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OGS1658

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PSF-TEF1-NH2-FLAG®-6HIS-EKT - N-TERMINAL FLAG® AND 6 HIS DUAL TAG YEAST 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

tag

6-His tagged
FLAG® tagged

form

buffered aqueous solution

mol wt

size 6772 bp

bacteria selection

kanamycin

Origin of replication

2Micron
pUC (500 copies)

Peptide cleavage

EKT

Peptide tag location

N-terminal

Promoter

Promoter name: TEF1
Promoter activity: constitutive
Promoter type: yeast

reporter gene

none

shipped in

ambient

storage temp.

−20°C

General description

This plasmid is designed to express tagged proteins in yeast cells (Saccharomyces cerevisiae). The plasmid contains an auxotrophic Uracil selection expression cassette (URA3) that allows for the positive selection of yeast that are deficient in the URA3 gene (YEL021W). This is typically achieved by growing the yeast in minimal media that is reconstituted with the essential amino acids and nucleotides but excluding Uracil.

About the Peptide Tag:This plasmid contains an n-terminal Hexa-Histidine (6His) affinity tag that can be fused to a gene of interest to allow protein detection and/or purification. The sequence of the tag is: HHHHHH. This plasmid also contains a secondary Flag protein tag. The sequence of this tag is: DYKDDDDKWe 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 Yeast Elongation Factor Alpha promoter (TEF1). This is the strongest of the yeast promoters that we sell. It is a constitutive promoter and requires no induction. If you are interested in weaker promoters levels than we also stock plasmids that contain the following promoters in order of decreasing strength TPI (strong), ADHI (medium), STE5 (weak). We also stock Galactose inducible promoter plasmids if inducible expression is required. Please contact us for further information.

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

FLAG is a registered trademark of Merck KGaA, Darmstadt, Germany
Oxford Genetics is a trademark of Oxford Genetics Ltd

related product

Product No.
Description
Pricing

Storage Class Code

12 - Non Combustible Liquids

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Geoffrey M Lynn et al.
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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.
Diana Romero et al.
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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
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Articles

A range of forward and reverse sequencing primers that allow you to sequence any insert that you make into a particular position within any plasmid. Where possible, the binding sites for each of these primers is conserved.

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