| 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
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