TABLE 2.
Oligonucleotide primers used in this study
| Name | Sequencea | Added site | Resulting constructb |
|---|---|---|---|
| APRE20 | 5′-AAAGAATTCGGATCAGCTTGTTGTTTGC-3′ | EcoRI | pRFP3RP |
| APRE1090R | 5′-AAGGAATTCCAAGATATGTTGCAGTGCTT-3′ | EcoRI | pRFP3RP |
| CGEAM490 | 5′-CCAGGATCCAACACTTGAGAGTGAAACA-3′ | BamHI | pCGEA8G |
| CGEA398R | 5′-GGACTCGAGGAAAAGAACGTAACGCTTTC-3′ | XhoI | pCGEA8G |
| COTA86 | 5′-ACAGGATCCTAAGTCACCATGGAGGAATG-3′ | BamHI | pCOTA8G |
| COTA1538R | 5′-TCGCTCGAGTTATGGGGATCAGTTATATCC-3′ | XhoI | pCOTA8G |
| COTE59 | 5′-AAAGGATCCCAATGCACCAACACCA-3′ | BamHI | pCOTE8G |
| COTE542R | 5′-CTTCTCGAGTCTTCAGGATCTCCCACTA-3′ | XhoI | pCOTE8G |
| COTT3 | 5′-GTTGGATCCTAGGATTACCCTTTGAATGAAC-3′ | BamHI | pCOTT8G |
| COTT246R | 5′-CCTCGAGTAACCGTAACCGTAACCTCCCCCAT-3′ | XhoI | pCOTT8G |
| DRED3 | 5′-CGCCTCGAGGGACAACACCGAGGACG-3′ | XhoI | pRFP3K |
| DRED681R | 5′-GTCAAGCTTGCTCTACTGGGAGCCGG-3′ | HindIII | pRFP3K |
| SIGE16 | 5′-ACTAAGCTTACGGTTGACGCACCTC-3′ | HindIII | pSIGE5E |
| SIGE208R | 5′-TATGGATCCAGACGCAAAATGCGTTC-3′ | BamHI | pSIGE5E |
| SIGF24 | 5′-TAAAAGCTTCGGCAAAAACGCTCAGCT-3′ | HindIII | pSIGF5E |
| SIGF207R | 5′-TTAGGATCCCGATGCAGCCGATCT-3′ | BamHI | pSIGF5E |
| SIGG121 | 5′-GAAAAGCTTGTAAACGGGAACTT-3′ | HindIII | pSIGG5E |
| SIGG315R | 5′-GTAGGATCCTGCGGATCTCTCCGA-3′ | BamHI | pSIGG5E |
| SIGK16 | 5′-AGGAAGCTTCGCAGCGCTCGGCTT-3′ | HindIII | pSIGK5E |
| SIGK228R | 5′-TCGCTCGAGAAGTTACCATTGTGGATTC-3′ | BamHI | pSIGK5E |
| SPCM189 | 5′-GCGAGTACTTGACAAATTATATGGTGATCTGTATAAT-3′ | ScaI | pKMR5 |
| SPC813R | 5′-GTTGATATCGTAAGCACCTGTTATTGCAA-3′ | EcoRV | pKMR5 |
| PMT353RD | 5′-AGCATTAGTGTATCAACAAGCTGGGG-3′ | Digoxigenin end-labeled | |
| PUC1697 | 5′-AAAGATATCTAGGTGAAGATCCTTTTT-3′ | EcoRV | pKMR5 |
| PUC723R | 5′-TTAAGTACTCATGAGCGGATACATATT-3′ | ScaI | pKMR5 |
| YABG69 | 5′-CGAAAGCTTGGAATAGAGCAAACAAGCAA-3′ | HindIII | pYABG5C |
| YABG220R | 5′-GAGGGATCCATTCATTCTGCTCTCATCT-3′ | BamHI | pYABG5C |
| YABG236 | 5′-AATGGATCCGCCAAGATTATAAGCTG-3′ | BamHI | pYABG8G |
| YABG869R | 5′-AACCTCGAGTTGGACTTATAAGGCATACC-3′ | XhoI | pYABG8G |
| YEEK7 | 5′-AATGGATCCGATACAAGACATATGTATGGC-3′ | BamHI | pYEEK8G |
| YEEK434R | 5′-CTTCTCGAGTTATAACCGTCTTTTCCCCA-3′ | XhoI | pYEEK8G, pYEEK3RP |
| YHCN51 | 5′-CCCGGATCCTATGACTGGCTGCGGTGT-3′ | BamHI | pYHCN8G |
| YHCN566R | 5′-TTCCTCGAGTCAGCGTTAGGGAATACAC-3′ | XhoI | pYHCN8G |
| YEEKM400 | 5′-GAAGGATCCATGGAAGCGGATGTTGATG-3′ | BamHI | pYEEK3RP |
| YEEK280RT7 | 5′-TAATACGACTCACTATAGGGCGACATCATTTTCATGGTGCATGC-3′ | yeeK RNA probe |
a
T7 promoter sequences are underlined.
b
PCR products were digested with restriction enzymes at primer-derived sequences and inserted into restriction enzyme-digested plasmids to generate the plasmids listed in Table 1.