagc cgatgacccg ccacccaccg tgcgtgtcgc 4080
tgtcactgcc gcgcggccgc cgcgcgccaa gccggccccg cgacggcgtg ctgcgcaacc 4140
ctccgacgct tcgccggccg cgcaggtgga tctacgcacg ctcggctaca gtcagcagca 4200
gcaagagaag atcaaaccga aggtgcgttc gacagtggcg cagcaccacg aggcactggt 4260
gggccatggg tttacacacg cgcacatcgt tgcgctcagc caacacccgg cagcgttagg 4320
gaccgtcgct gtcacgtatc agcacataat cacggcgttg ccagaggcga cacacgaaga 4380
catcgttggc gtcggcaaac agtggtccgg cgcacgcgcc ctggaggcct tgctcacgga 4440
tgcgggggag ttgagaggtc cgccgttaca gttggacaca ggccaacttg tgaagattgc 4500
aaaacgtggc ggcgtgaccg caatggaggc agtgcatgca tcgcgcaatg cactgacggg 4560
tgcccccctg aacctgaccc cggaccaagt ggtggctatc gccagcaaca ttggcggcaa 4620
gcaagcgctc gaaacggtgc agcggctgtt gccggtgctg tgccaggacc atggcctgac 4680
cccggaccaa gtggtggcta tcgccagcaa cggtggcggc aagcaagcgc tcgaaacggt 4740
gcagcggctg ttgccggtgc tgtgccagga ccatggcctg accccggacc aagtggtggc 4800
tatcgccagc aacattggcg gcaagcaagc gctcgaaacg gtgcagcggc tgttgccggt 4860
gctgtgccag gaccatggcc tgaccccgga ccaagtggtg gctatcgcca gcaacaatgg 4920
cggcaagcaa gcgctcgaaa cggtgcagcg gctgttgccg gtgctgtgcc aggaccatgg 4980
cctgaccccg gaccaagtgg tggctatcgc cagcaacatt ggcggcaagc aagcgctcga 5040
aacggtgcag cggctgttgc cggtgctgtg ccaggaccat ggcctgaccc cggaccaagt 5100
ggtggctatc gccagcaaca ttggcggcaa gcaagcgctc gaaacggtgc agcggctgtt 5160
gccggtgctg tgccaggacc atggcctgac tccggaccaa gtggtggcta tcgccagcca 5220
cgatggcggc aagcaagcgc tcgaaacggt gcagcggctg ttgccggtgc tgtgccagga 5280
ccatggcctg actccggacc aagtggtggc tatcgccagc cacgatggcg gcaagcaagc 5340
gctcgaaacg gtgcagcggc tgttgccggt gctgtgccag gaccatggcc tgaccccgga 5400
ccaagtggtg gctatcgcca gcaacaatgg cggcaagcaa gcgctcgaaa cggtgcagcg 5460
gctgttgccg gtgctgtgcc aggaccatgg cctgaccccg gaccaagtgg tggctatcgc 5520
cagcaacatt ggcggcaagc aagcgctcga aacggtgcag cggctgttgc cggtgctgtg 5580
ccaggaccat ggcctgaccc cggaccaagt ggtggctatc gccagcaacg gtggcggcaa 5640
gcaagcgctc gaaacggtgc agcggctgtt gccggtgctg tgccaggacc atggcctgac 5700
tccggaccaa gtggtggcta tcgccagcca cgatggcggc aagcaagcgc tcgaaacggt 5760
gcagcggctg ttgccggtgc tgtgccagga ccatggcctg actccggacc aagtggtggc 5820
tatcgccagc cacgatggcg gcaagcaagc gctcgaaacg gtgcagcggc tgttgccggt 5880
gctgtgccag gaccatggcc tgaccccgga ccaagtggtg gctatcgcca gcaacggtgg 5940
cggcaagcaa gcgctcgaaa cggtgcagcg gctgttgccg gtgctgtgcc aggaccatgg 6000
cctgactccg gaccaagtgg tggctatcgc cagccacgat ggcggcaagc aagcgctcga 6060
aacggtgcag cggctgttgc cggtgctgtg ccaggaccat ggcctgactc cggaccaagt 6120
ggtggctatc gccagccacg atggcggcaa gcaagcgctc gaaacggtgc agcggctgtt 6180
gccggtgctg tgccaggacc atggcctgac tccggaccaa gtggtggcta tcgccagcca 6240
cgatggcggc aagcaagcgc tcgaaacggt gcagcggctg ttgccggtgc tgtgccagga 6300
ccatggcctg accccggacc aagtggtggc tatcgccagc aacattggcg gcaagcaagc 6360
gctcgaaacg gtgcagcggc tgttgccggt gctgtgccag gaccatggcc tgaccccgga 6420
ccaagtggtg gctatcgcca gcaacggtgg cggcaagcaa gcgctcgaaa cggtgcagcg 6480
gctgttgccg gtgctgtgcc aggaccatgg cctgaccccg gaccaagtgg tggctatcgc 6540
cagcaacggt ggcggcaagc aagcgctcga aagcattgtg gcccagctga gccggcctga 6600
tccggcgttg gccgcgttga ccaacgacca cctcgtcgcc ttggcctgcc tcggcggacg 6660
tcctgccatg gatgcagtga aaaagggatt gccgcacgcg ccggaattga tcagaagagt 6720
caatcgccgt attggcgaac gcacgtccca tcgcgttgcc gactacgcgc aagtggttcg 6780
cgtgctggag tttttccagt gccactccca cccagcgtac gcatttgatg aggccatgac 6840
gcagttcggg atgagcagga acgggttggt acagctcttt cgcagagtgg gcgtcaccga 6900
actcgaagcc cgcggtggaa cgctcccccc agcctcgcag cgttgggacc gtatcctcca 6960
ggcatcaggg atgaaaaggg ccaaaccgtc ccctacttca gctcaaacac cggatcaggc 7020
gtctttgcat gcattcgccg attcgctgga gcgtgacctt gatgcgccca gcccaatgca 7080
cgagggagat cagacgcggg caagcagccg taaacggtcc cgatcggatc gtgctgtcac 7140
cggcccctcc gcacagcagg ctgtcgaggt gcgcgttccc gaacagcgcg atgcgctgca 7200
tttgcccctc agctggaggg taaaacgccc gcgtaccagg atctggggcg gcctcccggt 7260
taagtagtgc tttctacttt atgagtagtg ctttctactt tatgagtagt gctttctact 7320
ttatgagtag tgctttctac tttatgtaac caatgtgcag actactgtta accaatgtgc 7380
agactactgt taaccaatgt gcagactact gttaaccaat gtgcagacta ctgtgaaagc 7440
ggagtactgt cctccgagcg gagtactgtc ctccgagcgg agtactgtcc tccgagcgga 7500
gtactgtcct ccgagcggag ttctgtcctc cgagcggaga ctctagatat agaaccgatc 7560
ctcccattga attctaggcg tgtacggtgg gaggcctata taagcagagc tcgtttagtg 7620
aaccgtcaga tcgcctcgag tatagaaccg atcctcccat ttatagaacc gatcctccca 7680
tttatagaac cgatcctccc attactagaa gatcttgaat tcgccacctt gtcgaccatt 7740
cgtccgcgca ggccaagtcc tgcccgcgag cttctgcccg gaccccaacc ggatagggtt 7800
cagccgactg cagatcgtgg ggtgtctgcg cctgctggca gccctctgga tggcttgccc 7860
gctcggcgga cggtgtcccg gacccggctg ccatctcccc ctgcgccctc acctgcgttc 7920
tcggcgggca gcttcagcga tctgctccgt ccgttcgatc cgtcgcttct tgatacatcg 7980
cttcttgatt cgatgcctgc cgtcggcacg ccgcatacag cggctgcccc agcagagtgg 8040
gatgaggcgc aatcggctct gcgtgcagcc gatgacccgc cacccaccgt gcgtgtcgct 8100
gtcactgccg cgcggccgcc gcgcgccaag ccggccccgc gacggcgtgc tgcgcaaccc 8160
tccgacgctt cgccggccgc gcaggtggat ctacgcacgc tcggctacag tcagcagcag 8220
caagagaaga tcaaaccgaa ggtgcgttcg acagtggcgc agcaccacga ggcactggtg 8280
ggccatgggt ttacacacgc gcacatcgtt gcgctcagcc aacacccggc agcgttaggg 8340
accgtcgctg tcacgtatca gcacataatc acggcgttgc cagaggcgac acacgaagac 8400
atcgttggcg tcggcaaaca gtggtccggc gcacgcgccc tggaggcctt gctcacggat 8460
gcgggggagt tgagaggtcc gccgttacag ttggacacag gccaacttgt gaagattgca 8520
aaacgtggcg gcgtgaccgc aatggaggca gtgcatgcat cgcgcaatgc actgacgggt 8580
gcccccctga acctgacccc ggaccaagtg gtggctatcg ccagcaacat tggcggcaag 8640
caagcgctcg aaacggtgca gcggctgttg ccggtgctgt gccaggacca tggcctgact 8700
ccggaccaag tggtggctat cgccagccac gatggcggca agcaagcgct cgaaacggtg 8760
cagcggctgt tgccggtgct gtgccaggac catggcctga ctccggacca agtggtggct 8820
atcgccagcc acgatggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg 8880
ctgtgccagg accatggcct gactccggac caagtggtgg ctatcgccag ccacgatggc 8940
ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc 9000
ctgactccgg accaagtggt ggctatcgcc agccacgatg gcggcaagca agcgctcgaa 9060
acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg gcctgacccc ggaccaagtg 9120
gtggctatcg ccagcaacgg tggcggcaag caagcgctcg aaacggtgca gcggctgttg 9180
ccggtgctgt gccaggacca tggcctgact ccggaccaag tggtggctat cgccagccac 9240
gatggcggca agcaagcgct cgaaacggtg cagcggctgt tgccggtgct gtgccaggac 9300
catggcctga ccccggacca agtggtggct atcgccagca acggtggcgg caagcaagcg 9360
ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg accatggcct gactccggac 9420
caagtggtgg ctatcgccag ccacgatggc ggcaagcaag cgctcgaaac ggtgcagcgg 9480
ctgttgccgg tgctgtgcca ggaccatggc ctgactccgg accaagtggt ggctatcgcc 9540
agccacgatg gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc 9600
caggaccatg gcctgacccc ggaccaagtg gtggctatcg ccagcaacaa tggcggcaag 9660
caagcgctcg aaacggtgca gcggctgttg ccggtgctgt gccaggacca tggcctgact 9720
ccggaccaag tggtggctat cgccagccac gatggcggca agcaagcgct cgaaacggtg 9780
cagcggctgt tgccggtgct gtgccaggac catggcctga ccccggacca agtggtggct 9840
atcgccagca acggtggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg 9900
ctgtgccagg accatggcct gaccccggac caagtggtgg ctatcgccag caacggtggc 9960
ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc 10020
