Product Name: pSF-CMV-Ub_AscI
Product Code: OG399
Size (bp): 5595 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 / Human Ubiquitin Promoter
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.
Purpose: This plasmid contains two multiple cloning sites that allow the expression of two genes in mammalian cells one driven by the CMV promoter and one by the human Ubiquitin (Ub) promoter. The CMV promoter is significantly stronger than the Ub promoter (relative expression will vary with cell type) however the Ub promoter is reported to be less effected by methylation than the CMV promoter.
This plasmid contains two multiple cloning sites and allows the insertion of two genes, one downstream of the CMV promoter and one downstream of the human Ubiquitin promoter.
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.Second Multiple Cloning Site Notes: