The diploid strain with multiple heterozygous markers on chromosome XII (UCC8915) was generated by mating UCC8914 x UCC5179. UCC8914 was derived from UCC8913 by one-step integration of a PCR fragment carrying TRP1 into an intergenic region distal to the right end of the rDNA array (coordinates 486062-486189), which was generated using oligos RDNRF and RDNRR with pRS304 [64] as a template. Integration was verified by PCR from genomic DNA using oligos RDNRconF and RDNRconR. UCC8913 was derived from UCC5181 by one-step integration of a PCR fragment carrying KANMX into an intergenic region proximal to the left end of the rDNA array (coordinates 450191-450372), which was generated using oligos RDNLF and RDNLR with pRS400 [63] as a template. Integration was verified by PCR from genomic DNA using oligos RDNLconF and RDNLconR.
