TABLE 1.
Plasmids used in the study
| Plasmid | Descriptiona | Selective markerb | Repliconc | Reference or source |
|---|---|---|---|---|
| pBR332 | Cloning vector | Tc, Ap | pMB1 | 41 |
| pSU18 | Cloning vector | Cm | p15A | 4 |
| pSU19 | Cloning vector | Cm | p15A | 4 |
| cloneQ | Shuttle vector | Ap | oriR, oriC | 46 |
| pPM13 | Shuttle vector | Ap | oriR, oriC | 12, 48 |
| pGF1 | pGFIB PvuIIΩsuptRNACys | Ap | pMB1 | Promega |
| pGF2 | pGFIB PvuIIΩsuptRNAGly | Ap | pMB1 | Promega |
| pGF3 | pGFIB PvuIIΩsuptRNAGlu | Ap | pMB1 | Promega |
| pGF4 | pGFIB PvuIIΩsuptRNAHis | Ap | pMB1 | Promega |
| pGF5 | pGFIB PvuIIΩsuptRNAPhe | Ap | pMB1 | Promega |
| pGF6 | pGFIB PvuIIΩsuptRNAPro | Ap | pMB1 | Promega |
| pHMK1 | pSU18Δ(BamHI-EcoRI)Ω(9877-2502) | Cm | p15A | This study |
| pHMK2 | pSU19Δ(EcoRI-XbaI)Ω(2502-4863) | Cm | p15A | This study |
| pHMK3 | pSU18Δ(XbaI-SacI)Ω(4863-7173) | Cm | p15A | This study |
| pHMK4 | pSU19Δ(SacI-BamHI)Ω(7173-9877) | Cm | p15A | This study |
| pJB140 | pBR322 PstIΩPM2 | Ap | pMB1 | This study |
| pSM8 | pSM27 SmaIΩsuptRNACys | Ap (amber), Cm | oriR, oriC | This study |
| pSM9 | pSM27 SmaIΩsuptRNAGly | Ap (amber), Cm | oriR, oriC | This study |
| pSM10 | pSM27 SmaIΩsuptRNAGlu | Ap (amber), Cm | oriR, oriC | This study |
| pSM11 | pSM27 SmaIΩsuptRNAHis | Ap (amber), Cm | oriR, oriC | This study |
| pSM12 | pSM27 SmaIΩsuptRNAPhe | Ap (amber), Cm | oriR, oriC | This study |
| pSM13 | pSM27 SmaIΩsuptRNAPro | Ap (amber), Cm | oriR, oriC | This study |
| pSM25 | pPM13 with amber mutation in bla (Q37) | Ap (amber) | oriR, oriC | This study |
| pSM27 | pSM25Δ(NdeI-XbaI)Ωcat | Ap (amber), Cm | oriR, oriC | This study |
| pSM76 | pSM124 with amber mutation in gene X (H47) | Cm | p15A | This study |
| pSM93 | pBR322 BamHIΩsus2 | Ap | pMB1 | This study |
| pSM96 | pBR322 BamHIΩPM2V1 | Ap | pMB1 | This study |
| pSM123 | pSU18Δ(BamHI-XbaI)Ω(9877-4863) | Cm | p15A | This study |
| pSM124 | pSU18Δ(XbaI-BamHI)Ω(4863-9877) | Cm | p15A | This study |
a
bla, β-lactamase; cat, chloramphenicol transacetylase. The numbers refer to the PM2 genome coordinates (GenBank accession no. AF155037); Ω, insertion; Δ, deletion.
b
Cm, chloramphenicol; Ap, ampicillin; Tc, tetracycline.
c
oriR, origin of replication in P. haloplanktis TAC125; oriC, pMB1, and p15A, origin of replication in E. coli.