■ 基本信息
啟動子: | CMV |
復制子: | pUC |
終止子: | SV40 poly(A) signal |
質粒分類: | 哺乳細胞質粒；哺乳報告質粒；熒光定位質粒 |
質粒大小: | 4787bp |
質粒標簽: | C-mem，C-KillerRed |
原核抗性: | Kan |
真核抗性: | G418 |
克隆菌株: | DH5a |
培養(yǎng)條件: | 37度 |
表達宿主: | 293T等哺乳細胞 |
備注: | 哺乳細胞紅色熒光膜定位質粒 |
■ 質粒屬性
質粒宿主: | 哺乳細胞 |
質粒用途: | 定位 |
片段類型: | ORF |
片段物種: | 空載體 |
原核抗性: | Kan |
真核抗性: | G418
|
熒光標記: | 紅色
|
■ 質粒簡介
pKillerRed-mem質粒是一個哺乳細胞紅色熒光定位于細胞膜的質粒。pKillerRed-mem is a mammalian expression vector encoding membrane-targeted KillerRed. KillerRed localized on cellular membrane can be used for effective light-induced cell killing.KillerRed codon usage is optimized for high expression in mammalian cells (humanized) [Haas et al. 1996]. Membrane localization signal (MLS) of neuromodulin is linked to the KillerRed N-terminus. The MLS (Nterminal 20 amino acid residues of neuromodulin) contains a signal for posttranslational palmitoylation of cysteines 3 and 4 that targets KillerRed to cellular membranes [Skene and Virág 1989]. pKillerRed-mem vector can be used as a source of MLS-KillerRed hybrid sequence. The vector backbone contains unique restriction sites that permit its excision and further insertion into expression vector of choice.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.pKillerRed-mem vector can be transfected into mammalian cells by any known transfection method. CMV promoter provides strong, constitutive expression of memrane-targeted KillerRed in eukaryotic cells. If required, stable transformants can be selected using G418 [Gorman 1985].Suitable 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.
■ 質粒圖譜
■ 質粒序列
質粒序列請下載：
ZK531pKillerRed-Mem細胞膜定位質粒.txt
