TABLE 2.

Plasmids used

PlasmidRelevant propertiesSource or referencea
pTA131Integrative vector based on pBluescript, with pyrE2 marker3
pTA187Integrative vector based on pUC19, with hdrB marker3
pTA230Shuttle vector derived from pTA131, with pyrE2 marker and pHV2 replication origin3
pTA354Shuttle vector based on pBluescript, with pyrE2 marker and pHV1/4 replication origin31
pTA927Overexpression vector with pyrE2 marker and pHV2 origin, derived from pTA230 by insertion of 131-bp t.L11e terminator at KpnI site, 224-bp p.tnaA promoter at ApaI and ClaI sites, and 35-bp t.Syn terminator at NotI and BstXI sitesThis study; FN645893
pTA929Overexpression vector with 6×His tag, pyrE2 marker, and pHV2 origin, derived from pTA927 by insertion of 26-bp fragment containinga His tag (CAC)6 tract at the NdeI and ClaI sitesThis study; FN645892
pTA949Shuttle vector derived from pTA230, with pyrE2 and hdrB markers and pHV2 replication originThis study; FN645894
pTA962Overexpression vector with pyrE2 and hdrB markers and pHV2 origin, derived from pTA949 by insertion of t.L11e terminator, p.tnaA promoter and t.Syn terminator (see pTA927 for details)This study; FN645891
pTA963Overexpression vector with 6×His tag, pyrE2 and hdrB markers, and pHV2 origin, derived from pTA962 by insertion of a His tag (CAC)6 tract (see pTA929 for details)This study; FN645890
pTA1041pTA963 with insertion of 1.0-kb NcoI-BamHI radA+ fragment at the PciI and BamHI sites; for overexpression of 6×His-tagged RadAThis study
pTA1043pTA963 with insertion of 0.7-kb BspHI-BamHI radB+ fragment at the PciI and BamHI sites; for overexpression of 6×His-tagged RadBThis study
pTA1106pTA131 with 3.8-kb pitANph gene replacement construct, consisting of HindIII-EcoRI upstream ΔpitAHvo fragment, EcoRI-XbaI pitANph+ fragment, and XbaI-NotI downstream ΔpitAHvo fragment inserted at HindIII and NotI sitesThis study
pTA1150pTA131 with 1.3-kb Δmrr construct consisting of a KpnI-BamHI upstream Δmrr fragment and a BamHI-XbaI downstream Δmrr fragment inserted at KpnI and XbaI sitesThis study
pTA1180pTA131 with 878-bp Δcdc48d construct consisting of KpnI-BamHI upstream Δcdc48d fragment and BamHI-EcoRI downstream Δcdc48d fragment inserted at KpnI and EcoRI sitesThis study
pTA1182pTA963 with insertion of 1.0-kb NcoI-BamHI radA(A196V) fragment at PciI and BamHI sites; for overexpression of 6×His-tagged RadA(A196V)This study
pMM1231pTA131 with 2.0-kb ΔpitAHvo construct consisting of a HindIII-EcoRI upstream ΔpitAHvo fragment and an EcoRI-NotI downstream ΔpitAHvo fragment; inserted at HindIII and NotI sitesThis study
pMM1232pTA131 with 2.7-kb ΔpitAHvo::hdrB+ construct consisting of a HindIII-EcoRI upstream ΔpitAHvo fragment, an EcoRI-XbaI fragment of pTA187 with an hdrB marker, and an XbaI-NotI downstream ΔpitAHvo fragment; inserted at HindIII and NotI sitesThis study
  • a The accession number is given where appropriate.