Fig. 1.

Design of secreted Flag-tagged soluble protein. A Flag-tag (’MDYKDDDDK’) enabling N- or C-terminal tagged fusions to the gene in question was introduced into the polylinker of the pPiczαA vector via the introduced PmlI restriction site (CAC|GTG). Kex2 and Ste2 subsiding the α-factor signal sequence designate proteolytic cleavage sites that are cleaved during passage through the secretory pathway, resulting in mature secreted N- or C-terminal Flag-tagged protein. Abbreviations: α-ss, yeast α mating factor signal sequence; GT, glycosyltransferase; TMD, transmembrane domain, Soluble protein, SP