PSF-CMV-PGK - DUAL PROMOTER EXPRESSION PLASMID OGS387-5UG

This cloning vector contains two promoters that terminate transcription at the same poly-adenylation signal allowing the expression of two genes from one expression cassette. The second promoter (PGK) is approximately 10-fold weaker than the upstream CMV promoter.
Promoter Expression Level: PSF-CMV-PGK - DUAL PROMOTER EXPRESSION 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.
PSF-CMV-PGK - DUAL PROMOTER EXPRESSION PLASMID contains two multiple cloning sites and allows the insertion of two genes one downstream of the CMV promoter and one downstream of the PGK promoter. Both promoters terminate transcription at the same poly-adenylation signal
Multiple Cloning Site Notes: There is a start codon in the NcoI site that can be removed by digestion with KpnI if required. The MCS for gene insertions extends from NotI to XbaI but the Shine-Dalgarno sequences and KOZAK ribosomal entry site sequences are aligned with the start codon in the NcoI site. The PGK promoter has been inserted between the ClaI and BamHI sites and contains sites downstream to allow a second gene to be inserted.
The BsgI and BseRI restriction sites in the first MCS cleave within the stop codon in the XbaI site and allow the retrospective fusion of coding sequences.
Sequence
Analysis Note

form	buffered aqueous solution
mol wt	size 4747 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