■ 基本信息
啟動(dòng)子: | CMV IE promoter |
復(fù)制子: | pUC ori，F(xiàn)1 ori，SV40 ori |
終止子: | SV40 early polyA signals |
質(zhì)粒分類: | 哺乳細(xì)胞質(zhì)粒；哺乳熒光質(zhì)粒；哺乳紅色質(zhì)粒 |
質(zhì)粒大小: | 4698bp |
質(zhì)粒標(biāo)簽: | N-HcRed1 |
原核抗性: | Kan |
真核抗性: | G418 |
克隆菌株: | DH5a |
培養(yǎng)條件: | 37℃，5%CO2 |
表達(dá)宿主: | 293T等哺乳細(xì)胞 |
誘導(dǎo)方式: | 無需誘導(dǎo)，組成型表達(dá) |
5'測(cè)序引物: | CMV-F：CGCAAATGGGCGGTAGGCGTG |
3'測(cè)序引物
| Sv40-polyA-R：GAAATTTGTGATGCTATTGC
|
備注: | 高拷貝 |
■ 質(zhì)粒屬性
質(zhì)粒宿主: | 哺乳細(xì)胞 |
質(zhì)粒用途: | 蛋白表達(dá) |
片段類型: | ORF
|
片段物種: |
|
原核抗性: | Kan |
真核抗性: | G418
|
熒光標(biāo)記: |
|
■ 質(zhì)粒簡介
pHcRed1-C1質(zhì)粒是一個(gè)哺乳細(xì)胞紅色熒光蛋白表達(dá)載體，CMV IE啟動(dòng)子驅(qū)動(dòng)HcRed1紅色熒光蛋白基因和目的基因融合表達(dá)。
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 .
質(zhì)粒只保證關(guān)鍵序列正確，不保證表達(dá)效果。
■ 質(zhì)粒圖譜
■ 質(zhì)粒序列
質(zhì)粒序列請(qǐng)下載：
ZK449pHcRed1-C1哺乳熒光質(zhì)粒.txt