ctgactccgg accaagtggt ggctatcgcc agccacgatg gcggcaagca agcgctcgaa 10080
acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg gcctgacccc ggaccaagtg 10140
gtggctatcg ccagcaacgg tggcggcaag caagcgctcg aaagcattgt ggcccagctg 10200
agccggcctg atccggcgtt ggccgcgttg accaacgacc acctcgtcgc cttggcctgc 10260
ctcggcggac gtcctgccat ggatgcagtg aaaaagggat tgccgcacgc gccggaattg 10320
atcagaagag tcaatcgccg tattggcgaa cgcacgtccc atcgcgttgc cgactacgcg 10380
caagtggttc gcgtgctgga gtttttccag tgccactccc acccagcgta cgcatttgat 10440
gaggccatga cgcagttcgg gatgagcagg aacgggttgg tacagctctt tcgcagagtg 10500
ggcgtcaccg aactcgaagc ccgcggtgga acgctccccc cagcctcgca gcgttgggac 10560
cgtatcctcc aggcatcagg gatgaaaagg gccaaaccgt cccctacttc agctcaaaca 10620
ccggatcagg cgtctttgca tgcattcgcc gattcgctgg agcgtgacct tgatgcgccc 10680
agcccaatgc acgagggaga tcagacgcgg gcaagcagcc gtaaacggtc ccgatcggat 10740
cgtgctgtca ccggcccctc cgcacagcag gctgtcgagg tgcgcgttcc cgaacagcgc 10800
gatgcgctgc atttgcccct cagctggagg gtaaaacgcc cgcgtaccag gatctggggc 10860
ggcctcccgt aatacaaggc ggccgcaaat caacatcagt ctgataagct atcaacatca 10920
gtctgataag ctatcaacat cagtctgata agctatcaac atcagtctga taagctaaag 10980
cttcgaattc tgataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac 11040
ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg 11100
tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa 11160
gcattttttt cactgcgttc atcggagccc aggtttactc ccttaagtgg aaatttcttc 11220
ccccactccc tccttggctt tctccaagga gggaacccag gctactggaa agtccggctg 11280
gggcggggac tgtgggtttc agggtagaac tgcgtgtgga acgggacagg gagcggttag 11340
aagggtgggg ctattccggg aagtggtggg gggagggagc ccaaaactag cacctagtcc 11400
actcattatc cagccctctt atttctcggc cccgctctgc ttcagtggac ccggggaggg 11460
cggggaagtg gagtgggaga cctaggggtg ggcttcccga ccttgctgta caggacctcg 11520
acctagctgg ctttgttccc catccccacg ttagttgttg ccctgaggct aaaactagag 11580
cccaggggcc ccaagttcca gactgcccct cccccctccc ccggagccag ggagtggttg 11640
gtgaaagggg gaggccagct ggagaacaaa cgggtagtca gggggttgag cgattagagc 11700
ccttgtaccc tacccaggaa tggttgggga ggaggaggaa gaggtaggag gtaggggagg 11760
gggcggggtt ttgtcacctg tcacctgctc cggctgtgcc tagggcgggc gggcggggag 11820
tggggggacc ggtataaagc ggtaggcgcc tgtgcccgct ccacctctca agcagccagc 11880
gcctgcctga atctgttctg ccccctcccc acccatttca ccaccaccat gctgaagaag 11940
atcctgaaga tcgaggagct ggacgagaga gagctgatcg acatcgaggt gagcggcaac 12000
cacctgttct acgccaacga catcctgacc cacaacagca gtggccaagg cgattccctc 12060
catgagcaca tcgctaatct ggctggctcc cctgctatca aaaagggcat cctccagaca 12120
gtcaaagtcg tcgatgagct ggtcaaggtg atgggcaggc ataaacccga gaacattgtg 12180
attgagatgg ctagggagaa ccagaccacc cagaaaggcc agaaaaacag cagagaaaga 12240
atgaagagga tcgaggaggg catcaaagaa ctgggcagcc aaatcctcaa ggagcacccc 12300
gtcgaaaata cacaactcca gaacgaaaaa ctctacctct actatctgca gaacggcaga 12360
gacatgtacg tggaccagga actggacatc aacaggctct ccgattacga tgtggacgcc 12420
atcgtccctc agtcctttct gaaagatgat agcatcgaca acaaggtgct gaccaggtcc 12480
gacaagaata ggggcaagag cgataatgtg ccctccgagg aggtcgtcaa aaaaatgaaa 12540
aactactgga gacaactcct caacgctaag ctcatcaccc aaagaaagtt cgacaatctg 12600
accaaagccg agaggggcgg cctctccgaa ctggacaagg ccggcttcat caaaaggcaa 12660
ttggtggaaa ccaggcagat tacaaagcat gtcgctcaaa ttctcgatag caggatgaat 12720
accaaatatg acgagaacga caagctgatc agagaggtca aggtcatcac actcaagtcc 12780
aagctcgtga gcgacttcag aaaagatttc caattttata aagtcaggga gatcaacaat 12840
taccaccacg ctcacgacgc ttatctcaac gctgtcgtgg gaaccgccct gatcaaaaaa 12900
taccccaagc tggaaagcga gttcgtgtat ggcgattata aagtgtacga cgtgaggaag 12960
atgatcgcta aaagcgagca ggaaatcggc aaggctacag ccaagtactt tttctactcc 13020
aacattatga acttcttcaa gaccgagatt accctcgcca acggcgaaat taggaagagg 13080
cccctgattg aaacaaatgg agaaacaggc gaaatcgtct gggacaaggg cagggacttc 13140
gccacagtca gaaaagtgct gtccatgcct caagtcaaca tcgtcaaaaa gaccgaggtg 13200
cagaccggcg gctttagcaa agaaagcatc ctgcccaaga gaaactccga caagctcatc 13260
gctaggaaga aggactggga ccctaagaaa tacggaggat ttgactcccc taccgtcgcc 13320
tattccgtcc tcgtcgtcgc taaggtggag aagggcaaga gcaagaagct caagagcgtc 13380
aaggagctgc tgggaatcac catcatggag aggagctcct tcgaaaaaaa ccctattgat 13440
ttcctggagg ccaagggcta caaggaggtc aagaaggacc tcatcatcaa gctgcccaaa 13500
tacagcctct tcgaactgga aaatggcagg aagagaatgc tcgctagcgc cggcgagctc 13560
cagaaaggaa atgagctggc tctgcccagc aagtacgtca acttcctcta tctcgccagc 13620
cactatgaaa agctcaaggg cagccccgaa gacaatgagc agaagcagct cttcgtcgag 13680
cagcacaagc actacctcga tgaaatcatc gagcaaatca gcgagttttc caaaagggtg 13740
attctcgccg acgctaacct cgataaggtc ctctccgctt acaacaagca tagagacaag 13800
cccatcagag aacaggccga gaacatcatc cacctgttta cactcacaaa cctcggagcc 13860
cctgccgctt ttaaatactt cgatacaacc attgatagga agaggtacac ctccaccaag 13920
gaggtgctgg atgctaccct gattcatcaa tccatcacag gactctacga aacaaggatt 13980
gacctgtccc aactgggagg cgacactagt ggtggcggct caaagcgtcc tgctgctact 14040
aagaaagctg gtcaagctaa gaaaaagaaa tctagaatgg acgccaagag cctgaccgcc 14100
tggtcaagga ccctggtaac cttcaaagac gtttttgtgg acttcacaag ggaggaatgg 14160
aagttgctgg acaccgcaca gcagatcgtg tataggaacg tgatgttgga gaattacaag 14220
aacctggtta gcctgggcta tcagttgacc aagcccgacg taatccttag gctggaaaag 14280
ggcgaggaac cctggctggt ggagagggag attcatcagg aaacccaccc cgacagcgaa 14340
accgctttcg agatcaagtc atctgtgctc gagtaatcct caggtgcagg ctgcctatca 14400
gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa taccactgag atctttttcc 14460
ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt ctggctaata 14520
aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag 14580
gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt agagtttggc 14640
aacatatgcc catatgctgg ctgccatgaa caaaggttgg ctataaagag gtcatcagta 14700
tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga cttgaggtta 14760
gatttttttt atattttgtt ttgtgttatt tttttcttta acatccctaa aattttcctt 14820
acatgtttta ctagccagat ttttcctcct ctcctgacta ctcccagtca tagctgtccc 14880
tcttctctta tggagatc 14898
<210> 27
<211> 31000
<212> DNA
<213> Artificial
<220>
<223> 去除E1基因与部分E3基因5型腺病毒载体的序列
<400> 27
catcatcaat aatatacctt attttggatt gaagccaata tgataatgag ggggtggagt 60
ttgtgacgtg gcgcggggcg tgggaacggg gcgggtgacg tagtagtgtg gcggaagtgt 120
gatgttgcaa gtgtggcgga acacatgtaa gcgacggatg tggcaaaagt gacgtttttg 180
gtgtgcgccg gtgtacacag gaagtgacaa ttttcgcgcg gttttaggcg gatgttgtag 240
taaatttggg cgtaaccgag taagatttgg ccattttcgc gggaaaactg aataagagga 300
agtgaaatct gaataatttt gtgttactca tagcgcgtaa tatttgtcta gggccgcggg 360
gactttgacc gtttacgtgg agactcgccc aggtgttttt ctcaggtgtt ttctgcgttc 420
cgggtcaaag ttggcgtttt attattatag tcagtcgaac tgaaatgtgt gggcgtggct 480
taagggtggg aaagaatata taaggtgggg gtcttatgta gttttgtatc tgttttgcag 540
cagccgccgc cgccatgagc accaactcgt ttgatggaag cattgtgagc tcatatttga 600
caacgcgcat gcccccatgg gccggggtgc gtcagaatgt gatgggctcc agcattgatg 660
gtcgccccgt cctgcccgca aactctacta ccttgaccta cgagaccgtg tctggaacgc 720
cgttggagac tgcagcctcc gccgccgctt cagccgctgc agccaccgcc cgcgggattg 780
tgactgactt tgctttcctg agcccgcttg caagcagtgc agcttcccgt tcatccgccc 840
gcgatgacaa gttgacggct cttttggcac aattggattc tttgacccgg gaacttaatg 900
tcgtttctca gcagctgttg gatctgcgcc agcaggtttc tgccctgaag gcttcctccc 960
ctcccaatgc ggtttaaaac ataaataaaa aaccagactc tgtttggatt tggatcaagc 1020
