出品公司: | Clontech |
---|---|
载体名称: | pVSVG, pVSV-G, pVSV G |
质粒类型: | 哺乳动物细胞逆病毒表达载体 |
高拷贝/低拷贝: | 低拷贝 |
启动子: | CMV |
克隆方法: | 多克隆位点,限制性内切酶 |
载体大小: | 6507 bp |
5' 测序引物及序列: | CMV-F: CGCAAATGGGCGGTAGGCGTG |
3' 测序引物及序列: | -- |
载体标签: | -- |
载体抗性: | 氨苄 |
备注: | 载体使用Stbl3感受态细胞扩增质粒,可以减少载体基因重组几率。 |
产品目录号: | 631530 |
稳定性: | / |
组成型: | -- |
病毒/非病毒: | 慢病毒 |
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pVSVG载体基本信息
pVSVG载体质粒图谱和多克隆位点信息
pVSV-G载体特征
• CMV promoter: 1–768
• Rabbit β-globin IVS: 768–1432
• VSV-G envelope gene:
Start codon: 1450–1452; stop codon: 2983–2985
• β-globin poly A: 3288–3293
• Col E1 origin of replication
Site of replication initiation: 4087
• Ampicillin resistance gene (β-lactamase):
Start codon: 5712–5710; stop codon: 4853–4851
pVSVG载体简介
Description
pVSV-G expresses the G glycoprotein of the vesicular stomatitis virus (VSV-G) under the control of the CMV immediate-early promoter (1). VSV-G is used in pseudotyping of Moloney Murine Leukenia Virus (MMLV)-based retroviral vectors by mediating viral entry. VSV-G interacts with phospholipid components of the target cell membrane and fosters the fusion of viral and cellular membranes (2). VSV-G does not require a cell surface receptor and can serve as a surrogate viral envelope protein. pVSV-G Vector includes IVS, a synthetic intron known to enhance the stability of the mRNA (3), the Col E1 origin of replication and bacterial ampicillin rsistance (Ampr) gene for propagation and antibiotic selection in bacteria.
Use
As part of the Retro-X™ Universal Retroviral Expression System (Cat. No. 631530), pVSV-G is co-transfected with a retroviral expression vector into the GP2-293 Packaging Cell Line (4) to produce infectious, replication-incompetent retrovirus. The genes encoding the viral gag and pol proteins are stably integrated into GP2-293. Because the VSV-G envelope protein causes toxicity by fusing cellular membranes, it must be expressed transiently from pVSV-G during packaging (5). Although the resulting virus can infect target cell lines and transmit a gene-of-interest, it cannot replicate because target cell lines lack the viral structural and polymerase/integrase genes. The separate introduction and integration of the viral genes into the packaging cell line and the use of minimal viral sequences in the vector minimize the chance of producing replication-competent virus due to recombination events.
pVSVG载体序列
LOCUS pVSV-G 6507 bp DNA circular 23-NOV-2009 COMMENT This file is created by Vector NTI http://www.biovector.net/ COMMENT VNTAUTHORNAME|biovector.net| COMMENT Cat. No. 631457,631530,631460 FEATURES Location/Qualifiers promoter 1..768 /label=CMV\IE\Promoter intron 768..1432 /label=Rabbit\beta-globin\IVS polyA_signal 3288..3293 /label=Beta-globin\polyA CDS complement(4851..5711) /label=Amp(R) rep_origin 4087..4096 /label=pUC\origin CDS 1450..2985 /label=VSV-G BASE COUNT 1712 a 1513 c 1481 g 1801 t ORIGIN 1 gcggccgctc tagagagctt ggcccattgc atacgttgta tccatatcat aatatgtaca 61 tttatattgg ctcatgtcca acattaccgc catgttgaca ttgattattg actagttatt 121 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat 181 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa 241 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg 301 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc 361 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct 421 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga 481 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa 541 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc 601 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg 661 aggtctatat aagcagagct cgtttagtga accgtcagat cgcctggaga cgccatccac 721 gctgttttga cctccataga agacaccggg accgatccag cctccggtcg accgatcctg 781 agaacttcag ggtgagtttg gggacccttg attgttcttt ctttttcgct attgtaaaat 841 tcatgttata tggagggggc aaagttttca gggtgttgtt tagaatggga agatgtccct 901 tgtatcacca tggaccctca tgataatttt gtttctttca ctttctactc tgttgacaac 961 cattgtctcc tcttattttc ttttcatttt ctgtaacttt ttcgttaaac tttagcttgc 1021 atttgtaacg aatttttaaa ttcacttttg tttatttgtc agattgtaag tactttctct 1081 aatcactttt ttttcaaggc aatcagggta tattatattg tacttcagca cagttttaga 1141 gaacaattgt tataattaaa tgataaggta gaatatttct gcatataaat tctggctggc 1201 gtggaaatat tcttattggt agaaacaact acaccctggt catcatcctg cctttctctt 1261 tatggttaca atgatataca ctgtttgaga tgaggataaa atactctgag tccaaaccgg 1321 gcccctctgc taaccatgtt catgccttct tctctttcct acagctcctg ggcaacgtgc 1381 tggttgttgt gctgtctcat cattttggca aagaattcct cgacggatcc ctcgaggaat 1441 tctgacacta tgaagtgcct tttgtactta gcctttttat tcattggggt gaattgcaag 1501 ttcaccatag tttttccaca caaccaaaaa ggaaactgga aaaatgttcc ttctaattac 1561 cattattgcc cgtcaagctc agatttaaat tggcataatg acttaatagg cacagcctta 1621 caagtcaaaa tgcccaagag tcacaaggct attcaagcag acggttggat gtgtcatgct 1681 tccaaatggg tcactacttg tgatttccgc tggtatggac cgaagtatat aacacattcc 1741 atccgatcct tcactccatc tgtagaacaa tgcaaggaaa gcattgaaca aacgaaacaa 1801 ggaacttggc tgaatccagg cttccctcct caaagttgtg gatatgcaac tgtgacggat 1861 gccgaagcag tgattgtcca ggtgactcct caccatgtgc