pEYFP-Mem

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：4.8kb 

载体抗性：Kanamycin (卡那霉素)

筛选标记：新霉素（Neomycin）

pEYFP-Mem encodes a fusion protein consisting of the N-terminal 20 amino acids of neuromodulin, also called GAP-43, and a yellow-green fluorescent variant of the enhanced green fluorescent protein (EGFP). The neuromodulin fragment contains a signal for posttranslational palmitoylation of cysteines 3 and 4 that targets EYFP to membranes. The EYFP gene contains four amino acid substitutions previously published as GFP-10C. The fluorescence excitation maximum of EYFP is 513 nm, and the emission maximum is 527 nm (in the yellow-green region).

In addition to the chromophore mutations, EYFP contains >190 silent mutations that create an open reading frame comprised almost entirely of preferred human codons. Furthermore, upstream sequences flanking the EYFP-Mem fusion protein have been converted to a Kozak consensus translation initiation site. These changes increase the translational efficiency of the fusion protein and consequently its expression in mammalian cells.

Expression of EYFP-Mem is driven by the immediate early promoter of CMV (PCMV IE). The vector contains an SV40 origin of replication and a neomycin resistance (Neor) gene for selection in mammalian cells. A bacterial promoter upstream of this cassette (P) expresses kanamycin resistance in E. coli. The vector backbone also provides a pUC19 origin of replication for propagation in E. coli and an f1 origin for single-stranded DNA production.