出品公司: | Clontech |
---|---|
载体名称: | pLVX-IRES-mCherry |
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: | 8172 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-mCherry载体以双顺反子的形式同时表达mCherry荧光蛋白和目的基因; CMV启动子驱动目的基因的过表达; mCherry是第三代荧光蛋白,亮度高,适用于流式细胞仪。 |
产品目录号: | 631237 |
稳定性: | 稳表达 |
组成型: | 诱导型 |
病毒/非病毒: | 慢病毒 |
买家导航
pLVX-IRES-mCherry载体质粒基本信息
pLVX-IRES-mCherry质粒图谱载体图谱和pLVX-IRES-mCherry载体序列质粒序列多克隆位点信息
pLVX-IRES-mCherry质粒载体简介
Description
pLVX-IRES-mCherry is an HIV-1-based, lentiviral expression vector that allows the simultaneous expression of your protein of interest and mCherry in virtually any mammalian cell type, including primary cells. mCherry is a mutant fluorescent protein derived from the tetrameric Discosoma sp. red fluorescent protein, DsRed (1). The vector expresses the two proteins from a bicistronic mRNA transcript, allowing mCherry 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 mCherry, facilitates cap-independent translation of mCherry from an internal start site at the IRES/mCherry junction (1).
pLVX-IRES-mCherry 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-mCherry 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-mCherry is designed to constitutively coexpress your protein of interest and mCherry 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™ HT Packaging System (Cat. Nos. 632160 and 632161). 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).
The presence of mCherry allows transductants to be visualized by fluorescence microscopy and sorted by flow cytometry with standard FITC filter sets (mCherry has an excitation maximum of 587 nm and an emission maximum of 610 nm).
pLVX-IRES-mCherry质粒序列载体序列
LOCUS pLVX-IRES-mCherry 8172 bp DNA circular 28-MAY-2009 COMMENT Created by Clontech Laboratories Inc. COMMENT Cat. No. 631237 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_binding 1303..1536 /label=RRE promoter 2185..2787 /label=CMV\IE\Promoter misc_feature 2842..3416 /label=IRES misc_feature 4141..4732 /label=WPRE LTR 4935..5571 /label=3'\LTR CDS 3417..4127 /label=mCherry rep_origin complement(6040..6713) /label=pUC\ori CDS complement(6858..7854) /label=Amp(R) misc_feature 636..653 /label=PBS misc_feature 2028..2151 /label=cPPT/CTS\(DNA\flap) misc_feature 2803..2840 /label=MCS BASE COUNT 2133 a 1995 c 2099 g 1945 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 tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc 3481 acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc 3541 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg 3601 cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg 3661 ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga 3721 tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg 3781 agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc 3841 agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc 3901 tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg 3961 tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca 4021 tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg 4081 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtgaacg cgtctggaac 4141 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct 4201 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt 4261 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg 4321 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact 4381 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct 4441 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg 4501 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc 4561 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc 4621 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt 4681 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaattaat 4741 tctgcagtcg agacctagaa aaacatggag caatcacaag tagcaataca gcagctacca 4801 atgctgattg tgcctggcta gaagcacaag aggaggagga ggtgggtttt ccagtcacac 4861 ctcaggtacc tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa 4921 aagaaaagag gggactggaa gggctaattc actcccaacg aagacaagat atccttgatc 4981 tgtggatcta ccacacacaa ggctacttcc ctgattagca gaactacaca ccagggccag 5041 gggtcagata tccactgacc tttggatggt gctacaagct agtaccagtt gagccagata 5101 aggtagaaga ggccaataaa ggagagaaca ccagcttgtt acaccctgtg agcctgcatg 5161 ggatggatga cccggagaga gaagtgttag agtggaggtt tgacagccgc ctagcatttc 5221 atcacgtggc ccgagagctg catccggagt acttcaagaa ctgctgatat cgagcttgct 5281 acaagggact ttccgctggg gactttccag ggaggcgtgg cctgggcggg actggggagt 5341 ggcgagccct cagatcctgc atataagcag ctgctttttg cctgtactgg gtctctctgg 5401 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct 5461 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt 5521 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tagtagttca 5581 tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg 5641 ccttgacatt gctagcgttt accgtcgacc tctagctaga gcttggcgta atcatggtca 5701 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga 5761 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg 5821 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc 5881 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac 5941 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata 6001 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa 6061 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct 6121 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa 6181 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg 6241 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca 6301 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa 6361 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg 6421 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg 6481 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga 6541 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc 6601 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag 6661 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac 6721 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc 6781 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag 6841 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt 6901 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag 6961 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca 7021 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact 7081 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca 7141 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg 7201 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc 7261 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg 7321 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca 7381 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt 7441 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc 7501 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc 7561 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca 7621 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa 7681 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat 7741 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa 7801 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga 7861 tcgggagatc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac 7921 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat 7981 caatgtatct tatcatgtct ggatcaactg gataactcaa gctaaccaaa atcatcccaa 8041 acttcccacc ccatacccta ttaccactgc caattacctg tggtttcatt tactctaaac 8101 ctgtgattcc tctgaattat tttcatttta aagaaattgt atttgttaaa tatgtactac 8161 aaacttagta gt //
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pLVX-IRES-mCherry质粒载体应用举例
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