pmKate2-clathrin plasmid vector哺乳动物荧光表达质粒载体FP322 BioVector NTCC质粒载体菌种细胞基因保藏中心
- 价 格：¥49865
- 货 号：pmKate2-clathrin Cat.# FP322
- 产 地：北京
- BioVector NTCC典型培养物保藏中心
- 联系人：Dr.Xu, Biovector NTCC Inc.
电话：400-800-2947 工作QQ:1843439339 (微信同号)
邮件：Biovector@163.com
手机：18901268599
地址：北京
- 已注册
pmKate2-clathrin plasmid vector哺乳动物荧光表达质粒载体FP322
Vector type mammalian expression vectorReporter mKate2Reporter codon usage mammalianPromoter for mKate2 PCMV IEHost cells mammalianSelection prokaryotic – kanamycineukaryotic – neomycin (G418)Replication prokaryotic – pUC orieukaryotic – SV40 oriUse far-red fluorescent labeling of clathrin LCBVector descriptionpmKate2-clathrin is a mammalian expression vector encoding mKate2-clathrin fusion protein (see reporter description). The vector can be used for fluorescent labeling of clathrin LCB in living cells. C-terminal mKate2 fusion construct with human clathrin light chain expressed in human cervical adenocarcinoma cells (HeLa).Scale bar represents 10 μm. Image from Shcherbo et al., 2009.mKate2 codon usage is optimized for high expression in mammalian cells (humanized) [Haas et al., 1996]. Human clathrin LCB is fused to the mKate2 C-terminus. To increase mRNA translation efficiency, Kozak consensus translation initiation site is generated upstream of the mKate2-clathrin coding sequence [Kozak, 1987].pmKate2-clathrin vector can be used as a source of mKate2-clathrin hybrid sequence. The vector backbone contains unique restriction sites that permit its excision and further insertion into expression vector of choice.Note: The plasmid DNA was isolated from dam+-methylated E. coli. Therefore some restriction sites are blocked by methylation. If you wish to digest the vector using such sites you will need to transform the vector into a dam- host and make fresh DNA.The vector backbone contains immediate early promoter of cytomegalovirus (PCMV IE) for protein expression, SV40 origin for replication in mammalian cells expressing SV40 T-antigen, pUC origin of replication for propagation in E. coli, and f1 origin for single-stranded DNA production. SV40 polyadenylation signals (SV40 poly A) direct proper processing of the 3'-end of the reporter mRNA.SV40 early promoter (PSV40) provides neomycin resistance gene (Neor) expression to select stably transfected eukaryotic cells using G418. Bacterial promoter (P) provides kanamycin resistance gene expression (Kanr) in E. coli. Kanr/Neor gene is linked with herpes simplex virus (HSV) thymidine kinase (TK) polyadenylation signals.Expression in mammalian cellspmKate2-clathrin vector can be transfected into mammalian cells by any known transfection method. CMV promoter provides strong, constitutive expression of the mKate2-clathrin fusion in eukaryotic cells. If required, stable transformants can be selected using G418 [Gorman, 1985].Propagation in E. coliSuitable host strains for propagation in E. coli include DH5alpha, HB101, XL1-Blue, and other general purpose strains. Plasmid incompatibility group is pMB1/ColE1. The vector confers resistance to kanamycin (30 μg/ml) to E. coli hosts. Copy number in E. coli is about 500.Location of featuresPCMV IE: 1-589Enhancer region: 59-465TATA box: 554-560Transcription start point: 583Kozak consensus translation initiation site: 606-616mKate2-clathrin fusion: 613-1992mKate2: 613-1314Start codon (ATG): 613-615Last amino acid in mKate2: 1312-1314Clathrin LCB: 1360-1992Stop codon: 1993-1995SV40 early mRNA polyadenylation signalPolyadenylation signals: 2156-2161 & 2185-2190mRNA 3' ends: 2194 & 2206f1 single-strand DNA origin: 2253-2708Bacterial promoter for expression of Kanr gene-35 region: 2770-2775-10 region: 2793-2798Transcription start point: 2805SV40 origin of replication: 3049-3184SV40 early promoterEnhancer (72-bp tandem repeats): 2882-2953 & 2954-302521-bp repeats: 3029-3049, 3050-3070 & 3072-3092Early promoter element: 3105-3111Major transcription start points: 3101, 3139, 3145 & 3150Kanamycin/neomycin resistance geneNeomycin phosphotransferase coding sequences:Start codon (ATG): 3233-3235Stop codon: 4025-4027G->A mutation to remove Pst I site: 3415C->A (Arg to Ser) mutation to remove BssH II site: 3761Herpes simplex virus (HSV) thymidine kinase (TK) polyadenylation signalPolyadenylation signals: 4263-4268 & 4276-4281pUC plasmid replication origin: 4612-5255References:Gorman C. High efficiency gene transfer into mammalian cells. In DNA cloning: A Practical Approach, Vol. II. Ed. D. M. Glover. (IRL Press, Oxford, U.K.). 1985; 143-90.Haas J, Park EC, Seed B. Codon usage limitation in the expression of HIV-1 envelope glycoprotein. Curr Biol. 1996; 6 (3):315-24. / pmid: 8805248Kozak M. An analysis of 5'-noncoding sequences from 699 vertebrate messenger RNAs. Nucleic Acids Res. 1987; 15 (20):8125-48. / pmid: 3313277Shcherbo D, Murphy CS, Ermakova GV, Solovieva EA, Chepurnykh TV, Shcheglov AS, Verkhusha VV, Pletnev VZ, Hazelwood KL, Roche PM, Lukyanov S, Zaraisky AG, Davidson MW, Chudakov DM. Far-red fluorescent tags for protein imaging in living tissues. Biochem J. 2009; 418 (3):567-74. doi: 10.1042/BJ20081949 / pmid: 19143658
Supplier来源:BioVector NTCC Inc.
