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pLVX-Puro

pLVXpuro载体基本信息

pLVXpuro载体质粒图谱和多克隆位点信息

pLVXpuro载体简介

Description

pLVX-Puro is an HIV-1-based, lentiviral expression vector. Lentiviral particles derived from the vector allow you to express your gene of interest in virtually any cell type, even primary cells. Expression of your gene is driven by the constitutively active human cytomegalovirus immediate early promoter (PCMV IE), located just upstream of the multiple cloning site (MCS), allowing constitutive, high level expression of your protein of interest.

pLVX-Puro contains all of the viral processing elements necessary for the production of replication-incompetent lentivirus, as well as elements to improve viral titer, transgene expression, and overall vector function. The woodchuck hepatitis virus posttranscriptional regulatory element (WPRE) promotes RNA processing events and enhances nuclear export of viral and transgene RNA (1), leading to increased viral titers from packaging cells, and enhanced expression of your gene of interest in target cells. In addition, the vector includes a Rev-response element (RRE), which further increases viral titers by enhancing the transport of unspliced viral RNA out of the nucleus (2). Finally, pLVX-Puro also contains a central polypurine tract (cPPT) element that increases nuclear importation of the viral genome during target cell infection, resulting in improved vector integration and more effi cient transduction (3). In addition to lentiviral elements, pLVX-Puro contains a puromycin resistance gene (Puror) under the control of the murine phosphoglycerate kinase (PGK) promoter (PPGK) for the selection of stable transductants. The vector also contains a pUC origin of replication and an E. coli ampicillin resistance gene (Ampr) for propagation and selection in bacteria.

Use
pLVX-Puro constitutively expresses your gene of interest from PCMV IE when transduced into target cells. Before the vector can be transduced into cells, however, it must be transfected into 293T packaging cells with our Lenti-X™ HT Packaging System (Cat. Nos. 632160 and 632161). This packaging system allows you to safely produce high titer, infectious, replication-incompetent, VSV-G pseudotyped lentiviral particles that can infect a wide range of cell types, including non-dividing and primary cells (4).

