质粒类型：哺乳动物表达载体

启动子：CMV

克隆方法：Gateway

载体大小：7711 bp 

5' 测序引物及序列：T7 Fwd:5'd[TAATACGACTCACTATAGGG]3'

载体标签：V5 Epitope Tag

载体抗性：Ampicillin (氨苄青霉素)

筛选标记：Neomycin (新霉素)

        pcDNA6.2/EmGFP-Bsd/V5-DEST vector allows you to rapidly clone your gene using Gateway technology and simultaneously express it with Emerald Green Fluorescent Protein (EmGFP) for easy identification of transfected cells. The EmGFP is derived from well-characterized fluorescent proteins of the jellyfish Aequorea victoria and has been humanized for optimal mammalian expression. If cloned with the TAG (amber) codon, your gene will be expressed in its native configuration and fully compatible with Tag-On-Demand" technology. If cloned without a stop codon, a V5-epitope tag will be fused to the carboxy-terminal end of your protein of interest. In either case, expression of EmGFP provides a bright fluorescent marker to easily identify cells co-expressing your protein. This marker allows you to focus your functional analysis on specific cells, or to sort and enrich for transfected cells in a derived population.      

        pcDNA6.2/EmGFP-Bsd/V5-DEST vector provides:

（1）PGK promoter for high-level expression of EmGFP fused to the Blasticidin (bsd)-resistance marker.

（2）CMV promoter for high-level expression of your protein.

（3）V5-epitope tag option to fuse to the C-terminal end of your protein, if desired. 

        pCDNA6.2/EmGFP-Bsd/V5-DEST实际上是一个双表达框载体，PGK启动子带一个EmGFP荧光标记基因，CMV启动子调控外源基因的表达。不过，要把外源基因插入至CMV启动子后面，只能用LR重组的方法实现，所以，外源基因需要先构建至Entry载体上，例如pENTR/D-TOPO等。

        pCDNA6.2/EmGFP-Bsd/V5-DEST空载上，CMV启动子后有ccdB表达框，所以此空载体只能在DB3.1等专用的大肠杆菌中才能复制扩增，使用此载体时，请注意配套相应的DB3.1感受态（赢润生物HonorGene有售）。当外源基因成功插入，ccdB表达框将被置换掉，此时，重组质粒可以在DH5α、TOP10等常规大肠杆菌中扩增复制。