Table 2.
Oligonucleotide-primers used for A, amplification of DAS1/FLD1/TKL1 from chromosomal DNA; B, evaluation of DAS1/DAS2 transcription induction; C, the analysis of DAS1/FLD1/TKL1 overexpression; D, copy number determination of the reporter genes (via verification of the ZeoR copy number)
| A. | target | orientation | sequence 5'-3' |
|---|---|---|---|
| DAS1 | forward | AAAAGGCGCGCCGAAACGATGGCTAGAATTCCCAAAGCAG | |
| reverse | TTTGCGGCCGCTTACAACTTGTCATGCTTTGGTTTTC | ||
| FLD1 | forward | AACACTAGTATGTCTACCGAAGGTCAA | |
| reverse | AACGCGGCCGCTTAGTGCATAGTAATCAC | ||
| TKL1 | forward | AGAGAATTCGAAACGATGTCTGATCTCTTAGCTATCAACAC | |
| reverse | AGAGCGGCCGCCTACGCATGAACAGACTCAAAAG | ||
| B. | target | orientation | sequence 5'-3' |
| ARG4 | forward | TGCTGGCTACAGATCTTGCCGACT | |
| reverse | CTCGGCTTGTCTGACACATTCACCAG | ||
| AOX2 | forward | ATACTCATCCGAGGCCAGAGCTTACG | |
| reverse | ACCGTGAGCAAGACCAGCAGTCAA | ||
| DAS1 | forward | CTGAGAAACCAGCTAAAGGTGACGAGT | |
| reverse | TCTTGTCCCTCACGAGGGTACTCT | ||
| DAS2 | forward | CTGAAAAACCAGCCGAGGGTGATC | |
| reverse | TTCCTCACCTTCTTGAGGATAGTTCTTAACG | ||
| C. | target | orientation | sequence 5'-3' |
| ARG4 | forward | TGCTGGCTACAGATCTTGCCGACT | |
| reverse | CTCGGCTTGTCTGACACATTCACCAG | ||
| AOX2 | forward | ATACTCATCCGAGGCCAGAGCTTAC | |
| reverse | ACCGTGAGCAAGACCAGCAGTCAA | ||
| DAS1 | forward | CTGAGAAACCAGCTAAAGGTGACGAGT | |
| reverse | TCTTGTCCCTCACGAGGGTACTCT | ||
| FLD1 | forward | TGGATTATCTGTCATCCAAGGTGCAGTTTC | |
| reverse | GTCCGCCCATGCCTTCTTTGAATC | ||
| TKL1 | forward | GTCGCTACACATGACTCGATTGGTC | |
| reverse | CATGAGGTTTGGAAGAGCTCTCAAGTG | ||
| D. | target | orientation | sequence 5'-3' |
| ZeoR | forward | GACTCGGTTTCTCCCGTGACT | |
| reverse | CTGCGGAGATGAACAGGGTAA | ||
| ARG4 | forward | TCCTCCGGTGGCAGTTCTT | |
| reverse | TCCATTGACTCCCGTTTTGAG | ||