TABLE 2

Oligonucleotides

PrimerSequencePurpose
FS915′-CAGAGTTCTGTGCTTCGCTG-3′Score silent ARG4 markers
FS925′-GTATCCACGTTTCAGCGGTAG-3′
FS1055′-ACG-ACG-AGC-AGT-TAA-AGT-TTT-CAA-ATA-AGT-TGC-AAC-CAG-CAG- ACA-TGA-TAC-GTA-CGC-TGC-AGG-TCG-AC-3′Replace nucleotides 2190–2340 downstream of the FUS1 ORF ATG on chromosome III with KAN-MX4
FS1065′-TGT-GGC-GTT-TTA-CGT-GAA-AAT-TAC-GTA-AAG-AAA-AAG-ATC-CTG- GGG-TGC-CT-ATC-GAT-GAA-TTC-GAG-CTC-G-3′
FS1105′-GTA-ACT-CCG-GTT-TCA-AAG-CG-3′Confirm structure of KAN insertion
FS1115′-ACA-ATA-ATC-CAG-TAT-ACC-GC-3′
FS2285′-GCA-GCT-GAT-GGT-GCT-GAG-AGA-TAA-GGC-CAC-TGAAAG-GCC-CCG- TAC-GCT-GCA-GGT-CGA-C-3′Replace nucleotides 190–320 downstream of YCL033C ATG on chromosome III with NatMX4
FS2295′-ATT-AAA-GAA-TTG-TCA-CGA-TGA-TAT-GTG-ATG-GCT-CCA-GGG-GAT- CGA-TGA-ATT-CGA-GCT-CG-3′
FS2325′-TCC-TGG-CAA-TCT-TGC-AAG-CAC-AAT-TCC-GGC-3′Confirm NAT insertion
FS2335′-CCA-CGT-CCA-AGT-TCA-TCC-AGG-CAA-GGG-CG-3′
FS2805′-ATATT GTCAT GAACT ATACC ATATA CAACT TAGGA TAAAAATACA GGTAG CGTAC GCTGC AGGTC GAC-3′Delete TAM1
FS2815′-AACAG CAAAG AAAAG TTTTT TTTGG TTCAG ATGTAATATG GATAG CCCGT ATCGA TGAAT TCGAG CTCG -3′
FS2825′-GTTTC GTACT CAGTG ACGTA CCGGG-3′Confirm YFS644 and YFS645
FS2835′-AAATG CATTC CTACT AACGA ATCGG-3′
FS2845′-GAA-GGC-TTT-CCA-ACT-TAA-AAG-AGC-CTC-AAC-3′Replace MSH4 ORF of YFS0617 and YFS0618 with HphMX4 ORF
FS2855′-GTT-TTG-GTA-TGG-GAT-GAC-ATT-GTT-TTA-CGT-AG-3′
FS2885′-ATC-AAG-CAG-CAG-TAC-CGG-TAT-CTC-AAG-AGG-3′Confirm structure of HYG insertion
FS2945′-ACA-TTT-CAG-CAA-TAT-ATA-TAT-ATA-TAT-TTC-AAG-GAT-ATA-CCA- TTC-CGT-ACG-CTG-CAG-GTC-GAC-3′Amplify hygromycin-resistance gene for insertion at the native LEU2 site
FS2955′-TTC-ATT-TAT-AAA-GTT-TAT-GTA-CAA-ATA-TCA-TAA-AAA-AAG-AGA- ATC-ATC-GAT-GAA-TTC-GAG-CTC-G-3′
FS3005′-GAG-CTA-GGT-GGT-GTT-ACA-CTC-GGT-TCT-ATG-ACT-GCT-AAC-ATC- ACG-GCG-ACA-TTA-CTA-TAT-ATA-TAA-TAT-AGG-3′Introduce TRP1 upstream of ARG4 in vicinity of YHR032W
FS3015′-ACC-AAA-CAT-ACA-TCA-TTG-GCA-AGA-ACG-CCA-AGA-TGG-TGA-TCA- CAG-CCA-AAC-AAT-ACT-TAA-ATA-AAT-ACT-ACT-CAG-3′
FS3025′-GTG-AGT-ATA-CGT-GAT-TAA-GCA-CAC-AAA-GGC-AGC-TTG-GAG-TTC- GAC-AAC-CCT-TAA-TAT-AAC-TTC-G-3′Amplify bleomycin-resistance gene
FS3035′-TGC-ACA-AAC-AAT-ACT-TAA-ATA-AAT-ACT-ACT-CAG-TAA-TAA-CTC- GAC-AAC-CCT-TAA-TAT-AAC-TTC-G-3′