pLVX-tdTomato-N1

价格:2000元

联系方式:I47-825O-882O

买家导航

载体基本信息

出品公司: Clontech
载体名称: pLVX-tdTomato-N1
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: 9498 bp
5' 测序引物及序列: --
3' 测序引物及序列: --
载体标签: tdTomato (C-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: --
产品目录号: 632563
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-tdTomato-N1载体图谱



pLVX-tdTomato-N1多克隆位点

pLVX-tdTomato-N1载体特征

载体简介

pLVX-tdTomato-N1载体描述

pLVX-tdTomato-N1 is an HIV-1-based, lentiviral expression vector that allows you to express your gene of interest fused to tdTomato, a member of the family of fruit fluorescent proteins (1) derived from the Discosoma sp. red fluorescent protein, DsRed (2). Genes cloned into the multiple cloning site (MCS), located upstream of the tdTomato coding sequence, are expressed as N-terminal tdTomato fusion proteins. Expression of the fusion protein is driven by the constitutively active human cytomegalovirus immediate early promoter (PCMV IE), located just upstream of the MCS. Lentiviral particles derived from the vector allow the expression of tdTomato fusion proteins in virtually any cell type, including primary cells. The unmodified vector expresses tdTomato, and may be used to produce marker virus to optimize infection protocols.
pLVX-tdTomato-N1 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 (3), 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 (4). Finally, pLVX-tdTomato-N1 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 (5).

In addition to lentiviral elements, pLVX-tdTomato-N1 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.

To construct a fusion protein, the gene of interest must be cloned into pLVX-tdTomato-N1 so that it is in-frame with the tdTomato coding sequence. The inserted sequence should include an initiation codon (ATG) and lack in-frame stop codons.The fusion protein is constitutively expressed when pLVX-tdTomato-N1 is transduced into target cells. Before the vector can be transduced, however, 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 (6).

Selection of Stable Transfectants
 Selectable marker: plasmid confers resistance to 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: pUC
 Copy number: high

Excitation and emission maxima of tdTomato
 Excitation maximum = 554 nm
 Emission maximum = 581 nm

