CaLC2034: To generate a CAS5 heterozygous deletion mutant, the NAT flipper cassette (pLC49)60 was PCR amplified using primers oLC2017 and oLC2018 (4366 bp) and transformed into CaLC239 (SN95). NAT-resistant transformants were PCR tested with oLC275 in combination with oLC2034 for upstream integration and oLC274 in combination with oLC2035 for downstream integration. The SAP2 promoter was induced to drive expression of FLP recombinase to excise the NAT flipper cassette.