Table 2.
Primers used in this study.
| Primer | Description | Sequence |
|---|---|---|
| Primers for constructing YIPs to test new counterselectable markers alone in LEU2 locus | ||
| tag1.F | [tag1] forward | 5′-ACGCTATGAGGATTGATCGTAG-3′ |
| MS261 | [tag4] overlap pUG reverse | 5′-AGTCCTGGTCGTACTATGCTCGGAATTACTGCGTGTTTATAGGCCACTAGTGGATCTG-3′ |
|
| ||
| Primers for E. coli colony screening of YIP assemblies | ||
| MS147 | A. gossypii PTEF internal forward | 5′-TCACATCACATCCGAACATAAACAACCAT-3′ |
| MS268 | YIPin plasmid | 5′-TTGGCCGATTCATTAATGC-3′ |
| MS263 | PGPD internal reverse | 5′-CCGTCTTTATATACTTATTAGTCAAGTAGGGGAATAATTTC-3′ |
| MS123 | KlURA3 internal forward | 5′-TTAGCCAATCAAACCCTTCT-3′ |
|
| ||
| Primers for PCR amplification of YIPout fragments from the yeast genome | ||
| MS282 | [tag2] FCY1 YIPout forward | 5′-AAGGGACAACACCTCAGCCCGGCACATAGAAGTAACTTGGAATGCCAAAAATATCAT-3′ |
| MS283 | [tag3] FCY1 YIPout reverse | 5′-AATTGGGAGTTCTTCGAAGAGAGTGACACAGTCCTTGTTCAGGGTCTCCGGGT-3′ |
| MS328 | [tag2] HIS3 YIPout reverse | 5′-ATTGGGAGTTCTTCGAAGAGAGTGACACAGTCCTGTATATTCTTCGAAGAAATCACATTACTTTATATAATG-3′ |
| MS329 | [tag3] HIS3 YIPout forward | 5′-ACAACACCTCAGCCCGGCACATAGAAGTAACTTTAGCTTCTCGACGTGGGC-3′ |
| MS207 | [tag2] LEU2 YIPout forward | 5′-AACACCTCAGCCCGGCACATAGAAGTAACAACGCAAGGATTGATAATGTAA-3′ |
| MS208 | [tag3] LEU2 YIPout reverse | 5′-TTGGGAGTTCTTCGAAGAGAGTGACACAGTCCTTTGGTTGCTAAGAGATTCGAA-3′ |
| MS228 | [tag2] LYS5 YIPout forward | 5′-AAGGGACAACACCTCAGCCCGGCACATAGAAGTAACTCGAACAATGCCTTACGA-3′ |
| MS229 | [tag3] LYS5 YIPout reverse | 5′-AATTGGGAGTTCTTCGAAGAGAGTGACACAGTCCTTTTATTGAGGATTCTGGCTTGA-3′ |
|
| ||
| Primers for screening integrated yeast strains | ||
| MS127 | A. gossypii PTEF reverse | 5′-TTGTTTATGTTCGGATGTGAT-3′ |
| MS234 | KlURA3 internal reverse | 5′-CCAAAAGAGAATCAGCGCTCCCC-3′ |
| MS262 | PGPD internal forward | 5′-CATTCTCTTAGCGCAACTACAGAGAACAGG-3′ |
| MS203 | FCY1 locus screen “up” | 5′-AAGACCATAATTCGTGTATGACT-3′ |
| MS204 | FCY1 locus screen “down” | 5′-TGCCTATAAGGAAAATAAGGCA-3′ |
| MS320 | HIS3 locus screen “up” | 5′-AAGTTTCAGAGCAATGTCCTTAT-3′ |
| MS72 | HIS3 locus screen “down” | 5′-TTCAGAATGACACGTATAGAATGATGC-3′ |
| MS321 | LEU2 locus screen “up” | 5′-AGTCATCTAAATTAGTGGAAGCTGA-3′ |
| MS206 | LEU2 locus screen “down” | 5′-TAAAAGTGGAGCATCTTTAATGGAC-3′ |
| LYS5.A | LYS5 locus screen “up” *(Shoemaker, et al., 1996; Brachmann, et al., 1998) | 5′-TAAAAGTGGAGCATCTTTAATGGAC-3′ |
| LYS5.D | LYS5 locus screen “down” *(Shoemaker, et al., 1996; Brachmann, et al., 1998) | 5′-TATGCCAAACTGAACTAGTCACAAA-3′ |
*
These sequences were taken directly from the Yeast Genome Deletion Project primer set.