TABLE 3.
Oligonucleotides used in this study
fwd, forward; rev, reverse.
| No. | Sequence (5′–3′) | Use |
|---|---|---|
| 1 | GTTTTTCTCAGTTACTGGAACAGTGTGCTATGGTATGATTGAAATTTTCTTTGGTGACGGTGCTGGTTTA | (fwd); tagging Vtc5 at the C terminus; templates, pKT127 or pKT209 |
| 2 | GCATATTTTATATAGACTGTACACATGAAAAACTTTTATCGTCCAAAATAATCGATGAATTCGAGCTCG | (rev); tagging Vtc5 at the C terminus; templates, pKT127 or pKT209 |
| 3 | TGGCAAGTGTATGTCTGGGA | (fwd); control C-terminal tagging of Vtc5 |
| 4 | GGG ACA ACA CCA GTG AAT | (rev); control C-terminal tagging of Vtc5 |
| 5 | TATCTCCATCTCAGCCCTAGGTAAGGATAGATACGAAGGGTTGTACAATCATGCGTACGCTGCAGGTCGAC | (fwd); tagging Vtc5 N terminus; templates, pYM-NX plasmids |
| 6 | GTAGAATTTCCATTCAGGGATTGATTTATTCAATATGCGATCTTCAAATTTCATCGATGAATTCTCTGTCG | (rev); tagging Vtc5 N terminus; templates, pYM-NX plasmids |
| 7 | AAAGTTTGTGTCTCCCCTTG | (fwd); checking VTC5 deletion and N-terminal tagging |
| 8 | GTCTGCAGCGAGGAGCCGTA | (rev); checking integration of NAT cassette |
| 9 | TATCTCCATCTCAGCCCTAGGTAAGGATAGATACGAAGGGTTGTACAATCATGCGTACGCTGCAGGTCGAC | (fwd); deleting VTC5 through NAT; template, pFA6-natNT2 |
| 10 | GCATATTTTATATAGACTGTACACATGAAAAACTTTTATCGTCCAAAATAATTAATCGATGAATTCGAGCTCG | (rev); deleting VTC5 through NAT; template, pFA6-natNT2 |
| 11 | GTGTTTCTAATTCTCTCTATTTTATTTTCCTTTTTCTGCAACCTAGTTGCTAAACGTACGCTGCAGGTCGAC | (fwd); tagging Vtc4 C terminus; template, pYM22 |
| 12 | CTAATATGATTATTACTTAATTATACAGTAAAAAAAACACGCTGTGTATTCAATCGATGAATTCGAGCTCG | (rev); tagging Vtc4 C terminus; template, pYM22 |
| 13 | AAAAGTAGAAAGAACGACTACACCTCAACATAACGACACTTTTTTGACATGCGTACGCTGCAGGTCGAC | (fwd); deleting VTC2 through NAT; template, pFA6-natNT2 |
| 14 | GGTCTCAGTAGATAGAGTACATATTCTAATCACTGCTTGGCCCCATTAATCGATGAATTCGAGCTCG | (rev); used to delete VTC2 using the NAT marker (plasmid pFA6-natNT2 as a template) |
| 15 | CTATTAGAGCGAACAGCAGAATTTGTCCTTGGT TTTCAGAGTTTGAAAATGCGTACGCTGCAGGTCGAC | (fwd); deleting VTC3 through NAT; template, pFA6-natNT2 |
| 16 | AACTGGTACTTGTGTAATATATGTGTATATAAAAAATATACATGTTCTTAATCGTATGAATTCGTAGCTCG | (rev); deleting VTC3 through NAT; template, pFA6-natNT2 |
| 17 | GGCTAACAATCAAATCGGCCAATAAAAGAGCATAACAAGGCAGGAACAGCTATGCGTACGCTGCAGGTCGAC | (fwd); deleting VTC4 through NAT; template, pFA6-natNT2 |
| 18 | GATTATTACTTAATTATACAGTAAAAAAAACACGCTGTGTATTCATTTAATCGATGAATTCGAGCTCG | (rev); deleting VTC4 through NAT; template, pFA6-natNT2 |
| 19 | CAAGAATAATGACATTGAATCTTCCAGCCCATCTCAACTTCAACATGAAGCACGGATCCCCGGGTTAATTAA | (fwd); tagging Pho84 C terminus; template, pFA6a-GFP(S65T)-kanMX6 |
| 20 | GTTTTTGTATTATTTGTTCTAGTTTACAAGTTTTAGTGCATCTTTGAGGCTTGAATTCGAGCTCGTTTAAAC | (rev); tagging Pho84 C terminus; template, pFA6a-GFP(S65T)-kanMX6 |
| 21 | CCATAACTGTGCTAGAGACG | (fwd); check C-terminal tagging of Pho84 |
| 22 | GACTAGTATGAAATTTGAAGATCGCAT | (fwd); amplifying VTC5 |
| 23 | CCGCTCGAGTTAAAAGAAAATTTCAATCATACC | (rev) amplifying VTC5 |
| 24 | CGGAATTCGGTGACGGTGCTGGTTTA | (fwd); amplifying GFP; template, pKT127 |
| 25 | CCCAAGCTTTTATTTGTACAATTCATCCAT | (rev); amplifying GFP; template, pKT127 |
| 26 | CGAGCTCTGCATTATTTAGATCGGAAAAAGTC | (fwd); amplifying PHO4 and its endogenous promoter |
| 27 | GGAATTCCGTGCTCACGTTCTGCTGTAGGTG | (rev); amplifying PHO4 and its endogenous promoter |