PSF-CMV-EMCV - CMV IRES PLASMID OGS97-5UG

NACRES:

NA.85

This vector contains the internal ribosome entry site (IRES) from Encephalomyocarditis virus (EMCV) between the promoter and the multiple cloning site (MCS). It is designed to be used to express genes using cap-independent expression systems although the mRNAs produced from this vector contain a 5 prime cap. This vector is not designed for the expression of two proteins from the same mRNA. For this purpose we have designed pSF-CMV-EMCV-PciI where the PciI site contains start codons in the correct position for the IRES to express efficiently. In this vector there is an NcoI site (which contains an ATG start codon) in the correct position to enable efficient expression of a gene in the MCS. We have also developed this same vector with the T7 promoter upstream of the IRES to allow the expression of genes using T7 eukaryotic systems.
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.
Cloning in a gene: There is start codon within the NcoI restriction site that is positioned to allow efficient expression from the EMCV IRES. The main MCS for gene insertions in this vector therefore extends from the NcoI restriction site to the XbaI restriction site. The downstream sites (ClaI to NheI sites) have alternate functions and are used to insert other DNA sequences that we sell however they can be used to insert a gene if these functions are not required. These functions include adding peptide tags or downstream IRES elements. The BsgI and BseRI restriction sites cleave within the stop codon in the XbaI site and allow the retrospective fusion of coding sequences. These sites are normally only used on genes that we sell in the main multiple cloning site.
Sequence
Analysis Note

form	buffered aqueous solution
mol wt	size 4761 bp
Origin of replication	pUC (500 copies)
Peptide cleavage	no cleavage
Promoter	Promoter name: CMV Promoter activity: constitutive Promoter type: mammalian
bacteria selection	kanamycin
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