■ 基本信息
啟動(dòng)子: | CMV |
復(fù)制子: | pUC ori,f1 ori |
終止子: | SV40 poly(A) signal |
質(zhì)粒大小: | 5442bp |
質(zhì)粒標(biāo)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(dá)宿主: | 哺乳細(xì)胞293T |
誘導(dǎo)方式: | 無須誘導(dǎo)，瞬時(shí)表達(dá) |
5'測(cè)序引物:
| 根據(jù)序列設(shè)計(jì)引物 |
3'測(cè)序引物: | Sv40-polyA-R：GAAATTTGTGATGCTATTGC |
■ 質(zhì)粒屬性
質(zhì)粒宿主: | 哺乳細(xì)胞 |
質(zhì)粒用途: | 蛋白表達(dá) |
片段類型: | ORF
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片段物種: |
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原核抗性: | Kan |
真核抗性: | G418
|
熒光標(biāo)記: |
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■ 質(zhì)粒簡(jiǎn)介
ptdTomato-C1質(zhì)粒是一個(gè)哺乳細(xì)胞紅色熒光過表達(dá)質(zhì)粒，CMV啟動(dòng)子驅(qū)動(dòng)tdTomato紅色熒光蛋白和目的基因融合在一起表達(dá)。
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(zhì)粒圖譜
■ 質(zhì)粒序列
質(zhì)粒序列請(qǐng)下載：
ZK1703 ptdTomato-C1哺乳熒光質(zhì)粒.txt
