pEF4/V5-His A, B, and C are 5.7 kb vectors derived from pEF1/V5-His and designed for overproduction of recombinant proteins in mammalian cell lines. Features of the vectors allow purification and detection of expressed proteins. High-level stable and transient expression can be carried out in most mammalian cells. The vectors contain thefollowing elements:
- Human elongation factor 1a-subunit promoter (hEF-1α) promoter for high-level expression across a broad range of species and cell types (Goldman et al., 1996)(Mizushima and Nagata, 1990).
- Three reading frames to facilitate in-frame cloning with a C-terminal tag encoding the V5 epitope and a polyhistidine metal-binding peptide.
- Zeocin resistance gene for selection of stable cell lines (Mulsant et al., 1988).
- Episomal replication in cell lines that are latently infected with SV40 or that express the SV40 large T antigen(e.g. COS7)
The control plasmid, pEF4/V5-His/lacZ is included for use as a positive control for transfection, expression, purification, and detection in the cell line of choice.