全序列

1        TGGAAGGGCT AATTCACTCC CAAAGAAGAC AAGATATCCT TGATCTGTGG ATCTACCACA
61       CACAAGGCTA CTTCCCTGAT TAGCAGAACT ACACACCAGG GCCAGGGGTC AGATATCCAC
121      TGACCTTTGG ATGGTGCTAC AAGCTAGTAC CAGTTGAGCC AGATAAGGTA GAAGAGGCCA
181      ATAAAGGAGA GAACACCAGC TTGTTACACC CTGTGAGCCT GCATGGGATG GATGACCCGG
241      AGAGAGAAGT GTTAGAGTGG AGGTTTGACA GCCGCCTAGC ATTTCATCAC GTGGCCCGAG
301      AGCTGCATCC GGAGTACTTC AAGAACTGCT GATATCGAGC TTGCTACAAG GGACTTTCCG
361      CTGGGGACTT TCCAGGGAGG CGTGGCCTGG GCGGGACTGG GGAGTGGCGA GCCCTCAGAT
421      CCTGCATATA AGCAGCTGCT TTTTGCCTGT ACTGGGTCTC TCTGGTTAGA CCAGATCTGA
481      GCCTGGGAGC TCTCTGGCTA ACTAGGGAAC CCACTGCTTA AGCCTCAATA AAGCTTGCCT
541      TGAGTGCTTC AAGTAGTGTG TGCCCGTCTG TTGTGTGACT CTGGTAACTA GAGATCCCTC
601      AGACCCTTTT AGTCAGTGTG GAAAATCTCT AGCAGTGGCG CCCGAACAGG GACTTGAAAG
661      CGAAAGGGAA ACCAGAGGAG CTCTCTCGAC GCAGGACTCG GCTTGCTGAA GCGCGCACGG
721      CAAGAGGCGA GGGGCGGCGA CTGGTGAGTA CGCCAAAAAT TTTGACTAGC GGAGGCTAGA
781      AGGAGAGAGA TGGGTGCGAG AGCGTCAGTA TTAAGCGGGG GAGAATTAGA TCGCGATGGG
841      AAAAAATTCG GTTAAGGCCA GGGGGAAAGA AAAAATATAA ATTAAAACAT ATAGTATGGG
901      CAAGCAGGGA GCTAGAACGA TTCGCAGTTA ATCCTGGCCT GTTAGAAACA TCAGAAGGCT
961      GTAGACAAAT ACTGGGACAG CTACAACCAT CCCTTCAGAC AGGATCAGAA GAACTTAGAT
1021     CATTATATAA TACAGTAGCA ACCCTCTATT GTGTGCATCA AAGGATAGAG ATAAAAGACA
1081     CCAAGGAAGC TTTAGACAAG ATAGAGGAAG AGCAAAACAA AAGTAAGACC ACCGCACAGC
1141     AAGCGGCCGG CCGCTGATCT TCAGACCTGG AGGAGGAGAT ATGAGGGACA ATTGGAGAAG
1201     TGAATTATAT AAATATAAAG TAGTAAAAAT TGAACCATTA GGAGTAGCAC CCACCAAGGC
1261     AAAGAGAAGA GTGGTGCAGA GAGAAAAAAG AGCAGTGGGA ATAGGAGCTT TGTTCCTTGG
1321     GTTCTTGGGA GCAGCAGGAA GCACTATGGG CGCAGCGTCA ATGACGCTGA CGGTACAGGC
1381     CAGACAATTA TTGTCTGGTA TAGTGCAGCA GCAGAACAAT TTGCTGAGGG CTATTGAGGC
1441     GCAACAGCAT CTGTTGCAAC TCACAGTCTG GGGCATCAAG CAGCTCCAGG CAAGAATCCT
1501     GGCTGTGGAA AGATACCTAA AGGATCAACA GCTCCTGGGG ATTTGGGGTT GCTCTGGAAA
1561     ACTCATTTGC ACCACTGCTG TGCCTTGGAA TGCTAGTTGG AGTAATAAAT CTCTGGAACA
1621     GATTTGGAAT CACACGACCT GGATGGAGTG GGACAGAGAA ATTAACAATT ACACAAGCTT
1681     AATACACTCC TTAATTGAAG AATCGCAAAA CCAGCAAGAA AAGAATGAAC AAGAATTATT
1741     GGAATTAGAT AAATGGGCAA GTTTGTGGAA TTGGTTTAAC ATAACAAATT GGCTGTGGTA
1801     TATAAAATTA TTCATAATGA TAGTAGGAGG CTTGGTAGGT TTAAGAATAG TTTTTGCTGT
1861     ACTTTCTATA GTGAATAGAG TTAGGCAGGG ATATTCACCA TTATCGTTTC AGACCCACCT
1921     CCCAACCCCG AGGGGACCCG ACAGGCCCGA AGGAATAGAA GAAGAAGGTG GAGAGAGAGA
1981     CAGAGACAGA TCCATTCGAT TAGTGAACGG ATCTCGACGG TATCGCCTTT AAAAGAAAAG
