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网站首页 >> pEGFP-N1哺乳荧光载体质粒 货号：P2069
pEGFP-N1哺乳荧光载体质粒 货号：P2069

信息

别名 pEGFPN1
启动子 CMV
复制子 pUC
终止子 SV40 poly(A) signal
质粒分类 哺乳系列质粒；哺乳荧光质粒；哺乳绿色质粒
质粒大小 4733bp
质粒标签 C-EGFP
原核抗性 Kan
真核抗性 G418
克隆菌株 DH5α
培养条件 37℃
表达宿主 293T等哺乳细胞
培养条件 37℃，有氧，5%CO2
诱导方式 无须诱导，瞬时表达
5'测序引物 pEGFP-N-5：TGGGAGGTCTATATAAGCAGAG
3'测序引物 pEGFP-N-3: CGTCGCCGTCCAGCTCGACCAG
质粒宿主 哺乳细胞
质粒用途 蛋白表达
片段类型 基因CDS
片段物种 空载体
原核抗性 卡那霉素
荧光标记 绿色

简介
         pEGFP-N1 encodes a red-shifted variant of wild-type GFP  which has been optimized for brighter fluorescence and higher expression in mammalian cells. (Excitation maximum = 488 nm; emission maximum = 507 nm.) pEGFP-N1 encodes the GFPmut1 variant which contains the double-amino-acid substitution of Phe-64 to Leu and Ser-65 to Thr. The coding sequence of the EGFP gene contains more than 190 silent base changes which correspond to human codon-usage preferences. Sequences flanking EGFP have been converted to a Kozak consensus translation initiation site  to further increase the translation efficiency in eukaryotic cells. The MCS in pEGFP-N1 is between the immediate early promoter of CMV and the EGFP coding sequences. Genes cloned into the MCS will be expressed as fusions to the N-terminus of EGFP if they are in the same reading frame as EGFP and there are no intervening stop codons. SV40 polyadenylation signals downstream of the EGFP gene direct proper processing of the 3' end of the EGFP mRNA. The vector backbone also contains an SV40 origin for replication in mammalian cells expressing the SV40 T antigen. A neomycin-resistance cassette, 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 expresses kanamycin resistance in E. coli. The pEGFP-N1 backbone also provides a pUC origin of replication for propagation in E. coli and an f1 origin for single-stranded DNA production.
       Fusions to the N terminus of EGFP retain the fluorescent properties of the native protein allowing the localization of the fusion protein in vivo . The target gene should be cloned into pEGFP-N1 so that it is in frame with the EGFP coding sequences, with no intervening in-frame stop codons. The inserted gene should include the initiating ATG codon. The recombinant EGFP vector can be transfected into mammalian cells using any standard transfection method. If required, stable transformants can be selected using G418 . pEGFP-N1 can also be used simply to express EGFP in a cell line of interest (e.g., as a transfection marker).

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