■ 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)粒分類: | 哺乳細(xì)胞，熒光蛋白報(bào)告載體 |
質(zhì)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(dá)宿主: | 哺乳細(xì)胞 |
誘導(dǎo)方式: | 無須誘導(dǎo)，瞬時(shí)表達(dá) |
5'測(cè)序引物:
| CMV-F：CGCAAATGGGCGGTAGGCGTG
|
3'測(cè)序引物: | 根據(jù)序列設(shè)計(jì)引物 |
■ 質(zhì)粒屬性
質(zhì)粒宿主: | 哺乳細(xì)胞 |
質(zhì)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(biāo)記: |
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■ 質(zhì)粒簡(jiǎn)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(zhì)粒圖譜
■ 質(zhì)粒序列
質(zhì)粒序列請(qǐng)下載：
ZK2253pEGFP-Actin細(xì)胞骨架定位質(zhì)粒.txt