2041     GGGGGATTGG GGGGTACAGT GCAGGGGAAA GAATAGTAGA CATAATAGCA ACAGACATAC
2101     AAACTAAAGA ATTACAAAAA CAAATTACAA AAATTCAAAA TTTTCGGGTT TATTACAGGG
2161     ACAGCAGAGA TCCAGTTTAT CGATAAGCTT GGGAGTTCCG CGTTACATAA CTTACGGTAA
2221     ATGGCCCGCC TGGCTGACCG CCCAACGACC CCCGCCCATT GACGTCAATA ATGACGTATG
2281     TTCCCATAGT AACGCCAATA GGGACTTTCC ATTGACGTCA ATGGGTGGAG TATTTACGGT
2341     AAACTGCCCA CTTGGCAGTA CATCAAGTGT ATCATATGCC AAGTACGCCC CCTATTGACG
2401     TCAATGACGG TAAATGGCCC GCCTGGCATT ATGCCCAGTA CATGACCTTA TGGGACTTTC
2461     CTACTTGGCA GTACATCTAC GTATTAGTCA TCGCTATTAC CATGGTGATG CGGTTTTGGC
2521     AGTACATCAA TGGGCGTGGA TAGCGGTTTG ACTCACGGGG ATTTCCAAGT CTCCACCCCA
2581     TTGACGTCAA TGGGAGTTTG TTTTGGCACC AAAATCAACG GGACTTTCCA AAATGTCGTA
2641     ACAACTCCGC CCCATTGACG CAAATGGGCG GTAGGCGTGT ACGGTGGGAG GTCTATATAA
2701     GCAGAGCTCG TTTAGTGAAC CGTCAGATCG CCTGGAGACG CCATCCACGC TGTTTTGACC
2761     TCCATAGAAG ACACCGACTC TACTAGAGGA TCGCTAGCGC TACCGGACTC AGATCTCGAG
2821     CTCAAGCTTC GAATTCTGCA GTCGACGGTA CCGCGGGCCC GGGATCCCGC GACTCTAGAT
2881     AATTCTACCG GGTAGGGGAG GCGCTTTTCC CAAGGCAGTC TGGAGCATGC GCTTTAGCAG
2941     CCCCGCTGGG CACTTGGCGC TACACAAGTG GCCTCTGGCC TCGCACACAT TCCACATCCA
3001     CCGGTAGGCG CCAACCGGCT CCGTTCTTTG GTGGCCCCTT CGCGCCACCT TCTACTCCTC
3061     CCCTAGTCAG GAAGTTCCCC CCCGCCCCGC AGCTCGCGTC GTGCAGGACG TGACAAATGG
3121     AAGTAGCACG TCTCACTAGT CTCGTGCAGA TGGACAGCAC CGCTGAGCAA TGGAAGCGGG
3181     TAGGCCTTTG GGGCAGCGGC CAATAGCAGC TTTGCTCCTT CGCTTTCTGG GCTCAGAGGC
3241     TGGGAAGGGG TGGGTCCGGG GGCGGGCTCA GGGGCGGGCT CAGGGGCGGG GCGGGCGCCC
3301     GAAGGTCCTC CGGAGGCCCG GCATTCTGCA CGCTTCAAAA GCGCACGTCT GCCGCGCTGT
3361     TCTCCTCTTC CTCATCTCCG GGCCTTTCGA CCTGCAGCCC AAGCTTACCA TGACCGAGTA
3421     CAAGCCCACG GTGCGCCTCG CCACCCGCGA CGACGTCCCC AGGGCCGTAC GCACCCTCGC
3481     CGCCGCGTTC GCCGACTACC CCGCCACGCG CCACACCGTC GATCCGGACC GCCACATCGA
3541     GCGGGTCACC GAGCTGCAAG AACTCTTCCT CACGCGCGTC GGGCTCGACA TCGGCAAGGT
3601     GTGGGTCGCG GACGACGGCG CCGCGGTGGC GGTCTGGACC ACGCCGGAGA GCGTCGAAGC
3661     GGGGGCGGTG TTCGCCGAGA TCGGCCCGCG CATGGCCGAG TTGAGCGGTT CCCGGCTGGC
3721     CGCGCAGCAA CAGATGGAAG GCCTCCTGGC GCCGCACCGG CCCAAGGAGC CCGCGTGGTT
3781     CCTGGCCACC GTCGGCGTCT CGCCCGACCA CCAGGGCAAG GGTCTGGGCA GCGCCGTCGT
3841     GCTCCCCGGA GTGGAGGCGG CCGAGCGCGC CGGGGTGCCC GCCTTCCTGG AGACCTCCGC
3901     GCCCCGCAAC CTCCCCTTCT ACGAGCGGCT CGGCTTCACC GTCACCGCCG ACGTCGAGGT
3961     GCCCGAAGGA CCGCGCACCT GGTGCATGAC CCGCAAGCCC GGTGCCTGAC CGCGTCTGGA
4021     ACAATCAACC TCTGGATTAC AAAATTTGTG AAAGATTGAC TGGTATTCTT AACTATGTTG
