| 出品公司: | Clontech |
|---|---|
| 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。 |
| 产品目录号: | 631746 |
| 稳定性: | 稳表达 |
| 组成型/诱导型: | 组成型 |
| 病毒/非病毒: | 慢病毒 |
买家导航
载体基本信息
载体图谱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
//
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