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OGS1821

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PSF-TEF1-NH2-A-FACTORFL-6HIS-THR - ALPHA FACTOR SECRETION AND 6 HIS TAG 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

form

buffered aqueous solution

mol wt

size 7000 bp

bacteria selection

kanamycin

origin of replication

2Micron
pUC (500 copies)

peptide cleavage

thrombin

peptide tag location

N-terminal

promoter

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

reporter gene

none

secretion signal

alpha factor FL

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 full length Alpha Factor (A-FactorFL) secretory peptide to allow proteins to be exported from the cytosol. Alpha factor is an endogenous yeast protein that is naturally secreted into the supernatant. The wild-type Alpha Factor protein contains a series of repeating protein sequences that are cleaved out by cellular proteases that recognise a specific site. This produces a series of small peptides that contain the mature alpha peptide sequence. This plasmid contain the wild-type signal peptide from the full length Alpha factor gene but also the coding sequence that leads up to the first endogenous protease cleavage site. The addition of these sequences has been demonstrated to provide more consistent secretion of proteins in comparison to using just the signal peptide alone. All of the wild-type protein sequences will be removed during protein maturation and export by endogenous proteases. This plasmid also contains a secondary Hexa-Histidine (6His) protein tag. The sequence of this tag is: HHHHHHWe 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 Thrombin cleavage tag. The protein sequence of the cleavage tag is: LVPRGS. It cleaves preferentially between the Arg and Gly residues. Off target cleavage can often occur at non-specific sites normally from other contaminating proteases. To ensure maximal protein integrity the enzyme reagent must be highly pure.

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

To view sequence information for this product, please visit the product page

Analysis Note

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

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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