pDsRed2-ER哺乳細胞內質網(wǎng)定位質粒
基本信息
啟動子: CMV promoter
復制子: pUC ori,f1 ori
終止子: SV40 poly(A) signal
質粒分類: 哺乳系列質粒；哺乳熒光質粒；哺乳紅色質粒
質粒大小: 4757bp
質粒標簽: N-DsRed2
原核抗性: 卡那霉素Kan（50μg/ml）
篩選標記: 新霉素Neo/G418
克隆菌株: DH5α等大腸桿菌
培養(yǎng)條件: 37℃，有氧 LB
表達宿主: 293T等哺乳細胞
培養(yǎng)條件: 無須誘導，瞬時表達
誘導方式: 無需誘導，瞬時表達
5'測序引物: CMV-F（CGCAAATGGGCGGTAGGCGTG）
3'測序引物: SV40-polyA-R（GAAATTTGTGATGCTATTGC）
備注: 哺乳細胞內質網(wǎng)標記質粒
質粒簡介
pDsRed2-ER是一種哺乳動物表達載體，用于標記活細胞內質網(wǎng)。DsRed2基因是按人的密碼子優(yōu)化過的野生型DsRed變體，可以更快的成熟和較低的非特異性聚集。
The vector encodes a fusion consisting of Discosoma sp. red fluorescent protein; the endoplasmic reticulum (ER) targeting sequence of calreticulin , fused to the 5' end of DsRed2; and the ER retention sequence, KDEL, fused to the 3' end of DsRed2. DsRed2 is a human codon-optimized variant of wild-type DsRed that has been engineered for faster maturation and lower non-specific aggregation.To drive expression of DsRed2, this vector contains the immediate early promoter of cytomegalovirus (PCMV IE). SV40 polyadenylation signals downstream of the DsRed2 gene direct proper processing of the 3'-end of the DsRed2 mRNA transcript. The vector also contains an SV40 origin for replication in any mammalian cell line that expresses the SV40 Tantigen, a pUC origin of replication for propagation in E. coli, and an f1 origin for single-stranded DNA production. A neomycin resistance cassette—consisting of the SV40 early promoter (PSV40e), the neomycin/kanamycin resistance gene of Tn5 (Neor/Kanr), and polyadenylation signals from the herpes simplex virus thymidine kinase (HSV TK poly A) gene— allows stably transfected eukaryotic cells to be selected using G418 . A bacterial promoter (P) upstream of this cassette drives expression of the gene encoding kanamycin resistance in E. coli.pDsRed2-ER can be introduced into mammalian cells using any standard transfection method. If required, stable transformants can be selected using G418 .
質粒圖譜

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