载体序列

LOCUS       pLVX-tdTomato-N1   9498 bp    DNA   circular             7-OCT-2009
COMMENT     Created by Clontech Laboratories Inc.
COMMENT     Cat. No. 632563
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_feature    5466..6057
                     /label=WPRE
     CDS             complement(8184..9180)
                     /label=Amp(R)
     misc_binding    1303..1536
                     /label=RRE
     misc_binding    636..653
                     /label=PBS
     LTR             6260..6896
                     /label=3'\LTR
     promoter        4324..4832
                     /label=PGK\Promoter
     CDS             4853..5452
                     /label=Puro(R)
     CDS             2875..4305
                     /label=tdTomato
     misc_feature    2028..2151
                     /label=cPPT/CTS\(DNA\flap)
     promoter        2185..2787
                     /label=CMV\IE\Promoter
     rep_origin      complement(7366..8039)
                     /label=pUC\ori
     misc_feature    2815..2867
                     /label=MCS
BASE COUNT     2330 a      2523 c      2546 g      2099 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 gggatccacc ggtcatggtg 
     2881 agcaagggcg aggaggtcat caaagagttc atgcgcttca aggtgcgcat ggagggctcc 
     2941 atgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc 
     3001 cagaccgcca agctgaaggt gaccaagggc ggccccctgc ccttcgcctg ggacatcctg 
     3061 tccccccagt tcatgtacgg ctccaaggcg tacgtgaagc accccgccga catccccgat 
     3121 tacaagaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac 
     3181 ggcggtctgg tgaccgtgac ccaggactcc tccctgcagg acggcacgct gatctacaag 
     3241 gtgaagatgc gcggcaccaa cttccccccc gacggccccg taatgcagaa gaagaccatg 
     3301 ggctgggagg cctccaccga gcgcctgtac ccccgcgacg gcgtgctgaa gggcgagatc 
     3361 caccaggccc tgaagctgaa ggacggcggc cactacctgg tggagttcaa gaccatctac 
     3421 atggccaaga agcccgtgca actgcccggc tactactacg tggacaccaa gctggacatc 
     3481 acctcccaca acgaggacta caccatcgtg gaacagtacg agcgctccga gggccgccac 
     3541 cacctgttcc tggggcatgg caccggcagc accggcagcg gcagctccgg caccgcctcc 
     3601 tccgaggaca acaacatggc cgtcatcaaa gagttcatgc gcttcaaggt gcgcatggag 
     3661 ggctccatga acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag 
     3721 ggcacccaga ccgccaagct gaaggtgacc aagggcggcc ccctgccctt cgcctgggac 
     3781 atcctgtccc cccagttcat gtacggctcc aaggcgtacg tgaagcaccc cgccgacatc 
     3841 cccgattaca agaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc 
     3901 gaggacggcg gtctggtgac cgtgacccag gactcctccc tgcaggacgg cacgctgatc 
     3961 tacaaggtga agatgcgcgg caccaacttc ccccccgacg gccccgtaat gcagaagaag 
     4021 accatgggct gggaggcctc caccgagcgc ctgtaccccc gcgacggcgt gctgaagggc 
     4081 gagatccacc aggccctgaa gctgaaggac ggcggccact acctggtgga gttcaagacc 
     4141 atctacatgg ccaagaagcc cgtgcaactg cccggctact actacgtgga caccaagctg 
     4201 gacatcacct cccacaacga ggactacacc atcgtggaac agtacgagcg ctccgagggc 
     4261 cgccaccacc tgttcctgta cggcatggac gagctgtaca agtgagcggc cgcgactcta 
     4321 gataattcta ccgggtaggg gaggcgcttt tcccaaggca gtctggagca tgcgctttag 
     4381 cagccccgct gggcacttgg cgctacacaa gtggcctctg gcctcgcaca cattccacat 
     4441 ccaccggtag gcgccaaccg gctccgttct ttggtggccc cttcgcgcca ccttctactc 
     4501 ctcccctagt caggaagttc ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa 
     4561 tggaagtagc acgtctcact agtctcgtgc agatggacag caccgctgag caatggaagc 
     4621 gggtaggcct ttggggcagc ggccaatagc agctttgctc cttcgctttc tgggctcaga 
     4681 ggctgggaag gggtgggtcc gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg 
     4741 cccgaaggtc ctccggaggc ccggcattct gcacgcttca aaagcgcacg tctgccgcgc 
     4801 tgttctcctc ttcctcatct ccgggccttt cgacctgcag cccaagctta ccatgaccga 
     4861 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct 
     4921 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat 
     4981 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa 
     5041 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga 
     5101 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct 
     5161 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg 
     5221 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt 
     5281 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc 
     5341 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga 
     5401 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaccgcgtct 
     5461 ggaacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg 
     5521 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt 
     5581 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg 
     5641 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc 
     5701 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc 
     5761 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc 
     5821 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc 
     5881 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg 
     5941 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc 
     6001 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa 
     6061 ttaattctgc agtcgagacc tagaaaaaca tggagcaatc acaagtagca atacagcagc 
     6121 taccaatgct gattgtgcct ggctagaagc acaagaggag gaggaggtgg gttttccagt 
     6181 cacacctcag gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt 
     6241 tttaaaagaa aagaggggac tggaagggct aattcactcc caacgaagac aagatatcct 
     6301 tgatctgtgg atctaccaca cacaaggcta cttccctgat tagcagaact acacaccagg 
     6361 gccaggggtc agatatccac tgacctttgg atggtgctac aagctagtac cagttgagcc 
     6421 agataaggta gaagaggcca ataaaggaga gaacaccagc ttgttacacc ctgtgagcct 
     6481 gcatgggatg gatgacccgg agagagaagt gttagagtgg aggtttgaca gccgcctagc 
     6541 atttcatcac gtggcccgag agctgcatcc ggagtacttc aagaactgct gatatcgagc 
     6601 ttgctacaag ggactttccg ctggggactt tccagggagg cgtggcctgg gcgggactgg 
     6661 ggagtggcga gccctcagat cctgcatata agcagctgct ttttgcctgt actgggtctc 
     6721 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 
     6781 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 
     6841 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta 
     6901 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg 
     6961 agaggccttg acattgctag cgttttaccg tcgacctcta gctagagctt ggcgtaatca 
     7021 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga 
     7081 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt 
     7141 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga 
     7201 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc 
     7261 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg 
     7321 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc 
     7381 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc 
     7441 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga 
     7501 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc 
     7561 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat 
     7621 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg 
     7681 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc 
     7741 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga 
     7801 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact 
     7861 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt 
     7921 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag 
     7981 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg 
     8041 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa 
     8101 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata 
     8161 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg 
     8221 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata 
     8281 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg 
     8341 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct 
     8401 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt 
     8461 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc 
     8521 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga 
     8581 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt 
     8641 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc 
     8701 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa 
     8761 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca 
     8821 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca 
     8881 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct 
     8941 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc 
     9001 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa 
     9061 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt 
     9121 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc 
     9181 gacggatcgg gagatcaact tgtttattgc agcttataat ggttacaaat aaagcaatag 
     9241 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa 
     9301 actcatcaat gtatcttatc atgtctggat caactggata actcaagcta accaaaatca 
     9361 tcccaaactt cccaccccat accctattac cactgccaat tacctgtggt ttcatttact 
     9421 ctaaacctgt gattcctctg aattattttc attttaaaga aattgtattt gttaaatatg 
     9481 tactacaaac ttagtagt 
//

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