pLVX-AmCyan1-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-AmCyan1-C1, pLVX AmCyan1 C1
质粒类型: 慢病毒表达载体;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: 8757 bp
5' 测序引物及序列: --
3' 测序引物及序列: --
载体标签: AmCyan1(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-AmCyan1-C1载体表达N端AmCyan1融合蛋白;
青色荧光蛋白AmCyan1具有高度可溶性,高亮度,同时与其它荧光蛋白颜色区别大。
产品目录号: 632557
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-AmCyan1-C1载体图谱



pLVX-AmCyan1-C1多克隆位点

pLVX-AmCyan1-C1载体特征

载体简介

pLVX-AmCyan1-C1 is an HIV-1-based, lentiviral expression vector that allows you to express your gene of interest fused to AmCyan1, a human codon-optimized variant of the wild-type Anemonia majano cyan fluorescent protein (1). Genes cloned into the multiple cloning site (MCS), located at the C-terminal end of the AmCyan1 coding sequence, are expressed as C-terminal AmCyan1 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 AmCyan1 coding sequence. Lentiviral particles derived from the vector allow the expression of AmCyan1 fusion proteins in virtually any cell type, including primary cells. The unmodified vector expresses AmCyan1, and may be used to produce marker virus to optimize infection protocols.
pLVX-AmCyan1-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-AmCyan1-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-AmCyan1-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.
To construct a fusion protein, the gene of interest must be cloned into pLVX-AmCyan1-C1 so that it is in-frame with the AmCyan1 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-AmCyan1-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).

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 AmCyan1
 Excitation maximum = 458 nm
 Emission maximum = 489 nm

