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OGS140

Sigma-Aldrich

PSF-CMV-COOH-EKT-GST3 - C-TERMINAL GST PLASMID

plasmid vector for molecular cloning

Synonym(s):

cloning vector, expression vector, molecular cloning vector, plasmid, plasmid vector, snapfast vector, vector

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About This Item

UNSPSC Code:
12352200
NACRES:
NA.85
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tag

GST tagged

form

buffered aqueous solution

mol wt

size 4900 bp

bacteria selection

kanamycin

origin of replication

pUC (500 copies)

peptide cleavage

EKT

peptide tag location

C-terminal

promoter

Promoter name: CMV
Promoter activity: constitutive
Promoter type: mammalian

reporter gene

none

shipped in

ambient

storage temp.

−20°C

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.

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.

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 File

FASTA Vector Sequence File

Full Plasmid Map

Analysis Note

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

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.

Legal Information

Oxford Genetics is a trademark of Oxford Genetics Ltd

related product

Product No.
Description
Pricing

Storage Class

12 - Non Combustible Liquids

flash_point_f

Not applicable

flash_point_c

Not applicable


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Alexander C Cerny et al.
PLoS genetics, 11(10), e1005578-e1005578 (2015-10-29)
Recycling of signaling proteins is a common phenomenon in diverse signaling pathways. In photoreceptors of Drosophila, light absorption by rhodopsin triggers a phospholipase Cβ-mediated opening of the ion channels transient receptor potential (TRP) and TRP-like (TRPL) and generates the visual
Geoffrey M Lynn et al.
Nature biotechnology, 33(11), 1201-1210 (2015-10-27)
The efficacy of vaccine adjuvants such as Toll-like receptor agonists (TLRa) can be improved through formulation and delivery approaches. Here, we attached small molecule TLR-7/8a to polymer scaffolds (polymer-TLR-7/8a) and evaluated how different physicochemical properties of the TLR-7/8a and polymer
Diana Romero et al.
Carcinogenesis, 37(1), 18-29 (2015-10-28)
Dickkopf-3 (Dkk-3) is a secreted protein whose expression is downregulated in many types of cancer. Endogenous Dkk-3 is required for formation of acini in 3D cultures of prostate epithelial cells, where it inhibits transforming growth factor (TGF)-β/Smad signaling. Here, we
Jin-Gyoung Jung et al.
PLoS genetics, 10(10), e1004751-e1004751 (2014-10-31)
The Notch3 signaling pathway is thought to play a critical role in cancer development, as evidenced by the Notch3 amplification and rearrangement observed in human cancers. However, the molecular mechanism by which Notch3 signaling contributes to tumorigenesis is largely unknown.

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