Plasmids used in this study

PlasmidDescriptionResistance phenotypeaReference
R388IncW plasmidSu Tp42
R388::IS26R388 with IS26bSu TpThis study
R388::IS26-FS–RR388::IS26 frameshift mutantcSu TpThis study
pRMH7618.8-kb BamHI fragment of pRMH760 cloned into pUC19Ap Km Nm12
pRMH762pRMH761 Kms derivativedApThis study
pRMH962pRMH762 tnp26 FS-L mutanteApThis study
pRMH977pRMH762 derivativef,gApThis study
pRMH978pRMH977 tnp26 FS-L mutanteApThis study
pRMH979pRMH762 derivativef,hApThis study
pRMH987pRMH977 Tnp26 mutant D82LApThis study
pRMH988pRMH977 Tnp26 mutant D142LApThis study
pRMH989pRMH977 Tnp26 mutant E177LApThis study
  • a Ap, ampicillin; Km, kanamycin; Nm, neomycin; Su; sulfamethoxazole; Tp, trimethoprim.

  • b IS26 8-bp duplication of bases 26745 to 26752 in R388 (GenBank accession no. BR000038).

  • c Lacks 13 bp (bases 624 to 636 from the left end of IS26, as shown in Fig. 5).

  • d Loss of aphA1 and one IS26 after overnight growth without kanamycin selection.

  • e Frameshift generated by end filling the BsiWI site and duplicating 116 to 119 bp from the left end of IS26 as shown in Fig. 5.

  • f IS26 together with bases 119362 to 119454 and 122137 to 122225 from GenBank accession no. KF976462.

  • g Cloned insert in the same orientation as pRMH762, with tnp26 facing toward Plac in pUC19.

  • h Cloned insert in the opposite orientation to pRMH762, with tnp26 facing away from Plac in pUC19.