出品公司: | Clontech |
---|---|
载体名称: | pLVX-AmCyan1-N1, pLVX AmCyan1 N1 |
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 IE |
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(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: |
pLVX-AmCyan1-N1载体表达C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: | 632558 |
稳定性: | 稳表达 |
组成型: | 组成型 |
病毒/非病毒: | 慢病毒 |
买家导航
pLVX-AmCyan1-N1载体质粒基本信息
pLVX-AmCyan1-N1质粒图谱载体图谱和pLVX-AmCyan1-N1载体序列质粒序列多克隆位点信息
pLVX-AmCyan1-N1质粒载体简介
pLVX-AmCyan1-N1载体描述
pLVX-AmCyan1-N1 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 upstream of the AmCyan1 coding sequence, are expressed as N-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 MCS. 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-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 (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-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 (4).
In addition to lentiviral elements, pLVX-AmCyan1-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-AmCyan1-N1 so that it is in-frame with the AmCyan1 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-AmCyan1-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 (5).
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 AmCyan1
Excitation maximum = 458 nm
Emission maximum = 489 nm
pLVX-AmCyan1-N1质粒序列载体序列
LOCUS pLVX-AmCyan1-N1 8757 bp DNA circular 7-OCT-2009 COMMENT Created by Clontech Laboratories Inc. COMMENT Cat. No. 632558 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) CDS 2875..3564 /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 2815..2867 /label=MCS BASE COUNT 2163 a 2296 c 2300 g 1998 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 ggtcatggcc 2881 ctgtccaaca agttcatcgg cgacgacatg aagatgacct accacatgga cggctgcgtg 2941 aacggccact acttcaccgt gaagggcgag ggcagcggca agccctacga gggcacccag 3001 acctccacct tcaaggtgac catggccaac ggcggccccc tggccttctc cttcgacatc 3061 ctgtccaccg tgttcatgta cggcaaccgc tgcttcaccg cctaccccac cagcatgccc 3121 gactacttca agcaggcctt ccccgacggc atgtcctacg agagaacctt cacctacgag 3181 gacggcggcg tggccaccgc cagctgggag atcagcctga agggcaactg cttcgagcac 3241 aagtccacct tccacggcgt gaacttcccc gccgacggcc ccgtgatggc caagaagacc 3301 accggctggg acccctcctt cgagaagatg accgtgtgcg acggcatctt gaagggcgac 3361 gtgaccgcct tcctgatgct gcagggcggc ggcaactaca gatgccagtt ccacacctcc 3421 tacaagacca agaagcccgt gaccatgccc cccaaccacg tggtggagca ccgcatcgcc 3481 agaaccgacc tggacaaggg cggcaacagc gtgcagctga ccgagcacgc cgtggcccac 3541 atcacctccg tggtgccctt ctgagcggcc gcgactctag 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 //