aagtgtcttg ctgtctttat ttaggggttt tgcgcgcgcg gtaggcccgg gaccagcggt 1080
ctcggtcgtt gagggtcctg tgtatttttt ccaggacgtg gtaaaggtga ctctggatgt 1140
tcagatacat gggcataagc ccgtctctgg ggtggaggta gcaccactgc agagcttcat 1200
gctgcggggt ggtgttgtag atgatccagt cgtagcagga gcgctgggcg tggtgcctaa 1260
aaatgtcttt cagtagcaag ctgattgcca ggggcaggcc cttggtgtaa gtgtttacaa 1320
agcggttaag ctgggatggg tgcatacgtg gggatatgag atgcatcttg gactgtattt 1380
ttaggttggc tatgttccca gccatatccc tccggggatt catgttgtgc agaaccacca 1440
gcacagtgta tccggtgcac ttgggaaatt tgtcatgtag cttagaagga aatgcgtgga 1500
agaacttgga gacgcccttg tgacctccaa gattttccat gcattcgtcc ataatgatgg 1560
caatgggccc acgggcggcg gcctgggcga agatatttct gggatcacta acgtcatagt 1620
tgtgttccag gatgagatcg tcataggcca tttttacaaa gcgcgggcgg agggtgccag 1680
actgcggtat aatggttcca tccggcccag gggcgtagtt accctcacag atttgcattt 1740
cccacgcttt gagttcagat ggggggatca tgtctacctg cggggcgatg aagaaaacgg 1800
tttccggggt aggggagatc agctgggaag aaagcaggtt cctgagcagc tgcgacttac 1860
cgcagccggt gggcccgtaa atcacaccta ttaccggctg caactggtag ttaagagagc 1920
tgcagctgcc gtcatccctg agcagggggg ccacttcgtt aagcatgtcc ctgactcgca 1980
tgttttccct gaccaaatcc gccagaaggc gctcgccgcc cagcgatagc agttcttgca 2040
aggaagcaaa gtttttcaac ggtttgagac cgtccgccgt aggcatgctt ttgagcgttt 2100
gaccaagcag ttccaggcgg tcccacagct cggtcacctg ctctacggca tctcgatcca 2160
gcatatctcc tcgtttcgcg ggttggggcg gctttcgctg tacggcagta gtcggtgctc 2220
gtccagacgg gccagggtca tgtctttcca cgggcgcagg gtcctcgtca gcgtagtctg 2280
ggtcacggtg aaggggtgcg ctccgggctg cgcgctggcc agggtgcgct tgaggctggt 2340
cctgctggtg ctgaagcgct gccggtcttc gccctgcgcg tcggccaggt agcatttgac 2400
catggtgtca tagtccagcc cctccgcggc gtggcccttg gcgcgcagct tgcccttgga 2460
ggaggcgccg cacgaggggc agtgcagact tttgagggcg tagagcttgg gcgcgagaaa 2520
taccgattcc ggggagtagg catccgcgcc gcaggccccg cagacggtct cgcattccac 2580
gagccaggtg agctctggcc gttcggggtc aaaaaccagg tttcccccat gctttttgat 2640
gcgtttctta cctctggttt ccatgagccg gtgtccacgc tcggtgacga aaaggctgtc 2700
cgtgtccccg tatacagact tgagaggcct gtcctcgagc ggtgttccgc ggtcctcctc 2760
gtatagaaac tcggaccact ctgagacaaa ggctcgcgtc caggccagca cgaaggaggc 2820
taagtgggag gggtagcggt cgttgtccac tagggggtcc actcgctcca gggtgtgaag 2880
acacatgtcg ccctcttcgg catcaaggaa ggtgattggt ttgtaggtgt aggccacgtg 2940
accgggtgtt cctgaagggg ggctataaaa gggggtgggg gcgcgttcgt cctcactctc 3000
ttccgcatcg ctgtctgcga gggccagctg ttggggtgag tactccctct gaaaagcggg 3060
catgacttct gcgctaagat tgtcagtttc caaaaacgag gaggatttga tattcacctg 3120
gcccgcggtg atgcctttga gggtggccgc atccatctgg tcagaaaaga caatcttttt 3180
gttgtcaagc ttggtggcaa acgacccgta gagggcgttg gacagcaact tggcgatgga 3240
gcgcagggtt tggtttttgt cgcgatcggc gcgctccttg gccgcgatgt ttagctgcac 3300
gtattcgcgc gcaacgcacc gccattcggg aaagacggtg gtgcgctcgt cgggcaccag 3360
gtgcacgcgc caaccgcggt tgtgcagggt gacaaggtca acgctggtgg ctacctctcc 3420
gcgtaggcgc tcgttggtcc agcagaggcg gccgcccttg cgcgagcaga atggcggtag 3480
ggggtctagc tgcgtctcgt ccggggggtc tgcgtccacg gtaaagaccc cgggcagcag 3540
gcgcgcgtcg aagtagtcta tcttgcatcc ttgcaagtct agcgcctgct gccatgcgcg 3600
ggcggcaagc gcgcgctcgt atgggttgag tgggggaccc catggcatgg ggtgggtgag 3660
cgcggaggcg tacatgccgc aaatgtcgta aacgtagagg ggctctctga gtattccaag 3720
atatgtaggg tagcatcttc caccgcggat gctggcgcgc acgtaatcgt atagttcgtg 3780
cgagggagcg aggaggtcgg gaccgaggtt gctacgggcg ggctgctctg ctcggaagac 3840
tatctgcctg aagatggcat gtgagttgga tgatatggtt ggacgctgga agacgttgaa 3900
gctggcgtct gtgagaccta ccgcgtcacg cacgaaggag gcgtaggagt cgcgcagctt 3960
gttgaccagc tcggcggtga cctgcacgtc tagggcgcag tagtccaggg tttccttgat 4020
gatgtcatac ttatcctgtc cctttttttt ccacagctcg cggttgagga caaactcttc 4080
gcggtctttc cagtactctt ggatcggaaa cccgtcggcc tccgaacggt aagagcctag 4140
catgtagaac tggttgacgg cctggtaggc gcagcatccc ttttctacgg gtagcgcgta 4200
tgcctgcgcg gccttccgga gcgaggtgtg ggtgagcgca aaggtgtccc tgaccatgac 4260
tttgaggtac tggtatttga agtcagtgtc gtcgcatccg ccctgctccc agagcaaaaa 4320
gtccgtgcgc tttttggaac gcggatttgg cagggcgaag gtgacatcgt tgaagagtat 4380
ctttcccgcg cgaggcataa agttgcgtgt gatgcggaag ggtcccggca cctcggaacg 4440
gttgttaatt acctgggcgg cgagcacgat ctcgtcaaag ccgttgatgt tgtggcccac 4500
aatgtaaagt tccaagaagc gcgggatgcc cttgatggaa ggcaattttt taagttcctc 4560
gtaggtgagc tcttcagggg agctgagccc gtgctctgaa agggcccagt ctgcaagatg 4620
agggttggaa gcgacgaatg agctccacag gtcacgggcc attagcattt gcaggtggtc 4680
gcgaaaggtc ctaaactggc gacctatggc cattttttct ggggtgatgc agtagaaggt 4740
aagcgggtct tgttcccagc ggtcccatcc aaggttcgcg gctaggtctc gcgcggcagt 4800
cactagaggc tcatctccgc cgaacttcat gaccagcatg aagggcacga gctgcttccc 4860
aaaggccccc atccaagtat aggtctctac atcgtaggtg acaaagagac gctcggtgcg 4920
aggatgcgag ccgatcggga agaactggat ctcccgccac caattggagg agtggctatt 4980
gatgtggtga aagtagaagt ccctgcgacg ggccgaacac tcgtgctggc ttttgtaaaa 5040
acgtgcgcag tactggcagc ggtgcacggg ctgtacatcc tgcacgaggt tgacctgacg 5100
accgcgcaca aggaagcaga gtgggaattt gagcccctcg cctggcgggt ttggctggtg 5160
gtcttctact tcggctgctt gtccttgacc gtctggctgc tcgaggggag ttacggtgga 5220
tcggaccacc acgccgcgcg agcccaaagt ccagatgtcc gcgcgcggcg gtcggagctt 5280
gatgacaaca tcgcgcagat gggagctgtc catggtctgg agctcccgcg gcgtcaggtc 5340
aggcgggagc tcctgcaggt ttacctcgca tagacgggtc agggcgcggg ctagatccag 5400
gtgataccta atttccaggg gctggttggt ggcggcgtcg atggcttgca agaggccgca 5460
tccccgcggc gcgactacgg taccgcgcgg cgggcggtgg gccgcggggg tgtccttgga 5520
tgatgcatct aaaagcggtg acgcgggcga gcccccggag gtaggggggg ctccggaccc 5580
gccgggagag ggggcagggg cacgtcggcg ccgcgcgcgg gcaggagctg gtgctgcgcg 5640
cgtaggttgc tggcgaacgc gacgacgcgg cggttgatct cctgaatctg gcgcctctgc 5700
gtgaagacga cgggcccggt gagcttgaac ctgaaagaga gttcgacaga atcaatttcg 5760
gtgtcgttga cggcggcctg gcgcaaaatc tcctgcacgt ctcctgagtt gtcttgatag 5820
gcgatctcgg ccatgaactg ctcgatctct tcctcctgga gatctccgcg tccggctcgc 5880
tccacggtgg cggcgaggtc gttggaaatg cgggccatga gctgcgagaa ggcgttgagg 5940
cctccctcgt tccagacgcg gctgtagacc acgccccctt cggcatcgcg ggcgcgcatg 6000
accacctgcg cgagattgag ctccacgtgc cgggcgaaga cggcgtagtt tcgcaggcgc 6060
tgaaagaggt agttgagggt ggtggcggtg tgttctgcca cgaagaagta cataacccag 6120
cgtcgcaacg tggattcgtt gatatccccc aaggcctcaa ggcgctccat ggcctcgtag 6180
aagtccacgg cgaagttgaa aaactgggag ttgcgcgccg acacggttaa ctcctcctcc 6240
agaagacgga tgagctcggc gacagtgtcg cgcacctcgc gctcaaaggc tacaggggcc 6300
tcttcttctt cttcaatctc ctcttccata agggcctccc cttcttcttc ttctggcggc 6360
ggtgggggag gggggacacg gcggcgacga cggcgcaccg ggaggcggtc gacaaagcgc 6420
tcgatcatct ccccgcggcg acggcgcatg gtctcggtga cggcgcggcc gttctcgcgg 6480
gggcgcagtt ggaagacgcc gcccgtcatg tcccggttat gggttggcgg ggggctgcca 6540
tgcggcaggg atacggcgct aacgatgcat ctcaacaatt gttgtgtagg tactccgccg 6600
ccgagggacc tgagcgagtc cgcatcgacc ggatcggaaa acctctcgag aaaggcgtct 6660
aaccagtcac agtcgcaagg taggctgagc accgtggcgg gcggcagcgg gcggcggtcg 6720
gggttgtttc tggcggaggt gctgctgatg atgtaattaa agtaggcggt cttgagacgg 6780
cggatggtcg acagaagcac catgtccttg ggtccggcct gctgaatgcg caggcggtcg 6840
gccatgcccc aggcttcgtt ttgacatcgg cgcaggtctt tgtagtagtc ttgcatgagc 6900
ctttctaccg gcacttcttc ttctccttcc tcttgtcctg catctcttgc atctatcgct 6960
gcggcggcgg cggagtttgg ccgtaggtgg cgccctcttc ctcccatgcg tgtgaccccg 7020
