TABLE 2

Plasmids used and constructed during this study

PlasmidDescriptionReference or source
pBluescript II KS(+)General cloning vector68
pKC1132Cloning vector, conjugative (oriT from RK2)69
pSET152Cloning vector, conjugative (oriT from RK2), integrative (phiC31 attP)69
pTC192-KmSource of neo (kanamycin resistance gene)70
pGM1190pSG5 derivative, tsr, aac(3)IV, oriT, to terminator, PtipA, RBS, fd terminator40 (pGM1190, G. Muth, unpublished)
pESAC13PAC vector (P1 phage replicon) for genomic library construction; conjugative (oriT from RK2), integrative (phiC31 attP), tsr, neo, P1 rep, sacB27 (pESAC13, M. Sosio, unpublished)
pGUSgusA (β-glucuronidase gene, codon-optimized for streptomycetes) in pSET15238
pIJ10740pGUS derivative with ermE*p driving gusA transcriptionMorgan Feeney, unpublished
pIJ10257Expression vector for Streptomyces, with ermE*p, hyg, conjugative (oriT from RK2), integrative (phiC31 attP)71
pIJ12850Derivative of pGM1190 with JFC010/JFC009-neo-JFC011/JFC012 fragments; for deletion of cxmKThis work
pIJ12851Derivative of pKC1132 with JFC010/JFC009-neo-JFC011/JFC012 fragments; for deletion of cxmKThis work
pIJ12857Derivative of pKC1132 with JFC010/JFC009 fragment; for deletion of cxmKThis work
pIJ12858Derivative of pKC1132 with JFC011/JFC012 fragment; for deletion of cxmKThis work
pIJ12859Derivative of pKC1132 with JFC010/JFC009-JFC011/JFC012 fragment; for deletion of cxmKThis work
pIJ12860Derivative of pGM1190 with JFC010/JFC009-JFC011/JFC012 fragment; for deletion of cxmKThis work
pIJ12861pBluescript II KS(+) derivative with JFC026/JFC034 fragment; for cloning of cxmKThis work
pIJ12852pIJ10257 derivative with cxmK; for complementation of deletion mutantThis work
pIJ12853Derivative of pESAC13; contains 145 kb of S. leeuwenhoekii chromosome, including the chaxamycin biosynthesis gene clusterThis work