■ 基本信息
啟動子: | CMV |
復制子: | pUC ori,f1 ori |
終止子: | SV40 poly(A) signal |
質粒分類: | 哺乳細胞，熒光蛋白報告載體 |
質粒大小: | 5820bp |
原核抗性: | Kan |
真核抗性: | Neo |
克隆菌株: | DH5a |
培養(yǎng)條件: | 37℃，有氧，LB |
表達宿主: | 哺乳細胞 |
誘導方式: | 無須誘導，瞬時表達 |
5'測序引物:
| CMV-F：CGCAAATGGGCGGTAGGCGTG
|
3'測序引物: | 根據(jù)序列設計引物 |
■ 質粒屬性
質粒宿主: | 哺乳細胞 |
質粒用途: | 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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■ 質粒簡介
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.
■ 質粒圖譜
■ 質粒序列
質粒序列請下載：
ZK2253pEGFP-Actin細胞骨架定位質粒.txt
