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OGS1416

Sigma-Aldrich

PSF-CMV-PURO-NH2-INSULINSP-S-TAG-EKT - INSULIN SECRETION AND S-TAG TAG PLASMID

plasmid vector for molecular cloning

Synonym(s):
expression vector, molecular cloning vector, vector, snapfast vector, plasmid vector, cloning vector, plasmid
NACRES:
NA.85

conjugate

S-Tag

form

buffered aqueous solution

mol wt

size 6275 bp

Origin of replication

pUC (500 copies)

Peptide cleavage

EKT

Peptide tag location

N-terminal

Promoter

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

Secretion signal

human Insulin

bacteria selection

kanamycin

mammalian cells selection

puromycin

reporter gene

none

shipped in

ambient

storage temp.

−20°C

General description

This plasmid is designed to express tagged proteins in mammalian cells either by transient transfection or by creating stable cell lines. It contains a puromycin resistance expression cassette using the human Ubiquitin promoter to drive expression and allow for the selection of cells containing the plasmid.

About the Peptide Tag:This plasmid contains an Human Insulin (iSP) secretory signal peptide (SP) to allow proteins to be exported from the cytosol. During translocation from the cytosol the signal peptide is removed from the protein by endogenous proteases. This plasmid also contains a secondary S-Tag protein tag. The sequence of this tag is: KETAAAKFERQHMDSWe provide a range of dual peptide tag plasmids. This is because some peptide tags provide specific biological properties (e. g., small molecule affinity new epitopes solubility or protein secretion) that are not provided by others.

About the Cleavage Tag: This plasmid also encodes a protease cleavage site that is designed to be positioned between your gene of interest and the tag to allow the removal of the tag following protein purification or isolation. This plasmid contains a EKT cleavage tag. The protein sequence of the cleavage tag is: DDDDK. Enterokinase (EKT) protease cleaves after the Lysine residue. It can cleave at other basic residues but this is dependent on protein confirmation. If a proline follows the site it will not cut. None of our products contain a proline after the site.

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.

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

Certificate of Analysis

Certificate of Origin

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
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
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
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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