aagcccctca tcggctgaag cagggctagg tcggcgacaa cgcgctcggc taatatggcc 7080
tgctgcacct gcgtgagggt agactggaag tcatccatgt ccacaaagcg gtggtatgcg 7140
cccgtgttga tggtgtaagt gcagttggcc ataacggacc agttaacggt ctggtgaccc 7200
ggctgcgaga gctcggtgta cctgagacgc gagtaagccc tcgagtcaaa tacgtagtcg 7260
ttgcaagtcc gcaccaggta ctggtatccc accaaaaagt gcggcggcgg ctggcggtag 7320
aggggccagc gtagggtggc cggggctccg ggggcgagat cttccaacat aaggcgatga 7380
tatccgtaga tgtacctgga catccaggtg atgccggcgg cggtggtgga ggcgcgcgga 7440
aagtcgcgga cgcggttcca gatgttgcgc agcggcaaaa agtgctccat ggtcgggacg 7500
ctctggccgg tcaggcgcgc gcaatcgttg acgctctaga ccgtgcaaaa ggagagcctg 7560
taagcgggca ctcttccgtg gtctggtgga taaattcgca agggtatcat ggcggacgac 7620
cggggttcga gccccgtatc cggccgtccg ccgtgatcca tgcggttacc gcccgcgtgt 7680
cgaacccagg tgtgcgacgt cagacaacgg gggagtgctc cttttggctt ccttccaggc 7740
gcggcggctg ctgcgctagc ttttttggcc actggccgcg cgcagcgtaa gcggttaggc 7800
tggaaagcga aagcattaag tggctcgctc cctgtagccg gagggttatt ttccaagggt 7860
tgagtcgcgg gacccccggt tcgagtctcg gaccggccgg actgcggcga acgggggttt 7920
gcctccccgt catgcaagac cccgcttgca aattcctccg gaaacaggga cgagcccctt 7980
ttttgctttt cccagatgca tccggtgctg cggcagatgc gcccccctcc tcagcagcgg 8040
caagagcaag agcagcggca gacatgcagg gcaccctccc ctcctcctac cgcgtcagga 8100
ggggcgacat ccgcggttga cgcggcagca gatggtgatt acgaaccccc gcggcgccgg 8160
gcccggcact acctggactt ggaggagggc gagggcctgg cgcggctagg agcgccctct 8220
cctgagcggc acccaagggt gcagctgaag cgtgatacgc gtgaggcgta cgtgccgcgg 8280
cagaacctgt ttcgcgaccg cgagggagag gagcccgagg agatgcggga tcgaaagttc 8340
cacgcagggc gcgagctgcg gcatggcctg aatcgcgagc ggttgctgcg cgaggaggac 8400
tttgagcccg acgcgcgaac cgggattagt cccgcgcgcg cacacgtggc ggccgccgac 8460
ctggtaaccg catacgagca gacggtgaac caggagatta actttcaaaa aagctttaac 8520
aaccacgtgc gtacgcttgt ggcgcgcgag gaggtggcta taggactgat gcatctgtgg 8580
gactttgtaa gcgcgctgga gcaaaaccca aatagcaagc cgctcatggc gcagctgttc 8640
cttatagtgc agcacagcag ggacaacgag gcattcaggg atgcgctgct aaacatagta 8700
gagcccgagg gccgctggct gctcgatttg ataaacatcc tgcagagcat agtggtgcag 8760
gagcgcagct tgagcctggc tgacaaggtg gccgccatca actattccat gcttagcctg 8820
ggcaagtttt acgcccgcaa gatataccat accccttacg ttcccataga caaggaggta 8880
aagatcgagg ggttctacat gcgcatggcg ctgaaggtgc ttaccttgag cgacgacctg 8940
ggcgtttatc gcaacgagcg catccacaag gccgtgagcg tgagccggcg gcgcgagctc 9000
agcgaccgcg agctgatgca cagcctgcaa agggccctgg ctggcacggg cagcggcgat 9060
agagaggccg agtcctactt tgacgcgggc gctgacctgc gctgggcccc aagccgacgc 9120
gccctggagg cagctggggc cggacctggg ctggcggtgg cacccgcgcg cgctggcaac 9180
gtcggcggcg tggaggaata tgacgaggac gatgagtacg agccagagga cggcgagtac 9240
taagcggtga tgtttctgat cagatgatgc aagacgcaac ggacccggcg gtgcgggcgg 9300
cgctgcagag ccagccgtcc ggccttaact ccacggacga ctggcgccag gtcatggacc 9360
gcatcatgtc gctgactgcg cgcaatcctg acgcgttccg gcagcagccg caggccaacc 9420
ggctctccgc aattctggaa gcggtggtcc cggcgcgcgc aaaccccacg cacgagaagg 9480
tgctggcgat cgtaaacgcg ctggccgaaa acagggccat ccggcccgac gaggccggcc 9540
tggtctacga cgcgctgctt cagcgcgtgg ctcgttacaa cagcggcaac gtgcagacca 9600
acctggaccg gctggtgggg gatgtgcgcg aggccgtggc gcagcgtgag cgcgcgcagc 9660
agcagggcaa cctgggctcc atggttgcac taaacgcctt cctgagtaca cagcccgcca 9720
acgtgccgcg gggacaggag gactacacca actttgtgag cgcactgcgg ctaatggtga 9780
ctgagacacc gcaaagtgag gtgtaccagt ctgggccaga ctattttttc cagaccagta 9840
gacaaggcct gcagaccgta aacctgagcc aggctttcaa aaacttgcag gggctgtggg 9900
gggtgcgggc tcccacaggc gaccgcgcga ccgtgtctag cttgctgacg cccaactcgc 9960
gcctgttgct gctgctaata gcgcccttca cggacagtgg cagcgtgtcc cgggacacat 10020
acctaggtca cttgctgaca ctgtaccgcg aggccatagg tcaggcgcat gtggacgagc 10080
atactttcca ggagattaca agtgtcagcc gcgcgctggg gcaggaggac acgggcagcc 10140
tggaggcaac cctaaactac ctgctgacca accggcggca gaagatcccc tcgttgcaca 10200
gtttaaacag cgaggaggag cgcattttgc gctacgtgca gcagagcgtg agccttaacc 10260
tgatgcgcga cggggtaacg cccagcgtgg cgctggacat gaccgcgcgc aacatggaac 10320
cgggcatgta tgcctcaaac cggccgttta tcaaccgcct aatggactac ttgcatcgcg 10380
cggccgccgt gaaccccgag tatttcacca atgccatctt gaacccgcac tggctaccgc 10440
cccctggttt ctacaccggg ggattcgagg tgcccgaggg taacgatgga ttcctctggg 10500
acgacataga cgacagcgtg ttttccccgc aaccgcagac cctgctagag ttgcaacagc 10560
gcgagcaggc agaggcggcg ctgcgaaagg aaagcttccg caggccaagc agcttgtccg 10620
atctaggcgc tgcggccccg cggtcagatg ctagtagccc atttccaagc ttgatagggt 10680
ctcttaccag cactcgcacc acccgcccgc gcctgctggg cgaggaggag tacctaaaca 10740
actcgctgct gcagccgcag cgcgaaaaaa acctgcctcc ggcatttccc aacaacggga 10800
tagagagcct agtggacaag atgagtagat ggaagacgta cgcgcaggag cacagggacg 10860
tgccaggccc gcgcccgccc acccgtcgtc aaaggcacga ccgtcagcgg ggtctggtgt 10920
gggaggacga tgactcggca gacgacagca gcgtcctgga tttgggaggg agtggcaacc 10980
cgtttgcgca ccttcgcccc aggctgggga gaatgtttta aaaaaaaaaa aagcatgatg 11040
caaaataaaa aactcaccaa ggccatggca ccgagcgttg gttttcttgt attcccctta 11100
gtatgcggcg cgcggcgatg tatgaggaag gtcctcctcc ctcctacgag agtgtggtga 11160
gcgcggcgcc agtggcggcg gcgctgggtt ctcccttcga tgctcccctg gacccgccgt 11220
ttgtgcctcc gcggtacctg cggcctaccg gggggagaaa cagcatccgt tactctgagt 11280
tggcacccct attcgacacc acccgtgtgt acctggtgga caacaagtca acggatgtgg 11340
catccctgaa ctaccagaac gaccacagca actttctgac cacggtcatt caaaacaatg 11400
actacagccc gggggaggca agcacacaga ccatcaatct tgacgaccgg tcgcactggg 11460
gcggcgacct gaaaaccatc ctgcatacca acatgccaaa tgtgaacgag ttcatgttta 11520
ccaataagtt taaggcgcgg gtgatggtgt cgcgcttgcc tactaaggac aatcaggtgg 11580
agctgaaata cgagtgggtg gagttcacgc tgcccgaggg caactactcc gagaccatga 11640
ccatagacct tatgaacaac gcgatcgtgg agcactactt gaaagtgggc agacagaacg 11700
gggttctgga aagcgacatc ggggtaaagt ttgacacccg caacttcaga ctggggtttg 11760
accccgtcac tggtcttgtc atgcctgggg tatatacaaa cgaagccttc catccagaca 11820
tcattttgct gccaggatgc ggggtggact tcacccacag ccgcctgagc aacttgttgg 11880
gcatccgcaa gcggcaaccc ttccaggagg gctttaggat cacctacgat gatctggagg 11940
gtggtaacat tcccgcactg ttggatgtgg acgcctacca ggcgagcttg aaagatgaca 12000
ccgaacaggg cgggggtggc gcaggcggca gcaacagcag tggcagcggc gcggaagaga 12060
actccaacgc ggcagccgcg gcaatgcagc cggtggagga catgaacgat catgccattc 12120
gcggcgacac ctttgccaca cgggctgagg agaagcgcgc tgaggccgaa gcagcggccg 12180
aagctgccgc ccccgctgcg caacccgagg tcgagaagcc tcagaagaaa ccggtgatca 12240
aacccctgac agaggacagc aagaaacgca gttacaacct aataagcaat gacagcacct 12300
tcacccagta ccgcagctgg taccttgcat acaactacgg cgaccctcag accggaatcc 12360
gctcatggac cctgctttgc actcctgacg taacctgcgg ctcggagcag gtctactggt 12420
cgttgccaga catgatgcaa gaccccgtga ccttccgctc cacgcgccag atcagcaact 12480
ttccggtggt gggcgccgag ctgttgcccg tgcactccaa gagcttctac aacgaccagg 12540
ccgtctactc ccaactcatc cgccagttta cctctctgac ccacgtgttc aatcgctttc 12600
ccgagaacca gattttggcg cgcccgccag cccccaccat caccaccgtc agtgaaaacg 12660
ttcctgctct cacagatcac gggacgctac cgctgcgcaa cagcatcgga ggagtccagc 12720
gagtgaccat tactgacgcc agacgccgca cctgccccta cgtttacaag gccctgggca 12780
tagtctcgcc gcgcgtccta tcgagccgca ctttttgagc aagcatgtcc atccttatat 12840
cgcccagcaa taacacaggc tggggcctgc gcttcccaag caagatgttt ggcggggcca 12900
agaagcgctc cgaccaacac ccagtgcgcg tgcgcgggca ctaccgcgcg ccctggggcg 12960
