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pLVX-AcGFP1-N1载体质粒基本信息
pLVX-AcGFP1-N1质粒图谱载体图谱和pLVX-AcGFP1-N1载体序列质粒序列多克隆位点信息
pLVX-AcGFP1-N1质粒载体简介
Description
pLVX-AcGFP1-N1 is an HIV-1-based, lentiviral expression vector that allows you to express your gene of interest fused to AcGFP1, a green fl uorescent protein derived from Aequorea coerulescens. Genes cloned into the multiple cloning site (MCS), located upstream of the AcGFP1 coding sequence, are expressed as N-terminal fusions of the AcGFP1 protein. Expression of the fusion protein is driven by the constitutively active human cytomegalovirus immediate early promoter (PCMV IE) located just upstream of the MCS. Lentiviral particles derived from the vector allow the expression of AcGFP1 fusion proteins in virtually any cell type, including primary cells. The unmodifi ed vector expresses AcGFP1, and may be used to produce marker virus to optimize infection protocols.
pLVX-AcGFP1-N1 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-AcGFP1-N1 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 efficient transduction (3).
In addition to lentiviral elements, pLVX-AcGFP1-N1 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-AcGFP1-N1 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).
pLVX-AcGFP1-N1质粒序列载体序列
LOCUS pLVX-AcGFP1-N1 8787 bp DNA circular 9-NOV-2009
COMMENT Created by Clontech Laboratories Inc.
COMMENT Cat. No. 632154
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT VNTAUTHORNAME|biofeng.com|
FEATURES Location/Qualifiers
LTR 1..635
/label=5'\LTR
misc_feature 685..822
/label=Packaging\signal
misc_feature 2028..2151
/label=cPPT/CTS
promoter 2185..2787
/label=CMV\IE\Promoter
misc_feature 4755..5346
/label=WPRE
rep_origin complement(6655..7328)
/label=pUC\origin
CDS complement(7473..8469)
/label=Amp(R)
misc_binding 1303..1536
/label=RRE
misc_binding 636..653
/label=PBS
LTR 5549..6185
/label=3'\LTR
misc_feature 2796..2868
/label=MCS
CDS 2875..3591
/label=AcGFP1
promoter 3613..4121
/label=PGK\Promoter
CDS 4142..4741
/label=Puro(R)
BASE COUNT 2188 a 2273 c 2312 g 2014 t
ORIGIN
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 gggatccacc ggtcatggtg
2881 agcaagggcg ccgagctgtt caccggcatc gtgcccatcc tgatcgagct gaatggcgat
2941 gtgaatggcc acaagttcag cgtgagcggc gagggcgagg gcgatgccac ctacggcaag
3001 ctgaccctga agttcatctg caccaccggc aagctgcctg tgccctggcc caccctggtg
3061 accaccctga gctacggcgt gcagtgcttc tcacgctacc ccgatcacat gaagcagcac
3121 gacttcttca agagcgccat gcctgagggc tacatccagg agcgcaccat cttcttcgag
3181 gatgacggca actacaagtc gcgcgccgag gtgaagttcg agggcgatac cctggtgaat
3241 cgcatcgagc tgaccggcac cgatttcaag gaggatggca acatcctggg caataagatg
3301 gagtacaact acaacgccca caatgtgtac atcatgaccg acaaggccaa gaatggcatc
3361 aaggtgaact tcaagatccg ccacaacatc gaggatggca gcgtgcagct ggccgaccac
3421 taccagcaga atacccccat cggcgatggc cctgtgctgc tgcccgataa ccactacctg
3481 tccacccaga gcgccctgtc caaggacccc aacgagaagc gcgatcacat gatctacttc
3541 ggcttcgtga ccgccgccgc catcacccac ggcatggatg agctgtacaa gtgagcggcc
3601 gcgactctag ataattctac cgggtagggg aggcgctttt cccaaggcag tctggagcat
3661 gcgctttagc agccccgctg ggcacttggc gctacacaag tggcctctgg cctcgcacac
3721 attccacatc caccggtagg cgccaaccgg ctccgttctt tggtggcccc ttcgcgccac
3781 cttctactcc tcccctagtc aggaagttcc cccccgcccc gcagctcgcg tcgtgcagga
3841 cgtgacaaat ggaagtagca cgtctcacta gtctcgtgca gatggacagc accgctgagc
3901 aatggaagcg ggtaggcctt tggggcagcg gccaatagca gctttgctcc ttcgctttct
3961 gggctcagag gctgggaagg ggtgggtccg ggggcgggct caggggcggg ctcaggggcg
4021 gggcgggcgc ccgaaggtcc tccggaggcc cggcattctg cacgcttcaa aagcgcacgt
4081 ctgccgcgct gttctcctct tcctcatctc cgggcctttc gacctgcagc ccaagcttac
4141 catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt
4201 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga
4261 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga
4321 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga
4381 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg
4441 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga
4501 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg
4561 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct
4621 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc
4681 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg
4741 accgcgtctg gaacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
4801 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
4861 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
4921 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
4981 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
5041 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
5101 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct
5161 ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
5221 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
5281 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
5341 cgcctggaat taattctgca gtcgagacct agaaaaacat ggagcaatca caagtagcaa
5401 tacagcagct accaatgctg attgtgcctg gctagaagca caagaggagg aggaggtggg
5461 ttttccagtc acacctcagg tacctttaag accaatgact tacaaggcag ctgtagatct
5521 tagccacttt ttaaaagaaa agaggggact ggaagggcta attcactccc aacgaagaca
5581 agatatcctt gatctgtgga tctaccacac acaaggctac ttccctgatt agcagaacta
5641 cacaccaggg ccaggggtca gatatccact gacctttgga tggtgctaca agctagtacc
5701 agttgagcca gataaggtag aagaggccaa taaaggagag aacaccagct tgttacaccc
5761 tgtgagcctg catgggatgg atgacccgga gagagaagtg ttagagtgga ggtttgacag
5821 ccgcctagca tttcatcacg tggcccgaga gctgcatccg gagtacttca agaactgctg
5881 atatcgagct tgctacaagg gactttccgc tggggacttt ccagggaggc gtggcctggg
5941 cgggactggg gagtggcgag ccctcagatc ctgcatataa gcagctgctt tttgcctgta
6001 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
6061 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
6121 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
6181 gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat
6241 cagagagtga gaggccttga cattgctagc gttttaccgt cgacctctag ctagagcttg
6301 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
6361 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
6421 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
6481 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
6541 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
6601 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
6661 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
6721 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
6781 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
6841 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
6901 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
6961 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
7021 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
7081 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
7141 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
7201 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
7261 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
7321 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
7381 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
7441 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
7501 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
7561 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
7621 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
7681 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
7741 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
7801 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
7861 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
7921 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
7981 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
8041 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
8101 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
8161 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
8221 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
8281 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
8341 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
8401 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
8461 cctgacgtcg acggatcggg agatcaactt gtttattgca gcttataatg gttacaaata
8521 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
8581 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc aactggataa ctcaagctaa
8641 ccaaaatcat cccaaacttc ccaccccata ccctattacc actgccaatt acctgtggtt
8701 tcatttactc taaacctgtg attcctctga attattttca ttttaaagaa attgtatttg
8761 ttaaatatgt actacaaact tagtagt
//
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pLVX-AcGFP1-N1质粒载体应用举例
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