| 出品公司: | ZYbscience |
|---|---|
| 载体名称: | pLVX-IRES-mCherry |
| 质粒类型: | 慢病毒载体；哺乳动物细胞表达载体；双顺反子载体；荧光报告载体 |
| 高拷贝/低拷贝: | 高拷贝 |
| 启动子: | CMV |
| 克隆方法: | 多克隆位点，限制性内切酶 |
| 载体大小: | 8172 bp |
| 5' 测序引物及序列: | CMV-F: CGCAAATGGGCGGTAGGCGTG (Invitrogen) |
| 3' 测序引物及序列: | -- |
| 载体标签: | 无标签 |
| 载体抗性: | 氨苄青霉素 |
| 筛选标记: | 嘌呤霉素(Puromycin) |
| 克隆菌株: | Stbl3 |
| 备注: | pLVX-IRES-mCherry载体以双顺反子的形式同时表达mCherry荧光蛋白和目的基因； CMV启动子驱动目的基因的过表达； mCherry是第三代荧光蛋白，亮度高，适用于流式细胞仪。 |
| 产品目录号: | ZY631237 |
| 稳定性: | 稳表达 |
| 组成型: | 诱导型 |
| 病毒/非病毒: | 慢病毒 |
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- 详细信息
- 文献和实验
- 技术资料
- 保存条件：
-20℃低温保存
- 保质期：
三年
- 英文名：
pLVX-IRES-mCherry
- 库存：
20
- 供应商：
泽叶生物
载体基本信息
载体质粒图谱和多克隆位点信息
载体简介
Description
pLVX-IRES-mCherry is an HIV-1-based, lentiviral expression vector that allows the simultaneous expression of your protein of interest and mCherry in virtually any mammalian cell type, including primary cells. mCherry is a mutant fluorescent protein derived from the tetrameric Discosoma sp. red fluorescent protein, DsRed (1). The vector expresses the two proteins from a bicistronic mRNA transcript, allowing mCherry to be used as an indicator of transduction efficiency and a marker for selection by flow cytometry.
Expression of the bicistronic transcript is driven by the constitutively active human cytomegalovirus immediate early promoter (PCMV IE) located just upstream of the MCS. An encephalomyocarditis virus (EMCV) internal ribosome entry site (IRES), positioned between the MCS and mCherry, facilitates cap-independent translation of mCherry from an internal start site at the IRES/mCherry junction (1).
pLVX-IRES-mCherry 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 RNA (2), leading to increased viral titers from packaging 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-IRES-mCherry 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). The vector also contains a pUC origin of replication and an E. coli ampicillin resistance gene (Ampr) for propagation and selection in bacteria.
Use
pLVX-IRES-mCherry is designed to constitutively coexpress your protein of interest and mCherry from PCMV IE when transduced into mammalian cells. Before it can be transduced into target cells, the vector must be packaged into viral particles in HEK293T cells, using our Lenti-X™ HT Packaging System (Cat. Nos. 632160 and 632161). This packaging system allows the safe production of high titer, infectious, replication-incompetent, VSV-G pseudotyped lentiviral particles that can infect a wide range of cell types, including nondividing and primary cells (5).
The presence of mCherry allows transductants to be visualized by fluorescence microscopy and sorted by flow cytometry with standard FITC filter sets (mCherry has an excitation maximum of 587 nm and an emission maximum of 610 nm).
载体序列
LOCUS pLVX-IRES-mCherry 8172 bp DNA circular 28-MAY-2009
COMMENT Created by Clontech Laboratories Inc.
COMMENT Cat. No. 631237
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT VNTAUTHORNAME|biofeng.com|
FEATURES Location/Qualifiers
LTR 1..635
/label=5'\LTR
misc_feature 685..822
/label=Packaging\Signal
misc_binding 1303..1536
/label=RRE
promoter 2185..2787
/label=CMV\IE\Promoter
misc_feature 2842..3416
/label=IRES
misc_feature 4141..4732
/label=WPRE
LTR 4935..5571
/label=3'\LTR
CDS 3417..4127
/label=mCherry
rep_origin complement(6040..6713)
/label=pUC\ori
CDS complement(6858..7854)
/label=Amp(R)
misc_feature 636..653
/label=PBS
misc_feature 2028..2151
/label=cPPT/CTS\(DNA\flap)
misc_feature 2803..2840
/label=MCS
BASE COUNT 2133 a 1995 c 2099 g 1945 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 tctatttccg gtgaattcct cgagactagt
2821 tctagagcgg ccgcggatcc cgcccctctc cctccccccc ccctaacgtt actggccgaa
2881 gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt
2941 cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg
3001 gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc
3061 ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc
3121 ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa
3181 aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc
3241 tcacctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg
3301 gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaacg
3361 tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataatatgg
3421 tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc
3481 acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
3541 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg
3601 cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg
3661 ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga
3721 tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg
3781 agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc
3841 agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc
3901 tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg
3961 tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca
4021 tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg
4081 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtgaacg cgtctggaac
4141 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
4201 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
4261 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
4321 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
4381 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
4441 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
4501 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc
4561 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
4621 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
4681 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaattaat
4741 tctgcagtcg agacctagaa aaacatggag caatcacaag tagcaataca gcagctacca
4801 atgctgattg tgcctggcta gaagcacaag aggaggagga ggtgggtttt ccagtcacac
4861 ctcaggtacc tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa
4921 aagaaaagag gggactggaa gggctaattc actcccaacg aagacaagat atccttgatc
4981 tgtggatcta ccacacacaa ggctacttcc ctgattagca gaactacaca ccagggccag
5041 gggtcagata tccactgacc tttggatggt gctacaagct agtaccagtt gagccagata
5101 aggtagaaga ggccaataaa ggagagaaca ccagcttgtt acaccctgtg agcctgcatg
5161 ggatggatga cccggagaga gaagtgttag agtggaggtt tgacagccgc ctagcatttc
5221 atcacgtggc ccgagagctg catccggagt acttcaagaa ctgctgatat cgagcttgct
5281 acaagggact ttccgctggg gactttccag ggaggcgtgg cctgggcggg actggggagt
5341 ggcgagccct cagatcctgc atataagcag ctgctttttg cctgtactgg gtctctctgg
5401 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
5461 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
5521 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tagtagttca
5581 tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg
5641 ccttgacatt gctagcgttt accgtcgacc tctagctaga gcttggcgta atcatggtca
5701 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
5761 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
5821 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
5881 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
5941 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
6001 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
6061 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
6121 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
6181 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
6241 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
6301 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
6361 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
6421 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
6481 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
6541 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
6601 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
6661 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
6721 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
6781 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
6841 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
6901 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
6961 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
7021 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
7081 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
7141 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
7201 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
7261 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
7321 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
7381 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
7441 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
7501 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
7561 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
7621 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
7681 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
7741 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
7801 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
7861 tcgggagatc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
7921 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
7981 caatgtatct tatcatgtct ggatcaactg gataactcaa gctaaccaaa atcatcccaa
8041 acttcccacc ccatacccta ttaccactgc caattacctg tggtttcatt tactctaaac
8101 ctgtgattcc tctgaattat tttcatttta aagaaattgt atttgttaaa tatgtactac
8161 aaacttagta gt
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pLVX-IRES-mCherry载体
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