TABLE 2.
Plasmids and oligonucleotide primers used in this study
| Plasmid/primer | Description/sequence (5′-3′) | Source or location/use |
|---|---|---|
| Plasmids | ||
| pCKS01 | C-V5-H6-comW in plasmid pCRT7/CT TOPO | This study |
| pCKS02 | N-H6-Xa-comW in plasmid pET16b | This study |
| pCKS03 | comW in plasmid pEVP3 | This study |
| pCKS07 | C-V5-H6-comW in plasmid pEVP3 | This study |
| pEVP3 | Plasmid containing the Cmr-encoding gene and the promoterless lacZ reporter derived from pTV32 | 7 |
| pCR T7/CT TOPO | Plasmid for producing target proteins containing a C-terminal histidine tag | Invitrogen |
| pET16b | Plasmid for producing target proteins containing an N-terminal histidine tag | Novagen |
| pR410 | Plasmid containing the kan gene | 33 |
| Primers | ||
| CKS24 | TATCTAGAGGCTGGTATTTTAACAATTCA | Upstream of comW (CP1721) |
| CKS25 | ATGGATCCTCAACAAGAAATAAACCCCCGATTCA | In comW (CP1721) |
| CKS33 | ATGTTACAAAAAATTTATGAGCAGATGGC | In comW (pCKS01) |
| CKS34 | ACAAGAAATAAACCCCCGATTCATTACCAA | In comW (pCKS01) |
| CKS107 | ATTCTAGAGCAGATGGCTAATTTCTATGATAGT | In comW (CP1802) |
| CKS108 | ATGGATCCTCCGGATATAGTTCCTCCTTTCAG | In pCR T7/CT TOPO (CP1802) |
| CKS109 | GGCATATGATTATGTTACAAAAAATTTATGAGC | In comW (pCKS02) |
| CKS110 | ATGGATCCTCAACAAGAAATAAACCCCCGATTCA | In comW (pCKS02) |
| CKS116 | GATCTAGAAAGGAGATATACCATGGGCCATCA | In pET16b (CP1805) |
| CKS114 | ATGGGCCCTAGAAGTCTCAGCGAGCTCCATT | Upstream of comW (CP1805) |
| CKS115 | GCTCTAGATTATAAACTTATTCTAACAAAAAA | Upstream of comW (CP1805) |
| DAM303 | AAGGGCCCGTTTGATTTTTAATG | In pR410 (KANT) |
| DAM304 | AGGATCCATCGATACAAATTCCTC | In pR410 (KANT) |
| DAM497 | CAATTGACTATATTAGAGGCGAGACA | Upstream of comW (CP1805) |
| CKS113 | ATGGATCCTTCTTCTTAGAAAAGGCCGTTTTA | Upstream of comW (CP1805) |