pLVX-IRES-ZsGreen1

价格:1500元

联系方式:I47-825O-882O

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载体基本信息

出品公司: Clontech
载体名称: pLVX-IRES-ZsGreen1
质粒类型: 慢病毒载体;哺乳动物细胞表达载体;双顺反子载体;荧光报告载体
高拷贝/低拷贝: 高拷贝
启动子: CMV
克隆方法: 多克隆位点,限制性内切酶
载体大小: 8204 bp
5' 测序引物及序列: CMV-F: CGCAAATGGGCGGTAGGCGTG (Invitrogen)
3' 测序引物及序列: --
载体标签: 无标签
载体抗性: 氨苄青霉素
筛选标记: 无筛选标记
克隆菌株: Stbl3
备注: pLVX-IRES-ZsGreen1载体以双顺反子的形式同时表达ZsGreen1荧光蛋白和目的基因;
CMV启动子驱动目的基因的过表达;
ZsGreen1是第三代绿色荧光蛋白,是亮度最高的的荧光蛋白。
产品目录号: 632187
稳定性: 稳表达
组成型: 组成型
病毒/非病毒: 慢病毒

载体质粒图谱和多克隆位点信息

pLVX-IRES-ZsGreen1载体图谱



pLVX-IRES-ZsGreen1多克隆位点

pLVX-IRES-ZsGreen1载体特征

载体简介

Description
pLVX-IRES-ZsGreen1 is an HIV-1-based, lentiviral expression vector that allows the simultaneous expression of your protein of interest and ZsGreen1 in virtually any mammalian cell type, including primary cells. ZsGreen1 is a human codon-optimized variant of the reef coral Zoanthus sp. green fluorescent protein, ZsGreen. The vector expresses the two proteins from a bicistronic mRNA transcript, allowing ZsGreen1 to be used as an indicator of transduction efficiency and a marker for selection by flow cytometry.

Expression of the bicistronic transcript is driven by the constitutively active human cytomegalovirus immediate early promoter (PCMV IE) located just upstream of the MCS. An encephalomyocarditis virus (EMCV) internal ribosome entry site (IRES), positioned between the MCS and ZsGreen1, facilitates cap-independent translation of ZsGreen1 from an internal start site at the IRES/ZsGreen1 junction (1).

pLVX-IRES-ZsGreen1 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 RNA (2), leading to increased viral titers from packaging 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 (3). Finally, pLVX-IRES-ZsGreen1 also contains a central polypurine tract/central termination sequence element (cPPT/CTS). During target cell infection, this element creates a central DNA flap that increases nuclear import of the viral genome, resulting in improved vector integration and more efficient transduction (4). 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-IRES-ZsGreen1 is designed to constitutively coexpress your protein of interest and ZsGreen1 from PCMV IE when transduced into mammalian cells. Before it can be transduced into target cells, the vector must be packaged into viral particles in HEK293T cells, using our Lenti-X™ HTX Packaging System (Cat. Nos. 631247 and 631249). This packaging system allows the safe production of high titer, infectious, replication-incompetent, VSV-G pseudotyped lentiviral particles that can infect a wide range of cell types, including nondividing and primary cells (5).

ZsGreen1 is the brightest commercially available green fluorescent protein. The presence of this protein allows transductants to be visualized by fluorescence microscopy and sorted by flow cytometry with standard FITC filter sets (ZsGreen1 has an excitation maximum of 493 nm and an emission maximum of 505 nm).