cgcacaaacg cggccgcact gggcgcacca ccgtcgatga cgccatcgac gcggtggtgg 13020
aggaggcgcg caactacacg cccacgccgc caccagtgtc cacagtggac gcggccattc 13080
agaccgtggt gcgcggagcc cggcgctatg ctaaaatgaa gagacggcgg aggcgcgtag 13140
cacgtcgcca ccgccgccga cccggcactg ccgcccaacg cgcggcggcg gccctgctta 13200
accgcgcacg tcgcaccggc cgacgggcgg ccatgcgggc cgctcgaagg ctggccgcgg 13260
gtattgtcac tgtgcccccc aggtccaggc gacgagcggc cgccgcagca gccgcggcca 13320
ttagtgctat gactcagggt cgcaggggca acgtgtattg ggtgcgcgac tcggttagcg 13380
gcctgcgcgt gcccgtgcgc acccgccccc cgcgcaacta gattgcaaga aaaaactact 13440
tagactcgta ctgttgtatg tatccagcgg cggcggcgcg caacgaagct atgtccaagc 13500
gcaaaatcaa agaagagatg ctccaggtca tcgcgccgga gatctatggc cccccgaaga 13560
aggaagagca ggattacaag ccccgaaagc taaagcgggt caaaaagaaa aagaaagatg 13620
atgatgatga acttgacgac gaggtggaac tgctgcacgc taccgcgccc aggcgacggg 13680
tacagtggaa aggtcgacgc gtaaaacgtg ttttgcgacc cggcaccacc gtagtcttta 13740
cgcccggtga gcgctccacc cgcacctaca agcgcgtgta tgatgaggtg tacggcgacg 13800
aggacctgct tgagcaggcc aacgagcgcc tcggggagtt tgcctacgga aagcggcata 13860
aggacatgct ggcgttgccg ctggacgagg gcaacccaac acctagccta aagcccgtaa 13920
cactgcagca ggtgctgccc gcgcttgcac cgtccgaaga aaagcgcggc ctaaagcgcg 13980
agtctggtga cttggcaccc accgtgcagc tgatggtacc caagcgccag cgactggaag 14040
atgtcttgga aaaaatgacc gtggaacctg ggctggagcc cgaggtccgc gtgcggccaa 14100
tcaagcaggt ggcgccggga ctgggcgtgc agaccgtgga cgttcagata cccactacca 14160
gtagcaccag tattgccacc gccacagagg gcatggagac acaaacgtcc ccggttgcct 14220
cagcggtggc ggatgccgcg gtgcaggcgg tcgctgcggc cgcgtccaag acctctacgg 14280
aggtgcaaac ggacccgtgg atgtttcgcg tttcagcccc ccggcgcccg cgccgttcga 14340
ggaagtacgg cgccgccagc gcgctactgc ccgaatatgc cctacatcct tccattgcgc 14400
ctacccccgg ctatcgtggc tacacctacc gccccagaag acgagcaact acccgacgcc 14460
gaaccaccac tggaacccgc cgccgccgtc gccgtcgcca gcccgtgctg gccccgattt 14520
ccgtgcgcag ggtggctcgc gaaggaggca ggaccctggt gctgccaaca gcgcgctacc 14580
accccagcat cgtttaaaag ccggtctttg tggttcttgc agatatggcc ctcacctgcc 14640
gcctccgttt cccggtgccg ggattccgag gaagaatgca ccgtaggagg ggcatggccg 14700
gccacggcct gacgggcggc atgcgtcgtg cgcaccaccg gcggcggcgc gcgtcgcacc 14760
gtcgcatgcg cggcggtatc ctgcccctcc ttattccact gatcgccgcg gcgattggcg 14820
ccgtgcccgg aattgcatcc gtggccttgc aggcgcagag acactgatta aaaacaagtt 14880
gcatgtggaa aaatcaaaat aaaaagtctg gactctcacg ctcgcttggt cctgtaacta 14940
ttttgtagaa tggaagacat caactttgcg tctctggccc cgcgacacgg ctcgcgcccg 15000
ttcatgggaa actggcaaga tatcggcacc agcaatatga gcggtggcgc cttcagctgg 15060
ggctcgctgt ggagcggcat taaaaatttc ggttccaccg ttaagaacta tggcagcaag 15120
gcctggaaca gcagcacagg ccagatgctg agggataagt tgaaagagca aaatttccaa 15180
caaaaggtgg tagatggcct ggcctctggc attagcgggg tggtggacct ggccaaccag 15240
gcagtgcaaa ataagattaa cagtaagctt gatccccgcc ctcccgtaga ggagcctcca 15300
ccggccgtgg agacagtgtc tccagagggg cgtggcgaaa agcgtccgcg ccccgacagg 15360
gaagaaactc tggtgacgca aatagacgag cctccctcgt acgaggaggc actaaagcaa 15420
ggcctgccca ccacccgtcc catcgcgccc atggctaccg gagtgctggg ccagcacaca 15480
cccgtaacgc tggacctgcc tccccccgcc gacacccagc agaaacctgt gctgccaggc 15540
ccgaccgccg ttgttgtaac ccgtcctagc cgcgcgtccc tgcgccgcgc cgccagcggt 15600
ccgcgatcgt tgcggcccgt agccagtggc aactggcaaa gcacactgaa cagcatcgtg 15660
ggtctggggg tgcaatccct gaagcgccga cgatgcttct gatagctaac gtgtcgtatg 15720
tgtgtcatgt atgcgtccat gtcgccgcca gaggagctgc tgagccgccg cgcgcccgct 15780
ttccaagatg gctacccctt cgatgatgcc gcagtggtct tacatgcaca tctcgggcca 15840
ggacgcctcg gagtacctga gccccgggct ggtgcagttt gcccgcgcca ccgagacgta 15900
cttcagcctg aataacaagt ttagaaaccc cacggtggcg cctacgcacg acgtgaccac 15960
agaccggtcc cagcgtttga cgctgcggtt catccctgtg gaccgtgagg atactgcgta 16020
ctcgtacaag gcgcggttca ccctagctgt gggtgataac cgtgtgctgg acatggcttc 16080
cacgtacttt gacatccgcg gcgtgctgga caggggccct acttttaagc cctactctgg 16140
cactgcctac aacgccctgg ctcccaaggg tgccccaaat ccttgcgaat gggatgaagc 16200
tgctactgct cttgaaataa acctagaaga agaggacgat gacaacgaag acgaagtaga 16260
cgagcaagct gagcagcaaa aaactcacgt atttgggcag gcgccttatt ctggtataaa 16320
tattacaaag gagggtattc aaataggtgt cgaaggtcaa acacctaaat atgccgataa 16380
aacatttcaa cctgaacctc aaataggaga atctcagtgg tacgaaacag aaattaatca 16440
tgcagctggg agagtcctaa aaaagactac cccaatgaaa ccatgttacg gttcatatgc 16500
aaaacccaca aatgaaaatg gagggcaagg cattcttgta aagcaacaaa atggaaagct 16560
agaaagtcaa gtggaaatgc aatttttctc aactactgag gcagccgcag gcaatggtga 16620
taacttgact cctaaagtgg tattgtacag tgaagatgta gatatagaaa ccccagacac 16680
tcatatttct tacatgccca ctattaagga aggtaactca cgagaactaa tgggccaaca 16740
atctatgccc aacaggccta attacattgc ttttagggac aattttattg gtctaatgta 16800
ttacaacagc acgggtaata tgggtgttct ggcgggccaa gcatcgcagt tgaatgctgt 16860
tgtagatttg caagacagaa acacagagct ttcataccag cttttgcttg attccattgg 16920
tgatagaacc aggtactttt ctatgtggaa tcaggctgtt gacagctatg atccagatgt 16980
tagaattatt gaaaatcatg gaactgaaga tgaacttcca aattactgct ttccactggg 17040
aggtgtgatt aatacagaga ctcttaccaa ggtaaaacct aaaacaggtc aggaaaatgg 17100
atgggaaaaa gatgctacag aattttcaga taaaaatgaa ataagagttg gaaataattt 17160
tgccatggaa atcaatctaa atgccaacct gtggagaaat ttcctgtact ccaacatagc 17220
gctgtatttg cccgacaagc taaagtacag tccttccaac gtaaaaattt ctgataaccc 17280
aaacacctac gactacatga acaagcgagt ggtggctccc gggctagtgg actgctacat 17340
taaccttgga gcacgctggt cccttgacta tatggacaac gtcaacccat ttaaccacca 17400
ccgcaatgct ggcctgcgct accgctcaat gttgctgggc aatggtcgct atgtgccctt 17460
ccacatccag gtgcctcaga agttctttgc cattaaaaac ctccttctcc tgccgggctc 17520
atacacctac gagtggaact tcaggaagga tgttaacatg gttctgcaga gctccctagg 17580
aaatgaccta agggttgacg gagccagcat taagtttgat agcatttgcc tttacgccac 17640
cttcttcccc atggcccaca acaccgcctc cacgcttgag gccatgctta gaaacgacac 17700
caacgaccag tcctttaacg actatctctc cgccgccaac atgctctacc ctatacccgc 17760
caacgctacc aacgtgccca tatccatccc ctcccgcaac tgggcggctt tccgcggctg 17820
ggccttcacg cgccttaaga ctaaggaaac cccatcactg ggctcgggct acgaccctta 17880
ttacacctac tctggctcta taccctacct agatggaacc ttttacctca accacacctt 17940
taagaaggtg gccattacct ttgactcttc tgtcagctgg cctggcaatg accgcctgct 18000
tacccccaac gagtttgaaa ttaagcgctc agttgacggg gagggttaca acgttgccca 18060
gtgtaacatg accaaagact ggttcctggt acaaatgcta gctaactata acattggcta 18120
ccagggcttc tatatcccag agagctacaa ggaccgcatg tactccttct ttagaaactt 18180
ccagcccatg agccgtcagg tggtggatga tactaaatac aaggactacc aacaggtggg 18240
catcctacac caacacaaca actctggatt tgttggctac cttgccccca ccatgcgcga 18300
aggacaggcc taccctgcta acttccccta tccgcttata ggcaagaccg cagttgacag 18360
cattacccag aaaaagtttc tttgcgatcg caccctttgg cgcatcccat tctccagtaa 18420
ctttatgtcc atgggcgcac tcacagacct gggccaaaac cttctctacg ccaactccgc 18480
ccacgcgcta gacatgactt ttgaggtgga tcccatggac gagcccaccc ttctttatgt 18540
tttgtttgaa gtctttgacg tggtccgtgt gcaccagccg caccgcggcg tcatcgaaac 18600
cgtgtacctg cgcacgccct tctcggccgg caacgccaca acataaagaa gcaagcaaca 18660
tcaacaacag ctgccgccat gggctccagt gagcaggaac tgaaagccat tgtcaaagat 18720
cttggttgtg ggccatattt tttgggcacc tatgacaagc gctttccagg ctttgtttct 18780
ccacacaagc tcgcctgcgc catagtcaat acggccggtc gcgagactgg gggcgtacac 18840
tggatggcct ttgcctggaa cccgcactca aaaacatgct acctctttga gccctttggc 18900
