■ 基本信息
启动子: | CMV |
复制子: | pUC ori,f1 ori |
终止子: | SV40 poly(A) signal |
质粒大小: | 5442bp |
质粒标签: | N-tdTomato |
原核抗性: | Kan |
真核抗性: | G418 |
克隆菌株: | Stbl3 |
培养条件: | 37度 |
表达宿主: | 哺乳细胞293T |
诱导方式: | 无须诱导,瞬时表达 |
5'测序引物:
| 根据序列设计引物 |
3'测序引物: | Sv40-polyA-R:GAAATTTGTGATGCTATTGC |
■ 质粒属性
质粒宿主: | 哺乳细胞 |
质粒用途: | 蛋白表达 |
片段类型: | ORF
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片段物种: |
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原核抗性: | Kan |
真核抗性: | G418
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荧光标记: |
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■ 质粒简介
ptdTomato-C1质粒是一个哺乳细胞红色荧光过表达质粒,CMV启动子驱动tdTomato红色荧光蛋白和目的基因融合在一起表达。
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.
■ 质粒图谱
■ 质粒序列
质粒序列请下载:ZK1703 ptdTomato-C1哺乳荧光质粒.txt