载体序列

LOCUS       pLVX-AmCyan1-C1   8757 bp    DNA   circular             7-OCT-2009
COMMENT     Created by Clontech Laboratories Inc. 
COMMENT     Cat. No. 632557
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    4725..5316
                     /label=WPRE
     CDS             complement(7443..8439)
                     /label=Amp(R)
     misc_binding    1303..1536
                     /label=RRE
     misc_binding    636..653
                     /label=PBS
     LTR             5519..6155
                     /label=3'\LTR
     promoter        3583..4091
                     /label=PGK\Promoter
     CDS             4112..4711
                     /label=Puro(R)
     misc_feature    3568..3570
                     /label=Stop
     misc_feature    3572..3574
                     /label=Stop
     CDS             2807..3493
                     /label=AmCyan1
     misc_feature    2028..2151
                     /label=cPPT/CTS\(DNA\flap)
     promoter        2185..2787
                     /label=CMV\IE\Promoter
     rep_origin      complement(6625..7298)
                     /label=pUC\ori
     misc_feature    3576..3578
                     /label=Stop
     misc_feature    3507..3559
                     /label=MCS
BASE COUNT     2166 a      2296 c      2295 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 ccctgtccaa 
     2821 caagttcatc ggcgacgaca tgaagatgac ctaccacatg gacggctgcg tgaacggcca 
     2881 ctacttcacc gtgaagggcg agggcagcgg caagccctac gagggcaccc agacctccac 
     2941 cttcaaggtg accatggcca acggcggccc cctggccttc tccttcgaca tcctgtccac 
     3001 cgtgttcatg tacggcaacc gctgcttcac cgcctacccc accagcatgc ccgactactt 
     3061 caagcaggcc ttccccgacg gcatgtccta cgagagaacc ttcacctacg aggacggcgg 
     3121 cgtggccacc gccagctggg agatcagcct gaagggcaac tgcttcgagc acaagtccac 
     3181 cttccacggc gtgaacttcc ccgccgacgg ccccgtgatg gccaagaaga ccaccggctg 
     3241 ggacccctcc ttcgagaaga tgaccgtgtg cgacggcatc ttgaagggcg acgtgaccgc 
     3301 cttcctgatg ctgcagggcg gcggcaacta cagatgccag ttccacacct cctacaagac 
     3361 caagaagccc gtgaccatgc cccccaacca cgtggtggag caccgcatcg ccagaaccga 
     3421 cctggacaag ggcggcaaca gcgtgcagct gaccgagcac gccgtggccc acatcacctc 
     3481 cgtggtgccc ttctccggac tcagatctcg agctcaagct tcgaattctg cagtcgacgg 
     3541 taccgcgggc ccgggatcca ccggatctag ataactgatc ataattctac cgggtagggg 
     3601 aggcgctttt cccaaggcag tctggagcat gcgctttagc agccccgctg ggcacttggc 
     3661 gctacacaag tggcctctgg cctcgcacac attccacatc caccggtagg cgccaaccgg 
     3721 ctccgttctt tggtggcccc ttcgcgccac cttctactcc tcccctagtc aggaagttcc 
     3781 cccccgcccc gcagctcgcg tcgtgcagga cgtgacaaat ggaagtagca cgtctcacta 
     3841 gtctcgtgca gatggacagc accgctgagc aatggaagcg ggtaggcctt tggggcagcg 
     3901 gccaatagca gctttgctcc ttcgctttct gggctcagag gctgggaagg ggtgggtccg 
     3961 ggggcgggct caggggcggg ctcaggggcg gggcgggcgc ccgaaggtcc tccggaggcc 
     4021 cggcattctg cacgcttcaa aagcgcacgt ctgccgcgct gttctcctct tcctcatctc 
     4081 cgggcctttc gacctgcagc ccaagcttac catgaccgag tacaagccca cggtgcgcct 
     4141 cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta 
     4201 ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca 
     4261 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg 
     4321 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga 
     4381 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga 
     4441 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt 
     4501 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc 
     4561 ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt 
     4621 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac 
     4681 ctggtgcatg acccgcaagc ccggtgcctg accgcgtctg gaacaatcaa cctctggatt 
     4741 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg 
     4801 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct 
     4861 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc 
     4921 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca 
     4981 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac 
     5041 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt 
     5101 ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct 
     5161 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc 
     5221 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga 
     5281 cgagtcggat ctccctttgg gccgcctccc cgcctggaat taattctgca gtcgagacct 
     5341 agaaaaacat ggagcaatca caagtagcaa tacagcagct accaatgctg attgtgcctg 
     5401 gctagaagca caagaggagg aggaggtggg ttttccagtc acacctcagg tacctttaag 
     5461 accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa agaggggact 
     5521 ggaagggcta attcactccc aacgaagaca agatatcctt gatctgtgga tctaccacac 
     5581 acaaggctac ttccctgatt agcagaacta cacaccaggg ccaggggtca gatatccact 
     5641 gacctttgga tggtgctaca agctagtacc agttgagcca gataaggtag aagaggccaa 
     5701 taaaggagag aacaccagct tgttacaccc tgtgagcctg catgggatgg atgacccgga 
     5761 gagagaagtg ttagagtgga ggtttgacag ccgcctagca tttcatcacg tggcccgaga 
     5821 gctgcatccg gagtacttca agaactgctg atatcgagct tgctacaagg gactttccgc 
     5881 tggggacttt ccagggaggc gtggcctggg cgggactggg gagtggcgag ccctcagatc 
     5941 ctgcatataa gcagctgctt tttgcctgta ctgggtctct ctggttagac cagatctgag 
     6001 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt 
     6061 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca 
     6121 gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc 
     6181 agtatttata acttgcaaag aaatgaatat cagagagtga gaggccttga cattgctagc 
     6241 gttttaccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg 
     6301 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa 
     6361 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct 
     6421 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga 
     6481 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc 
     6541 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa 
     6601 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt 
     6661 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa 
     6721 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt 
     6781 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg 
     6841 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc 
     6901 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc 
     6961 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta 
     7021 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct 
     7081 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc 
     7141 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa 
     7201 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa 
     7261 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa 
     7321 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt 
     7381 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac 
     7441 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc 
     7501 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc 
     7561 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata 
     7621 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc 
     7681 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc 
     7741 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca 
     7801 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa 
     7861 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca 
     7921 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt 
     7981 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt 
     8041 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg 
     8101 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga 
     8161 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc 
     8221 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg 
     8281 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag 
     8341 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg 
     8401 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatcaactt 
     8461 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa 
     8521 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca 
     8581 tgtctggatc aactggataa ctcaagctaa ccaaaatcat cccaaacttc ccaccccata 
     8641 ccctattacc actgccaatt acctgtggtt tcatttactc taaacctgtg attcctctga 
     8701 attattttca ttttaaagaa attgtatttg ttaaatatgt actacaaact tagtagt 
//

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