pDsRED2-Peroxi

质粒类型：荧光蛋白报告载体

启动子：CMV

克隆方法：多克隆位点，限制性内切酶

载体大小：4.6kb 

载体抗性：Kanamycin (卡那霉素)

筛选标记：新霉素（Neomycin）

pDsRed2-Peroxi is a mammalian expression vector that encodes a fusion of Discosoma sp. red fluorescent protein (DsRed2) and the peroxisomal targeting signal 1 (PTS1). The PTS1 sequence is fused to the 3'-end of DsRed2, a DsRed variant engineered for faster maturation and lower non-specific aggregation. The PTS1 sequence encodes the tripeptide SKL, which targets the DsRed2-PTS1 fusion protein to the matrix of peroxisomes.

To drive expression of DsRed2-PTS1, this vector contains the immediate early promoter of cytomegalovirus (PCMV IE). SV40 polyadenylation signals downstream of the DsRed2 gene direct proper processing of the 3'-end of the DsRed2-PTS1 mRNA transcript. Because it encodes DsRed2, a gene variant that uses human-preferred codons, the DsRed2-PTS1 transcript is suited for efficient translation in mammalian cells. To further increase the translational efficiency of DsRed2-PTS1, upstream sequences have been converted to a Kozak consensus translation initiation site. The vector also contains an SV40 origin for replication in any mammalian cell line that expresses 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—consisting of the SV40 early promoter (PSV40e), the neomycin/kanamycin resistance gene of Tn5 (Neor/Kanr), and polyadenylation signals from the herpes simplex virus thymidine kinase (HSV TK poly A) gene—allow stably transfected eukaryotic cells to be selected using G418. A bacterial promoter (P) upstream of this cassette drives expression of the gene encoding kanamycin resistance in E. coli.