| 出品公司: | Clontech |
|---|---|
| 载体名称: | pLVX-DsRed-Monomer-C1 |
| 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: | 8753 bp |
| 5' 测序引物及序列: | CMV-F: CGCAAATGGGCGGTAGGCGTG |
| 3' 测序引物及序列: | -- |
| 载体标签: | DsRed-Monomer（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: |
pLVX-DsRed-Monomer-C1载体表达N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: | 632153 |
| 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-C1 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 at the C-terminal end of the DsRed-Monomer coding sequence, are expressed as C-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 DsRed-Monomer coding sequence. 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-C1 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 (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-C1 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-C1 contains a puromycin resistance gene (Puro) 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 (Amp) for propagation and selection in bacteria.
Use
pLVX-DsRed-Monomer-C1 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-C1 8753 bp DNA circular 9-NOV-2009
COMMENT Created by Clontech Laboratories Inc.
COMMENT Cat. No. 632153
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 4721..5312
/label=WPRE
rep_origin complement(6621..7294)
/label=pUC\origin
CDS complement(7439..8435)
/label=Amp(R)
misc_binding 1303..1536
/label=RRE
misc_binding 636..653
/label=PBS
LTR 5515..6151
/label=3'\LTR
CDS 2815..3570
/label=DsRed-Monomer
misc_feature 3564..3566
/label=Stop
misc_feature 3568..3570
/label=Stop
misc_feature 3572..3574
/label=Stop
misc_feature 3503..3556
/label=MCS
promoter 3579..4087
/label=PGK\Promoter
CDS 4108..4707
/label=Puro(R)
BASE COUNT 2170 a 2278 c 2314 g 1991 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 taccggtcgc caccatggac
2821 aacaccgagg acgtcatcaa ggagttcatg cagttcaagg tgcgcatgga gggctccgtg
2881 aacggccact acttcgagat cgagggcgag ggcgagggca agccctacga gggcacccag
2941 accgccaagc tgcaggtgac caagggcggc cccctgccct tcgcctggga catcctgtcc
3001 ccccagttcc agtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac
3061 atgaagctgt ccttccccga gggcttcacc tgggagcgct ccatgaactt cgaggacggc
3121 ggcgtggtgg aggtgcagca ggactcctcc ctgcaggacg gcaccttcat ctacaaggtg
3181 aagttcaagg gcgtgaactt ccccgccgac ggccccgtaa tgcagaagaa gactgccggc
3241 tgggagccct ccaccgagaa gctgtacccc caggacggcg tgctgaaggg cgagatctcc
3301 cacgccctga agctgaagga cggcggccac tacacctgcg acttcaagac cgtgtacaag
3361 gccaagaagc ccgtgcagct gcccggcaac cactacgtgg actccaagct ggacatcacc
3421 aaccacaacg aggactacac cgtggtggag cagtacgagc acgccgaggc ccgccactcc
3481 ggctcccagt ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc
3541 gcgggcccgg gatccaccgg atctagataa ctgatcataa ttctaccggg taggggaggc
3601 gcttttccca aggcagtctg gagcatgcgc tttagcagcc ccgctgggca cttggcgcta
3661 cacaagtggc ctctggcctc gcacacattc cacatccacc ggtaggcgcc aaccggctcc
3721 gttctttggt ggccccttcg cgccaccttc tactcctccc ctagtcagga agttcccccc
3781 cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc tcactagtct
3841 cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta ggcctttggg gcagcggcca
3901 atagcagctt tgctccttcg ctttctgggc tcagaggctg ggaaggggtg ggtccggggg
3961 cgggctcagg ggcgggctca ggggcggggc gggcgcccga aggtcctccg gaggcccggc
4021 attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc tcctcttcct catctccggg
4081 cctttcgacc tgcagcccaa gcttaccatg accgagtaca agcccacggt gcgcctcgcc
4141 acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc
4201 gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa
4261 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc
4321 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc
4381 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc
4441 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg
4501 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc
4561 gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac
4621 gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg
4681 tgcatgaccc gcaagcccgg tgcctgaccg cgtctggaac aatcaacctc tggattacaa
4741 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
4801 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
4861 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
4921 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
4981 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
5041 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
5101 ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat
5161 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
5221 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
5281 tcggatctcc ctttgggccg cctccccgcc tggaattaat tctgcagtcg agacctagaa
5341 aaacatggag caatcacaag tagcaataca gcagctacca atgctgattg tgcctggcta
5401 gaagcacaag aggaggagga ggtgggtttt ccagtcacac ctcaggtacc tttaagacca
5461 atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaagag gggactggaa
5521 gggctaattc actcccaacg aagacaagat atccttgatc tgtggatcta ccacacacaa
5581 ggctacttcc ctgattagca gaactacaca ccagggccag gggtcagata tccactgacc
5641 tttggatggt gctacaagct agtaccagtt gagccagata aggtagaaga ggccaataaa
5701 ggagagaaca ccagcttgtt acaccctgtg agcctgcatg ggatggatga cccggagaga
5761 gaagtgttag agtggaggtt tgacagccgc ctagcatttc atcacgtggc ccgagagctg
5821 catccggagt acttcaagaa ctgctgatat cgagcttgct acaagggact ttccgctggg
5881 gactttccag ggaggcgtgg cctgggcggg actggggagt ggcgagccct cagatcctgc
5941 atataagcag ctgctttttg cctgtactgg gtctctctgg ttagaccaga tctgagcctg
6001 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
6061 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
6121 cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta
6181 tttataactt gcaaagaaat gaatatcaga gagtgagagg ccttgacatt gctagcgttt
6241 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
6301 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
6361 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
6421 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
6481 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
6541 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
6601 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
6661 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
6721 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
6781 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
6841 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
6901 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
6961 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
7021 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
7081 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
7141 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
7201 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
7261 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
7321 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
7381 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
7441 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
7501 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
7561 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
7621 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
7681 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
7741 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
7801 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
7861 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
7921 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
7981 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
8041 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
8101 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
8161 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
8221 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
8281 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
8341 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
8401 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat caacttgttt
8461 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
8521 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
8581 tggatcaact ggataactca agctaaccaa aatcatccca aacttcccac cccataccct
8641 attaccactg ccaattacct gtggtttcat ttactctaaa cctgtgattc ctctgaatta
8701 ttttcatttt aaagaaattg tatttgttaa atatgtacta caaacttagt agt
//