载体序列

LOCUS       pLVX-IRES-ZsGreen1   8204 bp    DNA   circular             8-NOV-2009
COMMENT     Created by Clontech Laboratories Inc.
COMMENT     Cat. No. 632187
COMMENT     This file is created by Vector NTI
            http://www.biofeng.com/
COMMENT     VNTAUTHORNAME|biofeng.com|
FEATURES             Location/Qualifiers
     CDS             3417..4112
                     /label=ZsGreen1
     misc_feature    2842..3416
                     /label=IRES
     LTR             1..635
                     /label=5'\LTR
     misc_feature    685..822
                     /label=packaging\signal
     misc_feature    2028..2151
                     /label=cPPT/CTS\(DNA\flap)
     promoter        2185..2787
                     /label=CMV\IE\Promoter
     misc_feature    4126..4717
                     /label=WPRE
     rep_origin      complement(6025..6698)
                     /label=pUC\origin
     CDS             complement(6843..7839)
                     /label=Amp(R)
     misc_binding    1303..1536
                     /label=RRE
     misc_binding    636..653
                     /label=PBS
     LTR             4920..5556
                     /label=3'\LTR
     misc_feature    2803..2840
                     /label=MCS
BASE COUNT     2132 a      2010 c      2086 g      1976 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 tctatttccg gtgaattcct cgagactagt 
     2821 tctagagcgg ccgcggatcc cgcccctctc cctccccccc ccctaacgtt actggccgaa 
     2881 gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt 
     2941 cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg 
     3001 gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc 
     3061 ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc 
     3121 ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa 
     3181 aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc 
     3241 tcacctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg 
     3301 gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaacg 
     3361 tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataatatgg 
     3421 cccagtccaa gcacggcctg accaaggaga tgaccatgaa gtaccgcatg gagggctgcg 
     3481 tggacggcca caagttcgtg atcaccggcg agggcatcgg ctaccccttc aagggcaagc 
     3541 aggccatcaa cctgtgcgtg gtggagggcg gccccttgcc cttcgccgag gacatcttgt 
     3601 ccgccgcctt catgtacggc aaccgcgtgt tcaccgagta cccccaggac atcgtcgact 
     3661 acttcaagaa ctcctgcccc gccggctaca cctgggaccg ctccttcctg ttcgaggacg 
     3721 gcgccgtgtg catctgcaac gccgacatca ccgtgagcgt ggaggagaac tgcatgtacc 
     3781 acgagtccaa gttctacggc gtgaacttcc ccgccgacgg ccccgtgatg aagaagatga 
     3841 ccgacaactg ggagccctcc tgcgagaaga tcatccccgt gcccaagcag ggcatcttga 
     3901 agggcgacgt gagcatgtac ctgctgctga aggacggtgg ccgcttgcgc tgccagttcg 
     3961 acaccgtgta caaggccaag tccgtgcccc gcaagatgcc cgactggcac ttcatccagc 
     4021 acaagctgac ccgcgaggac cgcagcgacg ccaagaacca gaagtggcac ctgaccgagc 
     4081 acgccatcgc ctccggctcc gccttgccct gaacgcgtct ggaacaatca acctctggat 
     4141 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt 
     4201 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc 
     4261 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg 
     4321 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc 
     4381 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa 
     4441 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat 
     4501 tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc 
     4561 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt 
     4621 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag 
     4681 acgagtcgga tctccctttg ggccgcctcc ccgcctggaa ttaattctgc agtcgagacc 
     4741 tagaaaaaca tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct 
     4801 ggctagaagc acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa 
     4861 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aagaggggac 
     4921 tggaagggct aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca 
     4981 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac 
     5041 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca 
     5101 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg 
     5161 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag 
     5221 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg 
     5281 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat 
     5341 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga 
     5401 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 
     5461 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 
     5521 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt 
     5581 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggccttg acattgctag 
     5641 cgtttaccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg 
     5701 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa 
     5761 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct 
     5821 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga 
     5881 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc 
     5941 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa 
     6001 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt 
     6061 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa 
     6121 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt 
     6181 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg 
     6241 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc 
     6301 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc 
     6361 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta 
     6421 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct 
     6481 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc 
     6541 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa 
     6601 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa 
     6661 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa 
     6721 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt 
     6781 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac 
     6841 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc 
     6901 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc 
     6961 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata 
     7021 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc 
     7081 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc 
     7141 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca 
     7201 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa 
     7261 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca 
     7321 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt 
     7381 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt 
     7441 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg 
     7501 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga 
     7561 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc 
     7621 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg 
     7681 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag 
     7741 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg 
     7801 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatcaactt 
     7861 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa 
     7921 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca 
     7981 tgtctggatc aactggataa ctcaagctaa ccaaaatcat cccaaacttc ccaccccata 
     8041 ccctattacc actgccaatt acctgtggtt tcatttactc taaacctgtg attcctctga 
     8101 attattttca ttttaaagaa attgtatttg ttaaatatgt actacaaact tagtagtttt 
     8161 taaagaaatt gtatttgtta aatatgtact acaaacttag tagt 
//

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