tggttgatga atacacagga 1921 gaatgggttg attcacagtt catcaacgga aaatgcagca attacatatg ccccactgtc 1981 cataactcta caacctggca ttctgactat aaggtcaaag ggctatgtga ttctaacctc 2041 atttccatgg acatcacctt cttctcagag gacggagagc tatcatccct gggaaaggag 2101 ggcacagggt tcagaagtaa ctactttgct tatgaaactg gaggcaaggc ctgcaaaatg 2161 caatactgca agcattgggg agtcagactc ccatcaggtg tctggttcga gatggctgat 2221 aaggatctct ttgctgcagc cagattccct gaatgcccag aagggtcaag tatctctgct 2281 ccatctcaga cctcagtgga tgtaagtcta attcaggacg ttgagaggat cttggattat 2341 tccctctgcc aagaaacctg gagcaaaatc agagcgggtc ttccaatctc tccagtggat 2401 ctcagctatc ttgctcctaa aaacccagga accggtcctg ctttcaccat aatcaatggt 2461 accctaaaat actttgagac cagatacatc agagtcgata ttgctgctcc aatcctctca 2521 agaatggtcg gaatgatcag tggaactacc acagaaaggg aactgtggga tgactgggca 2581 ccatatgaag acgtggaaat tggacccaat ggagttctga ggaccagttc aggatataag 2641 tttcctttat acatgattgg acatggtatg ttggactccg atcttcatct tagctcaaag 2701 gctcaggtgt tcgaacatcc tcacattcaa gacgctgctt cgcaacttcc tgatgatgag 2761 agtttatttt ttggtgatac tgggctatcc aaaaatccaa tcgagcttgt agaaggttgg 2821 ttcagtagtt ggaaaagctc tattgcctct tttttcttta tcatagggtt aatcattgga 2881 ctattcttgg ttctccgagt tggtatccat ctttgcatta aattaaagca caccaagaaa 2941 agacagattt atacagacat agagatgaac cgacttggaa agtaactcaa atcctgcaca 3001 acagattctt catgtttgga ccaaatcaac ttgtgatacc atgctcaaag aggcctcaat 3061 tatatttgag tttttaattt ttatgaaaaa aaaaaaaaaa aacggaattc ctcgagggat 3121 ccgtcgagga attcactcct caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg 3181 ccaatgccct ggctcacaaa taccactgag atctttttcc ctctgccaaa aattatgggg 3241 acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt attttcattg 3301 caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc 3361 atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc catatgctgg 3421 ctgccatgaa caaaggttgg ctataaagag gtcatcagta tatgaaacag ccccctgctg 3481 tccattcctt attccataga aaagccttga cttgaggtta gatttttttt atattttgtt 3541 ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta ctagccagat 3601 ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctctta tggagatccc 3661 tcgacggatc ggccgcaatt cgtaatcatg tcatagctgt ttcctgtgtg aaattgttat 3721 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc 3781 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga 3841 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt 3901 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg 3961 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac 4021 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg 4081 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca 4141 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc 4201 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc 4261 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag 4321 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc 4381 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca 4441 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg 4501 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg 4561 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct 4621 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa 4681 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa 4741 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa 4801 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc 4861 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga 4921 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca 4981 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc 5041 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat 5101 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc 5161 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt 5221 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc 5281 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg 5341 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt 5401 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg 5461 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga 5521 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg 5581 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg 5641 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt 5701 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc 5761 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca 5821 tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa 5881 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata 5941 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac 6001 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc 6061 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa 6121 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg 6181 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg 6241 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc 6301 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat 6361 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt 6421 tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac gactcactat 6481 agggcgaatt ggagctccac cgcggtg
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