CaLC3209: To introduce a mutant allele of CAS5 carrying the S769E mutation, the CAS5 S769E-HA-HIS1 cassette was released from pLC791 with ApaI and transformed into CaLC2034. Transformants prototrophic for histidine were PCR tested for integration of the cassette with oLC3052 in combination with oLC2029 and oLC2164 in combination with oLC1645. The absence of a wildtype CAS5 allele was verified with oLC3052 in combination with oLC2164. The S769E mutation was sequence verified with oLC2029.