pEYFP-Tub

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：6045 bp 

载体抗性：Kanamycin (卡那霉素)

筛选标记：Neomycin（新霉素）

pEYFP-Tub encodes a fusion protein consisting of enhanced yellow fluorescent protein (EYFP) and human a-tubulin. EYFP is an enhanced yellow-green variant of the Aequorea victoria green fluorescent protein (GFP). The EYFP gene contains the four amino acid substitutions previously published as GFP-10C: Ser-65 to Gly; Val-68 to Leu; Ser-72 to Ala; and and Thr-203 to Tyr. It also contains the Phe-64 to Leu mutation of GFPmut1. The fluorescence excitation maximum of EYFP is 513 nm; the emission spectrum has a peak at 527 nm (in the yellow-green region). When excited at 513 nm, the Em of EYFP is 36,500 cm-1M-1 and the fluorescence quantum yield is 0.63, resulting in a bright fluorescent signal. The fluorescence observed is roughly equivalent to that from EGFP.

In addition to the chromophore mutations, pEYFP-Tub contains more than 190 silent base changes that correspond to human codon-usage preferences. SV40 polyadenylation signals downstream of the EYFP-Tub fusion direct proper processing of the 3' end of the mRNA. The vector backbone also contains an SV40 origin for replication in mammalian cells expressing the SV40 T-antigen. 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 drives expression of the gene encoding kanamycin resistance in E. coli. The pEYFP-Tub backbone also provides a pUC origin of replication for propagation in E. coli and an f1 origin for single-stranded DNA production.