TEL电话:400-800-2947
Website网址: http://www.biovector.net
Vector type mammalian expression vectorReporter mKate2Reporter codon usage mammalianPromoter for mKate2 PCMV IEHost cells mammalianSelection prokaryotic – kanamycineukaryotic – neomycin (G418)Replication prokaryotic – pUC orieukaryotic – SV40 oriUse far-red fluorescent labeling of clathrin LCBVector descriptionpmKate2-clathrin is a mammalian expression vector encoding mKate2-clathrin fusion protein (see reporter description). The vector can be used for fluorescent labeling of clathrin LCB in living cells. C-terminal mKate2 fusion construct with human clathrin light chain expressed in human cervical adenocarcinoma cells (HeLa).Scale bar represents 10 μm. Image from Shcherbo et al., 2009.mKate2 codon usage is optimized for high expression in mammalian cells (humanized) [Haas et al., 1996]. Human clathrin LCB is fused to the mKate2 C-terminus. To increase mRNA translation efficiency, Kozak consensus translation initiation site is generated upstream of the mKate2-clathrin coding sequence [Kozak, 1987].pmKate2-clathrin vector can be used as a source of mKate2-clathrin hybrid sequence. The vector backbone contains unique restriction sites that permit its excision and further insertion into expression vector of choice.Note: The plasmid DNA was isolated from dam+-methylated E. coli. Therefore some restriction sites are blocked by methylation. If you wish to digest the vector using such sites you will need to transform the vector into a dam- host and make fresh DNA.The vector backbone contains immediate early promoter of cytomegalovirus (PCMV IE) for protein expression, SV40 origin for replication in mammalian cells expressing SV40 T-antigen, pUC origin of replication for propagation in E. coli, and f1 origin for single-stranded DNA production. SV40 polyadenylation signals (SV40 poly A) direct proper processing of the 3'-end of the reporter mRNA.SV40 early promoter (PSV40) provides neomycin resistance gene (Neor) expression to select stably transfected eukaryotic cells using G418. Bacterial promoter (P) provides kanamycin resistance gene expression (Kanr) in E. coli. Kanr/Neor gene is linked with herpes simplex virus (HSV) thymidine kinase (TK) polyadenylation signals.Expression in mammalian cellspmKate2-clathrin vector can be transfected into mammalian cells by any known transfection method. CMV promoter provides strong, constitutive expression of the mKate2-clathrin fusion in eukaryotic cells. If required, stable transformants can be selected using G418 [Gorman, 1985].Propagation in E. coliSuitable host strains for propagation in E. coli include DH5alpha, HB101, XL1-Blue, and other general purpose strains. Plasmid incompatibility group is pMB1/ColE1. The vector confers resistance to kanamycin (30 μg/ml) to E. coli hosts. Copy number in E. coli is about 500.Location of featuresPCMV IE: 1-589Enhancer region: 59-465TATA box: 554-560Transcription start point: 583Kozak consensus translation initiation site: 606-616mKate2-clathrin fusion: 613-1992mKate2: 613-1314Start codon (ATG): 613-615Last amino acid in mKate2: 1312-1314Clathrin LCB: 1360-1992Stop codon: 1993-1995SV40 early mRNA polyadenylation signalPolyadenylation signals: 2156-2161 & 2185-2190mRNA 3' ends: 2194 & 2206f1 single-strand DNA origin: 2253-2708Bacterial promoter for expression of Kanr gene-35 region: 2770-2775-10 region: 2793-2798Transcription start point: 2805SV40 origin of replication: 3049-3184SV40 early promoterEnhancer (72-bp tandem repeats): 2882-2953 & 2954-302521-bp repeats: 3029-3049, 3050-3070 & 3072-3092Early promoter element: 3105-3111Major transcription start points: 3101, 3139, 3145 & 3150Kanamycin/neomycin resistance geneNeomycin phosphotransferase coding sequences:Start codon (ATG): 3233-3235Stop codon: 4025-4027G->A mutation to remove Pst I site: 3415C->A (Arg to Ser) mutation to remove BssH II site: 3761Herpes simplex virus (HSV) thymidine kinase (TK) polyadenylation signalPolyadenylation signals: 4263-4268 & 4276-4281pUC plasmid replication origin: 4612-5255References:Gorman C. High efficiency gene transfer into mammalian cells. In DNA cloning: A Practical Approach, Vol. II. Ed. D. M. Glover. (IRL Press, Oxford, U.K.). 1985; 143-90.Haas J, Park EC, Seed B. Codon usage limitation in the expression of HIV-1 envelope glycoprotein. Curr Biol. 1996; 6 (3):315-24. / pmid: 8805248Kozak M. An analysis of 5'-noncoding sequences from 699 vertebrate messenger RNAs. Nucleic Acids Res. 1987; 15 (20):8125-48. / pmid: 3313277Shcherbo D, Murphy CS, Ermakova GV, Solovieva EA, Chepurnykh TV, Shcheglov AS, Verkhusha VV, Pletnev VZ, Hazelwood KL, Roche PM, Lukyanov S, Zaraisky AG, Davidson MW, Chudakov DM. Far-red fluorescent tags for protein imaging in living tissues. Biochem J. 2009; 418 (3):567-74. doi: 10.1042/BJ20081949 / pmid: 19143658
Supplier来源:BioVector NTCC Inc.
TEL电话:400-800-2947
Website网址: http://www.biovector.net
- 公告/新闻
