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GST Tag Plasmids

OG139 - pSF-CMV-COOH-EKT-GST2

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Vector: pSF-CMV-COOH-EKT-GST2

Product Code: OG139

Quantity Provided:  5µg

Size (bp): 4899

Bacterial Antibiotic Selection: Kanamycin

Origin and Compatibility: pUC high copy, derived from pBR322 

Copy Number: 500-700 copies per cell

Promoter: Cytomegalovirus (CMV) immediate early promoter

Purpose: This vector adds a GST tag positioned at the 3’ end (C-terminus) of a gene (expressed protein) that is within the primary MCS (NotI-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.

 

Inserting a Gene: Each restriction site in this vector has a purpose and allows the insertion of specific pre-designed DNA sequences. To insert a gene, we recommend using any sites between (and including) the HindIII and XbaI sites. By doing this you will be able to insert most of our other products upstream, or downstream, of your gene. Click here to read more about the purpose of each restriction site.

Intellectual Property Status: According to our IP-friendly policy, this plasmid is sold free of reach-through rights on any derivatives you may create. 

Quality Sequencing Analysis: This vector has been fully sequenced using primers F1-F10. The sequences and details of these primers are available through our website.

Genetic Modifications to standard parts:

  • CMV promoter – An NcoI site has been ablated in the CMV promoter. This change does not reduce expression.
  • KanR cassette – This KanR resistance region is not a wild type sequence, it has been modified to remove all restriction sites that conflict with the SnapFast system and to retain a high level of antibiotic resistance. It has been validated in E.coli.
  • GST tag – The coding sequence of the GST tag has been modified to remove a BsgI site and a SwaI site. These changes conserved the amino acid sequence of the protein.

Restriction site notes:

  • Bgl2, KpnI, SwaI and PmeI each cut the vector at two sites positioned to flank the promoter, start codon, origin, and KanR, respectively.