质粒类型：逆转录病毒载体

高拷贝/低拷贝：高拷贝

克隆方法：多克隆位点，限制性内切酶

载体大小：5953bp 

3' 测序引物及序列：IRES-R:CCTCACATTGCCAAAAGACG

载体抗性：Ampicillin (氨苄青霉素)

备注：载体表达ZsGreen绿色荧光蛋白作为筛选标志，用来构建稳表达细胞系。

pRetroX-IRES-ZsGreen1 is a bicistronic, fl uorescent, retroviral vector that allows both a gene of interest and the ZsGreen1 gene to be translated from a single bicistronic mRNA. pRetroXIRES- ZsGreen1 is designed for effi cient delivery and selection (by fl ow cytometry or other methods) of stably transduced mammalian cells expressing the ZsGreen1 fl uorescent protein and the protein of interest. This vector can be used to obtain stable cell lines without timeconsuming drug and clonal selection. ZsGreen1 is derived from the reef coral Zoanthus sp. green fl uorescent protein (ZsGreen) and is easily detected with standard FITC fi lter sets (1).

Bicistronic expression from this vector is facilitated by the encephalomyocarditis virus (EMCV) internal ribosome entry site (IRES). This IRES facilitates cap-independent translation from an internal start site at the IRES/ZsGreen1 junction (2). ZsGreen1 is a human codon optimized ZsGreen variant that encodes the brightest commercially available green fl uorescent protein (1). This retroviral vector is derived from the pMIN series of vectors (3, 4). These optimized vectors have the ability to produce high viral titers, express genes at high levels, and, due to the absence of retroviral coding sequences, exhibit improved safety profi les. The multiple cloning site (MCS) in pRetroX-IRES-ZsGreen1 is between the 5’ MMLV LTR and the IRES sequence. Genes cloned into the MCS are expressed as a bicistronic message transcribed from the 5’ LTR.

pRetroX-IRES-ZsGreen1 contains all of the necessary viral RNA processing elements; these include the 5’ and 3’ LTRs, the packaging signal (ψ), and the tRNA primer-binding site. For safety reasons, however, the vector lacks the structural genes (gag, pol, and env) necessary for retroviral particle formation and replication. pRetroX-IRES-ZsGreen1 contains a ColE1 origin of replication, and an E. coli Ampr gene for propagation and selection in bacteria.

pRetroX-IRES-ZsGreen1(点击下载)

pRetroX-IRES-ZsGreen1质粒序列.txt