Analysis Note

To view the Certificate of Analysis for this product, please visit www.oxfordgenetics.com.

Application

Cloning in a gene: 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.

General description

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.

Legal Information

Oxford Genetics is a trademark of Oxford Genetics Ltd

Other Notes

Looking for more vector options to move your experiments forward faster? Consider a custom cloning vector designed and built by Oxford Genetics^™. Find out more at Oxford Genetics - Sigma's partner for cloning and expression vectors for molecular biology and synthetic biology applications.

Sequence

Please select the file type you require. For reference most cloning programs will import a .gb (Genbank) file and will show all of the plasmids features automatically when downloaded and imported.Genebank Vector Sequence FileFASTA Vector Sequence FileFull Plasmid Map

Quick-reference Plasmid Map