CaLC3693/CaLC3694/CaLC3695: To introduce a mutant allele of CAS5 carrying S462E, S464E, S472E, and S476E mutations, the CAS5S S462E/S464E/S472E/S476E -HA-HIS1 cassette was released from pLC857 with SacII and transformed into CaLC2034. Transformants prototrophic for histidine were PCR tested with oLC3052 in combination with oLC2029 and oLC2164 in combination with oLC1645. The absence of an untagged wild type CAS5 allele was verified with oLC3052 in combination with oLC2164. The S462A/S464A/S472A/S476A mutations were sequence verified with oLC3371.