Table 1.

Primers Used in This Study

5′ to 3′ primer sequenceApplication
ATGCTACGTACCTGTTTAACTCTTC Amplification of cyh2R (N37E) allele, set 1.
GTTAATTCTGTGGTGATGTTCACCACCGGCCATACCTCTAC Amplification ofcyh2R (N37E) allele, set 1.
TATGTTTATATATGGATTTTGAAA Amplification ofcyh2R (N37E) allele, set 2.
GTAGAGGTATGGCCGGTGGTGAACATCACCACAGAATTAAC Amplification ofcyh2R (N37E) allele, set 2.
CCACCGCGGTGGCGGCCGCTCTAGAACTAGTGGATCCCCCATGCTACGTA CCTGTTTAACTCTTC Amplification of CYH2 gene for recombinational subcloning intoSmaI site of pRS316.
CGACGGTATCGATAAGCTTGATATCGAATTCCTGCAGCCCCTATGTTTAT ATATGGATTTTGAAA Amplification of CYH2 gene for recombinational subcloning intoSmaI site of pRS316.
TTATTTTTATAGCACGTGATGAAAAGGACCGCGGCCGCAAGGGGTTCGCG TTGGCCGATT Amplification of the CEN-ARS, URA3 region of pRS316 for recombinational subcloning into SrfI-digested pBeloBAC11.
TTCAATTTAATTATATCAGTTATTACCCTGCGTCGACCAATTCTCATGTT TGACAGCTTA Amplification of the CEN-ARS, URA3 region of pRS316 for recombinational subcloning into SrfI-digested pBeloBAC11
CGATTCATTAATGCAGGCCCGGGCATGCTACGTACCTGTTTAACTCTTC Amplification of CYH2 for recombinational cloning intoNotI-digested pCRG4.
TATAGCATACATTATACGAAGTTATATTCGATGCGGCCGCCTATGTTTAT ATATGGATTTTGAAA Amplification of CYH2 for recombinational cloning intoNotI-digested pCRG4.
AACAGGTACGTAGCATGCCCGGGCCTGCATTAATGAATCGGCCAACGCG Amplification of the AmpR, colE1 ori region of pRS316 for recombinational subcloning into NotI-digested pCRG4.
GCGGCCGCGCCCGGGCACGTCAGGTGGCACTTTTCGGGGAA Amplification of the AmpR, colE1 ori region of pRS316 for recombinational subcloning into NotI-digested pCRG4.
TTTCCCCGAAAAGTGCCACCTGACGTGCCCGGGCGGCCGCGGTCCTTTTC ATCACGTGCTATAAAAATAATTATAATTTA Top strand of recombination linker used to join the AmpR, colE1 fragment to NotI-digested pCRG4.
TAAATTATAATTATTTTTATAGCACGTGATGAAAAGGACCGCGGCCGCCC GGGCACGTCAGGTGGCACTTTTCGGGGAAA Bottom strand of recombination linker used to join the AmpR, colE1 fragment to NotI-digested pCRG4.
TCATCGAATTTCTGCCATTCATCCG Amplification of CmRfragment from pBeloBAC11.
ACTTTCACCATAATGAAATAAGATC Amplification of CmR fragment from pBeloBAC11.
CCACCGCGGTGGCGGCCGCTCTAGAACTAGTGGATCCCCCAATTTCTGCC ATTCATCCGCTTATTATCACTTATTCAGGC Top strand of linker set 1 used for recombinational subcloning of the CmR fragment into pRS316.
GCCTGAATAAGTGATAATAAGCGGATGAATGGCAGAAATTGGGGGATCCA CTAGTTCTAGAGCGGCCGCCACCGCGGTGG Bottom strand of linker set 1 used for recombinational subcloning of the CmR fragment into pRS316.
TCCTGAAAATCTCGATAACTCAAAAAATACGCCCGGTAGTGGGCTGCAGG AATTCGATATCAAGCTTATCGATACCGTCG Top strand of linker set 2 used for recombinational subcloning of the CmR fragment into pRS316.
CGACGGTATCGATAAGCTTGATATCGAATTCCTGCAGCCCACTACCGGGC GTATTTTTTGAGTTATCGAGATTTTCAGGA Bottom strand of linker set 2 used for recombinational subcloning of the CmR fragment into pRS316.
GGTGCGGGCCTCTTCGCTATTACGC Amplification of 450 bp “repair fragment” from pRS316.
CCGCGCGTTGGCCGATTCATTAATG Amplification of 450 bp “repair fragment” from pRS316.
CGAGCTCATCGCTAATAACTTCGTA Vector-specific PCR primer used to detect recombinant BAC subclones.
TATAGCACGTGATGAAAAGGACCGC Vector-specific PCR primer used to detect recombinant BAC subclones.
TATAGCATACATTATACGAAGTTATATTCGATGCGGCCGCNNNNNNNNNN NNNNNNNNNNNNNNNNNNNNNNNNNNNNNN Top strand of “generic” linker set 1 (left side) used to subclone BACs (N=BAC-specific sequence).
NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNGCGGCCGCAT CGAATATAACTTCGTATAATGTATGCTATA Bottom strand of “generic” linker set 1 (left side) used to subclone BACs (N=BAC-specific sequence).
NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNGCGGCCGCGG TCCTTTTCATCACGTGCTATAAAAATAATT Top strand of “generic” linker set 2 (right side) used to subclone BACs (N=BAC-specific sequence).
AATTATTTTTATAGCACGTGATGAAAAGGACCGCGGCCGCNNNNNNNNNN NNNNNNNNNNNNNNNNNNNNNNNNNNNNNN Bottom strand of “generic” linker set 2 (right side) used to subclone BACs (N=BAC-specific sequence).
CAAAAAGTTCTGTTGCTTCTGCAGC RPCI-11-98D12L-specific primer used to detect recombinant BAC subclones.
AATCTCTAGTACCAAAGAGAATTAT RPCI-11-98D12L-specific primer used to detect recombinant BAC subclones.
TGAAAGGGATCACTTGTATGATCTG RPCI-11-98D12R-specific primer used to detect recombinant BAC subclones.
CTTTTATTACCCTGAGTTCAGAATT RPCI-11-98D12R-specific primer used to detect recombinant BAC subclones.
AAAAGTATTTTACTAAGGCCAGAAC RPCI-11-100L10-specific primer used to detect recombinant BAC subclones.
TCTACATCCTCCAAGAGATTCACCT RPCI-11-100L10-specific primer used to detect recombinant BAC subclones.