■ 基本信息
啟動子: | CMV |
復(fù)制子: | pUC |
終止子: | SV40 poly(A) signal |
質(zhì)粒分類: | 哺乳系列質(zhì)粒；哺乳熒光質(zhì)粒；哺乳紅色質(zhì)粒 |
質(zhì)粒大小: | 4757bp |
質(zhì)粒標簽: | N-DsRed2 |
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 |
誘導(dǎo)方式: | 無需誘導(dǎo)，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(nèi)質(zhì)網(wǎng)標記質(zhì)粒 |
■ 質(zhì)粒屬性
質(zhì)粒宿主: | 哺乳細胞 |
質(zhì)粒用途: | 定位 |
片段類型: |
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片段物種: |
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原核抗性: | Kan |
真核抗性: | G418
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熒光標記: |
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■ 質(zhì)粒簡介
pDsRed2-ER是一種哺乳動物表達載體，用于標記活細胞內(nèi)質(zhì)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(zhì)粒圖譜
■ 質(zhì)粒序列
質(zhì)粒序列請下載：
ZK525pDsRed2-ER內(nèi)質(zhì)網(wǎng)定位質(zhì)粒.txt
