PSF-TEF1-NH2-A-FACTORFL - FULL LENGTH ALPHA FACTOR SECRETION PLASMID OGS1875-5UG

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.
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
Analysis Note

form	buffered aqueous solution
mol wt	size 6964 bp
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
yeast selection	uracil

RIDADR	NONH for all modes of transport
Flash Point F	Not applicable
Flash Point C	Not applicable