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
pLVX-DsRed-Monomer-C1载体质粒基本信息
pLVX-DsRed-Monomer-C1质粒图谱载体图谱和pLVX-DsRed-Monomer-C1载体序列质粒序列多克隆位点信息
pLVX-DsRed-Monomer-C1质粒载体简介
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).
pLVX-DsRed-Monomer-C1质粒序列载体序列
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 //