载体基本信息 载体名称: | pCDH-EF1-MCS-T2A-Puro | 质粒类型: | 慢病毒表达载体；cDNA克隆载体 | 克隆方法: | 多克隆位点，限制性内切酶 | 启动子: | EF1α | 载体大小: | 7095 bp | 5' 测序引物及序列: | 5’-CTCCACGCTTTGCCTGACCCTGCTT-3’ | 载体抗性: | 氨苄青霉素（Ampicillin） | 筛选标记: | 嘌呤霉素 | 宿主细胞（系）: | 大部分细胞类型（分裂与非分裂细胞）；初级分化细胞 | 备注: | pCDH-EF1-MCS-T2A-Puro慢病毒表达载体是基于HIV的慢病毒载体；
用于cDNA表达和克隆；高效转染细胞，建立稳定细胞系；
单一启动子，由EF1a 启动子驱动目的基因和与抗性基因转录为双顺反子；
二者之间含一段T2A肽，能够在翻译后水平上自我剪切，保证目的基因与抗性基因翻译水平相同。 | 稳定性: | 稳表达 | 组成型/诱导型: | 组成型 | 病毒/非病毒: | 慢病毒（HIV) |
全序列 1 ACGCGTGTAG TCTTATGCAA TACTCTTGTA GTCTTGCAAC ATGGTAACGA TGAGTTAGCA
61 ACATGCCTTA CAAGGAGAGA AAAAGCACCG TGCATGCCGA TTGGTGGAAG TAAGGTGGTA
121 CGATCGTGCC TTATTAGGAA GGCAACAGAC GGGTCTGACA TGGATTGGAC GAACCACTGA
181 ATTGCCGCAT TGCAGAGATA TTGTATTTAA GTGCCTAGCT CGATACAATA AACGGGTCTC
241 TCTGGTTAGA CCAGATCTGA GCCTGGGAGC TCTCTGGCTA ACTAGGGAAC CCACTGCTTA
301 AGCCTCAATA AAGCTTGCCT TGAGTGCTTC AAGTAGTGTG TGCCCGTCTG TTGTGTGACT
361 CTGGTAACTA GAGATCCCTC AGACCCTTTT AGTCAGTGTG GAAAATCTCT AGCAGTGGCG
421 CCCGAACAGG GACCTGAAAG CGAAAGGGAA ACCAGAGCTC TCTCGACGCA GGACTCGGCT
481 TGCTGAAGCG CGCACGGCAA GAGGCGAGGG GCGGCGACTG GTGAGTACGC CAAAAATTTT
541 GACTAGCGGA GGCTAGAAGG AGAGAGATGG GTGCGAGAGC GTCAGTATTA AGCGGGGGAG
601 AATTAGATCG CGATGGGAAA AAATTCGGTT AAGGCCAGGG GGAAAGAAAA AATATAAATT
661 AAAACATATA GTATGGGCAA GCAGGGAGCT AGAACGATTC GCAGTTAATC CTGGCCTGTT
721 AGAAACATCA GAAGGCTGTA GACAAATACT GGGACAGCTA CAACCATCCC TTCAGACAGG
781 ATCAGAAGAA CTTAGATCAT TATATAATAC AGTAGCAACC CTCTATTGTG TGCATCAAAG
841 GATAGAGATA AAAGACACCA AGGAAGCTTT AGACAAGATA GAGGAAGAGC AAAACAAAAG
901 TAAGACCACC GCACAGCAAG CGGCCACTGA TCTTCAGACC TGGAGGAGGA GATATGAGGG
961 ACAATTGGAG AAGTGAATTA TATAAATATA AAGTAGTAAA AATTGAACCA TTAGGAGTAG
1021 CACCCACCAA GGCAAAGAGA AGAGTGGTGC AGAGAGAAAA AAGAGCAGTG GGAATAGGAG
1081 CTTTGTTCCT TGGGTTCTTG GGAGCAGCAG GAAGCACTAT GGGCGCAGCC TCAATGACGC
1141 TGACGGTACA GGCCAGACAA TTATTGTCTG GTATAGTGCA GCAGCAGAAC AATTTGCTGA
1201 GGGCTATTGA GGCGCAACAG CATCTGTTGC AACTCACAGT CTGGGGCATC AAGCAGCTCC
1261 AGGCAAGAAT CCTGGCTGTG GAAAGATACC TAAAGGATCA ACAGCTCCTG GGGATTTGGG
1321 GTTGCTCTGG AAAACTCATT TGCACCACTG CTGTGCCTTG GAATGCTAGT TGGAGTAATA
1381 AATCTCTGGA ACAGATTGGA ATCACACGAC CTGGATGGAG TGGGACAGAG AAATTAACAA
1441 TTACACAAGC TTAATACACT CCTTAATTGA AGAATCGCAA AACCAGCAAG AAAAGAATGA
