TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Relevant characteristics | Reference/source |
|---|---|---|
| Plasmids | ||
| pTNS3 | Apr; helper plasmid encoding the site-specific TnsABCD Tn7 transposition pathway | 32 |
| pUC18-mini-Tn7T-GM-LAC | Apr Gmr; mini-Tn7 vector containing lacIq and tac promoter | 18 |
| pUC18-mini-Tn7T-GM-Plac-HA | Apr Gmr; mini-Tn7 vector containing lacIq and lac promoter | This study |
| pUC18-mini-Tn7T-GM-Ptac-HA | Apr Gmr; mini-Tn7 vector containing lacIq and tac promoter | This study |
| pUC18-mini-Tn7T-GM-Ptac-sfgfp | Apr Gmr; mini-Tn7 vector containing sfGFP under the control of the tac promoter | This study |
| pUC18-mini-Tn7T-GM-Plac-sfgfp | Apr Gmr; mini-Tn7 vector containing sfGFP under the control of the lac promoter | This study |
| pNTPS138/139 | Kmr; Suicide vector containing oriT and sacB | D. Alley |
| pMEH0125 | Suicide plasmid for allelic exchange of tetRA locus | Fuqua laboratory |
| pNTPS139 ΔtetRA::a-attTn7 | Suicide plasmid for allelic exchange of tetRA locus with artificial attTn7 site | This study |
| pSRKKm | Kmr; broad-host-range vector containing lacIq and lac promoter | 17 |
| pSRKKm-Plac-HA | pSRKKm vector containing lacIq and lac promoter with an HA tag | This study |
| pSRKKm-Plac-sfgfp | pSRKKm vector containing lacIq and lac promoter with sfGFP | This study |
| pSRKKm-Ptac-sfgfp | pSRKKm vector containing lacIq and tac promoter with sfGFP | This study |
| pNTPS138Δrem | Kmr Sucs; deletion plasmid for rem | Fuqua laboratory |
| pSRKKm-Plac-rem-HA | Kmr; complementation vector containing rem-HA under the control of the lac promoter | This study |
| pUC18-mini-Tn7T-GM-Plac-rem-HA | Apr Gmr; mini-Tn7 vector containing rem-HA under the control of the lac promoter | This study |
| pNTPS138ΔuppX | Kmr Sucs; deletion plasmid for uppX | Fuqua laboratory |
| pSRKKm-Plac-uppX-HA | Kmr; complementation vector containing uppX-HA under the control of lac promoter | This study |
| pUC18-mini-Tn7T-GM-Plac-uppX-HA | Apr Gmr; mini-Tn7 vector containing uppX-HA under the control of the lac promoter | This study |
| pJEH005 (pNTPS138ΔctA) | Kmr Sucs; deletion plasmid for ctrA | Fuqua laboratory |
| pUC18-mini-Tn7T-GM-Plac-ctrA | Apr Gmr; mini-Tn7 vector containing ctrA under the control of the lac promoter | This study |
| pUC18-mini-Tn7T-GM-Ptac-ctrA | Apr Gmr; mini-Tn7 vector containing ctrA under the control of the tac promoter | This study |
| pKC129 | Source of sfGFP | Huang laboratory |
| pRVGFPC-2 | Source of multiple-cloning-site fragment | 50 |
| E. coli strains | ||
| DH5α | Cloning strain | Life Technologies |
| S17-1 | Smr; RP4-2 Tc::Mu Km-Tn7; for plasmid mobilization | 51 |
| A. tumefaciens strains | ||
| C58 | Nopaline type strain; pTiC58; pAtC58 | 52 |
| C58 pSRKKm-Plac-HA | C58 with empty pSRKKm plasmid | This study |
| C58ΔtetRA::a-attTn7 | Replacement of the tetRA locus with an artificial attTn7 site | This study |
| C58ΔtetRA::a-attTn7 pSRKKm-Plac-HA | C58ΔtetRA::a-attTn7 with empty pSRKm-Plac-HA plasmid | This study |
| C58ΔtetRA::a-attTn7 pSRKKm-Plac-sfgfp | C58ΔtetRA::a-attTn7 with pSRKm expressing sfGFP under the control of the lac promoter | This study |
| C58ΔtetRA::a-attTn7 pSRKKm-Ptac-sfgfp | C58ΔtetRA::a-attTn7 with pSRKm expressing sfGFP under the control of the tac promoter | This study |
| C58ΔtetRA::mini-Tn7T-GM-LAC | Mini-Tn7T-GM-LAC inserted into a-attTn7 site | This study |
| C58ΔtetRA::mini-Tn7T-GM-Plac-sfgfp | Mini-Tn7T-GM-Plac-sfgfp inserted into a-attTn7 site to express sfGFP under the lac promoter | This study |
| C58ΔtetRA::mini-Tn7T-GM-Ptac-sfgfp | Mini-Tn7T-GM-Ptac-sfgfp inserted into a-attTn7 site to express sfGFP under the tac promoter | This study |
| C58ΔtetRA::a-attTn7 Δrem | Δrem | This study |
| C58ΔtetRA::mini-Tn7T-GM-Plac-rem-HA Δrem | Chromosome-based complementation of Δrem with Rem-HA | This study |
| C58ΔtetRA::a-attTn7 Δrem pSRKKm-Plac-HA | Empty pSRKKm-Plac-HA plasmid in Δrem | This study |
| C58ΔtetRA::a-attTn7 Δrem pSRKKm-Plac-rem-HA | Plasmid-based complementation of Δrem with Rem-HA | This study |
| C58ΔtetRA::a-attTn7 ΔuppX | ΔuppX | This study |
| C58ΔtetRA::mini-Tn7T-GM-Plac-uppX-HA ΔuppX | Chromosome-based complementation of ΔuppX with UppX-HA | This study |
| C58ΔtetRA::a-attTn7 ΔuppX pSRKKm-Plac-HA | Empty pSRKKm-Plac-HA plasmid in ΔuppX | This study |
| C58ΔtetRA::a-attTn7 ΔuppX pSRKKm-Plac-uppX-HA | Plasmid-based complementation of ΔuppX with UppX-HA | This study |
| C58ΔtetRA::mini-Tn7T-GM-Plac-ctrA | Mini-Tn7T-GM-Ptac-ctrA inserted into a-attTn7 site to express CtrA under the lac promoter | This study |
| C58ΔtetRA::mini-Tn7T-GM-Ptac-ctrA | Mini-Tn7T-GM-Ptac-ctrA inserted into a-attTn7 site to express CtrA under the tac promoter | This study |
| C58ΔtetRA::mini-Tn7T-GM-Ptac-ctrA ΔctrA | Chromosome-based complementation of ΔctrA with CtrA | This study |