Table 3.
Oligonucleotides for the synthesis of kanMX4 cassettes | |
Primer | Sequence 5' to 3' |
YGR038w F | AGG AAC TGC TGA GGC GGC TTC TAC CTC GTA TAG TCG AAA TCA AAC CGT ACG CTG CAG GTC GAC |
YGR038w R | TGT TCA ACT AAT TTG GGC GCG ACC TGT GAT ACC TGG GAT AGA AAT ATC GAT GAA TTC GAG CTC G |
YLR350w F | ATC ATG ATT GAC CGC ACT AAA AAC GAA TCT CCA GCT TTT GCG TAC GCT GCA GGT CGA C |
YLR350w R | CTA ACT AAT TTG AGC ACG GCC CGT AAT ACC AGG GAT GGA TAT CGA TGA ATT CGA GCT CG |
Oligonucleotides for gene integrity and gene deletion verification | |
Verification of ORM1 gene integrity | |
A1 (F) | GTT TAG GAA CGG ATT ATA GA |
A2 (R) | CTG GTC ATT TTC ATC TTC AA |
Verification of ORM1 substitution | |
A1 (F) | GTT TAG GAA CGG ATT ATA GA |
KAN5 (R) | GTT CGG ATG TGA TGT GAG |
Verification of ORM2 gene integrity | |
YLR350w up400 (F) | GCG TAT TTT GAT TGC TCA TA |
YLR350w ver (R) | TGG TTT CAG GTT AGA CAC AT |
Verification of ORM2 substitution | |
YLR350w up400 (F) | GCG TAT TTT GAT TGC TCA TA |
KAN5 (R) | GTT CGG ATG TGA TGT GAG |
Oligonucleotides in bold are 18-19 nucleotide homology regions to pFA6a MCS sequences.