■ 基本信息
启动子: | CMV IE promoter |
复制子: | pUC ori，F1 ori，SV40 ori |
终止子: | SV40 early polyA signals |
质粒分类: | 哺乳细胞质粒；哺乳荧光质粒；哺乳红色质粒 |
质粒大小: | 4698bp |
质粒标签: | N-HcRed1 |
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: | 无需诱导，组成型表达 |
5'测序引物: | CMV-F：CGCAAATGGGCGGTAGGCGTG |
3'测序引物
| Sv40-polyA-R：GAAATTTGTGATGCTATTGC
|
备注: | 高拷贝 |
■ 质粒属性
质粒宿主: | 哺乳细胞 |
质粒用途: | 蛋白表达 |
片段类型: | ORF
|
片段物种: |
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原核抗性: | Kan |
真核抗性: | G418
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荧光标记: |
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■ 质粒简介
pHcRed1-C1质粒是一个哺乳细胞红色荧光蛋白表达载体，CMV IE启动子驱动HcRed1红色荧光蛋白基因和目的基因融合表达。
pHcRed1-C1 is a mammalian expression vector designed to express a protein of interest fused to the C-terminus of the far-red fluorescent protein HcRed1. pHcRed1-C1 can be used to monitor gene expression and protein localization in vivo or as a cotransfection marker; the unmodified vector will express HcRed1 in mammalian cells. HcRed1 was generated by mutagenesis of a non-fluorescent chromoprotein from the reef coral Heteractis crispa . The coding sequence for HcRed1 is human codon-optimized for higher expression in mammalian cells . Genes cloned into the multiple cloning site (MCS) downstream of the HcRed1 coding sequence are expressed as fusions to the C-terminus of HcRed1. The MCS in pHcRed1-C1 is positioned between the HcRed1 coding sequence and the SV40 polyadenylation signal (SV40 poly A). Genes cloned into the MCS will be expressed as fusions to the C-terminus of HcRed1 if they are in the same reading frame as HcRed1 and there are no intervening stop codons. A Kozak consensus sequence upstream of HcRed1 increases the translation efficiency in eukaryotic cells . SV40 poly A signals downstream of the MCS direct proper processing of the 3' end of mRNA transcripts. The vector also contains an SV40 origin for replication in mammalian cells expressing the SV40 T-antigen, a pUC origin of replication for propagation in E. coli, and an f1 origin for single-stranded DNA production. 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. pHcRed1-C1 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：
ZK449pHcRed1-C1哺乳荧光质粒.txt
