Table 2.
PCR primers used in the homologous recombination cloning.
| Namea | Primer sequence | Sequence detectedb |
|---|---|---|
| seAGT1prom_F | 5΄-GCTATGACCATGATTACGCCAGAATTTAAAGGCGCGCGT -3΄ | YEplac195 flanking sequence 5΄ of HindIII underlined, −1900 to −1878 of seAGT1 promoter |
| seAGT1promR | 5΄- AGCGAAAGTATATTTTTCATATTCTGATATTTCGTAACTCTTTCTGATTA -3΄ | seAGT1 gene 1–20 underlined, seAGT1 promoter from −1 to −30 |
| seAGT1_F | 5΄- TATCAGAATATGAAAAATATACTTTCGCTGGTAGGAAG -3΄ | SeAGT1 prom from −1 to −10 underlined, seAGT1 gene from 1 to 29 |
| PGKterm_R | 5΄-TAGAGTCGACCTGCAGGC -’3΄ | YEplac195 flanking sequence 3΄ of HindIII |
| scMALx1prom_F | 5΄- GCTATGACCATGATTACGCC ATCAGAAATAGTCATTTATGTATTTTAGTTACGCT -3΄ | YEplac195 flanking sequence 5΄ of HindIII underlined, −800 to −765 of seMALx1 promoter |
| scMALx1_prom_R | 5΄- ATGAGGATAATCCCTTCAT AGTTAATTAATAGTCTTGGATGTAATTCTTATTGT -3΄ | scMALx1 gene underlined, scMALx1prom from −1 to −36 |
| scMALx1-mCherry-PGKterm_F | 5΄- TTAATTAACT ATGAAGGGATTATCCTCAT TAATAAACAGA -3΄ | scMALx1 promoter from −1 to −10, scMALx1 gene |
| scAGT1prom_F | 5΄- GATCCTCTAGAGTCGAGTTCTAGAAGCCTCCGGCAAA -3΄ | YEplac195 flanking sequence underlined (SalI site), scAGT1prom |
| scAGT1_prom_R | 5΄- CAATGAAATGATATTTTTCATATTATAATATTTTTTTAGTT GTTTGATGTTCTTCTATGTA -3΄ | scAGT1 gene underlined from 1 to 21,scAGT1prom from −1 to −38 |
| scAGT1_F | 5΄- CAACTAAAAAAATATTATAATATGAAAAATATCATTTCATTGGTAAGCAAG -3΄ | scAGT1prom from −1 to −21, scAGT1 gene from 1 to 30 |
| TDH3_R | 5΄-AAAGCTTGCATGCCTGCAGGTCGACATGTAGATGATACTGACTGCACG-3΄ | Yeplac195 flanking sequence underlined (SalI), TDH3 term |
| seMALx1_prom_F | 5΄-CCTCTAGAGTCGACCTGCAGG TTCTGTACGCGATTTGGTTATGTG-3΄ | YEplac195 flanking sequence underlined (SphI site), seMALx1prom |
| seMALx1_prom_R | 5΄-CATTGAGGATAGACCCTTCATAGTTGCCTAATAGCTTTTGAAATTTTTT-3΄ | seMALx1 gene underlined from 1 to 21, seMALx1 promoter from −1 to −28 |
| seMALx1_F | 5΄-AAATTTCAAAAGCTATTAGGCAACTATGAAGGGTCTATCCTCAATGATAAAT-3΄ | seMALx1 prom underlined from −1 to −25, seMALx1 from 1 to 27 |
| seMALx1_R | 5΄-AAATTCTGCGTTCGTTAAAGCTTG CATGTAGATGATACTGACTGCACG-3΄ | YEplac195 flanking sequence underlined (SphI site), TDH3 terminator |
| TDH3prom_FRW(SalI) | 5΄-ACCCGGGGATCCTCTAGAGTCGA CACGCTTTTTCAGTTCGAGT-3΄ | Yeplac195 flanking SalI sequence underlined, TDH3promoter sequence |
| TDH3promREV_AGT1 | 5΄-AATGAAATGATATTTTTCATTTTGTTTGTTTATGTGTGTTTATTCGA-3΄ | scAGT1 sequence underlined from 1 to 20, TDH3 prom |
| ScAGT1mCherry_FRW_TDH3p | 5΄-TAAACAAACAAAATGAAAAATATCATTTCATTGGTAAG-3΄ | TDH3 prom underlined, scAGT1 from 1 to 26 |
| ScAGT1mCherry_REV_TDH3t | 5΄-AAGATTTAAAGTAAATTCACTACAGCTCGTCCATGCC -3΄ | TDH3term underlined, mCherry |
| TDH3term_FRW_mCherry | 5΄-ATGGACGAGCTGTAGTGAATTTACTTTAAATCTTGCATTTAAAT-3΄ | mCherry underlined, TDH3 term |
| TDH3term_REV_plasmidi (SalI site) | 5΄- AAAGCTTGCATGCCTGCAGG CATGTAGATGATACTGACTGCACG-3΄ | Plasmid 3΄ SalI site underlined, TDH3 term |
| TDH3p_FRW_SphI_site | 5΄-CCTCTAGAGTCGACCTGCAGGCACGCTTTTTCAGTTCGAGT -3΄ | Plasmid 5΄ SphI site underlined, TDH3 prom |
| TDH3p_REV_SeMALx1 | 5΄-TAGACCCTTCATTTTGTTTGTTTATGTGTGTTTATTCGA-3΄ | seMALx1, TDH3prom |
| SeMALx1_FRW_TDH3prom | 5΄-TAAACAAACAAA ATGAAGGGTCTATCCTCAATG-3΄ | TDH3prom underlined, seMALx1 |
| SbMALx1_REV_GFP | 5΄-CGCCGCACTAGTTCATCTAATCATGAGAGATGGG-3΄ | linker underlined, seMALx1 |
| GFP_FRW(linker_SbMALx1) | 5΄-TGAACTAGTGCGGCGATGGTGAGCAAGGGCGA-3΄ | linker_seMALx1, GFP |
| GFP_REV (TDH3term) | 5΄-AAGTAAATTCACTTACTTGTACAGCTCGTCCATGC-3΄ | TDH3term, GFP |
| TDH3Term-FRW(Gfp) | 5΄-CTGTACAAGTAA GTGAATTTACTTTAAATCTTGCATTTAAAT -3΄ | GFP underlined, TDH3 term |
| TDH3term-REV(plasm) | 5΄- TCTGCGTTCGTTAAAGCTT GCATGTAGATGATACTGACTGCACG-3΄ | Yeplac195 flanking sequence underlined (SphI), TDH3 term |
aF, forward; R, reverse.
bThe numbering is from the first nucleotide of the translational start.