4081     CTCCTTTTAC GCTATGTGGA TACGCTGCTT TAATGCCTTT GTATCATGCT ATTGCTTCCC
4141     GTATGGCTTT CATTTTCTCC TCCTTGTATA AATCCTGGTT GCTGTCTCTT TATGAGGAGT
4201     TGTGGCCCGT TGTCAGGCAA CGTGGCGTGG TGTGCACTGT GTTTGCTGAC GCAACCCCCA
4261     CTGGTTGGGG CATTGCCACC ACCTGTCAGC TCCTTTCCGG GACTTTCGCT TTCCCCCTCC
4321     CTATTGCCAC GGCGGAACTC ATCGCCGCCT GCCTTGCCCG CTGCTGGACA GGGGCTCGGC
4381     TGTTGGGCAC TGACAATTCC GTGGTGTTGT CGGGGAAGCT GACGTCCTTT CCATGGCTGC
4441     TCGCCTGTGT TGCCACCTGG ATTCTGCGCG GGACGTCCTT CTGCTACGTC CCTTCGGCCC
4501     TCAATCCAGC GGACCTTCCT TCCCGCGGCC TGCTGCCGGC TCTGCGGCCT CTTCCGCGTC
4561     TTCGCCTTCG CCCTCAGACG AGTCGGATCT CCCTTTGGGC CGCCTCCCCG CCTGGAATTA
4621     ATTCTGCAGT CGAGACCTAG AAAAACATGG AGCAATCACA AGTAGCAATA CAGCAGCTAC
4681     CAATGCTGAT TGTGCCTGGC TAGAAGCACA AGAGGAGGAG GAGGTGGGTT TTCCAGTCAC
4741     ACCTCAGGTA CCTTTAAGAC CAATGACTTA CAAGGCAGCT GTAGATCTTA GCCACTTTTT
4801     AAAAGAAAAG AGGGGACTGG AAGGGCTAAT TCACTCCCAA CGAAGACAAG ATATCCTTGA
4861     TCTGTGGATC TACCACACAC AAGGCTACTT CCCTGATTAG CAGAACTACA CACCAGGGCC
4921     AGGGGTCAGA TATCCACTGA CCTTTGGATG GTGCTACAAG CTAGTACCAG TTGAGCCAGA
4981     TAAGGTAGAA GAGGCCAATA AAGGAGAGAA CACCAGCTTG TTACACCCTG TGAGCCTGCA
5041     TGGGATGGAT GACCCGGAGA GAGAAGTGTT AGAGTGGAGG TTTGACAGCC GCCTAGCATT
5101     TCATCACGTG GCCCGAGAGC TGCATCCGGA GTACTTCAAG AACTGCTGAT ATCGAGCTTG
5161     CTACAAGGGA CTTTCCGCTG GGGACTTTCC AGGGAGGCGT GGCCTGGGCG GGACTGGGGA
5221     GTGGCGAGCC CTCAGATCCT GCATATAAGC AGCTGCTTTT TGCCTGTACT GGGTCTCTCT
5281     GGTTAGACCA GATCTGAGCC TGGGAGCTCT CTGGCTAACT AGGGAACCCA CTGCTTAAGC
5341     CTCAATAAAG CTTGCCTTGA GTGCTTCAAG TAGTGTGTGC CCGTCTGTTG TGTGACTCTG
5401     GTAACTAGAG ATCCCTCAGA CCCTTTTAGT CAGTGTGGAA AATCTCTAGC AGTAGTAGTT
5461     CATGTCATCT TATTATTCAG TATTTATAAC TTGCAAAGAA ATGAATATCA GAGAGTGAGA
5521     GGCCTTGACA TTGCTAGCGT TTTACCGTCG ACCTCTAGCT AGAGCTTGGC GTAATCATGG
5581     TCATAGCTGT TTCCTGTGTG AAATTGTTAT CCGCTCACAA TTCCACACAA CATACGAGCC
5641     GGAAGCATAA AGTGTAAAGC CTGGGGTGCC TAATGAGTGA GCTAACTCAC ATTAATTGCG
5701     TTGCGCTCAC TGCCCGCTTT CCAGTCGGGA AACCTGTCGT GCCAGCTGCA TTAATGAATC
5761     GGCCAACGCG CGGGGAGAGG CGGTTTGCGT ATTGGGCGCT CTTCCGCTTC CTCGCTCACT
5821     GACTCGCTGC GCTCGGTCGT TCGGCTGCGG CGAGCGGTAT CAGCTCACTC AAAGGCGGTA
5881     ATACGGTTAT CCACAGAATC AGGGGATAAC GCAGGAAAGA ACATGTGAGC AAAAGGCCAG
5941     CAAAAGGCCA GGAACCGTAA AAAGGCCGCG TTGCTGGCGT TTTTCCATAG GCTCCGCCCC
6001     CCTGACGAGC ATCACAAAAA TCGACGCTCA AGTCAGAGGT GGCGAAACCC GACAGGACTA