1501 ACAAGAATTA TTGGAATTAG ATAAATGGGC AAGTTTGTGG AATTGGTTTA ACATAACAAA
1561 TTGGCTGTGG TATATAAAAT TATTCATAAT GATAGTAGGA GGCTTGGTAG GTTTAAGAAT
1621 AGTTTTTGCT GTACTTTCTA TAGTGAATAG AGTTAGGCAG GGATATTCAC CATTATCGTT
1681 TCAGACCCAC CTCCCAACCC CGAGGGGACC CGACAGGCCC GAAGGAATAG AAGAAGAAGG
1741 TGGAGAGAGA GACAGAGACA GATCCATTCG ATTAGTGAAC GGATCTCGAC GGTTAACTTT
1801 TAAAAGAAAA GGGGGGATTG GGGGGTACAG TGCAGGGGAA AGAATAGTAG ACATAATAGC
1861 AACAGACATA CAAACTAAAG AATTACAAAA ACAAATTACA AAAATTCAAA ATTTTATCGA
1921 TACTAGTAAG GATCTGCGAT CGCTCCGGTG CCCGTCAGTG GGCAGAGCGC ACATCGCCCA
1981 CAGTCCCCGA GAAGTTGGGG GGAGGGGTCG GCAATTGAAC GGGTGCCTAG AGAAGGTGGC
2041 GCGGGGTAAA CTGGGAAAGT GATGTCGTGT ACTGGCTCCG CCTTTTTCCC GAGGGTGGGG
2101 GAGAACCGTA TATAAGTGCA GTAGTCGCCG TGAACGTTCT TTTTCGCAAC GGGTTTGCCG
2161 CCAGAACACA GCTGAAGCTT CGAGGGGCTC GCATCTCTCC TTCACGCGCC CGCCGCCCTA
2221 CCTGAGGCCG CCATCCACGC CGGTTGAGTC GCGTTCTGCC GCCTCCCGCC TGTGGTGCCT
2281 CCTGAACTGC GTCCGCCGTC TAGGTAAGTT TAAAGCTCAG GTCGAGACCG GGCCTTTGTC
2341 CGGCGCTCCC TTGGAGCCTA CCTAGACTCA GCCGGCTCTC CACGCTTTGC CTGACCCTGC
2401 TTGCTCAACT CTACGTCTTT GTTTCGTTTT CTGTTCTGCG CCGTTACAGA TCCAAGCTGT
2461 GACCGGCGCC TACTCTAGAG CTAGCGAATT CGAATTTAAA TCGGATCCGC GGCCGCTGAG
2521 GGCAGAGGAA GTCTTCTAAC ATGCGGTGAC GTGGAGGAGA ATCCCGGCCC TTCCGGGATG
2581 ACCGAGTACA AGCCCACGGT GCGCCTCGCC ACCCGCGACG ACGTCCCCAG GGCCGTACGC
2641 ACCCTCGCCG CCGCGTTCGC CGACTACCCC GCCACGCGCC ACACCGTCGA TCCGGACCGC
2701 CACATCGAGC GGGTCACCGA GCTGCAAGAA CTCTTCCTCA CGCGCGTCGG GCTCGACATC
2761 GGCAAGGTGT GGGTCGCGGA CGACGGCGCC GCGGTGGCGG TCTGGACCAC GCCGGAGAGC
2821 GTCGAAGCGG GGGCGGTGTT CGCCGAGATC GGCCCGCGCA TGGCCGAGTT GAGCGGTTCC
2881 CGGCTGGCCG CGCAGCAACA GATGGAAGGC CTCCTGGCGC CGCACCGGCC CAAGGAGCCC
2941 GCGTGGTTCC TGGCCACCGT CGGCGTCTCG CCCGACCACC AGGGCAAGGG TCTGGGCAGC
3001 GCCGTCGTGC TCCCCGGAGT GGAGGCGGCC GAGCGCGCCG GGGTGCCCGC CTTCCTGGAG
3061 ACCTCCGCGC CCCGCAACCT CCCCTTCTAC GAGCGGCTCG GCTTCACCGT CACCGCCGAC
3121 GTCGAGGTGC CCGAAGGACC GCGCACCTGG TGCATGACCC GCAAGCCCGG TGCCTGAATC
3181 TAGGTCGACA ATCAACCTCT GGATTACAAA ATTTGTGAAA GATTGACTGG TATTCTTAAC
3241 TATGTTGCTC CTTTTACGCT ATGTGGATAC GCTGCTTTAA TGCCTTTGTA TCATGCTATT
3301 GCTTCCCGTA TGGCTTTCAT TTTCTCCTCC TTGTATAAAT CCTGGTTGCT GTCTCTTTAT
