pSF-CMV-COOH-EKT-GST1 (OG310) C-Terminal GST Tag Plasmid

Product Code: OG310R1
Product Code: OG310C1
Product Code: OG310G1

Product Description

C-terminal GST affinity/solubility tag plasmid (Frame 1). Adds a GST tag to the end of your gene of interest and also has an EKT cleavage tag.

Plasmid Information

Product Name: pSF-CMV-COOH-EKT-GST1

Product Code: OG310

Size (bp): 4889 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

Plasmid Purpose:

This vector adds a Glutathione-S-transferase (GST) purification tag that allows proteins to be purified by binding to glutathione. The GST tag will be positioned at the 3 prime end (C terminus) of a gene (expressed protein) in the primary MCS (NotI-XbaI but XbaI is ablated in this vector because of the stop codon in the site). The GST tag coding sequence is 219 amino acids in length. There is an enterokinase cleavage site immediately upstream of this tag (DDDDK) that can be used to remove the tag from a purified protein it cleaves after the lysine residue.

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.

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Transcription Termination:

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.

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.

Intellectual Property Status

This product is part of our SnapFastâ„¢ plasmid range, for more information on the intellectual property status of this plasmid please click here. For more information on the terms of our licences please click here.