出品公司: | Clontech |
---|---|
载体名称: | pLVX-DsRed-Monomer-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 |
克隆方法: | 多克隆位点,限制性内切酶 |
载体大小: | 8751 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-Monomer(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-DsRed-Monomer-N1载体表达C端DsRed-Monomer融合蛋白; DsRed-Monomer是DsRed的单体突变体,与DsRed相比,编码序列经过了人工优化,适用于在哺乳动物细胞中的高水平表达; CMV启动子是过表达启动子。 |
产品目录号: | 632152 |
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-DsRed-Monomer-N1载体质粒基本信息
pLVX-DsRed-Monomer-N1质粒图谱载体图谱和pLVX-DsRed-Monomer-N1载体序列质粒序列多克隆位点信息
pLVX-DsRed-Monomer-N1质粒载体简介
Description
pLVX-DsRed-Monomer-N1 is an HIV-1-based, lentiviral expression vector that allows you to express your gene of interest fused to DsRed-Monomer, a monomeric mutant of the Discosoma sp. red fluorescent protein. Genes cloned into the multiple cloning site (MCS),located upstream of the DsRed-Monomer coding sequence, are expressed as N-terminal fusions of the DsRed-Monomer protein. 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 DsRed-Monomer fusion proteins in virtually any cell type, including primary cells. The unmodified vector expresses DsRed-Monomer, and may be used to produce marker virus to
optimize infection protocols.
pLVX-DsRed-Monomer-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 nhances nuclear export of viral and transgene RNA (1), 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 (2). Finally, pLVX-DsRed-Monomer-N1 also contains a central polypurine tract (cPPT) element that increases nuclear importation of the viral genome during target cell infection, resulting in improved vector integration and more efficient transduction (3).
In addition to lentiviral elements, pLVX-DsRed-Monomer-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.
Use
pLVX-DsRed-Monomer-N1 constitutively expresses your gene of interest from PCMV IE when transduced into target cells. Before the vector can be transduced into cells, however, it must be transfected into 293T packaging cells with our Lenti-X™ HTX Packaging System (Cat. Nos. 631247 and 631249). 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 (4).
pLVX-DsRed-Monomer-N1质粒序列载体序列
LOCUS pLVX-DsRed-Monomer-N1 8751 bp DNA circular 9-NOV-2009 COMMENT Created by Clontech Laboratories Inc. COMMENT Cat. No. 632152 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 2028..2151 /label=cPPT/CTS promoter 2185..2787 /label=CMV\IE\Promoter misc_feature 4720..5311 /label=WPRE rep_origin complement(6619..7292) /label=pUC\origin CDS complement(7437..8433) /label=Amp(R) misc_binding 1303..1536 /label=RRE misc_binding 636..653 /label=PBS LTR 5514..6150 /label=3'\LTR misc_feature 2814..2868 /label=MCS CDS 2881..3558 /label=DsRed-Monomer promoter 3578..4086 /label=PGK\Promoter CDS 4107..4706 /label=Puro(R) BASE COUNT 2168 a 2279 c 2317 g 1987 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 ggtcgccacc 2881 atggacaaca ccgaggacgt catcaaggag ttcatgcagt tcaaggtgcg catggagggc 2941 tccgtgaacg gccactactt cgagatcgag ggcgagggcg agggcaagcc ctacgagggc 3001 acccagaccg ccaagctgca ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc 3061 ctgtcccccc agttccagta cggctccaag gcctacgtga agcaccccgc cgacatcccc 3121 gactacatga agctgtcctt ccccgagggc ttcacctggg agcgctccat gaacttcgag 3181 gacggcggcg tggtggaggt gcagcaggac tcctccctgc aggacggcac cttcatctac 3241 aaggtgaagt tcaagggcgt gaacttcccc gccgacggcc ccgtaatgca gaagaagact 3301 gccggctggg agccctccac cgagaagctg tacccccagg acggcgtgct gaagggcgag 3361 atctcccacg ccctgaagct gaaggacggc ggccactaca cctgcgactt caagaccgtg 3421 tacaaggcca agaagcccgt gcagctgccc ggcaaccact acgtggactc caagctggac 3481 atcaccaacc acaacgagga ctacaccgtg gtggagcagt acgagcacgc cgaggcccgc 3541 cactccggct cccagtagag cggccgcgac tctagataat tctaccgggt aggggaggcg 3601 cttttcccaa ggcagtctgg agcatgcgct ttagcagccc cgctgggcac ttggcgctac 3661 acaagtggcc tctggcctcg cacacattcc acatccaccg gtaggcgcca accggctccg 3721 ttctttggtg gccccttcgc gccaccttct actcctcccc tagtcaggaa gttccccccc 3781 gccccgcagc tcgcgtcgtg caggacgtga caaatggaag tagcacgtct cactagtctc 3841 gtgcagatgg acagcaccgc tgagcaatgg aagcgggtag gcctttgggg cagcggccaa 3901 tagcagcttt gctccttcgc tttctgggct cagaggctgg gaaggggtgg gtccgggggc 3961 gggctcaggg gcgggctcag gggcggggcg ggcgcccgaa ggtcctccgg aggcccggca 4021 ttctgcacgc ttcaaaagcg cacgtctgcc gcgctgttct cctcttcctc atctccgggc 4081 ctttcgacct gcagcccaag cttaccatga ccgagtacaa gcccacggtg cgcctcgcca 4141 cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg 4201 ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac 4261 tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg 4321 cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg 4381 gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc 4441 tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc 4501 ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg 4561 agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg 4621 agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt 4681 gcatgacccg caagcccggt gcctgaccgc gtctggaaca atcaacctct ggattacaaa 4741 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac 4801 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc 4861 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt 4921 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc 4981 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc 5041 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg 5101 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt 5161 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc 5221 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt 5281 cggatctccc tttgggccgc ctccccgcct ggaattaatt ctgcagtcga gacctagaaa 5341 aacatggagc aatcacaagt agcaatacag cagctaccaa tgctgattgt gcctggctag 5401 aagcacaaga ggaggaggag gtgggttttc cagtcacacc tcaggtacct ttaagaccaa 5461 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagagg ggactggaag 5521 ggctaattca ctcccaacga agacaagata tccttgatct gtggatctac cacacacaag 5581 gctacttccc tgattagcag aactacacac cagggccagg ggtcagatat ccactgacct 5641 ttggatggtg ctacaagcta gtaccagttg agccagataa ggtagaagag gccaataaag 5701 gagagaacac cagcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag 5761 aagtgttaga gtggaggttt gacagccgcc tagcatttca tcacgtggcc cgagagctgc 5821 atccggagta cttcaagaac tgctgatatc gagcttgcta caagggactt tccgctgggg 5881 actttccagg gaggcgtggc ctgggcggga ctggggagtg gcgagccctc agatcctgca 5941 tataagcagc tgctttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg 6001 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg 6061 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc 6121 ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat 6181 ttataacttg caaagaaatg aatatcagag agtgagaggc cttgacattg ctagcgttta 6241 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat 6301 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg 6361 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag 6421 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt 6481 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg 6541 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg 6601 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag 6661 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga 6721 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct 6781 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc 6841 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg 6901 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc 6961 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca 7021 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag 7081 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct 7141 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc 7201 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga 7261 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca 7321 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat 7381 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac 7441 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt 7501 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt 7561 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag 7621 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct 7681 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt 7741 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc 7801 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt 7861 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg 7921 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg 7981 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct 8041 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc 8101 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt 8161 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt 8221 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg 8281 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat 8341 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg 8401 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatca acttgtttat 8461 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt 8521 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg 8581 gatcaactgg ataactcaag ctaaccaaaa tcatcccaaa cttcccaccc cataccctat 8641 taccactgcc aattacctgt ggtttcattt actctaaacc tgtgattcct ctgaattatt 8701 ttcattttaa agaaattgta tttgttaaat atgtactaca aacttagtag t //