pLVX-DsRed-Express2-C1

价格:2500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-DsRed-Express2-C1
质粒类型: 慢病毒载体;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: 8745 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: DsRed-Express2(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: pLVX-DsRed-Express2-C1载体表达N端DsRed-Express2融合蛋白;
DsRed-Express2是DsRed的突变体,
与DsRed相比,DsRed-Express2细胞毒性低,尤其适用于原代细胞和干细胞的荧光标记
产品目录号: 632559
稳定性: 稳表达
组成型: 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-DsRed-Express2-C1载体图谱



pLVX-DsRed-Express2-C1多克隆位点

pLVX-DsRed-Express2-C1载体特征

载体简介

Description
pLVX-DsRed-Express2-C1 is an HIV-1-based, lentiviral expression vector that allows you to express your gene of interest fused to DsRed-Express2, a mutant fluorescent protein derived from the tetrameric Discosoma sp. red fluorescent protein, DsRed (1). Genes cloned into the multiple cloning site (MCS), located at the C-terminal end of the DsRed-Express2 coding sequence, are expressed as C-terminal DsRed-Express2 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 DsRed-Express2 coding sequence. Lentiviral particles derived from the vector allow the expression of DsRed-Express2 fusion proteins in virtually any cell type, including primary cells. The unmodified vector expresses DsRed-Express2, and may be used to produce marker virus to optimize infection protocols.

pLVX-DsRed-Express2-C1 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-DsRed-Express2-C1 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-DsRed-Express2-C1 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.


Use
To construct a fusion protein, the gene of interest must be cloned into pLVX-DsRed-Express2-C1 so that it is in-frame with the DsRed-Express2 coding sequence. The inserted sequence does not require an initiation codon (ATG) or a stop codon (TAA, TAG, TGA); however, if you don't want to use the stop codons downstream of the MCS (see map), you can add a stop codon to the end of your gene of interest.

The fusion protein is constitutively expressed when pLVX-DsRed-Express2-C1 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 (5).

