TABLE 1

Primers used for gene overexpression, quantitative real-time PCR, and mutation construction

Gene and primersSequencea
Cloning of promoters and terminators
    BAT
        BAT-5′-sp15′-TCAACCATCCTTCCTCCTTATTCCATTC-3′
        BAT-5′-sp25′-AGGGCTTGGGGGTGGTATTCTTGGTG-3′
        BAT-5′-sp35′-GCTGAGGCTGCGCGGNNNNNNNNNCTGAAG-3′
        BAT-5′-sp45′-TGCCCAGCATGTTGGCAGAG-3′
        BAT-3′-sp15′-TATTACCCCACCGGCTTCAAGGCCGTC-3′
        BAT-3′-sp25′-CCACCAGCAGAACCTGTGGCTCTTC-3′
        BAT-3′-sp35′-TCGGAGCCGGTACCGNNNNNNNNNTCAGCC-3′
        BAT-3′-sp45′-CTCTTGATGGCACCATTCTC-3′
    ARO8-2
        ARO8-2-5′-sp15′-ACTCTGGCCGTCTCGGAGATCGTGCTTG-3′
        ARO8-2-5′-sp25′-AGTATTCACTCAGAGGAAGCCCTCCTCC-3′
        ARO8-2-5′-sp35′-TTGAAGGCACCCTTGNNNNNNNNNCCAGTC-3′
        ARO8-2-5′-sp45′-ATAACTTAGGCGATCAGTCC-3′
        ARO8-2-3′-sp15′-TATCCGCATGGACTCCTTCAGCAAAG-3′
        ARO8-2-3′-sp25′-CCTCAGCCAGATTGCCCTCTTCAAAC-3′
        ARO8-2-3′-sp35′-TTGTAAGTAACCAATNNNNNNNNNCACGTT-3′
        ARO8-2-3′-sp45′-GAGCGTCACCTTCCCAAGAG-3′
    PDC
        PDC-5′-sp15′-TCCAACACCTGCATAGACACCGCCATAG-3′
        PDC-5′-sp25′-TCTTGTCAGAAAGCGCAAGAACCTCGTC-3′
        PDC-5′-sp35′-AAGACGACATCCACTNNNNNNNNNTCCTTC-3′
        PDC-5′-sp45′-ATCCAGCATTCGCGCAGAGC-3′
        PDC-3′-sp15′-ACTGCGGGTTTCTCATACCGAACATC-3′
        PDC-3′-sp25′-AGGCGTGTTGCGAAAGCTAACCAGCAC-3′
        PDC-3′-sp35′-CATCTGGGACACCAANNNNNNNNNGGACGT-3′
        PDC-3′-sp45′-CAGCTCGCCAACAATTACCC-3′
    STR3
        STR3-5′-sp15′-AGATCATCGCCGGTGATGACCTCGTCAC-3′
        STR3-5′-sp25′-TTGGCAAGATGGCGTTCTAGATGGGTTC-3′
        STR3-5′-sp35′-TCATACTCAGCCTGTNNNNNNNNNGCGCTG-3′
        STR3-5′-sp45′-ATGCTCGAAGCACCATATTG-3′
        STR3-3′-sp15′-TCACCCAATGACTCATTCCTTCTCATG-3′
        STR3-3′-sp25′-AGTTCGATATCCCGGACTTAAAAGCCA-3′
        STR3-3′-sp35′-CAAGAAGAGAAAGACNNNNNNNNNAAGATA-3′
        STR3-3′-sp45′-GAATCGTTGAGGCGGCAAGACTG-3′
Overexpression of Met catabolism genes
    BAT
        BAT-OE-F5′-GAATTCGAGCTCGGTACCCGGGGGGAAATGATGGCAGGACCTTGACAAG-3′
        BAT-OE-R5′-CGATCGGGGAAATTCGAGCTGGCCTCACAGCGTCTCTACTTGGGCTC-3′
    ARO8-2
        ARO8-2-OE-F5′-GAATTCGAGCTCGGTACCCGGGAGAACAAGCTGCAAGCCATTCAGGTAG-3′
        ARO8-2-OE-R5′-CGATCGGGGAAATTCGAGCTGGAAGTGTACACATGAATACTCATGAC-3′
    PDC
        PDC-OE-F5′-GAATTCGAGCTCGGTACCCGGGTCGAGGGCGCAGCAGGGCGATCTGGTG-3′
