Table 2.
Oligonucleotides
| Primer | Sequence (5′ to 3′) | Use |
|---|---|---|
| ICL1-5′ | CCCTCGAGGTCATGGAATCG | Cloning of ICL1 promoter (XhoI site underlined) |
| ICL1-3′ | GGAAGCTTTTATTAATGTTTATTC | Cloning of ICL1 promoter (HindIII site underlined) |
| PYK5′-2 | CCCTCGAGTATAGAAATTGGGTTGC | Cloning of PYK1 promoter (XhoI site underlined) |
| PYK3′-3 | AAACTGCAGTGATGTAGTTTGTAGG | Cloning of PYK1 promoter (PstI site underlined) |
| PFK2-5′ | CCGCTCGAGATTCTATGACGTGGGT | Cloning of PFK2 promoter (XhoI site underlined) |
| PFK2-3′ | CCCAAGCTTGGTGGTTGTTTTTCGT | Cloning of PFK2 promoter (HindIII site underlined) |
| ICL1-L1 | TCCGGATGGCAATGTTCTTCTACTGCTTCCACTTCTAACGAACCATCTCCAGATTTGGCTCCAGGGTTTTCCCAGTCACG | PCR amplification of icl1 disruption cassettes (homology to pLHL and pLUL plasmids underlined) |
| ICL1-L2B | CTTGATGTAAGTTTCTTGTTCGTCAGCTGGCATGGCCTTGTTCCAGTTGAAAGATGGAGACTCACTAAAGGGAACAAAAGC | PCR amplification of icl1 disruption cassettes (homology to pLHL and pLUL plasmids underlined) |
| ICL1-IP | TTCTGGGTTAGTGGCACCAA | Diagnosis of icl1 alleles, internal 3’ primer |
| ICL1-5′-DIAG | CAGGTGAAGAGTCACTTCTA | Diagnosis of icl1 alleles, upstream primer |
| PCK-L1 | AAATAATTATCAATCATGGCTCCTCCTACTGCTGTTGAATCTTCAATCAATTTCGGAGGTCACCCAGGGTTTTCCCAGTCACG | PCR amplification of pck1 disruption cassettes (homology to pLHL and pLUL plasmids underlined) |
| PCK1-L2 | GACAATTCACCAGAGTGGATAGCATCCAAGATAGCTCTGGTGTATTTCAATGGACATCTCACTAAAGGGAACAAAAGC | PCR amplification of pck1 disruption cassettes (homology to pLHL and pLUL plasmids underlined) |
| PCK-5′-DIAG | GACATCACCTTTTTCAACC | Diagnosis of pck1 alleles, internal 3’ primer |
| PCK-IP | CAACAATACCGTCAACTC | Diagnosis of pck1 alleles, upstream primer |
| PYK-L1 | CTACATCAACAATGTCTCACTCATCTTTATCTTGGTTATCCAACTTCAATGTTGAAACTGTTCCAGGGTTTTCCCAGTCACG | PCR amplification of pyk1 disruption cassettes (homology to pLHL and pLUL plasmids underlined) |
| PYK-L2 | ATCAAGGCTTCTTTTCCACTTAAGCTTGGACGATTCTAACAGTGTTAGAGTGACCAGAACCTCACTAAAGGGAACAAAAGC | PCR amplification of pyk1 disruption cassettes (homology to pLHL and pLUL plasmids underlined) |
| PYK-IP | TCCTTACCTTCTTCACC | Diagnosis of pyk1 alleles, internal 3’ primer |
| PYK-5′-DIAG | TTAGAAAGGAAACAAAGGG | Diagnosis of pyk1 alleles, upstream primer |
| HIS1-REV-DIAG | CACAAGAAGCCTCAACGG | Diagnosis of LHL alleles, downstream primer |
| URA3-REV-DIAG | TAGTGTTACGAATCAATGGC | Diagnosis of LUL alleles, downstream primer |
| ICL1-CIp20-5′ | CCATCGATCAAAAACTTAATTTAACCAGCG | Cloning of ICL1 locus (ClaI site underlined) |
| ICL1-CIp20-3′ | CGACGCGTTAAAGGCTCAAATTGTTCCCG | Cloning of ICL1 locus (MluI site underlined) |
| PYK-CIp20-5′ | CCATCGATTATAGAAATTGGGTTGC | Cloning of PYK1 locus (ClaI site underlined) |
| PYK-CIp20-3′ | CGACGCGTGCAGAATTAACGTATG | Cloning of PYK1 locus (MluI site underlined) |
| PCK-CIp20-5′ | CCATCGATTTGGTCCATTGCCGT | Cloning of PCK1 locus (ClaI site underlined) |
| PCK-CIp20-3′ | CGACGCGTCCATAGGATTATGAGAC | Cloning of PCK1 locus (MluI site underlined) |