| 出品公司: | Clontech |
|---|---|
| 载体名称: | pLVX-MetLuc |
| 质粒类型: | 慢病毒载体；无启动子载体；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: | 无 |
| 载体大小: | 8088 bp |
| 5' 测序引物及序列: | -- |
| 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没有启动子元件； MetLuc是已知最小分子量的荧光素酶； 荧光素酶报告基因可以用来定性定量分析启动子及其它顺式转录元件的活 性。 |
| 产品目录号: | 631746 |
| 稳定性: | 稳表达 |
| 组成型/诱导型: | -- |
| 病毒/非病毒: | 慢病毒 |
买家导航
pLVX-MetLuc载体质粒基本信息
pLVX-MetLuc质粒图谱载体图谱和pLVX-MetLuc载体序列质粒序列多克隆位点信息
pLVX-MetLuc质粒载体简介
pLVX-MetLuc载体描述
pLVX-MetLuc is a promoterless, HIV-1-based, lentiviral vector that allows the functional analysis of different promoters and promoter/enhancer combinations inserted into its multiple cloning site (MCS). The vector encodes Metridia luciferase (MetLuc), a secreted reporter protein that can be easily detected in the medium surrounding the cells.
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). When a functional promoter is cloned into the MCS, located immediately upstream of the reporter, Metridia luciferase is expressed and secreted into the surrounding medium, where it is easily detected by the addition of a chemiluminescent substrate.
Lentiviral particles derived from pLVX-MetLuc allow you to monitor the activity of your promoter of interest in virtually any cell type, even primary cells. pLVX-MetLuc 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 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 contains a puromycin resistance gene (Puror) under the control of the murine phosphoglycerate kinase promoter (PPGK) for 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 vector, available as part of the Lenti-X Ready-To-Glow Secreted Luciferase Reporter System (Cat. No. 631746), can be used to monitor promoter activity in live cells as well as in vivo. A promoter must be cloned into the MCS, located upstream of the Metridia luciferase coding sequence. Without the addition of a functional promoter, the vector will not express MetLuc. Lentiviral particles derived from the vector allow you to monitor your promoter of interest in virtually any cell type, even primary cells. 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.
pLVX-MetLuc质粒序列载体序列
LOCUS pLVX-MetLuc_Reporter 8088 bp DNA circular 6-NOV-2009
COMMENT Created by Clontech Laboratories Inc. http://www.clontech.com
COMMENT Cat. No. 631747,631746
FEATURES Location/Qualifiers
LTR 1..635
/label=5'\LTR
misc_feature 685..822
/label=packaging\signal
misc_feature 4056..4647
/label=WPRE
rep_origin complement(5956..6629)
/label=pUC\origin
CDS complement(6774..7770)
/label=Amp(R)
misc_binding 1303..1536
/label=RRE
misc_binding 636..653
/label=PBS
misc_feature 2028..2151
/label=cPPT/CTS
LTR 4850..5486
/label=3'\LTR
misc_feature 2914..3422
/label=PGK\promoter
misc_feature 3443..4042
/label=Puro(R)
misc_feature 2195..2234
/label=MCS
CDS 2247..2903
/label=Metridia\luciferase
BASE COUNT 2019 a 2085 c 2157 g 1827 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 cgtcgagctc aagcttcgaa ttctgcagtc gacggtaccg
2221 cgggcccggg atcccgcgac tctagaatgg acatcaaggt ggtgttcacc ctggtgttca
2281 gcgccctggt gcaggccaag agcaccgagt tcgaccccaa catcgacatc gtgggcctgg
2341 aaggcaagtt cggcatcacc aacctggaaa ccgacctgtt caccatctgg gagaccatgg
2401 aagtgatgat caaggccgac atcgccgaca ccgaccgggc cagcaacttc gtggccaccg
2461 agaccgacgc caaccggggc aagatgcccg gcaagaagct gcccctggcc gtcatcatgg
2521 aaatggaagc caacgccttc aaggccggct gcacccgggg ctgcctgatc tgcctgagca
2581 agatcaagtg caccgccaag atgaaggtgt acatccccgg caggtgccac gactacggcg
2641 gcgacaagaa aaccggccag gccggcatcg tgggcgccat cgtggacatc cccgagatca
2701 gcggcttcaa agaaatggcc cccatggaac agttcatcgc ccaggtggac agatgcgcca
2761 gctgcaccac cggctgcctg aagggcctgg ccaacgtgaa gtgcagcgag ctgctgaaga
2821 agtggctgcc cgaccgctgc gccagcttcg ccgacaagat ccagaaagag gtgcacaaca
2881 tcaagggcat ggccggcgac aggtagtcta gataattcta ccgggtaggg gaggcgcttt
2941 tcccaaggca gtctggagca tgcgctttag cagccccgct gggcacttgg cgctacacaa
3001 gtggcctctg gcctcgcaca cattccacat ccaccggtag gcgccaaccg gctccgttct
3061 ttggtggccc cttcgcgcca ccttctactc ctcccctagt caggaagttc ccccccgccc
3121 cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact agtctcgtgc
3181 agatggacag caccgctgag caatggaagc gggtaggcct ttggggcagc ggccaatagc
3241 agctttgctc cttcgctttc tgggctcaga ggctgggaag gggtgggtcc gggggcgggc
3301 tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc ccggcattct
3361 gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct ccgggccttt
3421 cgacctgcag cccaagctta ccatgaccga gtacaagccc acggtgcgcc tcgccacccg
3481 cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
3541 gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt
3601 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
3661 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
3721 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
3781 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga
3841 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
3901 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg
3961 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
4021 gacccgcaag cccggtgcct gaccgcgtct ggaacaatca acctctggat tacaaaattt
4081 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
4141 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
4201 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
4261 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
4321 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
4381 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
4441 tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc
4501 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
4561 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
4621 tctccctttg ggccgcctcc ccgcctggaa ttaattctgc agtcgagacc tagaaaaaca
4681 tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct ggctagaagc
4741 acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa gaccaatgac
4801 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aagaggggac tggaagggct
4861 aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca cacaaggcta
4921 cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac tgacctttgg
4981 atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca ataaaggaga
5041 gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg agagagaagt
5101 gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag agctgcatcc
5161 ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg ctggggactt
5221 tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat cctgcatata
5281 agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga gcctgggagc
5341 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
5401 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
5461 agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat
5521 aacttgcaaa gaaatgaata tcagagagtg agaggccttg acattgctag cgttttaccg
5581 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
5641 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
5701 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
5761 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
5821 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
5881 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
5941 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
6001 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
6061 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
6121 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
6181 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
6241 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
6301 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
6361 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
6421 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
6481 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
6541 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
6601 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
6661 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
6721 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
6781 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
6841 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
6901 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
6961 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
7021 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
7081 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
7141 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
7201 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
7261 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
7321 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
7381 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
7441 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
7501 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
7561 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
7621 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
7681 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
7741 acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatcaact tgtttattgc
7801 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
7861 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat
7921 caactggata actcaagcta accaaaatca tcccaaactt cccaccccat accctattac
7981 cactgccaat tacctgtggt ttcatttact ctaaacctgt gattcctctg aattattttc
8041 attttaaaga aattgtattt gttaaatatg tactacaaac ttagtagt
//
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pLVX-MetLuc质粒载体应用举例
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