Product Name: pSF-CMV-PGK
Product Code: OG387
Size (bp): 4747 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 / Mouse Phosphoglycerate Kinase (PGK) Promoter
This plasmid 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:
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 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.