3361 GAGGAGTTGT GGCCCGTTGT CAGGCAACGT GGCGTGGTGT GCACTGTGTT TGCTGACGCA
3421 ACCCCCACTG GTTGGGGCAT TGCCACCACC TGTCAGCTCC TTTCCGGGAC TTTCGCTTTC
3481 CCCCTCCCTA TTGCCACGGC GGAACTCATC GCCGCCTGCC TTGCCCGCTG CTGGACAGGG
3541 GCTCGGCTGT TGGGCACTGA CAATTCCGTG GTGTTGTCGG GGAAATCATC GTCCTTTCCT
3601 TGGCTGCTCG CCTGTGTTGC CACCTGGATT CTGCGCGGGA CGTCCTTCTG CTACGTCCCT
3661 TCGGCCCTCA ATCCAGCGGA CCTTCCTTCC CGCGGCCTGC TGCCGGCTCT GCGGCCTCTT
3721 CCGCGTCTTC GCCTTCGCCC TCAGACGAGT CGGATCTCCC TTTGGGCCGC CTCCCCGCCT
3781 GGTACCTTTA AGACCAATGA CTTACAAGGC AGCTGTAGAT CTTAGCCACT TTTTAAAAGA
3841 AAAGGGGGGA CTGGAAGGGC TAATTCACTC CCAACGAAAA TAAGATCTGC TTTTTGCTTG
3901 TACTGGGTCT CTCTGGTTAG ACCAGATCTG AGCCTGGGAG CTCTCTGGCT AACTAGGGAA
3961 CCCACTGCTT AAGCCTCAAT AAAGCTTGCC TTGAGTGCTT CAAGTAGTGT GTGCCCGTCT
4021 GTTGTGTGAC TCTGGTAACT AGAGATCCCT CAGACCCTTT TAGTCAGTGT GGAAAATCTC
4081 TAGCAGTAGT AGTTCATGTC ATCTTATTAT TCAGTATTTA TAACTTGCAA AGAAATGAAT
4141 ATCAGAGAGT GAGAGGAACT TGTTTATTGC AGCTTATAAT GGTTACAAAT AAAGCAATAG
4201 CATCACAAAT TTCACAAATA AAGCATTTTT TTCACTGCAT TCTAGTTGTG GTTTGTCCAA
4261 ACTCATCAAT GTATCTTATC ATGTCTGGCT CTAGCTATCC CGCCCCTAAC TCCGCCCAGT
4321 TCCGCCCATT CTCCGCCCCA TGGCTGACTA ATTTTTTTTA TTTATGCAGA GGCCGAGGCC
4381 GCCTCGGCCT CTGAGCTATT CCAGAAGTAG TGAGGAGGCT TTTTTGGAGG CCTAGACTTT
4441 TGCAGAGACG GCCCAAATTC GTAATCATGG TCATAGCTGT TTCCTGTGTG AAATTGTTAT
4501 CCGCTCACAA TTCCACACAA CATACGAGCC GGAAGCATAA AGTGTAAAGC CTGGGGTGCC
4561 TAATGAGTGA GCTAACTCAC ATTAATTGCG TTGCGCTCAC TGCCCGCTTT CCAGTCGGGA
4621 AACCTGTCGT GCCAGCTGCA TTAATGAATC GGCCAACGCG CGGGGAGAGG CGGTTTGCGT
4681 ATTGGGCGCT CTTCCGCTTC CTCGCTCACT GACTCGCTGC GCTCGGTCGT TCGGCTGCGG
4741 CGAGCGGTAT CAGCTCACTC AAAGGCGGTA ATACGGTTAT CCACAGAATC AGGGGATAAC
4801 GCAGGAAAGA ACATGTGAGC AAAAGGCCAG CAAAAGGCCA GGAACCGTAA AAAGGCCGCG
4861 TTGCTGGCGT TTTTCCATAG GCTCCGCCCC CCTGACGAGC ATCACAAAAA TCGACGCTCA
4921 AGTCAGAGGT GGCGAAACCC GACAGGACTA TAAAGATACC AGGCGTTTCC CCCTGGAAGC
4981 TCCCTCGTGC GCTCTCCTGT TCCGACCCTG CCGCTTACCG GATACCTGTC CGCCTTTCTC
5041 CCTTCGGGAA GCGTGGCGCT TTCTCATAGC TCACGCTGTA GGTATCTCAG TTCGGTGTAG
5101 GTCGTTCGCT CCAAGCTGGG CTGTGTGCAC GAACCCCCCG TTCAGCCCGA CCGCTGCGCC
5161 TTATCCGGTA ACTATCGTCT TGAGTCCAAC CCGGTAAGAC ACGACTTATC GCCACTGGCA
5221 GCAGCCACTG GTAACAGGAT TAGCAGAGCG AGGTATGTAG GCGGTGCTAC AGAGTTCTTG