ttttctgacc agcgactcaa gcaggtttac cagtttgagt acgagtcact cctgcgccgt 18960
agcgccattg cttcttcccc cgaccgctgt ataacgctgg aaaagtccac ccaaagcgta 19020
caggggccca actcggccgc ctgtggacta ttctgctgca tgtttctcca cgcctttgcc 19080
aactggcccc aaactcccat ggatcacaac cccaccatga accttattac cggggtaccc 19140
aactccatgc tcaacagtcc ccaggtacag cccaccctgc gtcgcaacca ggaacagctc 19200
tacagcttcc tggagcgcca ctcgccctac ttccgcagcc acagtgcgca gattaggagc 19260
gccacttctt tttgtcactt gaaaaacatg taaaaataat gtactagaga cactttcaat 19320
aaaggcaaat gcttttattt gtacactctc gggtgattat ttacccccac ccttgccgtc 19380
tgcgccgttt aaaaatcaaa ggggttctgc cgcgcatcgc tatgcgccac tggcagggac 19440
acgttgcgat actggtgttt agtgctccac ttaaactcag gcacaaccat ccgcggcagc 19500
tcggtgaagt tttcactcca caggctgcgc accatcacca acgcgtttag caggtcgggc 19560
gccgatatct tgaagtcgca gttggggcct ccgccctgcg cgcgcgagtt gcgatacaca 19620
gggttgcagc actggaacac tatcagcgcc gggtggtgca cgctggccag cacgctcttg 19680
tcggagatca gatccgcgtc caggtcctcc gcgttgctca gggcgaacgg agtcaacttt 19740
ggtagctgcc ttcccaaaaa gggcgcgtgc ccaggctttg agttgcactc gcaccgtagt 19800
ggcatcaaaa ggtgaccgtg cccggtctgg gcgttaggat acagcgcctg cataaaagcc 19860
ttgatctgct taaaagccac ctgagccttt gcgccttcag agaagaacat gccgcaagac 19920
ttgccggaaa actgattggc cggacaggcc gcgtcgtgca cgcagcacct tgcgtcggtg 19980
ttggagatct gcaccacatt tcggccccac cggttcttca cgatcttggc cttgctagac 20040
tgctccttca gcgcgcgctg cccgttttcg ctcgtcacat ccatttcaat cacgtgctcc 20100
ttatttatca taatgcttcc gtgtagacac ttaagctcgc cttcgatctc agcgcagcgg 20160
tgcagccaca acgcgcagcc cgtgggctcg tgatgcttgt aggtcacctc tgcaaacgac 20220
tgcaggtacg cctgcaggaa tcgccccatc atcgtcacaa aggtcttgtt gctggtgaag 20280
gtcagctgca acccgcggtg ctcctcgttc agccaggtct tgcatacggc cgccagagct 20340
tccacttggt caggcagtag tttgaagttc gcctttagat cgttatccac gtggtacttg 20400
tccatcagcg cgcgcgcagc ctccatgccc ttctcccacg cagacacgat cggcacactc 20460
agcgggttca tcaccgtaat ttcactttcc gcttcgctgg gctcttcctc ttcctcttgc 20520
gtccgcatac cacgcgccac tgggtcgtct tcattcagcc gccgcactgt gcgcttacct 20580
cctttgccat gcttgattag caccggtggg ttgctgaaac ccaccatttg tagcgccaca 20640
tcttctcttt cttcctcgct gtccacgatt acctctggtg atggcgggcg ctcgggcttg 20700
ggagaagggc gcttcttttt cttcttgggc gcaatggcca aatccgccgc cgaggtcgat 20760
ggccgcgggc tgggtgtgcg cggcaccagc gcgtcttgtg atgagtcttc ctcgtcctcg 20820
gactcgatac gccgcctcat ccgctttttt gggggcgccc ggggaggcgg cggcgacggg 20880
gacggggacg acacgtcctc catggttggg ggacgtcgcg ccgcaccgcg tccgcgctcg 20940
ggggtggttt cgcgctgctc ctcttcccga ctggccattt ccttctccta taggcagaaa 21000
aagatcatgg agtcagtcga gaagaaggac agcctaaccg ccccctctga gttcgccacc 21060
accgcctcca ccgatgccgc caacgcgcct accaccttcc ccgtcgaggc acccccgctt 21120
gaggaggagg aagtgattat cgagcaggac ccaggttttg taagcgaaga cgacgaggac 21180
cgctcagtac caacagagga taaaaagcaa gaccaggaca acgcagaggc aaacgaggaa 21240
caagtcgggc ggggggacga aaggcatggc gactacctag atgtgggaga cgacgtgctg 21300
ttgaagcatc tgcagcgcca gtgcgccatt atctgcgacg cgttgcaaga gcgcagcgat 21360
gtgcccctcg ccatagcgga tgtcagcctt gcctacgaac gccacctatt ctcaccgcgc 21420
gtacccccca aacgccaaga aaacggcaca tgcgagccca acccgcgcct caacttctac 21480
cccgtatttg ccgtgccaga ggtgcttgcc acctatcaca tctttttcca aaactgcaag 21540
atacccctat cctgccgtgc caaccgcagc cgagcggaca agcagctggc cttgcggcag 21600
ggcgctgtca tacctgatat cgcctcgctc aacgaagtgc caaaaatctt tgagggtctt 21660
ggacgcgacg agaagcgcgc ggcaaacgct ctgcaacagg aaaacagcga aaatgaaagt 21720
cactctggag tgttggtgga actcgagggt gacaacgcgc gcctagccgt actaaaacgc 21780
agcatcgagg tcacccactt tgcctacccg gcacttaacc taccccccaa ggtcatgagc 21840
acagtcatga gtgagctgat cgtgcgccgt gcgcagcccc tggagaggga tgcaaatttg 21900
caagaacaaa cagaggaggg cctacccgca gttggcgacg agcagctagc gcgctggctt 21960
caaacgcgcg agcctgccga cttggaggag cgacgcaaac taatgatggc cgcagtgctc 22020
gttaccgtgg agcttgagtg catgcagcgg ttctttgctg acccggagat gcagcgcaag 22080
ctagaggaaa cattgcacta cacctttcga cagggctacg tacgccaggc ctgcaagatc 22140
tccaacgtgg agctctgcaa cctggtctcc taccttggaa ttttgcacga aaaccgcctt 22200
gggcaaaacg tgcttcattc cacgctcaag ggcgaggcgc gccgcgacta cgtccgcgac 22260
tgcgtttact tatttctatg ctacacctgg cagacggcca tgggcgtttg gcagcagtgc 22320
ttggaggagt gcaacctcaa ggagctgcag aaactgctaa agcaaaactt gaaggaccta 22380
tggacggcct tcaacgagcg ctccgtggcc gcgcacctgg cggacatcat tttccccgaa 22440
cgcctgctta aaaccctgca acagggtctg ccagacttca ccagtcaaag catgttgcag 22500
aactttagga actttatcct agagcgctca ggaatcttgc ccgccacctg ctgtgcactt 22560
cctagcgact ttgtgcccat taagtaccgc gaatgccctc cgccgctttg gggccactgc 22620
taccttctgc agctagccaa ctaccttgcc taccactctg acataatgga agacgtgagc 22680
ggtgacggtc tactggagtg tcactgtcgc tgcaacctat gcaccccgca ccgctccctg 22740
gtttgcaatt cgcagctgct taacgaaagt caaattatcg gtacctttga gctgcagggt 22800
ccctcgcctg acgaaaagtc cgcggctccg gggttgaaac tcactccggg gctgtggacg 22860
tcggcttacc ttcgcaaatt tgtacctgag gactaccacg cccacgagat taggttctac 22920
gaagaccaat cccgcccgcc taatgcggag cttaccgcct gcgtcattac ccagggccac 22980
attcttggcc aattgcaagc catcaacaaa gcccgccaag agtttctgct acgaaaggga 23040
cggggggttt acttggaccc ccagtccggc gaggagctca acccaatccc cccgccgccg 23100
cagccctatc agcagcagcc gcgggccctt gcttcccagg atggcaccca aaaagaagct 23160
gcagctgccg ccgccaccca cggacgagga ggaatactgg gacagtcagg cagaggaggt 23220
tttggacgag gaggaggagg acatgatgga agactgggag agcctagacg aggaagcttc 23280
cgaggtcgaa gaggtgtcag acgaaacacc gtcaccctcg gtcgcattcc cctcgccggc 23340
gccccagaaa tcggcaaccg gttccagcat ggctacaacc tccgctcctc aggcgccgcc 23400
ggcactgccc gttcgccgac ccaaccgtag atgggacacc actggaacca gggccggtaa 23460
gtccaagcag ccgccgccgt tagcccaaga gcaacaacag cgccaaggct accgctcatg 23520
gcgcgggcac aagaacgcca tagttgcttg cttgcaagac tgtgggggca acatctcctt 23580
cgcccgccgc tttcttctct accatcacgg cgtggccttc ccccgtaaca tcctgcatta 23640
ctaccgtcat ctctacagcc catactgcac cggcggcagc ggcagcaaca gcagcggcca 23700
cacagaagca aaggcgaccg gatagcaaga ctctgacaaa gcccaagaaa tccacagcgg 23760
cggcagcagc aggaggagga gcgctgcgtc tggcgcccaa cgaacccgta tcgacccgcg 23820
agcttagaaa caggattttt cccactctgt atgctatatt tcaacagagc aggggccaag 23880
aacaagagct gaaaataaaa aacaggtctc tgcgatccct cacccgcagc tgcctgtatc 23940
acaaaagcga agatcagctt cggcgcacgc tggaagacgc ggaggctctc ttcagtaaat 24000
actgcgcgct gactcttaag gactagtttc gcgccctttc tcaaatttaa gcgcgaaaac 24060
tacgtcatct ccagcggcca cacccggcgc cagcacctgt tgtcagcgcc attatgagca 24120
aggaaattcc cacgccctac atgtggagtt accagccaca aatgggactt gcggctggag 24180
ctgcccaaga ctactcaacc cgaataaact acatgagcgc gggaccccac atgatatccc 24240
gggtcaacgg aatacgcgcc caccgaaacc gaattctcct ggaacaggcg gctattacca 24300
ccacacctcg taataacctt aatccccgta gttggcccgc tgccctggtg taccaggaaa 24360
gtcccgctcc caccactgtg gtacttccca gagacgccca ggccgaagtt cagatgacta 24420
actcaggggc gcagcttgcg ggcggctttc gtcacagggt gcggtcgccc gggcagggta 24480
taactcacct gacaatcaga gggcgaggta ttcagctcaa cgacgagtcg gtgagctcct 24540
cgcttggtct ccgtccggac gggacatttc agatcggcgg cgccggccgc tcttcattca 24600
cgcctcgtca ggcaatccta actctgcaga cctcgtcctc tgagccgcgc tctggaggca 24660
ttggaactct gcaatttatt gaggagtttg tgccatcggt ctactttaac cccttctcgg 24720
gacctcccgg ccactatccg gatcaattta ttcctaactt tgacgcggta aaggactcgg 24780
