| 出品公司: | Clontech |
|---|---|
| 载体名称: | pLVX-mCherry-Actin |
| 质粒类型: | 慢病毒载体；荧光报告载体 |
| 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: | 9893 bp |
| 5' 测序引物及序列: | CMV Forward： CGCAAATGGGCGGTAGGCGTG |
| 3' 测序引物及序列: | -- |
| 载体标签: | mCherry (N-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: | 慢病毒载体pLVX-mCherry-Actin组成型表达红色荧光标记的肌动蛋白。 |
| 产品目录号: | 631078 |
| 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-mCherry-Actin载体描述
pLVX-mCherry-Actin is an HIV-1-based, lentiviral expression vector that expresses human b-actin fused to mCherry, a mutant fluorescent protein derived from the tetrameric Discosoma sp. red fluorescent protein, DsRed (1). The excitation and emission maxima of the native mCherry protein are 587 nm and 610 nm, respectively. Expression of the mCherry-actin fusion protein is driven by the constitutively active human cytomegalovirus immediate early promoter (PCMV IE), located just upstream of the mCherry coding sequence. Lentiviral particles derived from the vector allow the expression of the mCherry-actin fusion protein in virtually any cell type, including primary cells.
pLVX-mCherry-Actin 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-mCherry-Actin 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-mCherry-Actin 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.
pLVX-mCherry-Actin constitutively expresses an mCherry-actin fusion protein when transduced into target cells, where the fusion protein is incorporated into actin filaments, allowing the visualization of actin-containing subcellular structures in both live and fixed cells (5, 6). Note: pLVX-mCherry-Actin is not designed to be used as a cloning vector.
Before the vector can be transduced, it must be transfected into 293T packaging cells with our Lenti-X HT Packaging System (Cat. Nos. 632160 and 632161). 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 (7). If required, stable transfectants can be selected using puromycin.
Propagation in E. coli
Suitable host strains: DH5α, DH10B and other general purpose strains.
Selectable marker: plasmid confers resistance to ampicillin (100 μg/ml) in E. coli hosts.
E. coli replication origin: ColE1
Copy number: high
Excitation and emission maxima of mCherry
Excitation maximum = 587 nm
Emission maximum = 610 nm
载体序列
LOCUS pLVX-mCherry-Actin 9899 bp DNA circular 21-DEC-2009
COMMENT Created by Clontech Laboratories Inc. http://www.clontech.com
COMMENT Cat. No. 631076,631078
FEATURES Location/Qualifiers
LTR 1..635
/label=5'\LTR
misc_feature 685..822
/label=packaging\signal
misc_feature 2028..2151
/label=cPPT/CTS\(DNA\flap)
promoter 2185..2787
/label=CMV\IE\Promoter
misc_feature 5867..6458
/label=WPRE
rep_origin complement(7767..8440)
/label=pUC\origin
CDS complement(8585..9581)
/label=Amp(R)
misc_binding 1303..1536
/label=RRE
misc_binding 636..653
/label=PBS
misc_feature 2031..2046
/label=cPPT
LTR 6661..7297
/label=LTR
misc_feature 4725..5233
/label=PGK\promoter
misc_feature 5254..5853
/label=Puro(R)
CDS 2861..3568
/label=mCherry
CDS 3590..4714
/label=Actin
BASE COUNT 2413 a 2641 c 2639 g 2206 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 atggtgagca agggcgagga
2881 ggataacatg gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt
2941 gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca
3001 gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc
3061 ccctcagttc atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta
3121 cttgaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg
3181 cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt
3241 gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg
3301 ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa
3361 gcagaggctg aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa
3421 ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac
3481 ctcccacaac gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc
3541 caccggcggc atggacgagc tgtacaagtc cggactcaga tctcgagcta tggatgatga
3601 tatcgccgcg ctcgtcgtcg acaacggctc cggcatgtgc aaggccggct tcgcgggcga
3661 cgatgccccc cgggccgtct tcccctccat cgtggggcgc cccaggcacc agggcgtgat
3721 ggtgggcatg ggtcagaagg attcctatgt gggcgacgag gcccagagca agagaggcat
3781 cctcaccctg aagtacccca tcgagcacgg catcgtcacc aactgggacg acatggagaa
3841 aatctggcac cacaccttct acaatgagct gcgtgtggct cccgaggagc accccgtgct
3901 gctgaccgag gcccccctga accccaaggc caaccgcgag aagatgaccc agatcatgtt
3961 tgagaccttc aacaccccag ccatgtacgt tgctatccag gctgtgctat ccctgtacgc
4021 ctctggccgt accactggca tcgtgatgga ctccggtgac ggggtcaccc acactgtgcc
4081 catctacgag gggtatgccc tcccccatgc catcctgcgt ctggacctgg ctggccggga
4141 cctgactgac tacctcatga agatcctcac cgagcgcggc tacagcttca ccaccacggc
4201 cgagcgggaa atcgtgcgtg acattaagga gaagctgtgc tacgtcgccc tggacttcga
4261 gcaagagatg gccacggctg cttccagctc ctccctggag aagagctacg agctgcctga
4321 cggccaggtc atcaccattg gcaatgagcg gttccgctgc cctgaggcac tcttccagcc
4381 ttccttcctg ggcatggagt cctgtggcat ccacgaaact accttcaact ccatcatgaa
4441 gtgtgacgtg gacatccgca aagacctgta cgccaacaca gtgctgtctg gcggcaccac
4501 catgtaccct ggcattgccg acaggatgca gaaggagatc actgccctgg cacccagcac
4561 aatgaagatc aagatcattg ctcctcctga gcgcaagtac tccgtgtgga tcggcggctc
4621 catcctggcc tcgctgtcca ccttccagca gatgtggatc agcaagcagg agtatgacga
4681 gtccggcccc tccatcgtcc accgcaaatg cttctagtct agataattct accgggtagg
4741 ggaggcgctt ttcccaaggc agtctggagc atgcgcttta gcagccccgc tgggcacttg
