pLVX-mCherry-Actin

价格:3500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-mCherry-Actin
质粒类型: 慢病毒载体;荧光报告载体
高拷贝/低拷贝: 高拷贝
克隆方法: 限制性内切酶,多克隆位点
启动子: CMV
载体大小: 9893 bp
5' 测序引物及序列: CMV Forward: CGCAAATGGGCGGTAGGCGTG 
3' 测序引物及序列: --
载体标签: mCherry (N-端)
载体抗性: 氨苄青霉素
筛选标记: 嘌呤霉素(Puromycin)
克隆菌株: Stbl3
宿主细胞(系): 常规细胞系(293、CV-1、CHO等)
备注: 慢病毒载体pLVX-mCherry-Actin组成型表达红色荧光标记的肌动蛋白。
产品目录号: 631078
稳定性: 稳表达
组成型/诱导型: 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-mCherry-Actin载体图谱



pLVX-mCherry-Actin 载体特征

载体简介

pLVX-mCherry-Actin载体描述

pLVX-mCherry-Actin is an HIV-1-based, lentiviral expression vector that expresses human b-actin fused to mCherry, a mutant fluorescent protein derived from the tetrameric Discosoma sp. red fluorescent protein, DsRed (1). The excitation and emission maxima of the native mCherry protein are 587 nm and 610 nm, respectively. Expression of the mCherry-actin fusion protein is driven by the constitutively active human cytomegalovirus immediate early promoter (PCMV IE), located just upstream of the mCherry coding sequence. Lentiviral particles derived from the vector allow the expression of the mCherry-actin fusion protein in virtually any cell type, including primary cells.
pLVX-mCherry-Actin 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 (2), 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 (3). Finally, pLVX-mCherry-Actin 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).
In addition to lentiviral elements, pLVX-mCherry-Actin contains a puromycin resistance gene (Puror) 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.

pLVX-mCherry-Actin constitutively expresses an mCherry-actin fusion protein when transduced into target cells, where the fusion protein is incorporated into actin filaments, allowing the visualization of actin-containing subcellular structures in both live and fixed cells (5, 6). Note: pLVX-mCherry-Actin is not designed to be used as a cloning vector.
Before the vector can be transduced, 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 (7). If required, stable transfectants can be selected using puromycin.

Propagation in E. coli
 Suitable host strains: DH5α, DH10B and other general purpose strains.
 Selectable marker: plasmid confers resistance to ampicillin (100 μg/ml) in E. coli hosts.
 E. coli replication origin: ColE1
 Copy number: high

Excitation and emission maxima of mCherry
 Excitation maximum = 587 nm
 Emission maximum = 610 nm

