■ 基本信息
启动子: | CMV |
复制子: | pUC ori,f1 ori |
终止子: | SV40 poly(A) signal |
质粒分类: | 哺乳细胞，荧光蛋白报告载体 |
质粒大小: | 5820bp |
原核抗性: | Kan |
真核抗性: | Neo |
克隆菌株: | DH5a |
培养条件: | 37℃，有氧，LB |
表达宿主: | 哺乳细胞 |
诱导方式: | 无须诱导，瞬时表达 |
5'测序引物:
| CMV-F：CGCAAATGGGCGGTAGGCGTG
|
3'测序引物: | 根据序列设计引物 |
■ 质粒属性
质粒宿主: | 哺乳细胞 |
质粒用途: | 定位 |
片段类型: |
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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
pEGFP-Actin encodes a fusion protein consisting of the red-shifted, human codon-optimized variant of green fluorescent protein (EGFP; 1–3) and the gene encoding human cytoplasmic b-actin (4). EGFP, a derivative of the GFPmut1 variant (5), has been optimized for brighter fluorescence and higher expression in mammalian cells. (Excitation maximum = 488 nm; emission maximum = 507 nm.) This variant contains the double-amino-acid substitution of Phe-64 to Leu and Ser-65 to Thr, and also contains more than 190 silent base changes which correspond to human codon-usage preferences (6). SV40 polyadenylation signals downstream of the EGFP-Actin fusion direct proper processing of the 3' end of the EGFP mRNA. The vector backbone also contains an SV40 origin for replication in any mammalian cell line that expresses the SV40 T-antigen. A neomycin resistance cassette (Neor), consisting of the SV40 early promoter, the neomycin/kanamycin resistance gene of Tn5, and polyadenylation signals from the herpes simplex virus thymidine kinase (HSV-TK) gene, allows stably transfected eukaryotic cells to be selected using G418. A bacterial promoter upstream of this cassette drives expression of the gene encoding kanamycin resistance in E. coli. The pEGFPActin backbone also provides a pUC origin of replication for propagation in E. coli and an f1 origin for single-stranded DNA production.
■ 质粒图谱
■ 质粒序列
质粒序列请下载：
ZK2253pEGFP-Actin细胞骨架定位质粒.txt