6061     TAAAGATACC AGGCGTTTCC CCCTGGAAGC TCCCTCGTGC GCTCTCCTGT TCCGACCCTG
6121     CCGCTTACCG GATACCTGTC CGCCTTTCTC CCTTCGGGAA GCGTGGCGCT TTCTCATAGC
6181     TCACGCTGTA GGTATCTCAG TTCGGTGTAG GTCGTTCGCT CCAAGCTGGG CTGTGTGCAC
6241     GAACCCCCCG TTCAGCCCGA CCGCTGCGCC TTATCCGGTA ACTATCGTCT TGAGTCCAAC
6301     CCGGTAAGAC ACGACTTATC GCCACTGGCA GCAGCCACTG GTAACAGGAT TAGCAGAGCG
6361     AGGTATGTAG GCGGTGCTAC AGAGTTCTTG AAGTGGTGGC CTAACTACGG CTACACTAGA
6421     AGAACAGTAT TTGGTATCTG CGCTCTGCTG AAGCCAGTTA CCTTCGGAAA AAGAGTTGGT
6481     AGCTCTTGAT CCGGCAAACA AACCACCGCT GGTAGCGGTG GTTTTTTTGT TTGCAAGCAG
6541     CAGATTACGC GCAGAAAAAA AGGATCTCAA GAAGATCCTT TGATCTTTTC TACGGGGTCT
6601     GACGCTCAGT GGAACGAAAA CTCACGTTAA GGGATTTTGG TCATGAGATT ATCAAAAAGG
6661     ATCTTCACCT AGATCCTTTT AAATTAAAAA TGAAGTTTTA AATCAATCTA AAGTATATAT
6721     GAGTAAACTT GGTCTGACAG TTACCAATGC TTAATCAGTG AGGCACCTAT CTCAGCGATC
6781     TGTCTATTTC GTTCATCCAT AGTTGCCTGA CTCCCCGTCG TGTAGATAAC TACGATACGG
6841     GAGGGCTTAC CATCTGGCCC CAGTGCTGCA ATGATACCGC GAGACCCACG CTCACCGGCT
6901     CCAGATTTAT CAGCAATAAA CCAGCCAGCC GGAAGGGCCG AGCGCAGAAG TGGTCCTGCA
6961     ACTTTATCCG CCTCCATCCA GTCTATTAAT TGTTGCCGGG AAGCTAGAGT AAGTAGTTCG
7021     CCAGTTAATA GTTTGCGCAA CGTTGTTGCC ATTGCTACAG GCATCGTGGT GTCACGCTCG
7081     TCGTTTGGTA TGGCTTCATT CAGCTCCGGT TCCCAACGAT CAAGGCGAGT TACATGATCC
7141     CCCATGTTGT GCAAAAAAGC GGTTAGCTCC TTCGGTCCTC CGATCGTTGT CAGAAGTAAG
7201     TTGGCCGCAG TGTTATCACT CATGGTTATG GCAGCACTGC ATAATTCTCT TACTGTCATG
7261     CCATCCGTAA GATGCTTTTC TGTGACTGGT GAGTACTCAA CCAAGTCATT CTGAGAATAG
7321     TGTATGCGGC GACCGAGTTG CTCTTGCCCG GCGTCAATAC GGGATAATAC CGCGCCACAT
7381     AGCAGAACTT TAAAAGTGCT CATCATTGGA AAACGTTCTT CGGGGCGAAA ACTCTCAAGG
7441     ATCTTACCGC TGTTGAGATC CAGTTCGATG TAACCCACTC GTGCACCCAA CTGATCTTCA
7501     GCATCTTTTA CTTTCACCAG CGTTTCTGGG TGAGCAAAAA CAGGAAGGCA AAATGCCGCA
7561     AAAAAGGGAA TAAGGGCGAC ACGGAAATGT TGAATACTCA TACTCTTCCT TTTTCAATAT
7621     TATTGAAGCA TTTATCAGGG TTATTGTCTC ATGAGCGGAT ACATATTTGA ATGTATTTAG
7681     AAAAATAAAC AAATAGGGGT TCCGCGCACA TTTCCCCGAA AAGTGCCACC TGACGTCGAC
7741     GGATCGGGAG ATCAACTTGT TTATTGCAGC TTATAATGGT TACAAATAAA GCAATAGCAT
7801     CACAAATTTC ACAAATAAAG CATTTTTTTC ACTGCATTCT AGTTGTGGTT TGTCCAAACT
7861     CATCAATGTA TCTTATCATG TCTGGATCAA CTGGATAACT CAAGCTAACC AAAATCATCC
7921     CAAACTTCCC ACCCCATACC CTATTACCAC TGCCAATTAC CTGTGGTTTC ATTTACTCTA
7981     AACCTGTGAT TCCTCTGAAT TATTTTCATT TTAAAGAAAT TGTATTTGTT AAATATGTAC
8041     TACAAACTTA GTAGTTTTTA AAGAAATTGT ATTTGTTAAA TATGTACTAC AAACTTAGTA
8101     GT