pLVX-MetLuc Control

保存条件 ：-20℃

保质期 ：6个月

英文名 ：pLVX-MetLuc Control

库存 ：大量

供应商 ：钰博生物

规格 ：1ug/5ug

pLVX-MetLuc Control

载体基本信息

出品公司:	Ybscience
载体名称:	pLVX-MetLuc Control
质粒类型:	慢病毒载体；荧光素酶报告载体
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:	8768 bp
5' 测序引物及序列:	CMV Forward： CGCAAATGGGCGGTAGGCGTG
3' 测序引物及序列:	--
载体标签:	--
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:	慢病毒载体pLVX-MetLuc Control组成型表达荧光素酶MetLuc，作为pLVX-MetLuc载体的阳性对照； MetLuc是已知最小分子量的荧光素酶。
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

载体简介

 

pLVX-MetLuc Control is an HIV-1-based, lentiviral vector that constitutively expresses Metridia luciferase (MetLuc), a secreted reporter protein that can be easily detected in the medium surrounding the cells. The vector can be used as a control to monitor the constitutive expression and secretion of Metridia luciferase in your cell type of interest.Metridia luciferase is a naturally secreted luciferase from the marine copepod Metridia longa. The human codon-optimized Metridia luciferase gene encodes a 219 amino acid (24 kDa) polypeptide that includes a 17 amino acid N-terminal signal peptide necessary for secretion (1). Expression of Metridia luciferase is driven by the constitutively active human cytomegalovirus immediate early promoter (PCMV IE), located just upstream of the Metridia luciferase coding sequence.pLVX-MetLuc Control 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-MetLuc Control 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-MetLuc Control contains a puromycin resistance gene (Puror) under the control of the murine phosphoglycerate kinase 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.The pLVX-MetLuc Control vector is available as part of the Lenti-X Ready-To-Glow Secreted Luciferase Reporter System (Cat. No. 631746). Lentiviral particles derived from the vector allow you to monitor the constitutive expression and secretion of Metridia luciferase in your cell type of interest. Secretion of Metridia luciferase can be monitored by adding chemiluminescent substrate to a sample of the cell medium. The vector can also be used to monitor the inhibitory effect of drugs, like Brefeldin A, on protein secretion. If required, stable transfectants can be selected using puromycin.Propagation in E. coli Recommended host strains: DH5α and other general purpose strains. Selectable marker: plasmid confers resistance to ampicillin (100 μg/ml) in E. coli hosts. E. coli replication origin: pUCReferences1. Markova, S.V. et al. (2004) J. Bio. Chem. 279(5):3212-3217.2. Zufferey, R. et al. (1999) J. Virol. 73(4):2886–2892.3. Cochrane, A. W. et al. (1990) Proc. Natl. Acad. Sci. USA 87(3):1198–1202.4. Zennou, V. et al. (2000) Cell 101(2):173–185.

 

