■ 基本信息
啟動子: | CMV |
復制子: | pUC ori,f1 ori |
終止子: | SV40 poly(A) signal |
質粒大小: | 5442bp |
質粒標簽: | N-tdTomato |
原核抗性: | Kan |
真核抗性: | G418 |
克隆菌株: | Stbl3 |
培養(yǎng)條件: | 37度 |
表達宿主: | 哺乳細胞293T |
誘導方式: | 無須誘導，瞬時表達 |
5'測序引物:
| 根據序列設計引物 |
3'測序引物: | Sv40-polyA-R：GAAATTTGTGATGCTATTGC |
■ 質粒屬性
質粒宿主: | 哺乳細胞 |
質粒用途: | 蛋白表達 |
片段類型: | ORF
|
片段物種: |
|
原核抗性: | Kan |
真核抗性: | G418
|
熒光標記: |
|
■ 質粒簡介
ptdTomato-C1質粒是一個哺乳細胞紅色熒光過表達質粒，CMV啟動子驅動tdTomato紅色熒光蛋白和目的基因融合在一起表達。
ptdTomato-C1 is a mammalian expression vector designed to express a protein of interest fused to the C-terminus of tdTomato. td Tomato is a member of the family of fruit fluorescent proteins (1) derived from the Discosoma sp. red fluorescent protein, DsRed (2). Because the Tomato protein has a tendency to dimerize, the vector was designed with two copies of the Tomato coding region linked together to allow intramolecular dimerization. As a result, each tdTomato RNA transcript encodes a tandem dimer of the Tomato protein (excitation and emission maxima equal 554nm and 581nm, respectively). Expression of td Tomato as a tandem dimer prevents the fused protein of interest from being forced into a dimeric complex. Fusions that retain the fluorescence properties of tdTomato can be monitored by flow cytometry and localized by fluorescence microscopy. The multiple cloning site (MCS) in ptdTomato-C1 is positioned downstream of the tdTomato coding sequence. A Kozak consensus sequence, located immediately upstream of the tdTomato coding sequence, enhances translational efficiency in eukaryotic cells (3). SV40 polyadenylation signals downstream of the tdTomato coding sequence and the MCS direct proper processing of the 3' end of the tdTomato (or fusion gene) mRNA. The vector backbone contains an SV40 origin for replication in mammalian cells expressing the SV40 large T antigen, a pUC origin of replication for propagation in E. coli, and an f1 origin for single-stranded DNA production. This vector also has a neomycin resistance cassette (Neor) that allows G418 selection of stably transfected eukaryotic cells. This cassette consists of the SV40 early promoter, the Tn5 neomycin/kanamycin resistance gene, and polyadenylation signals from the herpes simplex virus thymidine kinase (HSV TK) gene. A bacterial promoter upstream of the cassette confers kanamycin resistance in E. Coli.
■ 質粒圖譜
■ 質粒序列
質粒序列請下載：
ZK1703 ptdTomato-C1哺乳熒光質粒.txt
