Plasmid allows the expression of one poly-protein that is self-cleaved into two separate proteins using the FMDV P2A peptide sequence (Frame 3).
Product Name: pSF-CMV-COOH-P2A-3
Product Code: OG65
Size (bp): 4280 bp
Bacterial Antibiotic Selection: KanR
Origin and Compatibility: pUC high copy derived from pBR322
Bacterial Copy Number: 500-700 per cell
Promoter: Cytomegalovirus (CMV) immediate early promoter
This vector adds a Foot and Mouth Disease P2A peptide between the primary MCS (NotI-XbaI but XbaI is ablated in this vector) and the fusion MCS (ClaI-NheI). The P2A peptide coding sequence is: GSGATNFSLLKQAGDVEENPGP. The peptide self-cleaves between the penultimate glycine residue and the final proline residue. This leaves the first (upstream) protein in the polypeptide with the majority of the amino acids of the P2A peptide on the C-terminus whilst the last (downstream) protein has a proline remaining on the N-terminus. We have not confirmed if secretory proteins are able to detach from cytosolic proteins when fused using this method. The intracellular trafficking of your proteins should therefore be considered.Promoter Expression Level:
This plasmid contains the mammalian CMV promoter to drive gene expression. We have tested all of our mammalian promoters in a range of cell types and CMV is consistently the strongest in those we have studied. However there are many reports of the CMV promoter demonstrating silencing by methylation in long-term culture. For this reason we stock a range of other promoters that are compatible with this plasmid and are available on request.
This plasmid contains three alternative transcription terminators for mammalian bacterial and bacteriophage (T7) expression. This means that only the promoter needs to be changed to alter the expression system you are using. We sell multiple promoters that can be used in each of these systems. The presence of each terminator does not reduce expression in the alternative systems.
There is a start codon in the NcoI site can be removed by digestion with KpnI if required. The MCS for gene insertions in most SnapFast vectors extends from NotI to XbaI.