cggacggcta cgactgaatg ttaagtggag aggcagagca actgcgcctg aaacacctgg 24840
tccactgtcg ccgccacaag tgctttgccc gcgactccgg tgagttttgc tactttgaat 24900
tgcccgagga tcatatcgag ggcccggcgc acggcgtccg gcttaccgcc cagggagagc 24960
ttgcccgtag cctgattcgg gagtttaccc agcgccccct gctagttgag cgggacaggg 25020
gaccctgtgt tctcactgtg atttgcaact gtcctaaccc tggattacat caagatcttt 25080
gttgccatct ctgtgctgag tataataaat acagaaatta aaatatactg gggctcctat 25140
cgccatcctg taaacgccac cgtcttcacc cgcccaagca aaccaaggcg aaccttacct 25200
ggtactttta acatctctcc ctctgtgatt tacaacagtt tcaacccaga cggagtgagt 25260
ctacgagaga acctctccga gctcagctac tccatcagaa aaaacaccac cctccttacc 25320
tgccgggaac gtacgagtgc gtcaccggcc gctgcaccac acctaccgcc tgaccgtaaa 25380
ccagactttt tccggacaga cctcaataac tctgtttacc agaacaggag gtgagcttag 25440
aaaaccctta gggtattagg ccaaaggcgc agctactgtg gggtttatga acaattcaag 25500
caactctacg ggctattcta attcaggttt ctctagaaat ggacggaatt attacagagc 25560
agcgcctgct agaaagacgc agggcagcgg ccgagcaaca gcgcatgaat caagagctcc 25620
aagacatggt taacttgcac cagtgcaaaa ggggtatctt ttgtctggta aagcaggcca 25680
aagtcaccta cgacagtaat accaccggac accgccttag ctacaagttg ccaaccaagc 25740
gtcagaaatt ggtggtcatg gtgggagaaa agcccattac cataactcag cactcggtag 25800
aaaccgaagg ctgcattcac tcaccttgtc aaggacctga ggatctctgc acccttatta 25860
agaccctgtg cggtctcaaa gatcttattc cctttaacta ataaaaaaaa ataataaagc 25920
atcacttact taaaatcagt tagcaaattt ctgtccagtt tattcagcag cacctccttg 25980
ccctcctccc agctctggta ttgcagcttc ctcctggctg caaactttct ccacaatcta 26040
aatggaatgt cagtttcctc ctgttcctgt ccatccgcac ccactatctt catgttgttg 26100
cagatgaagc gcgcaagacc gtctgaagat accttcaacc ccgtgtatcc atatgacacg 26160
gaaaccggtc ctccaactgt gccttttctt actcctccct ttgtatcccc caatgggttt 26220
caagagagtc cccctggggt actctctttg cgcctatccg aacctctagt tacctccaat 26280
ggcatgcttg cgctcaaaat gggcaacggc ctctctctgg acgaggccgg caaccttacc 26340
tcccaaaatg taaccactgt gagcccacct ctcaaaaaaa ccaagtcaaa cataaacctg 26400
gaaatatctg cacccctcac agttacctca gaagccctaa ctgtggctgc cgccgcacct 26460
ctaatggtcg cgggcaacac actcaccatg caatcacagg ccccgctaac cgtgcacgac 26520
tccaaactta gcattgccac ccaaggaccc ctcacagtgt cagaaggaaa gctagccctg 26580
caaacatcag gccccctcac caccaccgat agcagtaccc ttactatcac tgcctcaccc 26640
cctctaacta ctgccactgg tagcttgggc attgacttga aagagcccat ttatacacaa 26700
aatggaaaac taggactaaa gtacggggct cctttgcatg taacagacga cctaaacact 26760
ttgaccgtag caactggtcc aggtgtgact attaataata cttccttgca aactaaagtt 26820
actggagcct tgggttttga ttcacaaggc aatatgcaac ttaatgtagc aggaggacta 26880
aggattgatt ctcaaaacag acgccttata cttgatgtta gttatccgtt tgatgctcaa 26940
aaccaactaa atctaagact aggacagggc cctcttttta taaactcagc ccacaacttg 27000
gatattaact acaacaaagg cctttacttg tttacagctt caaacaattc caaaaagctt 27060
gaggttaacc taagcactgc caaggggttg atgtttgacg ctacagccat agccattaat 27120
gcaggagatg ggcttgaatt tggttcacct aatgcaccaa acacaaatcc cctcaaaaca 27180
aaaattggcc atggcctaga atttgattca aacaaggcta tggttcctaa actaggaact 27240
ggccttagtt ttgacagcac aggtgccatt acagtaggaa acaaaaataa tgataagcta 27300
actttgtgga ccacaccagc tccatctcct aactgtagac taaatgcaga gaaagatgct 27360
aaactcactt tggtcttaac aaaatgtggc agtcaaatac ttgctacagt ttcagttttg 27420
gctgttaaag gcagtttggc tccaatatct ggaacagttc aaagtgctca tcttattata 27480
agatttgacg aaaatggagt gctactaaac aattccttcc tggacccaga atattggaac 27540
tttagaaatg gagatcttac tgaaggcaca gcctatacaa acgctgttgg atttatgcct 27600
aacctatcag cttatccaaa atctcacggt aaaactgcca aaagtaacat tgtcagtcaa 27660
gtttacttaa acggagacaa aactaaacct gtaacactaa ccattacact aaacggtaca 27720
caggaaacag gagacacaac tccaagtgca tactctatgt cattttcatg ggactggtct 27780
ggccacaact acattaatga aatatttgcc acatcctctt acactttttc atacattgcc 27840
caagaataaa gaatcgtttg tgttatgttt caacgtgttt atttttcaat tgcagaaaat 27900
ttcgaatcat ttttcattca gtagtatagc cccaccacca catagcttat acagatcacc 27960
gtaccttaat caaactcaca gaaccctagt attcaacctg ccacctccct cccaacacac 28020
agagtacaca gtcctttctc cccggctggc cttaaaaagc atcatatcat gggtaacaga 28080
catattctta ggtgttatat tccacacggt ttcctgtcga gccaaacgct catcagtgat 28140
attaataaac tccccgggca gctcacttaa gttcatgtcg ctgtccagct gctgagccac 28200
aggctgctgt ccaacttgcg gttgcttaac gggcggcgaa ggagaagtcc acgcctacat 28260
gggggtagag tcataatcgt gcatcaggat agggcggtgg tgctgcagca gcgcgcgaat 28320
aaactgctgc cgccgccgct ccgtcctgca ggaatacaac atggcagtgg tctcctcagc 28380
gatgattcgc accgcccgca gcataaggcg ccttgtcctc cgggcacagc agcgcaccct 28440
gatctcactt aaatcagcac agtaactgca gcacagcacc acaatattgt tcaaaatccc 28500
acagtgcaag gcgctgtatc caaagctcat ggcggggacc acagaaccca cgtggccatc 28560
ataccacaag cgcaggtaga ttaagtggcg acccctcata aacacgctgg acataaacat 28620
tacctctttt ggcatgttgt aattcaccac ctcccggtac catataaacc tctgattaaa 28680
catggcgcca tccaccacca tcctaaacca gctggccaaa acctgcccgc cggctataca 28740
ctgcagggaa ccgggactgg aacaatgaca gtggagagcc caggactcgt aaccatggat 28800
catcatgctc gtcatgatat caatgttggc acaacacagg cacacgtgca tacacttcct 28860
caggattaca agctcctccc gcgttagaac catatcccag ggaacaaccc attcctgaat 28920
cagcgtaaat cccacactgc agggaagacc tcgcacgtaa ctcacgttgt gcattgtcaa 28980
agtgttacat tcgggcagca gcggatgatc ctccagtatg gtagcgcggg tttctgtctc 29040
aaaaggaggt agacgatccc tactgtacgg agtgcgccga gacaaccgag atcgtgttgg 29100
tcgtagtgtc atgccaaatg gaacgccgga cgtagtcata tttcctgaag caaaaccagg 29160
tgcgggcgtg acaaacagat ctgcgtctcc ggtctcgccg cttagatcgc tctgtgtagt 29220
agttgtagta tatccactct ctcaaagcat ccaggcgccc cctggcttcg ggttctatgt 29280
aaactccttc atgcgccgct gccctgataa catccaccac cgcagaataa gccacaccca 29340
gccaacctac acattcgttc tgcgagtcac acacgggagg agcgggaaga gctggaagaa 29400
ccatgttttt ttttttattc caaaagatta tccaaaacct caaaatgaag atctattaag 29460
tgaacgcgct cccctccggt ggcgtggtca aactctacag ccaaagaaca gataatggca 29520
tttgtaagat gttgcacaat ggcttccaaa aggcaaacgg ccctcacgtc caagtggacg 29580
taaaggctaa acccttcagg gtgaatctcc tctataaaca ttccagcacc ttcaaccatg 29640
cccaaataat tctcatctcg ccaccttctc aatatatctc taagcaaatc ccgaatatta 29700
agtccggcca ttgtaaaaat ctgctccaga gcgccctcca ccttcagcct caagcagcga 29760
atcatgattg caaaaattca ggttcctcac agacctgtat aagattcaaa agcggaacat 29820
taacaaaaat accgcgatcc cgtaggtccc ttcgcagggc cagctgaaca taatcgtgca 29880
ggtctgcacg gaccagcgcg gccacttccc cgccaggaac catgacaaaa gaacccacac 29940
tgattatgac acgcatactc ggagctatgc taaccagcgt agccccgatg taagcttgtt 30000
gcatgggcgg cgatataaaa tgcaaggtgc tgctcaaaaa atcaggcaaa gcctcgcgca 30060
aaaaagaaag cacatcgtag tcatgctcat gcagataaag gcaggtaagc tccggaacca 30120
ccacagaaaa agacaccatt tttctctcaa acatgtctgc gggtttctgc ataaacacaa 30180
aataaaataa caaaaaaaca tttaaacatt agaagcctgt cttacaacag gaaaaacaac 30240
ccttataagc ataagacgga ctacggccat gccggcgtga ccgtaaaaaa actggtcacc 30300
gtgattaaaa agcaccaccg acagctcctc ggtcatgtcc ggagtcataa tgtaagactc 30360
ggtaaacaca tcaggttgat tcacatcggt cagtgctaaa aagcgaccga aatagcccgg 30420
gggaatacat acccgcaggc gtagagacaa cattacagcc cccataggag gtataacaaa 30480
attaatagga gagaaaaaca cataaacacc tgaaaaaccc tcctgcctag gcaaaatagc 30540
accctcccgc tccagaacaa catacagcgc ttccacagcg gcagccataa cagtcagcct 30600
taccagtaaa aaagaaaacc tattaaaaaa acaccactcg acacggcacc agctcaatca 30660
gtcacagtgt aaaaaagggc caagtgcaga gcgagtatat ataggactaa aaaatgacgt 30720
