PSF-CMV-COOH-P2A-3 - FMDV P2A PLASMID OGS65-5UG

NACRES:

NA.85

PSF-CMV-COOH-P2A-3 - FMDV P2A plasmid 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.
This vector is designed to allow two genes to be inserted one on either site of the P2A self-cleaving tag. This can be achieved using standard cloning methodologies.
Multiple Cloning Site Notes: 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 however in this vector the XbaI site has been ablated to remove the stop codon in the site and allow read-through into the P2A coding sequence.
Sequence
Analysis Note

form	buffered aqueous solution
mol wt	size 4280 bp
conjugate	P2A tagged
Peptide cleavage	no cleavage
reporter gene	none
shipped in	ambient
storage temp.	−20°C

RIDADR	NONH for all modes of transport
Flash Point F	Not applicable
Flash Point C	Not applicable