Table 3.
DNA primers used in this study and their application.
| Primera | Application | Sequence (5′ – 3′)b |
|---|---|---|
| KanMXuF | amplify uptag | ATGGATGTCCACGAGGTCTCT |
| KanMXuR | amplify uptag | CGCACCTGATTGCCCGACAT |
| KanMXuseq | sequence uptag | GCGCACGTCAAGACTGTCAAG |
| KanMXdF | amplify downtag | CTCACCGGATTCAGTCGTC |
| KanMXdR | amplify downtag | ACGGTGTCGGTCTCGTAG |
| KanMXdseq | sequence downtag | TCATCTGCCCAGATGCGAAG |
| YCp50F | sequence | CGCTTCGCTACTTGGAGCCACT |
| YCp50R | sequence | GCGCCAGCAACCGCACCTGT |
| ScTSC10u74F | amplify, sequence | CTCTGCATTTACTGGAAGCGA |
| ScTSC10d46R | amplify, sequence | ATAAAGAGAGTCACGGGGAAT |
| CgFEN1-TRP1F | fen1Δ | ATATCCTTTGCTGTAATTTCACTGATCAATAAGAACTGATAATCACTTATAAGTACTATGTCTGTTATTAATTTCACAGG |
| CgFEN1-TRP1R | fen1Δ | TTAGATGCAAACTCCGAGTATAAGAGAAAAAGAGGTTATCACTTGACCTCAACGAGCCTATTTCTTAGCATTTTTGACGA |
| CgFEN1u248F | fen1Δ screen | CACCAACGTATAAGGACTGTA |
| CgFEN1c201R | fen1Δ screen | AGGAAGTTCACCAGCGACAA |
| ScTRP1c345R | fen1Δ screen | TGGCAAACCGAGGAACTCTT |
| CgCKA2-URA3F | cka2Δ | AAGCTGTGAGGGATAGCCAGGAGGGGGAGCAGCAAGAGAGACCAAGCCAAGCGATGTCGAAAGCTACATATAAG |
| CgCKA2-URA3R | cka2Δ | GCAAATCCTTTCTGATAATTGCGTGCTTGGAGATAATTCAGAATGTACATATATGTGTTTAGTTTTGCTGGCCGCATC |
| CgCKA2u418F | cka2Δ screen | CCTGGTTATCAACTTCGACC |
| CgCKA2c507R | cka2Δ screen | GCTTCACGTCCCTGTGCAT |
| ScURA3c349R | cka2Δ screen | CTGCCCATTCTGCTATTCTG |
| CgFEN1u481F | amplify | CCCTTGCTCAATGACACATC |
| CgFEN1d192R | amplify | TCCCAACATGTGAATAGGTCT |
| CgFEN1u361F | sequence | CCTCATCGCAAAAGGCAAAC |
| CgFEN1c378F | sequence | TGACTCCAATGATCTACCATC |
| CgSUR4u340F | amplify | GATTTGTCCTTGTCGTCGTC |
| CgSUR4d91R | amplify | AAAGCCACATCTAACTGTCAG |
| CgSUR4c495R | sequence | GATAGCCCAGAACAAACCGT |
| CgSUR4c355F | sequence | ATCAACGCCTCTCCAATGCA |
| CgIFA38u280F | amplify | ACTGTAGAACCTGCATTGGC |
| CgIFA38d137R | amplify | GACATAGGGTAAGGACCACT |
| CgIFA38u134F | sequence | TCAAGTATCGGCTGCTAGCT |
| CgIFA38c400F | sequence | TCCCTTTGTGATGACCTTCC |
| CgPHS1u178F | amplify | ACATAACCGAGCACGATTGTA |
| CgPHS1d185R | amplify | CTTAGCCAGCAAAGTCCGTA |
| CgPHS1u136F | sequence | CTCTAGCAAGAGACTGTGAG |
| CgTSC13u276F | amplify | CGAAGCTCATCTCATCGCAA |
| CgTSC13d111R | amplify | GGTAGAACACATGATAAGGGA |
| CgTSC13u158F | sequence | GGGATATCCAATTTCCTGGG |
| CgTSC13c359F | sequence | TCCACAGAGACCCATTCTTG |
| CgSUR2u483F | amplify | GGATATGGGAACTTCTCAATG |
| CgSUR2d72R | amplify | CGCTCAATAATTATCGGAGCA |
| CgSUR2c241F | sequence | GAGATCGAGAAGCGTAACAG |
| CgSUR2c436R | sequence | CGTAGTAGACAGCAGCATCT |
| pGRB-CgFEN1u587F | gap-repair | CGAGGTCGACGGTATCGATAAGCTTGATATCGAATTCCTGACAAGTCTTCTGTCGAAACC |
| pGRB-CgFEN1d193R | gap-repair | GGAGCTCCACCGCGGTGGCGGCCGCTCTAGAACTAGTGATCCCAACATGTGAATAGGTC |
| CgFEN1c141R | screen | AGGAAGTTCACCAGCGACAA |
| T7 | screen | TAATACGACTCACTATAGGG |
a
Numbers in primer names correspond to location upstream (u) or within the coding region (c) relative to the start codon, or downstream (d) relative to the stop codon.
b
Underlined regions of deletion primers correspond to S. cerevisiae TRP1 or URA3 coding sequences; underlined regions of gap-repair cloning primers correspond to C. glabrata FEN1 upstream or downstream sequences.