PSF-TEF1-NH2-A-FACTORFL-10HIS - ALPHA FACTOR SECRETION AND 10HIS TAG PLASMID OGS1867-5UG

NACRES:

NA.85

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 Deca-Histidine (10His) protein tag. The sequence of this tag is: HHHHHHHHHHWe 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.
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,
Quick-reference Plasmid Map,
Analysis Note
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.
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form	buffered aqueous solution
mol wt	size 6994 bp
conjugate	10-His tagged
Origin of replication	2Micron, pUC (500 copies)
Peptide cleavage	no cleavage
Peptide tag location	N-terminal
Promoter	Promoter name: TEF1 Promoter activity: constitutive Promoter type: yeast
Secretion signal	alpha factor FL
bacteria selection	kanamycin
reporter gene	none
shipped in	ambient
storage temp.	20°C

RIDADR

NONH for all modes of transport