TABLE 1.

Plasmids used in this study for functional analysis.

PlasmidRelevant featuresaReference or source
Rms149IncP-6, Cbr Gmr Smr Spr Sur Mob+ 37
pUB307IncP-1, deletion derivative of RP1, Kmr Tcr Tra+ 4
pGEM-T EasyCloning vector for PCR products, received linearized PnrPromega
pCR-4Blunt-TOPOCloning vector for shotgun sequencing, received linearized, Kmr PnrInvitrogen
pAH149-1Clone of Rms149 bases 54514 to 56668 (repA) in pCR-4Blunt-TOPO, Kmr Pnr Rep+This work
pAH149-2Clone of Rms149 bases 55315 to 349 (parABC) in pCR-4Blunt-TOPO, Kmr Pnr RepThis work
pAH149-3Clone of Rms149 bases 55923 to 753 (parABC) in pCR-4Blunt-TOPO, Kmr Pnr RepThis work
pAH149-4Clone of Rms149 bases 53432 to 55556 (′repA) in pCR-4Blunt-TOPO, Kmr Pnr RepThis work
pAH149-5Clone of Rms149 bases 53922 to 56224 (repA) in pCR-4Blunt-TOPO, insert orientation is opposite pAH149-1 to 4, Kmr Pnr Rep+This work
pAH149-6Clone of Rms149 bases 4961 to 7198 [intI1 aac(3)-I] in pCR-4Blunt-TOPO, Gmr Kmr PnrThis work
pAH149-7Clone of Rms149 bases 4125 to 6601 [intI1′ aac(3)-I aadA5] in pCR-4Blunt-TOPO, Gmr Kmr Pnr Smr SprThis work
pAH149-12Derivative of pAH149-1 and pAH149-2 carrying Rms149 bases 54514 to 349, Kmr PnrThis work
pAH149-13Derivative of pAH149-1 and pAH149-3 carrying Rms149 bases 54514 to 753, Kmr PnrThis work
pAH149-1rDeletion derivative of pAH149-1 without vector replicon or penicillin resistance, KmrThis work
pAH149-12rDeletion derivative of pAH149-12 without vector replicon or penicillin resistance, KmrThis work
pAH149-13rDeletion derivative of pAH149-13 without vector replicon or penicillin resistance, KmrThis work
pGEM-149mobPCR product of mobilisation region, bases 30665 to 35361 cloned into pGEM-T Easy, Pnr Mob+This work
  • a Antibiotic resistances: Cb, carbenicillin; Gm, gentamicin; Km, kanamycin; Pn, penicillin; Sm, streptomycin; Sp, spectinomycin; Su, sulfonamide. Other phenotypic traits: Mob+, mobilizable; Tra+, conjugative. For pAH149-1 to 5, Rep+ and Rep indicate ability to replicate in P. putida from a replicon on the Rms149 insert.