Fig. 5
Vector map of a general ARS plasmid carrying a carbon marker cassette. Besides an origin of replication and an Ampicillin resistance cassette for cloning in E. coli, the episomal plasmids used in this study harbor the PARS1 sequence for the replication in K. phaffii, a reporter cassette with the de-repressable PDC promoter driving CalB expression, whose secretion is facilitated by the D-alpha signal sequence, a deletion variant of the S. cerevisiae mating factor alpha signal peptide. Selective pressure in K. phaffii is provided by either GUT1 or TPI1 genes with one of the 5 tested promoters placed upstream