4801 gcgctacaca agtggcctct ggcctcgcac acattccaca tccaccggta ggcgccaacc
4861 ggctccgttc tttggtggcc ccttcgcgcc accttctact cctcccctag tcaggaagtt
4921 cccccccgcc ccgcagctcg cgtcgtgcag gacgtgacaa atggaagtag cacgtctcac
4981 tagtctcgtg cagatggaca gcaccgctga gcaatggaag cgggtaggcc tttggggcag
5041 cggccaatag cagctttgct ccttcgcttt ctgggctcag aggctgggaa ggggtgggtc
5101 cgggggcggg ctcaggggcg ggctcagggg cggggcgggc gcccgaaggt cctccggagg
5161 cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg ctgttctcct cttcctcatc
5221 tccgggcctt tcgacctgca gcccaagctt accatgaccg agtacaagcc cacggtgcgc
5281 ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac
5341 taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg
5401 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac
5461 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc
5521 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg
5581 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc
5641 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag
5701 gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc
5761 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc
5821 acctggtgca tgacccgcaa gcccggtgcc tgaccgcgtc tggaacaatc aacctctgga
5881 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
5941 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
6001 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
6061 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
6121 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
6181 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
6241 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
6301 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
6361 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
6421 gacgagtcgg atctcccttt gggccgcctc cccgcctgga attaattctg cagtcgagac
6481 ctagaaaaac atggagcaat cacaagtagc aatacagcag ctaccaatgc tgattgtgcc
6541 tggctagaag cacaagagga ggaggaggtg ggttttccag tcacacctca ggtaccttta
6601 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagagggga
6661 ctggaagggc taattcactc ccaacgaaga caagatatcc ttgatctgtg gatctaccac
6721 acacaaggct acttccctga ttagcagaac tacacaccag ggccaggggt cagatatcca
6781 ctgacctttg gatggtgcta caagctagta ccagttgagc cagataaggt agaagaggcc
6841 aataaaggag agaacaccag cttgttacac cctgtgagcc tgcatgggat ggatgacccg
6901 gagagagaag tgttagagtg gaggtttgac agccgcctag catttcatca cgtggcccga
6961 gagctgcatc cggagtactt caagaactgc tgatatcgag cttgctacaa gggactttcc
7021 gctggggact ttccagggag gcgtggcctg ggcgggactg gggagtggcg agccctcaga
7081 tcctgcatat aagcagctgc tttttgcctg tactgggtct ctctggttag accagatctg
7141 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
7201 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
7261 cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat
7321 tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggcctt gacattgcta
7381 gcgttttacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
7441 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
7501 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
7561 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
7621 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
7681 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
7741 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
7801 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
7861 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
7921 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
7981 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
8041 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
8101 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
8161 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
8221 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
8281 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
8341 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
8401 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
8461 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
8521 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
8581 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
8641 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
8701 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
8761 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
8821 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
8881 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
8941 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
9001 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
9061 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
9121 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
9181 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
9241 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
9301 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
9361 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
9421 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
9481 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
9541 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatcaac
9601 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
9661 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
9721 catgtctgga tcaactggat aactcaagct aaccaaaatc atcccaaact tcccacccca
9781 taccctatta ccactgccaa ttacctgtgg tttcatttac tctaaacctg tgattcctct
9841 gaattatttt cattttaaag aaattgtatt tgttaaatat gtactacaaa cttagtagt
//
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