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pLVX-AcGFP1-C1载体质粒基本信息
pLVX-AcGFP1-C1质粒图谱载体图谱和pLVX-AcGFP1-C1载体序列质粒序列多克隆位点信息
pLVX-AcGFP1-C1质粒载体简介
Description
pLVX-AcGFP1-C1 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 at the C-terminal end of the AcGFP1 coding sequence, are expressed as C-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 AcGFP1 coding sequence. Lentiviral particles derived from the vector allow the expression of AcGFP1 fusion proteins in virtually any cell type, including primary cells. The unmodified vector expresses AcGFP1, and may be used to produce marker virus to optimize infection protocols.
pLVX-AcGFP1-C1 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-C1 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-C1 contains a puromycin resistance gene (Puromycin resistance) 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-C1 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-C1质粒序列载体序列
LOCUS pLVX-AcGFP1-C1 8787 bp DNA circular 9-NOV-2009 COMMENT Created by Clontech Laboratories Inc. COMMENT Cat. No. 632155 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 splicing_signal 744..745 /label=SD misc_binding 636..653 /label=PBS LTR 5549..6185 /label=3'\LTR CDS 2807..3604 /label=AcGFP1 misc_feature 3537..3591 /label=MCS promoter 3613..4121 /label=PGK\Promoter CDS 4142..4741 /label=Puro(R) misc_feature 3598..3600 /label=Stop misc_feature 3602..3604 /label=Stop misc_feature 3602..3604 /label=Stop BASE COUNT 2191 a 2273 c 2307 g 2016 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 tctaccggtc gccaccatgg tgagcaaggg 2821 cgccgagctg ttcaccggca tcgtgcccat cctgatcgag ctgaatggcg atgtgaatgg 2881 ccacaagttc agcgtgagcg gcgagggcga gggcgatgcc acctacggca agctgaccct 2941 gaagttcatc tgcaccaccg gcaagctgcc tgtgccctgg cccaccctgg tgaccaccct 3001 gagctacggc gtgcagtgct tctcacgcta ccccgatcac atgaagcagc acgacttctt 3061 caagagcgcc atgcctgagg gctacatcca ggagcgcacc atcttcttcg aggatgacgg 3121 caactacaag tcgcgcgccg aggtgaagtt cgagggcgat accctggtga atcgcatcga 3181 gctgaccggc accgatttca aggaggatgg caacatcctg ggcaataaga tggagtacaa 3241 ctacaacgcc cacaatgtgt acatcatgac cgacaaggcc aagaatggca tcaaggtgaa 3301 cttcaagatc cgccacaaca tcgaggatgg cagcgtgcag ctggccgacc actaccagca 3361 gaataccccc atcggcgatg gccctgtgct gctgcccgat aaccactacc tgtccaccca 3421 gagcgccctg tccaaggacc ccaacgagaa gcgcgatcac atgatctact tcggcttcgt 3481 gaccgccgcc gccatcaccc acggcatgga tgagctgtac aagtccggac tcagatctcg 3541 agctcaagct tcgaattctg cagtcgacgg taccgcgggc ccgggatcca ccggatctag 3601 ataactgatc 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-C1质粒载体应用举例
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