Table 4.
Primers and oligonucleotides used in strain constructions
| Primer | Sequencea | Note |
|---|---|---|
| Orf19.681UP-1-ApaI | 5′-AAGAGGGCCCCAGGATGAGGCTTTCACCAT-3′ | ApaI, forward, p19.681 5′f UP |
| Orf19.681UP-2-XhoI | 5′-TATTCTCGAGGCCGATTACTCGCTGATTTC-3′ | XhoI, reverse, p19.681 5′f UP |
| Orf19.681DOWN-1-SacII | 5′-TAATCCGCGGTTTCTTTTCTTTTCGTTTTGAATTG-3′ | SacII, forward, pSFS1A-35fCaHap43 |
| Orf19.681DOWN-2-SacI | 5′-CTATGAGCTCTGTTGTCAAGATTGTCTTGCATT-3′ | SacI, reverse, pSFS1A-35fCaHap43 |
| CaHap43-ORF-2-XhoI | 5′-AATGCTCGAGCTAATTATATGCTCTTCT-3′ | XhoI, reverse, pSFS2A-35fCaHAp43-ORF |
| CaSfu1up-1-ApaI | 5′-AATAGGGCCCAATGCTTTGGCATGCTTT-3′ | ApaI, forward, pSFS1A-5fCaSfu1 |
| CaSfu1up-2-XhoI | 5′-AATGCTCGAGTGGATTGGAAATTGGAGGAA-3′ | XhoI, reverse, pSFS1A-5fCaSfu1 |
| CaSfu1down-1-SacII | 5′-TTATCCGCGGAGTTAATATCAAGGGTTG-3′ | SacII, forward, pSFS1A-35fCaSfu1 |
| CaSfu1down-2-SacI | 5′-CTATGAGCTCGCAGCTGGTTCAACTGGACT-3′ | SacI, reverse, pSFS1A-35fCaSfu1 |
| CaSfu1-ORF-2-XhoI | 5′-AATGCTCGAGCTATCCATTTAACAACTTCCCAATAG-3′ | XhoI, reverse, pSFS1A-35fCaSfu1-ORF |
| SaLexA-HindIII-1 | 5′-GTCAAAGCTTAAAATGAGAGAATTAACAAAACG-3′ | HindIII, forward, pCIplexA-F1 |
| SaLexA-MluI-2 | 5′-ACCAACGCGTACCCATTTCGCGGTACAAACCAATTAC-3′ | MluI, reverse, pCIplexA-F1 |
| CaGcn4ORF-1-MluI | 5′-GGTCCACGCGTGGTGGAGGTCCAGGTGGACCTGCTACTACTCCTACTAT-3′ | MluI, polylinker, forward, pCIplexA-F-CaGCN4-21 |
| CaGcn4ORF-2-PstI | 5′-GCCCGCCTGCAGTCACTAAAATTGAATACCATT-3′ | PstI, reverse, pCIplexA-F-CaGCN4-21 |
| CaNrg1ORF-1-MluI | 5′-GGTCCACGCGTGGTGGAGGTCCAGGTGGACTTTATCAACAATCATATCC-3′ | MluI, polylinker, forward, pCIplexA-F-CaNRG1-18 |
| CaNrg1ORF-2-PstI | 5′-GCCCGCCTGCAGTCACTATACTAGGCTCTTGGT-3′ | PstI, reverse, pCIplexA-F-CaNRG1-18 |
| CaHap43ORF-1-MluI | 5′-GGTCCACGCGTGGTGGAGGTCCAGGTGGAATGCCCGCAAAAGGTCCTA-3′ | MluI, polylinker, forward, pCIplexA-F-CaHap43-3 |
| CaHap43ORF-2-PstI | 5′-GCCCGCCTGCAGTCACTAATTATATGCTCTTCTATCTAATTC-3′ | PstI, reverse, pCIplexA-F-CaHap43-3 |
| SaLexA-2(2) | 5′-GCAACTGTCAAACGC-3′ | Forward, integration of pCIplexA F1 derivatives |
| RPS1-1 | 5′-CGTATTCACTTAATCCCACAC-3′ | Reverse, integration of pCIplexA F1 derivatives |
| ade2hisG-1 | 5′-CTTTCTCATGCGTTCATGCACCAC-3′ | Forward, integration of pCR-lacZ derivatives |
| pCR-LacZ-vector-2 | 5′-CTGGTCTATAGTGTCACCT-3′ | Reverse, integration of pCR-lacZ derivatives |
| OPGCRE-1 | 5′-TCGACATACGAACAAATATTCGCAATGACTCTGACTCTGACTCTGACTCTGACTCCTGCA-3′ | SalI, SalexA operator, (GCRE)5, PstI, top strand, pCR-OPGCRElacZ |
| OPGCRE-2 | 5′-GGAGTCAGAGTCAGAGTCAGAGTCAGAGTCATTGCGAATATTTGTTCGTATG-3′ | PstI, (GCRE)5, SalexA operator, SalI, down strand, pCR-OPGCRElacZ |
| CaGFP-ORF-HindIII-1 | 5′-GTCAAAGCCTAAAATGAGTAAGGGAGAAGAACT-3′ | HindIII, forward, CIpGFP-F-CaHap43-1 |
| CaGFP-ORF-MluI-2 | 5′-ACCAACGCGTACCTTTGTATAGTTCATCCA-3′ | MluI, reverse, CIpGFP-F-CaHap43-1 |
a
Restriction sites introduced into the primers are underlined and described in the note. S. aureus lexA operator sequences are in boldface. The (GCRE)5 is in italics.