        PDC-OE-R5′-CGATCGGGGAAATTCGAGCTGCCTGTGCTATAGGATCATCATCGTCG-3′
    STR3
        STR3-OE-F5′-GAATTCGAGCTCGGTACCCGGGCAGAAAAGGGCCTCGGCTGGAGGAAC-3′
        STR3-OE-R5′-CGATCGGGGAAATTCGAGCTGGATCCACTTAACGTTACTGAAATCATC-3′
Identification of overexpressing strains
    HPT-F5′-CTATTTCTTTGCCCTCGGACGAGTGC-3′
    BAT-R5′-TTTGGAAGACACGTAGCTCCACTCG-3′
    ARO8-2-R5′-GAGCCTCTCCCCACCGCCTGATGGCC-3′
    PDC-R5′-TGCCGGCATCCTTGGCTGCAAGTGC-3′
    STR3-OE-R5′-TTTGGCTGTGCGTGGAGTTCGTCC-3′
Location of promoter regulatory regions
    ARO8-2
        ARO8-2-promoter-F15′-CGGCCGCTCTAGAACTAGTGAGAACAAGCTGCAAGCCATTCAGGTAG-3′
        ARO8-2-promoter-F25′-CGGCCGCTCTAGAACTAGTGCAGTGCATGATCCTGAAGCAGATCTTC-3′
        ARO8-2-promoter-F35′-CGGCCGCTCTAGAACTAGTGTTGAGCCGAGTTTCGCCTGCGGAAAG-3′
        ARO8-2-promoter-F45′-CGGCCGCTCTAGAACTAGTGCCCCACTACCAGACAAGGTTGCCAGC-3′
        ARO8-2-promoter-R5′-GAGTTCAGGCTTTTTCATATTTTCCGATATTTTTATAAATTCAATTC-3′
    PDC
        PDC-promoter-F15′-CGGCCGCTCTAGAACTAGTGTCGAGGGCGCAGCAGGGCGATCTGGTG-3′
        PDC-promoter-F25′-CGGCCGCTCTAGAACTAGTGGGATATCGGGCAACTCAATCATGGCAC-3′
        PDC-promoter-F35′-CGGCCGCTCTAGAACTAGTGGATACACCCCCCGTGTCCGGGATGGATC-3′
        PDC-promoter-F45′-CGGCCGCTCTAGAACTAGTGTTCGTAAGGTATAAATTAGCTTTCCTC-3′
        PDC-promoter-R5′-GAGTTCAGGCTTTTTCATCTCCGAATAAGCACCGGCAACGCCATTG-3′
    STR3
        STR3-promoter-F5′-CGGCCGCTCTAGAACTAGTGCAGAAAAGGGCCTCGGCTGGAGGAAC-3′
        STR3-promoter-F15′-CGGCCGCTCTAGAACTAGTCGGAGGAGTGGTGAGCATGAAGCAAG-3′
        STR3-promoter-F25′-CGGCCGCTCTAGAACTAGTAATCCGAAATGACGACTTTTTTGGCAG-3′
        STR3-promoter-F35′-CGGCCGCTCTAGAACTAGTCATGCCTTTTTTCTATTCAGTGGCTG-3′
        STR3-promoter-R5′-GAGTTCAGGCTTTTTCATGACGGAGGATCGCAAGGGAAGAAGAG-3′
        pCB301hpt-F5′-ATGAAAAAGCCTGAACTCACCGCGAC-3′
        pCB301hpt-R5′-TCTGCAATGCCTATTTCTTTGCCCTCGGACGAGTGC-3′
        TrpC-terminator-F5′-AAAGAAATAGGCATTGCAGATGAGCTGTATCTGGAAG-3′
        TrpC-terminator-R5′-CGAGGTCGACGGTATCGATAGATCCACTTAACGTTACTGAAATCATC-3′
Quantitative real-time PCR
    18S rRNA
        18S rRNA-F5′-CTTCGGGGCTCTCTGGTGAT-3′
        18S rRNA-R5′-TGCTGCCTTCCTTGGATGTAGT-3′
    BAT
        BAT-Real-F5′-AAATCATCTGCCCGCATCGC-3′
        BAT-Real-R5′-ATCATGGTTGGGCGCAGGTA-3′
    ARO8-2
        ARO8-2-Real-F5′-GGTATCCGCCCAGACAGCTT-3′
        ARO8-2-Real-R5′-GGGTTCTGTCCCGTGGGAAT-3′
    PDC
