| 出品公司: | Clontech |
|---|---|
| 载体名称: | pLVX-DsRed-Monomer-N1 |
| 质粒类型: | 慢病毒表达载体；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: | 8751 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: | DsRed-Monomer（C-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: |
pLVX-DsRed-Monomer-N1载体表达C端DsRed-Monomer融合蛋白； DsRed-Monomer是DsRed的单体突变体，与DsRed相比，编码序列经过了人工优化，适用于在哺乳动物细胞中的高水平表达； CMV启动子是过表达启动子。 |
| 产品目录号: | 632152 |
| 稳定性: | 稳表达 |
| 组成型: | 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
Description
pLVX-DsRed-Monomer-N1 is an HIV-1-based, lentiviral expression vector that allows you to express your gene of interest fused to DsRed-Monomer, a monomeric mutant of the Discosoma sp. red fluorescent protein. Genes cloned into the multiple cloning site (MCS),located upstream of the DsRed-Monomer coding sequence, are expressed as N-terminal fusions of the DsRed-Monomer protein. Expression of the fusion protein is driven by the constitutively active human cytomegalovirus immediate early promoter (PCMV IE) located just upstream of the MCS. Lentiviral particles derived from the vector allow the expression of DsRed-Monomer fusion proteins in virtually any cell type, including primary cells. The unmodified vector expresses DsRed-Monomer, and may be used to produce marker virus to
optimize infection protocols.
pLVX-DsRed-Monomer-N1 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 nhances nuclear export of viral and transgene RNA (1), 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 (2). Finally, pLVX-DsRed-Monomer-N1 also contains a central polypurine tract (cPPT) element that increases nuclear importation of the viral genome during target cell infection, resulting in improved vector integration and more efficient transduction (3).
In addition to lentiviral elements, pLVX-DsRed-Monomer-N1 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.
Use
pLVX-DsRed-Monomer-N1 constitutively expresses your gene of interest from PCMV IE when transduced into target cells. Before the vector can be transduced into cells, however, it must be transfected into 293T packaging cells with our Lenti-X™ HTX Packaging System (Cat. Nos. 631247 and 631249). 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 (4).
载体序列
LOCUS pLVX-DsRed-Monomer-N1 8751 bp DNA circular 9-NOV-2009
COMMENT Created by Clontech Laboratories Inc.
COMMENT Cat. No. 632152
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_feature 2028..2151
/label=cPPT/CTS
promoter 2185..2787
/label=CMV\IE\Promoter
misc_feature 4720..5311
/label=WPRE
rep_origin complement(6619..7292)
/label=pUC\origin
CDS complement(7437..8433)
/label=Amp(R)
misc_binding 1303..1536
/label=RRE
misc_binding 636..653
/label=PBS
LTR 5514..6150
/label=3'\LTR
misc_feature 2814..2868
/label=MCS
CDS 2881..3558
/label=DsRed-Monomer
promoter 3578..4086
/label=PGK\Promoter
CDS 4107..4706
/label=Puro(R)
BASE COUNT 2168 a 2279 c 2317 g 1987 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 gggatccacc ggtcgccacc
2881 atggacaaca ccgaggacgt catcaaggag ttcatgcagt tcaaggtgcg catggagggc
2941 tccgtgaacg gccactactt cgagatcgag ggcgagggcg agggcaagcc ctacgagggc
3001 acccagaccg ccaagctgca ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc
3061 ctgtcccccc agttccagta cggctccaag gcctacgtga agcaccccgc cgacatcccc
3121 gactacatga agctgtcctt ccccgagggc ttcacctggg agcgctccat gaacttcgag
3181 gacggcggcg tggtggaggt gcagcaggac tcctccctgc aggacggcac cttcatctac
3241 aaggtgaagt tcaagggcgt gaacttcccc gccgacggcc ccgtaatgca gaagaagact
3301 gccggctggg agccctccac cgagaagctg tacccccagg acggcgtgct gaagggcgag
3361 atctcccacg ccctgaagct gaaggacggc ggccactaca cctgcgactt caagaccgtg
3421 tacaaggcca agaagcccgt gcagctgccc ggcaaccact acgtggactc caagctggac
3481 atcaccaacc acaacgagga ctacaccgtg gtggagcagt acgagcacgc cgaggcccgc
3541 cactccggct cccagtagag cggccgcgac tctagataat tctaccgggt aggggaggcg
3601 cttttcccaa ggcagtctgg agcatgcgct ttagcagccc cgctgggcac ttggcgctac
3661 acaagtggcc tctggcctcg cacacattcc acatccaccg gtaggcgcca accggctccg
3721 ttctttggtg gccccttcgc gccaccttct actcctcccc tagtcaggaa gttccccccc
3781 gccccgcagc tcgcgtcgtg caggacgtga caaatggaag tagcacgtct cactagtctc
3841 gtgcagatgg acagcaccgc tgagcaatgg aagcgggtag gcctttgggg cagcggccaa
3901 tagcagcttt gctccttcgc tttctgggct cagaggctgg gaaggggtgg gtccgggggc
3961 gggctcaggg gcgggctcag gggcggggcg ggcgcccgaa ggtcctccgg aggcccggca
4021 ttctgcacgc ttcaaaagcg cacgtctgcc gcgctgttct cctcttcctc atctccgggc
4081 ctttcgacct gcagcccaag cttaccatga ccgagtacaa gcccacggtg cgcctcgcca
4141 cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg
4201 ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac
4261 tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg
4321 cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg
4381 gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc
4441 tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc
4501 ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg
4561 agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg
4621 agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt
4681 gcatgacccg caagcccggt gcctgaccgc gtctggaaca atcaacctct ggattacaaa
4741 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
4801 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
4861 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
4921 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
4981 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
5041 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
5101 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt
5161 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
5221 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
5281 cggatctccc tttgggccgc ctccccgcct ggaattaatt ctgcagtcga gacctagaaa
5341 aacatggagc aatcacaagt agcaatacag cagctaccaa tgctgattgt gcctggctag
5401 aagcacaaga ggaggaggag gtgggttttc cagtcacacc tcaggtacct ttaagaccaa
5461 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagagg ggactggaag
5521 ggctaattca ctcccaacga agacaagata tccttgatct gtggatctac cacacacaag
5581 gctacttccc tgattagcag aactacacac cagggccagg ggtcagatat ccactgacct
5641 ttggatggtg ctacaagcta gtaccagttg agccagataa ggtagaagag gccaataaag
5701 gagagaacac cagcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag
5761 aagtgttaga gtggaggttt gacagccgcc tagcatttca tcacgtggcc cgagagctgc
5821 atccggagta cttcaagaac tgctgatatc gagcttgcta caagggactt tccgctgggg
5881 actttccagg gaggcgtggc ctgggcggga ctggggagtg gcgagccctc agatcctgca
5941 tataagcagc tgctttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg
6001 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
6061 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
6121 ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat
6181 ttataacttg caaagaaatg aatatcagag agtgagaggc cttgacattg ctagcgttta
6241 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
6301 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
6361 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
6421 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
6481 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
6541 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
6601 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
6661 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
6721 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
6781 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
6841 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
6901 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
6961 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
7021 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
7081 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
7141 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
7201 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
7261 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
7321 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
7381 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
7441 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
7501 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
7561 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
7621 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
7681 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
7741 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
7801 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
7861 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
7921 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
7981 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
8041 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
8101 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
8161 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
8221 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
8281 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
8341 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
8401 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatca acttgtttat
8461 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
8521 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
8581 gatcaactgg ataactcaag ctaaccaaaa tcatcccaaa cttcccaccc cataccctat
8641 taccactgcc aattacctgt ggtttcattt actctaaacc tgtgattcct ctgaattatt
8701 ttcattttaa agaaattgta tttgttaaat atgtactaca aacttagtag t
//