载体序列

LOCUS       pLVX-mCherry-Actin   9899 bp    DNA   circular            21-DEC-2009
COMMENT     Created by Clontech Laboratories Inc. http://www.clontech.com
COMMENT     Cat. No. 631076,631078
FEATURES             Location/Qualifiers
     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    5867..6458
                     /label=WPRE
     rep_origin      complement(7767..8440)
                     /label=pUC\origin
     CDS             complement(8585..9581)
                     /label=Amp(R)
     misc_binding    1303..1536
                     /label=RRE
     misc_binding    636..653
                     /label=PBS
     misc_feature    2031..2046
                     /label=cPPT
     LTR             6661..7297
                     /label=LTR
     misc_feature    4725..5233
                     /label=PGK\promoter
     misc_feature    5254..5853
                     /label=Puro(R)
     CDS             2861..3568
                     /label=mCherry
     CDS             3590..4714
                     /label=Actin
BASE COUNT     2413 a      2641 c      2639 g      2206 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 tcgctagcgc taccggactc agatctcgag 
     2821 ctcaagcttc gaattctgca gtcgacggta ccgcgggccc atggtgagca agggcgagga 
     2881 ggataacatg gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt 
     2941 gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca 
     3001 gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc 
     3061 ccctcagttc atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta 
     3121 cttgaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg 
     3181 cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt 
     3241 gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg 
     3301 ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa 
     3361 gcagaggctg aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa 
     3421 ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac 
     3481 ctcccacaac gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc 
     3541 caccggcggc atggacgagc tgtacaagtc cggactcaga tctcgagcta tggatgatga 
     3601 tatcgccgcg ctcgtcgtcg acaacggctc cggcatgtgc aaggccggct tcgcgggcga 
     3661 cgatgccccc cgggccgtct tcccctccat cgtggggcgc cccaggcacc agggcgtgat 
     3721 ggtgggcatg ggtcagaagg attcctatgt gggcgacgag gcccagagca agagaggcat 
     3781 cctcaccctg aagtacccca tcgagcacgg catcgtcacc aactgggacg acatggagaa 
     3841 aatctggcac cacaccttct acaatgagct gcgtgtggct cccgaggagc accccgtgct 
     3901 gctgaccgag gcccccctga accccaaggc caaccgcgag aagatgaccc agatcatgtt 
     3961 tgagaccttc aacaccccag ccatgtacgt tgctatccag gctgtgctat ccctgtacgc 
     4021 ctctggccgt accactggca tcgtgatgga ctccggtgac ggggtcaccc acactgtgcc 
     4081 catctacgag gggtatgccc tcccccatgc catcctgcgt ctggacctgg ctggccggga 
     4141 cctgactgac tacctcatga agatcctcac cgagcgcggc tacagcttca ccaccacggc 
     4201 cgagcgggaa atcgtgcgtg acattaagga gaagctgtgc tacgtcgccc tggacttcga 
     4261 gcaagagatg gccacggctg cttccagctc ctccctggag aagagctacg agctgcctga 
     4321 cggccaggtc atcaccattg gcaatgagcg gttccgctgc cctgaggcac tcttccagcc 
     4381 ttccttcctg ggcatggagt cctgtggcat ccacgaaact accttcaact ccatcatgaa 
     4441 gtgtgacgtg gacatccgca aagacctgta cgccaacaca gtgctgtctg gcggcaccac 
     4501 catgtaccct ggcattgccg acaggatgca gaaggagatc actgccctgg cacccagcac 
     4561 aatgaagatc aagatcattg ctcctcctga gcgcaagtac tccgtgtgga tcggcggctc 
     4621 catcctggcc tcgctgtcca ccttccagca gatgtggatc agcaagcagg agtatgacga 
     4681 gtccggcccc tccatcgtcc accgcaaatg cttctagtct agataattct accgggtagg 
     4741 ggaggcgctt ttcccaaggc agtctggagc atgcgcttta gcagccccgc tgggcacttg 
     4801 gcgctacaca agtggcctct ggcctcgcac acattccaca tccaccggta ggcgccaacc 
     4861 ggctccgttc tttggtggcc ccttcgcgcc accttctact cctcccctag tcaggaagtt 
     4921 cccccccgcc ccgcagctcg cgtcgtgcag gacgtgacaa atggaagtag cacgtctcac 
     4981 tagtctcgtg cagatggaca gcaccgctga gcaatggaag cgggtaggcc tttggggcag 
     5041 cggccaatag cagctttgct ccttcgcttt ctgggctcag aggctgggaa ggggtgggtc 
     5101 cgggggcggg ctcaggggcg ggctcagggg cggggcgggc gcccgaaggt cctccggagg 
     5161 cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg ctgttctcct cttcctcatc 
     5221 tccgggcctt tcgacctgca gcccaagctt accatgaccg agtacaagcc cacggtgcgc 
     5281 ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac 
     5341 taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg 
     5401 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac 
     5461 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc 
     5521 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg 
     5581 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc 
     5641 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag 
     5701 gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc 
     5761 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc 
     5821 acctggtgca tgacccgcaa gcccggtgcc tgaccgcgtc tggaacaatc aacctctgga 
     5881 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg 
     5941 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt 
     6001 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag 
     6061 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc 
     6121 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga 
     6181 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa 
     6241 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac 
     6301 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct 
     6361 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca 
     6421 gacgagtcgg atctcccttt gggccgcctc cccgcctgga attaattctg cagtcgagac 
     6481 ctagaaaaac atggagcaat cacaagtagc aatacagcag ctaccaatgc tgattgtgcc 
     6541 tggctagaag cacaagagga ggaggaggtg ggttttccag tcacacctca ggtaccttta 
     6601 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagagggga 
     6661 ctggaagggc taattcactc ccaacgaaga caagatatcc ttgatctgtg gatctaccac 
     6721 acacaaggct acttccctga ttagcagaac tacacaccag ggccaggggt cagatatcca 
     6781 ctgacctttg gatggtgcta caagctagta ccagttgagc cagataaggt agaagaggcc 
     6841 aataaaggag agaacaccag cttgttacac cctgtgagcc tgcatgggat ggatgacccg 
     6901 gagagagaag tgttagagtg gaggtttgac agccgcctag catttcatca cgtggcccga 
     6961 gagctgcatc cggagtactt caagaactgc tgatatcgag cttgctacaa gggactttcc 
     7021 gctggggact ttccagggag gcgtggcctg ggcgggactg gggagtggcg agccctcaga 
     7081 tcctgcatat aagcagctgc tttttgcctg tactgggtct ctctggttag accagatctg 
     7141 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc 
     7201 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct 
     7261 cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat 
     7321 tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggcctt gacattgcta 
     7381 gcgttttacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg 
     7441 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta 
     7501 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg 
     7561 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga 
     7621 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg 
     7681 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag 
     7741 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc 
     7801 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca 
     7861 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt 
     7921 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc 
     7981 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc 
     8041 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc 
     8101 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact 
     8161 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg 
     8221 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta 
     8281 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca 
     8341 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa 
     8401 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg 
     8461 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc 
     8521 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg 
     8581 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat 
     8641 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg 
     8701 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa 
     8761 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca 
     8821 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc 
     8881 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt 
     8941 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa 
     9001 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat 
     9061 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct 
     9121 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga 
     9181 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag 
     9241 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga 
     9301 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca 
     9361 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg 
     9421 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc 
     9481 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag 
     9541 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatcaac 
     9601 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat 
     9661 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat 
     9721 catgtctgga tcaactggat aactcaagct aaccaaaatc atcccaaact tcccacccca 
     9781 taccctatta ccactgccaa ttacctgtgg tttcatttac tctaaacctg tgattcctct 
     9841 gaattatttt cattttaaag aaattgtatt tgttaaatat gtactacaaa cttagtagt 
//

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