■ 基本信息
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: | 4151bp |
质粒标签: | C-EGFP |
原核抗性: | Kan |
真核抗性: | G418 |
克隆菌株: | DH5a |
培养条件: | 37℃，5%CO2 |
表达宿主: | 293T等哺乳细胞 |
诱导方式: | 无须诱导，瞬时表达 |
5'测序引物:
| EGFP-N（CGTCGCCGTCCAGCTCGACCAG）
|
3'测序引物: | EGFP-F (CCAGCAACGCGGCCTTTTTA) |
■ 质粒属性
质粒宿主: | 哺乳细胞 |
质粒用途: | 信号报告 |
片段类型: | Promoter
|
片段物种: | 空载体
|
原核抗性: | Kan |
真核抗性: | G418 |
荧光标记: |
|
■ 质粒简介
pEGFP-1编码野生型GFP（1-3）的红移型变体，其已经针对更明亮的荧光和哺乳动物细胞中更高的表达进行了优化。（激发最大值= 488nm;发射最大值= 507nm）pEGFP-1骨架还提供了用于在大肠杆菌中繁殖的pUC起始点和用于单链DNA生产的f1起源。
pEGFP-1 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-1 encodes the GFPmut1 variant (4) which contains the double-aminoacid 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. pEGFP-1 is a promoterless EGFP vector which can be used to monitor transcription from different promoters and promoter/enhancer combinations inserted into the MCS located upstream of the EGFP coding sequence. 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) allows stably transfected eukaryotic cells to be selected using G418. The Neor cassette consists 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. A bacterial promoter upstream of this cassette confers kanamycin resistance in E. coli. The pEGFP-1 backbone also provides a pUC origin of replication for propagation in E. coli and an f1 origin for single-stranded DNA production.
EGFP can be used as an in vivo reporter of gene expression . Promoters should be cloned into the pEGFP-1 MCS upstream from the EGFP coding sequences. Without the addition of a functional promoter, this vector will not express EGFP. The recombinant EGFP vector can be transfected into mammalian cells using any standard transfection method. If required, stable transformants can be selected using G418.
质粒只保证关键序列正确，不保证表达效果。
■ 质粒图谱
■ 质粒序列
质粒序列请下载：
ZK410pEGFP-1哺乳启动子检测质粒.txt