        PDC-Real-F5′-ACAGAGACAGGCACTGCCAA-3′
        PDC-Real-R5′-GGCATCCTTGGCTGCAAGTG-3′
    STR3
        STR3-Real-F5′-GGCGGACAGGCTGAGTATGA-3′
        STR3-Real-R5′-ATCACGTCGAGTGCACCCAT-3′
    hpt
        HPT-Real-F5′-ATGTCCTGCGGGTAAATAGC-3′
        HPT-Real-R5′-GGTCAGGCTCTCGCTAAACT-3′
    S. cerevisiae 18S rRNA
        Sc18S rRNA-F5′-GAAACGGCTACCACATCCAAGGAAG-3′
        Sc18S rRNA-R5′-GTTCCTCGTTAAGGTATTTACATTG-3′
    S. cerevisiae STR3
        ScSTR3-F5′-GTCTTACTTAACGGCACTGACAACCATAC-3′
        ScSTR3-R5′-GAAGTGTCCACATGAACAGAGACTGCATG-3′
    S. cerevisiae CYS3
        ScCYS3-F5′-GTCTCTATCGGTGATGTGTACGGTGGTAC-3′
        ScCYS3-R5′-CTTCAAAGTTGGGTTGGTTGGGGTTTCGA-3′
Mutation and deletion of TFBSs
    ARO8-2
        ARO8-2-RPN4-M-R5′-CACCCCCGTGGCTTCCAGCTTCCATCGATACCCGAACCCGGGATATC-3′
        ARO8-2-RPN4-F5′-GAAGCCACGGGGGTGGACAGCTCGC-3′
        ARO8-2-RPN4-D-R5′-CACCCCCGTGGCTTCCATCGATACCCGAACCCGGGATATC-3′
        ARO8-2-MSN2-M-R5′-CGACGCCTGCCAACCCATATTCAAGGGAGGAGTCGACCTCTTCAC-3′
        ARO8-2-MSN2-F5′-GGTTGGCAGGCGTCGTAGTGGTGGC-3′
        ARO8-2-MSN2-D-R5′-CGACGCCTGCCAACCTCAAGGGAGGAGTCGACCTCTTCAC-3′
    PDC
        PDC-MSN2-M-R5′-GTGAGTTGGGTATTTTGGTCCTTGCTATACTAATAAACAACAAACGTGG-3′
        PDC-MSN2-F5′-ACCAAAATACCCAACTCACACCAATC-3′
        PDC-MSN2-D-R5′-GTGAGTTGGGTATTTTGGTCTATACTAATAAACAACAAACGTGG-3′
        PDC-ASG1-M-R5′-TCCCGAGCGCTGGTTCCCGCTTCATTCTGCTACAATGCCTGGATTGAGCA-3′
        PDC-ASG1-F5′-GCGGGAACCAGCGCTCGGGATTGTTG-3′
        PDC-ASG1-D-R5′-TCCCGAGCGCTGGTTCCCGCCTGCTACAATGCCTGGATTGAGCA-3′
        PDC-ARG80-M-R5′-CGTGGGATGGCAAAACAGTGGCCTGACATAAAACTTGCCCAGAGCCACATAA-3′
        PDC-ARG80-F5′-CACTGTTTTGCCATCCCACGTTGCAG-3′
        PDC-ARG80-D-R5′-CGTGGGATGGCAAAACAGTGCATAAAACTTGCCCAGAGCCACATAA-3′
        PDC-GLN3-M-R5′-GACACGGGGGGTGTATCCGTTTTCTCATCTATCCCTGGCTGCCTCACCAC-3′
        PDC-GLN3-F5′-ACGGATACACCCCCCGTGTCCGGGAT-3′
        PDC-GLN3-D-R5′-GACACGGGGGGTGTATCCGTCATCTATCCCTGGCTGCCTCACCAC-3′
    STR3
        STR3-GLN3-M-R5′-CGCCCAGATGCTTGTTGTGTTTCTTAATTGGCTATACAGCCACATCCCTTG-3′
        STR3-GLN3-F5′-CACAACAAGCATCTGGGCGAGGGAG-3′
        STR3-GLN3-D-F5′-CGCCCAGATGCTTGTTGTGTAATTGGCTATACAGCCACATCCCTTG-3′
Southern blot analysis
    hpt
        Hptprobe-F5′-GGTCAGGCTCTCGCTAAACT-3′
        Hptprobe-R5′-ATGAAAAAGCCTGAACTCACCGCGAC-3′
  • a Homologous sequences are underlined.