TABLE 1.
Plasmids and strains
| Plasmid or strain | Genotype or phenotype | Remarks and reference |
|---|---|---|
| Plasmids | ||
| pUC19 | Apr; 2.7 kb | General cloning vector (35) |
| pBluescript SK+ | Apr; 3.0 kb | General cloning vector (Stratagene) |
| pRN5542 | Cmr; 3 kb | Staphylococcal cloning vector, derivative of pC194 (25) |
| pJIM2246 | Cmr; 6.0 kb | Shuttle vector (27) |
| pGO621 | Apr; 5.2 kb | mecR1-mecI ligated into puc19 at EcoRI-BamHI sites (this study) |
| pGO621C | Apr Cmr; 8.2 kb | pGO621 with pSK265 ligated at BamHI site (this study) |
| pGO630 | Spr Tcr Emr ts; 16 kb | Promoterless lacZ gene from E. coli fused to first 50 bp of mecA, intact mecR1-mecI ligated into pSK950 (31) |
| pSK950 | Emr Spr Tcr ts; 10.4 kb | pCL84 with pE194ts at PstI site (24); integration vector |
| pGO640 | Apr; 5.2 kb | PstI-EaeI fragment containing intact blaZ, blaR1, and truncated blaI from pCH2278 ligated into PstI-NotI sites of pBluescript KS+ |
| pGO641 | Apr Emr; 9.5 kb | pE194 ligated into ClaI site of pGO640 (this study) |
| pCH2278 | Cmr; 6.2 kb | 3.2-kb EcoRI-XhoI fragment containing bla locus ligated into EcoRI-SalI sites of pRN5542 (10) |
| pCH1988 | Cmr; 6.2 kb | Same as pCH2278; contains linker insertion mutation in blaR1 (10) |
| pGO630ΔR1 | Spr Tcr Emr ts; 16 kb | mecR1 mutant of pGO630 created by filling in ClaI site present in mecR1 (this study) |
| pGO681 | Cmr; 8.0 kb | 2.0-kb BamHI-PstI fragment containing first 50 bp of mecA, intact mecR1, and mecI truncated at PstI site ligated into BamHI-PstI sites of pJIM2246 (this study) |
| pGO682 | Cmr; 8.8 kb | 2.8-kb PstI-EaeI fragment containing intact blaZ, blaR1, and blaI truncated at EaeI site ligated into PstI-NotI sites of pJIM2246 (this study) |
| pACT2 | Apr; 8.1 kb | Activation domain hybrid cloning vector (Clonetech) |
| pAS1 | Apr; 7.3 kb | DNA-binding domain hybrid vector (Clonetech) |
| S. aureus | ||
| RN450 | Cms Ems Gms Tcsbla | ATCC 8235-4 |
| RN4220 | Cms Ems Gms Tcsbla | Restriction-deficient mutagenized RN450 shuttle plasmid host (22) |
| GO450TF | Cms Ems Gms Tcsbla | Provided by Tim Foster |
| GO450TF::630 | Cms Emr Gms Tcrbla geh | GO450TF with integrated pGO630 (this study) |
| GO450TF::630ΔI | Cms Emr Gms Tcrbla geh | mecR1 mecI mutant derivative of GO450TF::630 created by allelic replacement (this study) |
| GO450TF::630ΔRI | Cms Emr Gms Tcrbla geh | mecR1 mecI+ derivative of GO450TF::630 created by allelic replacement (this study) |
| E. coli | ||
| DH5α | λ−φ80dlacZΔM15 Δ(lacZYA-argF)U169 deoR recA1 endA1 phoA hsdR17(r−K m+K) supE441 thi-1 gyrA96 relA91 glnV44 (7) | |
| Able C | lac (LacZw−) (Kanr McrA− McrB− McrF− Mrr− | |
| hsdR(rK− mK−) (F′ proAB laclqZΔM15 Tn10(Tetr) (Stratagene) | ||
| S. cerevisiae | ||
| Y190 | MATa ura3-52 his3-200 ade2-101 lys2-801 trp1-901 leu2-3 gal4Δ gal80Δ cyhr2 LYS::GAL1uas−HIS3tata−HIS3 URA3::GAL1uas−GAL1tata−lacZ (Clontech) |