货号：	HZ0452
供应商：	沪震生物
数量：	200
英文名：	pEGFP-N3
保质期：	半年
保存条件：	-20℃
规格：	5μl

pEGFP-N3

产品信息

产品货号	产品名称	产品规格	优惠价
HZ0452	pEGFP-N3	20μl	¥请询价

使用说明

沪震质粒平台的各批次质粒菌株发货前均经过严格的多重验证，如存在质量问题，请在收到产品的三个月内通知我司。收到质粒后请短暂离心，取2μl转化至对应感受态中，挑取单克隆重新提取质粒后使用。

基本信息

启动子:	CMV
复制子:	pUC 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:	4729bp
原核抗性:	Kan
筛选标记:	Neo
克隆菌株:	DH5α
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

质粒简介

pEGFP-N3 encodes a red-shifted variant of wild-type GFP (1–3) which has been optimized for brighter fluorescence and higher expression in mammalian cells. (Excitation maximum = 488 nm; emission maximum = 507 nm.) pEGFP-N3 encodes the GFPmut1 variant (4) 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 (5). Sequences flanking EGFP have been converted to a Kozak consensus translation initiation site (6) to further increase the translation efficiency in eukaryotic cells. The MCS in pEGFP-N3 is between the immediate early promoter of CMV (PCMV IE) 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 (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 expresses kanamycin resistance in E. coli. The pEGFP-N3 backbone also provides a pUC origin of replication for propagation in E. coli and an f1 origin for single-stranded DNA production.