aacggttaaa gtccacaaaa aacacccaga aaaccgcacg cgaacctacg cccagaaacg 30780
aaagccaaaa aacccacaac ttcctcaaat cgtcacttcc gttttcccac gttacgtcac 30840
ttcccatttt aagaaaacta caattcccaa cacatacaag ttactccgcc ctaaaaccta 30900
cgtcacccgc cccgttccca cgccccgcgc cacgtcacaa actccacccc ctcattatca 30960
tattggcttc aatccaaaat aaggtatatt attgatgatg 31000
Claims (46)
1.一种针对于胃癌的表达系统，其特征在于，包括:
第一核酸分子，所述第一核酸分子含有细胞特异性启动子；
第二核酸分子，所述第二核酸分子与所述第一核酸分子可操作地连接，所述第二核酸分子编码转录激活因子；
第三核酸分子，所述第三核酸分子含有所述转录激活因子的第一识别序列；
第四核酸分子，所述第四核酸分子与所述第三核酸分子可操作地连接，所述第四核酸分子含有第一启动子和第一调控元件；
第五核酸分子，所述第五核酸分子与第四核酸分子可操作地连接，所述第五核酸分子编码第一调控蛋白和目的蛋白，所述目的蛋白与所述第一调控蛋白是以融合蛋白的形式表达的，并且所述目的蛋白与所述第一调控蛋白之间通过可切割的连接肽连接的，并且所述目的蛋白包括选自病毒复制包装蛋白、免疫效应因子的至少之一；
第六核酸分子，所述第六核酸分子含有所述转录激活因子的第二识别序列；
第七核酸分子，所述第七核酸分子与所述第六核酸分子可操作地连接，所述第七核酸分子含有第二启动子和第二调控元件；
第八核酸分子，所述第八核酸分子与第七核酸分子可操作地连接，并且所述第八核酸分子编码第二调控蛋白；
第九核酸分子，所述第九核酸分子与所述第五核酸分子可操作地连接，所述第九核酸分子被配置为条件性抑制所述第一调控蛋白的表达；以及
第十核酸分子，所述第十核酸分子与所述第八核酸分子可操作地连接，所述第十核酸分子被配置为条件性抑制所述第二调控蛋白的表达，
其中，
所述第一调控元件适于通过结合所述第二调控蛋白抑制所述第一启动子的功能，所述第二调控元件适于通过结合所述第一调控蛋白抑制所述第二启动子的功能；
所述第九核酸分子与所述第十核酸分子分别独立地借助RNA干扰抑制所述第一调控蛋白或所述第二调控蛋白的表达；
所述第九核酸分子含有被第一microRNA特异性识别的核酸序列，所述第十核酸分子含有被第二microRNA特异性识别的核酸序列，所述第一microRNA为正常细胞特异性microRNA，所述第二microRNA为异常细胞特异性microRNA；
所述第一核酸分子与所述第二核酸分子负载在第一表达载体上，所述第三核酸分子、所述第四核酸分子、所述第五核酸分子以及所述第九核酸分子负载在第二表达载体上，所述第六核酸分子、所述第七核酸分子、所述第八核酸分子以及所述第十核酸分子负载在第三表达载体上；
所述细胞特异性启动子为CEA205，所述第一microRNA为mir-135a-5p，所述第二microRNA为mir-21，所述第一识别序列与所述第二识别序列为5×UAS；或
所述细胞特异性启动子为CEA368，所述第一microRNA为mir-135a-5p，所述第二microRNA为mir-21，所述第一识别序列与所述第二识别序列为5×UAS；或
所述细胞特异性启动子为CXCR4，所述第一microRNA为mir-135a-5p，所述第二microRNA为mir-21，所述第一识别序列与所述第二识别序列为5×UAS；或
所述细胞特异性启动子为Survivin1，所述第一microRNA为mir-135a-5p，所述第二microRNA为mir-21，所述第一识别序列与所述第二识别序列为5×UAS；或
所述细胞特异性启动子为CEA368，所述第一microRNA为mir-143-3p，所述第二microRNA为mir-21，所述第一识别序列与所述第二识别序列为5×UAS；或
所述细胞特异性启动子为CEA205，所述第一microRNA为mir-135a-5p，所述第二microRNA为mir-21，所述第一识别序列与所述第二识别序列为4×UAS；或
所述细胞特异性启动子为CEA368，所述第一microRNA为mir-135a-5p，所述第二microRNA为mir-21，所述第一识别序列与所述第二识别序列为4×UAS；或
所述细胞特异性启动子为CXCR4，所述第一microRNA为mir-135a-5p，所述第二microRNA为mir-21，所述第一识别序列与所述第二识别序列为4×UAS；或
所述细胞特异性启动子为Survivin1，所述第一microRNA为mir-135a-5p，所述第二microRNA为mir-21，所述第一识别序列与所述第二识别序列为4×UAS；或
所述细胞特异性启动子为CEA368，所述第一microRNA为mir-143-3p，所述第二microRNA为mir-21，所述第一识别序列与所述第二识别序列为4×UAS。
2.根据权利要求1所述的表达系统，其特征在于，所述转录激活因子为选自Gal4VP16、Gal4vp64、dCas9-VPR、dCas9-VP64、dCas9-VP16、dCas9-VTR以及rtTA至少之一。
3.根据权利要求1所述的表达系统，其特征在于，所述第一启动子与所述第二启动子分别独立地选自miniCMV、TATA box。
4.根据权利要求1所述的表达系统，其特征在于，所述第一调控蛋白和第二调控蛋白分别独立地选自Lacl、tetR、zinc finger、TALE、KRAB、tetR-KRAB、TALE-KRAB、dCas9-KRAB、miniCas9-KRAB、分割dCas9-KRAB至少之一。
5.根据权利要求1所述的表达系统，其特征在于，所述第一调控元件和所述第二调控元件分别独立地选自tetO、LacO、锌指蛋白靶标序列、TALE蛋白靶标序列、dCas9的靶标序列、miniCas9的靶标序列至少之一。
6.根据权利要求1所述的表达系统，其特征在于所述第一调控蛋白是LacI，所述第二调控元件包括多个重复的LacO序列，所述多个重复的LacO序列的至少之一设置在所述第二启动子的下游。
7.根据权利要求1所述的表达系统，其特征在于所述第二调控蛋白是tetR-KRAB所述第一调控元件包括多个重复的tetO序列，所述多个重复的tetO序列的至少之一设置在所述第一启动子的下游。
8.根据权利要求1所述的表达系统，其特征在于所述病毒复制包装相关蛋白包括选自腺病毒E1基因、E1A基因、E1B基因、E2基因和E4基因至少之一。
9.根据权利要求1所述的表达系统，其特征在于，所述免疫效应因子包括选自拮抗PD-1基因的抑制序列、拮抗PD-L1基因的抑制序列、拮抗CTLA4基因的抑制序列、拮抗Tim-3基因的抑制序列、GM-CSF、IL-2、IL-12、IL-15至少之一。
10.根据权利要求1所述的表达系统，其特征在于，所述第一表达载体、第二表达载体和第三表达载体分别独立地选自下列的至少之一：
质粒、病毒、纳米材料、脂质体、分子耦联载体、裸露DNA、染色体载体、多聚物。
11.根据权利要求10所述的表达系统，其特征在于，所述病毒包括选自腺病毒、牛痘病毒、疱疹病毒、逆转录病毒的至少之一。
12.根据权利要求1所述的表达系统，其特征在于，所述第一表达载体、第二表达载体和第三表达载体负载在同一个载体上。
13.根据权利要求12所述的表达系统，其特征在于，所述同一个载体为腺病毒载体。
14.根据权利要求13所述的表达系统，其特征在于，所述腺病毒载体如SEQ ID NO：1、3、5、7、9、11、13、15、17或19的核苷酸序列所示。
15.根据权利要求14所述的表达系统，其特征在于，所述腺病毒是通过如下方式获得的：
所述的腺病毒载体去除了与腺病毒复制包装相关的E1基因和部分E3基因，E1A基因通过逐级Golden Gate的方法构建到基因线路中，最终通过Gateway或Gibson的方式将基因线路插入到腺病毒载体中。
16.根据权利要求14所述的表达系统，其特征在于，所述腺病毒载体为去除E1基因与部分E3基因的B、C亚型腺病毒。
17.根据权利要求14所述的表达系统，其特征在于，所述腺病毒载体为去除E1基因与部分E3基因的5、11、12、34或35型腺病毒。
18.根据权利要求17所述的表达系统，其特征在于，SEQ ID NO:1、3、5、7、9、11、13、15、17、19任一所示的核苷酸序列在所述5型腺病毒载体的插入位点为E1基因区域、E3基因区域或E4基因区域。
19.一种针对于胰腺癌的表达系统，其特征在于，包括:
第一核酸分子，所述第一核酸分子含有细胞特异性启动子；
第二核酸分子，所述第二核酸分子与所述第一核酸分子可操作地连接，所述第二核酸分子编码转录激活因子；
第三核酸分子，所述第三核酸分子含有所述转录激活因子的第一识别序列；
第四核酸分子，所述第四核酸分子与所述第三核酸分子可操作地连接，所述第四核酸分子含有第一启动子和第一调控元件；
第五核酸分子，所述第五核酸分子与第四核酸分子可操作地连接，所述第五核酸分子编码第一调控蛋白和目的蛋白，所述目的蛋白与所述第一调控蛋白是以融合蛋白的形式表达的，并且所述目的蛋白与所述第一调控蛋白之间通过可切割的连接肽连接的，并且所述目的蛋白包括选自病毒复制包装蛋白、免疫效应因子的至少之一；
第六核酸分子，所述第六核酸分子含有所述转录激活因子的第二识别序列；
第七核酸分子，所述第七核酸分子与所述第六核酸分子可操作地连接，所述第七核酸分子含有第二启动子和第二调控元件；
第八核酸分子，所述第八核酸分子与第七核酸分子可操作地连接，并且所述第八核酸分子编码第二调控蛋白；
第九核酸分子，所述第九核酸分子与所述第五核酸分子可操作地连接，所述第九核酸分子被配置为条件性抑制所述第一调控蛋白的表达；以及
第十核酸分子，所述第十核酸分子与所述第八核酸分子可操作地连接，所述第十核酸分子被配置为条件性抑制所述第二调控蛋白的表达，
其中，
所述第一调控元件适于通过结合所述第二调控蛋白抑制所述第一启动子的功能，所述第二调控元件适于通过结合所述第一调控蛋白抑制所述第二启动子的功能；
所述第九核酸分子与所述第十核酸分子分别独立地借助RNA干扰抑制所述第一调控蛋白或所述第二调控蛋白的表达；
所述第九核酸分子含有被第一microRNA特异性识别的核酸序列，所述第十核酸分子含有被第二microRNA特异性识别的核酸序列，所述第一microRNA为正常细胞特异性microRNA，所述第二microRNA为异常细胞特异性microRNA；
所述第一核酸分子与所述第二核酸分子负载在第一表达载体上，所述第三核酸分子、所述第四核酸分子、所述第五核酸分子以及所述第九核酸分子负载在第二表达载体上，所述第六核酸分子、所述第七核酸分子、所述第八核酸分子以及所述第十核酸分子负载在第三表达载体上；
所述细胞特异性启动子为hMuc1，所述第一microRNA为mir-199a-3p，所述第二microRNA为mir-21，所述第一识别序列与所述第二识别序列为4×UAS。
20.根据权利要求19所述的表达系统，其特征在于，所述转录激活因子为选自Gal4VP16、Gal4vp64、dCas9-VPR、dCas9-VP64、dCas9-VP16、dCas9-VTR以及rtTA至少之一。
21.根据权利要求19所述的表达系统，其特征在于，所述第一启动子与所述第二启动子分别独立地选自miniCMV、TATA box。
22.根据权利要求19所述的表达系统，其特征在于，所述第一调控蛋白和第二调控蛋白分别独立地选自Lacl、tetR、zinc finger、TALE、KRAB、tetR-KRAB、TALE-KRAB、dCas9-KRAB、miniCas9-KRAB、分割dCas9-KRAB至少之一。
23.根据权利要求19所述的表达系统，其特征在于，所述第一调控元件和所述第二调控元件分别独立地选自tetO、LacO、锌指蛋白靶标序列、TALE蛋白靶标序列、dCas9的靶标序列、miniCas9的靶标序列至少之一。
24.根据权利要求19所述的表达系统，其特征在于，所述第一调控蛋白是LacI，所述第二调控元件包括多个重复的LacO序列，所述多个重复的LacO序列的至少之一设置在所述第二启动子的下游。
25.根据权利要求19所述的表达系统，其特征在于，所述第二调控蛋白是tetR-KRAB所述第一调控元件包括多个重复的tetO序列，所述多个重复的tetO序列的至少之一设置在所述第一启动子的下游。
26.根据权利要求19所述的表达系统，其特征在于，所述病毒复制包装相关蛋白包括选自腺病毒E1基因、E1A基因、E1B基因、E2基因和E4基因至少之一。
27.根据权利要求19所述的表达系统，其特征在于，所述免疫效应因子包括选自拮抗PD-1基因的抑制序列、拮抗PD-L1基因的抑制序列、拮抗CTLA4基因的抑制序列、拮抗Tim-3基因的抑制序列、GM-CSF、IL-2、IL-12、IL-15至少之一。
28.根据权利要求19所述的表达系统，其特征在于，所述第一表达载体、第二表达载体和第三表达载体分别独立地选自下列的至少之一：
质粒、病毒、纳米材料、脂质体、分子耦联载体、裸露DNA、染色体载体、多聚物。
29.根据权利要求28所述的表达系统，其特征在于，所述病毒包括选自腺病毒、牛痘病毒、疱疹病毒、逆转录病毒的至少之一。
30.根据权利要求19所述的表达系统，其特征在于，所述第一表达载体、第二表达载体和第三表达载体负载在同一个载体上。
31.根据权利要求30所述的表达系统，其特征在于，所述同一个载体为腺病毒载体。
32.根据权利要求31所述的表达系统，其特征在于，所述腺病毒载体携带具有SEQ IDNO：21-22任一所示核苷酸序列的核酸。
33.根据权利要求29所述的表达系统，其特征在于，所述腺病毒是通过如下方式获得的：
所述的腺病毒载体去除了与腺病毒复制包装相关的E1基因和部分E3基因，E1A基因通过逐级Golden Gate的方法构建到基因线路中，最终通过Gateway或Gibson的方式将基因线路插入到腺病毒载体中。
34.根据权利要求29所述的表达系统，其特征在于，所述腺病毒载体为去除E1基因与部分E3基因的B、C亚型腺病毒。
35.根据权利要求29所述的表达系统，其特征在于，所述腺病毒载体为去除E1基因与部分E3基因的5、11、12、34或35型腺病毒。
36.根据权利要求32所述的表达系统，其特征在于，SEQ ID NO:21-22任一所示的核苷酸序列在所述5型腺病毒载体的插入位点为E1基因区域、E3基因区域或E4基因区域。
37.一种重组细胞，其特征在于，含有权利要求1～36任一项所述的表达系统。
38.根据权利要求37所述的重组细胞，其特征在于，所述表达系统的至少一部分整合于所述重组细胞的基因组中。
39.权利要求1～36任一项所述的表达系统、权利要求37～38任一项所述的重组细胞在制备药物中的用途，所述药物用于治疗胃癌。
40.根据权利要求39所述的用途，其特征在于，所述药物用于治疗胰腺癌。
41.一种药物组合物，其特征在于，包含权利要求37～38任一项所述的重组细胞。
42.根据权利要求41所述的药物组合物，其特征在于，进一步包括药学上可接受的辅料。
43.根据权利要求41所述的药物组合物，其特征在于，进一步包括其他治疗胃癌的药物。
44.根据权利要求43所述的药物组合物，其特征在于，所述其他治疗胃癌的药物包括选自Pembrolizumab、Ogivri、Everolimus、Lanreotide、Ramucirumab、Apatinib、Trastuzumab的至少之一。
45.根据权利要求41所述的药物组合物，其特征在于，进一步包括其他治疗胰腺癌的药物。
46.根据权利要求45所述的药物组合物，其特征在于，所述其他治疗胰腺癌的药物包括选自Lanreotide、Abraxane、Olaparib、Afinitor、Erlotinib、Everolimus、5-FU、Gemzar、Sunitinib、Onivyde、Gemzar的至少之一。
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| CN109576231A (zh) * | 2017-09-28 | 2019-04-05 | 杭州康万达医药科技有限公司 | 分离的重组溶瘤腺病毒、药物组合物及其在治疗肿瘤和/或癌症的药物中的用途 |
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