CaLC3932: To generate a GLC7 heterozygous deletion mutant, the NAT flipper cassette (pLC49)60 was PCR amplified using primers oLC3490 in combination with oLC3491 and transformed into CaLC239 (SN95). NAT-resistant transformants were PCR tested with oLC275 in combination with oLC3558 and oLC274 in combination with oLC3561. The SAP2 promoter was induced to drive expression of FLP recombinase to excise the NAT flipper cassette. To regulate the expression of GLC7, the tetracycline-repressible transactivator, the tetO promoter, and the NAT flipper cassette were PCR amplified from pLC60561 using primers oLC3793 in combination with oLC3492 and transformed into GLC7/glc7::FRT. NAT-resistant transformants were PCR tested with oLC3794 in combination with oLC534 and oLC274 in combination with oLC3495. The absence of a wildtype GLC7 promoter was verified with oLC3794 in combination with oLC3495, and the presence of a deleted allele was verified with oLC3794 in combination with oLC3494. The SAP2 promoter was induced to drive expression of FLP recombinase to excise the NAT flipper cassette.