pLVX-MetLuc Control

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: pUC

References
1. 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-2009
COMMENT     Created by Clontech Laboratories Inc. http://www.clontech.com
COMMENT     Cat. No. 631746,631747
FEATURES             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\luciferase
BASE 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