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 V5 epitope tag that can be fused to a gene of interest to allow protein detection and/or purification. The sequence of the tag is: GKPIPNPLLGLDST. 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 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.
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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