Table 5.
Primers used in this study
| Purpose | Primer # | Sequence | Product sizes |
|---|---|---|---|
| Amplify PGAL4-DEG1-SEC62-HIS3:natMX4 cassette for integration at DOA10 locus in SGA query strain from pVJ490 | VJR264 | 5′ TAGCCAAGAGTACCACTAATTGAATCAAAGAGACTAGAAGTGTGAAAGTCcgcgcaattaaccctcacta 3′ | 4338 bp |
| VJR265 | 5′ TATATGTAAATATGCTAGCATTCATTTTAAATGTAAGGAAGAAAACGCCTggcggcgttagtatcgaat 3′ | ||
| Confirm integration of PGAL4-DEG1-SEC62-HIS3:natMX4 cassette at DOA10 locus (5′ end) | VJR46 | 5′ CATGGTACCGAGCTCCTTGT 3′ | WT DOA10–842 bp doa10Δ::PGAL4-DEG1-SEC62-HIS3:natMX4 – 1073 bp |
| VJR82 | 5′ AATCGGTAGCGCGTATGACT 3′ | ||
| VJR260 | 5′ CGTGGTTAATTCTGGAGTTGC 3′ | ||
| Confirm integration of PGAL4-DEG1-SEC62-HIS3:natMX4 cassette at DOA10 locus (3′ end) | VJR11 | 5′ ATTCCCAACATGGACAAGGA 3′ | WT DOA10–550 bp doa10Δ::PGAL4-DEG1-SEC62-HIS3:natMX4 – 746 bp |
| VJR107 | 5′ GTTTGGGATGAGGGCAGAG 3′ | ||
| VJR249 | 5′ ACCTCTGGCTGGAGGTCAC 3′ | ||
| Genotyping HRD1 locus (HRD1versushrd1Δ::kanMX4) | VJR70 | 5′ TGCAAAAAGGAAACGCTTGT 3′ | WT HRD1 – 780 bp hrd1Δ::kanMX4 – 987 bp |
| VJR163 | 5′ ATTGGCCATTAGAGGTGACG 3′ | ||
| VJR259 (Alias kanB) | 5′ CTGCAGCGAGGAGCCGTAAT 3′ | ||
| Genotyping INO4 locus (INO4versusino4Δ::kanMX4) | VJR371 | 5′ GAAAAGGGTTGCAGTTAAGCA 3′ | WT INO4 – 450 bp ino4Δ::kanMX4 – 655 bp |
| VJR372 | 5′ TCTTCTTAGACCTGCGTTTCC 3′ | ||
| VJR259 (Alias kanB) | 5′ CTGCAGCGAGGAGCCGTAAT 3′ | ||