C-Terminal Glutathione S Transferase Solubility and Affinity Peptide Tag plasmid (Frame 2). Contains the CMV promoter and Kanamycin resistance.
Product Name: pSF-CMV-COOH-EKT-GST2
Product Code: OG139
Size (bp): 4899 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
This vector adds a GST tag positioned at the 3 prime end (C-terminus) of a gene (expressed protein) that is within the primary MCS (NotI to XbaI). The tag can be used to purify a protein by binding to glutathione and the tag can also be used to increase the solubility of recombinant protein produced in bacterial cells. For reference the coding sequence of the GST tag is one base pair out of frame with the TAG stop codon that is found within the upstream XbaI site. There is an enterokinase cleavage site immediately upstream of this tag (DDDDK) that can be used to remove the tag from a purified protein if required.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 can be removed by digestion with KpnI if required. The MCS for gene insertions extends from NotI to XbaI. There are built in Shine-Dalgarno and KOZAK sequences aligned with the start codon within the NcoI restriction site. If these are removed you will need to add a ribosomal entry site to the 5 prime end of your gene for optimal expression.