载体序列

LOCUS       pLVX-MetLuc_Control   8721 bp    DNA   circular             6-NOV-2009COMMENT     Created by Clontech Laboratories Inc. http://www.clontech.comCOMMENT     Cat. No. 631746,631747FEATURES             Location/Qualifiers     LTR             1..635                     /label=5'\LTR     misc_feature    685..822                     /label=packaging\signal     promoter        2185..2787                     /label=CMV\IE\Promoter     misc_feature    4689..5280                     /label=WPRE     rep_origin      complement(6589..7262)                     /label=pUC\origin     CDS             complement(7407..8403)                     /label=Amp(R)     misc_binding    1303..1536                     /label=RRE     misc_binding    636..653                     /label=PBS     misc_feature    2028..2151                     /label=cPPT/CTS     LTR             5483..6119                     /label=3'\LTR     misc_feature    3547..4055                     /label=PGK\promoter     misc_feature    4076..4675                     /label=Puro(R)     CDS             2880..3536                     /label=Metridia\luciferaseBASE COUNT     2175 a      2250 c      2311 g      1985 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 gggatcccgc gactctagaa      2881 tggacatcaa ggtggtgttc accctggtgt tcagcgccct ggtgcaggcc aagagcaccg      2941 agttcgaccc caacatcgac atcgtgggcc tggaaggcaa gttcggcatc accaacctgg      3001 aaaccgacct gttcaccatc tgggagacca tggaagtgat gatcaaggcc gacatcgccg      3061 acaccgaccg ggccagcaac ttcgtggcca ccgagaccga cgccaaccgg ggcaagatgc      3121 ccggcaagaa gctgcccctg gccgtcatca tggaaatgga agccaacgcc ttcaaggccg      3181 gctgcacccg gggctgcctg atctgcctga gcaagatcaa gtgcaccgcc aagatgaagg      3241 tgtacatccc cggcaggtgc cacgactacg gcggcgacaa gaaaaccggc caggccggca      3301 tcgtgggcgc catcgtggac atccccgaga tcagcggctt caaagaaatg gcccccatgg      3361 aacagttcat cgcccaggtg gacagatgcg ccagctgcac caccggctgc ctgaagggcc      3421 tggccaacgt gaagtgcagc gagctgctga agaagtggct gcccgaccgc tgcgccagct      3481 tcgccgacaa gatccagaaa gaggtgcaca acatcaaggg catggccggc gacaggtagt      3541 ctagataatt ctaccgggta ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt      3601 tagcagcccc gctgggcact tggcgctaca caagtggcct ctggcctcgc acacattcca      3661 catccaccgg taggcgccaa ccggctccgt tctttggtgg ccccttcgcg ccaccttcta      3721 ctcctcccct agtcaggaag ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac      3781 aaatggaagt agcacgtctc actagtctcg tgcagatgga cagcaccgct gagcaatgga      3841 agcgggtagg cctttggggc agcggccaat agcagctttg ctccttcgct ttctgggctc      3901 agaggctggg aaggggtggg tccgggggcg ggctcagggg cgggctcagg ggcggggcgg      3961 gcgcccgaag gtcctccgga ggcccggcat tctgcacgct tcaaaagcgc acgtctgccg      4021 cgctgttctc ctcttcctca tctccgggcc tttcgacctg cagcccaagc ttaccatgac      4081 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac      4141 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca      4201 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg      4261 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt      4321 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg      4381 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc      4441 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc      4501 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac      4561 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt      4621 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaccgcg      4681 tctggaacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact      4741 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg      4801 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg      4861 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa      4921 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc      4981 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg      5041 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat      5101 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt      5161 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc      5221 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg      5281 gaattaattc tgcagtcgag acctagaaaa acatggagca atcacaagta gcaatacagc      5341 agctaccaat gctgattgtg cctggctaga agcacaagag gaggaggagg tgggttttcc      5401 agtcacacct caggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca      5461 ctttttaaaa gaaaagaggg gactggaagg gctaattcac tcccaacgaa gacaagatat      5521 ccttgatctg tggatctacc acacacaagg ctacttccct gattagcaga actacacacc      5581 agggccaggg gtcagatatc cactgacctt tggatggtgc tacaagctag taccagttga      5641 gccagataag gtagaagagg ccaataaagg agagaacacc agcttgttac accctgtgag      5701 cctgcatggg atggatgacc cggagagaga agtgttagag tggaggtttg acagccgcct      5761 agcatttcat cacgtggccc gagagctgca tccggagtac ttcaagaact gctgatatcg      5821 agcttgctac aagggacttt ccgctgggga ctttccaggg aggcgtggcc tgggcgggac      5881 tggggagtgg cgagccctca gatcctgcat ataagcagct gctttttgcc tgtactgggt      5941 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc      6001 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg      6061 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta      6121 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga      6181 gtgagaggcc ttgacattgc tagcgtttta ccgtcgacct ctagctagag cttggcgtaa      6241 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata      6301 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta      6361 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa      6421 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg      6481 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag      6541 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa      6601 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc      6661 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca      6721 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg      6781 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct      6841 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt      6901 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag      6961 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc      7021 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac      7081 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga      7141 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc      7201 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg      7261 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca      7321 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt      7381 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca      7441 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg      7501 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca      7561 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt      7621 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt      7681 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca      7741 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca      7801 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga      7861 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact      7921 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga      7981 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg      8041 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc      8101 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga      8161 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat      8221 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt      8281 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt      8341 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac      8401 gtcgacggat cgggagatca acttgtttat tgcagcttat aatggttaca aataaagcaa      8461 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc      8521 caaactcatc aatgtatctt atcatgtctg gatcaactgg ataactcaag ctaaccaaaa      8581 tcatcccaaa cttcccaccc cataccctat taccactgcc aattacctgt ggtttcattt      8641 actctaaacc tgtgattcct ctgaattatt ttcattttaa agaaattgta tttgttaaat      8701 atgtactaca aacttagtag t