| 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 |
买家导航
载体基本信息
载体图谱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-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
//
