| 出品公司: | ZYbscience |
|---|---|
| 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: | 嘌呤霉素(Puromycin) |
| 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: | ZY632187 |
| 稳定性: | 稳表达 |
| 组成型: | 组成型 |
| 病毒/非病毒: | 慢病毒 |
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保存条件 ：-20℃低温保存
保质期 ：三年
英文名 ：pLVX-IRES-ZsGreen1
库存 ：20
供应商 ：泽叶生物
载体基本信息
载体质粒图谱和多克隆位点信息
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-2009COMMENT Created by Clontech Laboratories Inc.COMMENT Cat. No. 632187COMMENT 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=MCSBASE 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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