载体序列

LOCUS       pLVX-DsRed-Express2-C1   8745 bp    DNA   circular             7-OCT-2009
COMMENT     Created by Clontech Laboratories Inc. 
COMMENT     Cat. No. 632559
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    4713..5304
                     /label=WPRE
     CDS             complement(7431..8427)
                     /label=Amp(R)
     misc_binding    1303..1536
                     /label=RRE
     misc_binding    636..653
                     /label=PBS
     LTR             5507..6143
                     /label=3'\LTR
     promoter        3571..4079
                     /label=PGK\Promoter
     CDS             4100..4699
                     /label=Puro(R)
     misc_feature    3556..3558
                     /label=Stop
     misc_feature    3560..3562
                     /label=Stop
     CDS             2807..3481
                     /label=DsRed-Express2
     misc_feature    2028..2151
                     /label=cPPT/CTS\(DNA\flap)
     promoter        2185..2787
                     /label=CMV\IE\Promoter
     rep_origin      complement(6613..7286)
                     /label=pUC\ori
     misc_feature    3564..3566
                     /label=Stop
     misc_feature    3495..3547
                     /label=MCS
BASE COUNT     2163 a      2270 c      2312 g      2000 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 tctaccggtc gccaccatgg atagcactga 
     2821 gaacgtcatc aagcccttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca 
     2881 cgagttcgag atcgagggcg agggcgaggg caagccctac gagggcaccc agaccgccaa 
     2941 gctgcaggtg accaagggcg gccccctgcc cttcgcctgg gacatcctgt ccccccagtt 
     3001 ccagtacggc tccaaggtgt acgtgaagca ccccgccgac atccccgact acaagaagct 
     3061 gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt 
     3121 gaccgtgacc caggactcct ccctgcagga cggcaccttc atctaccacg tgaagttcat 
     3181 cggcgtgaac ttcccctccg acggccccgt aatgcagaag aagactctgg gctgggagcc 
     3241 ctccaccgag cgcctgtacc cccgcgacgg cgtgctgaag ggcgagatcc acaaggcgct 
     3301 gaagctgaag ggcggcggcc actacctggt ggagttcaag tcaatctaca tggccaagaa 
     3361 gcccgtgaag ctgcccggct actactacgt ggactccaag ctggacatca cctcccacaa 
     3421 cgaggactac accgtggtgg agcagtacga gcgcgccgag gcccgccacc acctgttcca 
     3481 gtccggactc agatctcgag ctcaagcttc gaattctgca gtcgacggta ccgcgggccc 
     3541 gggatccacc ggatctagat aactgatcat aattctaccg ggtaggggag gcgcttttcc 
     3601 caaggcagtc tggagcatgc gctttagcag ccccgctggg cacttggcgc tacacaagtg 
     3661 gcctctggcc tcgcacacat tccacatcca ccggtaggcg ccaaccggct ccgttctttg 
     3721 gtggcccctt cgcgccacct tctactcctc ccctagtcag gaagttcccc cccgccccgc 
     3781 agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga 
     3841 tggacagcac cgctgagcaa tggaagcggg taggcctttg gggcagcggc caatagcagc 
     3901 tttgctcctt cgctttctgg gctcagaggc tgggaagggg tgggtccggg ggcgggctca 
     3961 ggggcgggct caggggcggg gcgggcgccc gaaggtcctc cggaggcccg gcattctgca 
     4021 cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg ggcctttcga 
     4081 cctgcagccc aagcttacca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga 
     4141 cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg 
     4201 ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct 
     4261 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc 
     4321 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg 
     4381 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc 
     4441 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca 
     4501 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc 
     4561 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct 
     4621 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac 
     4681 ccgcaagccc ggtgcctgac cgcgtctgga acaatcaacc tctggattac aaaatttgtg 
     4741 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt 
     4801 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata 
     4861 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg 
     4921 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc 
     4981 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct 
     5041 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt 
     5101 cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg 
     5161 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc 
     5221 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct 
     5281 ccctttgggc cgcctccccg cctggaatta attctgcagt cgagacctag aaaaacatgg 
     5341 agcaatcaca agtagcaata cagcagctac caatgctgat tgtgcctggc tagaagcaca 
     5401 agaggaggag gaggtgggtt ttccagtcac acctcaggta cctttaagac caatgactta 
     5461 caaggcagct gtagatctta gccacttttt aaaagaaaag aggggactgg aagggctaat 
     5521 tcactcccaa cgaagacaag atatccttga tctgtggatc taccacacac aaggctactt 
     5581 ccctgattag cagaactaca caccagggcc aggggtcaga tatccactga cctttggatg 
     5641 gtgctacaag ctagtaccag ttgagccaga taaggtagaa gaggccaata aaggagagaa 
     5701 caccagcttg ttacaccctg tgagcctgca tgggatggat gacccggaga gagaagtgtt 
     5761 agagtggagg tttgacagcc gcctagcatt tcatcacgtg gcccgagagc tgcatccgga 
     5821 gtacttcaag aactgctgat atcgagcttg ctacaaggga ctttccgctg gggactttcc 
     5881 agggaggcgt ggcctgggcg ggactgggga gtggcgagcc ctcagatcct gcatataagc 
     5941 agctgctttt tgcctgtact gggtctctct ggttagacca gatctgagcc tgggagctct 
     6001 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag 
     6061 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt 
     6121 cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag tatttataac 
     6181 ttgcaaagaa atgaatatca gagagtgaga ggccttgaca ttgctagcgt tttaccgtcg 
     6241 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat 
     6301 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc 
     6361 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga 
     6421 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt 
     6481 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg 
     6541 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac 
     6601 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg 
     6661 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca 
     6721 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc 
     6781 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc 
     6841 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag 
     6901 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc 
     6961 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca 
     7021 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg 
     7081 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg 
     7141 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct 
     7201 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa 
     7261 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa 
     7321 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa 
     7381 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc 
     7441 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga 
     7501 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca 
     7561 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc 
     7621 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat 
     7681 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc 
     7741 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt 
     7801 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc 
     7861 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg 
     7921 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt 
     7981 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg 
     8041 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga 
     8101 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg 
     8161 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg 
     8221 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt 
     8281 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc 
     8341 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca 
     8401 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atcaacttgt ttattgcagc 
     8461 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc 
     8521 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatcaa 
     8581 ctggataact caagctaacc aaaatcatcc caaacttccc accccatacc ctattaccac 
     8641 tgccaattac ctgtggtttc atttactcta aacctgtgat tcctctgaat tattttcatt 
     8701 ttaaagaaat tgtatttgtt aaatatgtac tacaaactta gtagt 
//

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