5281 AAGTGGTGGC CTAACTACGG CTACACTAGA AGGACAGTAT TTGGTATCTG CGCTCTGCTG
5341 AAGCCAGTTA CCTTCGGAAA AAGAGTTGGT AGCTCTTGAT CCGGCAAACA AACCACCGCT
5401 GGTAGCGGTG GTTTTTTTGT TTGCAAGCAG CAGATTACGC GCAGAAAAAA AGGATCTCAA
5461 GAAGATCCTT TGATCTTTTC TACGGGGTCT GACGCTCAGT GGAACGAAAA CTCACGTTAA
5521 GGGATTTTGG TCATGAGATT ATCAAAAAGG ATCTTCACCT AGATCCTTTT AAATTAAAAA
5581 TGAAGTTTTA AATCAATCTA AAGTATATAT GAGTAAACTT GGTCTGACAG TTACCAATGC
5641 TTAATCAGTG AGGCACCTAT CTCAGCGATC TGTCTATTTC GTTCATCCAT AGTTGCCTGA
5701 CTCCCCGTCG TGTAGATAAC TACGATACGG GAGGGCTTAC CATCTGGCCC CAGTGCTGCA
5761 ATGATACCGC GAGACCCACG CTCACCGGCT CCAGATTTAT CAGCAATAAA CCAGCCAGCC
5821 GGAAGGGCCG AGCGCAGAAG TGGTCCTGCA ACTTTATCCG CCTCCATCCA GTCTATTAAT
5881 TGTTGCCGGG AAGCTAGAGT AAGTAGTTCG CCAGTTAATA GTTTGCGCAA CGTTGTTGCC
5941 ATTGCTACAG GCATCGTGGT GTCACGCTCG TCGTTTGGTA TGGCTTCATT CAGCTCCGGT
6001 TCCCAACGAT CAAGGCGAGT TACATGATCC CCCATGTTGT GCAAAAAAGC GGTTAGCTCC
6061 TTCGGTCCTC CGATCGTTGT CAGAAGTAAG TTGGCCGCAG TGTTATCACT CATGGTTATG
6121 GCAGCACTGC ATAATTCTCT TACTGTCATG CCATCCGTAA GATGCTTTTC TGTGACTGGT
6181 GAGTACTCAA CCAAGTCATT CTGAGAATAG TGTATGCGGC GACCGAGTTG CTCTTGCCCG
6241 GCGTCAATAC GGGATAATAC CGCGCCACAT AGCAGAACTT TAAAAGTGCT CATCATTGGA
6301 AAACGTTCTT CGGGGCGAAA ACTCTCAAGG ATCTTACCGC TGTTGAGATC CAGTTCGATG
6361 TAACCCACTC GTGCACCCAA CTGATCTTCA GCATCTTTTA CTTTCACCAG CGTTTCTGGG
6421 TGAGCAAAAA CAGGAAGGCA AAATGCCGCA AAAAAGGGAA TAAGGGCGAC ACGGAAATGT
6481 TGAATACTCA TACTCTTCCT TTTTCAATAT TATTGAAGCA TTTATCAGGG TTATTGTCTC
6541 ATGAGCGGAT ACATATTTGA ATGTATTTAG AAAAATAAAC AAATAGGGGT TCCGCGCACA
6601 TTTCCCCGAA AAGTGCCACC TGACGTCTAA GAAACCATTA TTATCATGAC ATTAACCTAT
6661 AAAAATAGGC GTATCACGAG GCCCTTTCGT CTCGCGCGTT TCGGTGATGA CGGTGAAAAC
6721 CTCTGACACA TGCAGCTCCC GGAGACGGTC ACAGCTTGTC TGTAAGCGGA TGCCGGGAGC
6781 AGACAAGCCC GTCAGGGCGC GTCAGCGGGT GTTGGCGGGT GTCGGGGCTG GCTTAACTAT
6841 GCGGCATCAG AGCAGATTGT ACTGAGAGTG CACCATATGC GGTGTGAAAT ACCGCACAGA
6901 TGCGTAAGGA GAAAATACCG CATCAGGCGC CATTCGCCAT TCAGGCTGCG CAACTGTTGG
6961 GAAGGGCGAT CGGTGCGGGC CTCTTCGCTA TTACGCCAGC TGGCGAAAGG GGGATGTGCT
7021 GCAAGGCGAT TAAGTTGGGT AACGCCAGGG TTTTCCCAGT CACGACGTTG TAAAACGACG
7081 GCCAGTGCCA AGCTG
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