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Table 1 Strains and plasmids

From: SimReg1 is a master switch for biosynthesis and export of simocyclinone D8 and its precursors

Bacterial strains and plasmids Description Source or reference
E. coli DH5α supE44 ΔlacU169(φ80lacZΔM15) Hanahan 1985
E. coli BL21 (DE3) pLysS Host for the heterologous expression of His6 -tagged simReg1 Novagen
E. coli ET12567/pUB307 hsdR17 recA1endA1gyrA96 thi-1 relA1 dam-13::Tn9(Cmr) dcm-6 hsd M; harbors conjugative plasmid pUB307; Cmr, Kmr Flett et al. 1997
S. antibioticus Tü6040 Simocyclinone D8 producing strain Trefzer et al. 2002
S. antibioticus Derivative of S. antibioticus Tü6040 with This work
ΔsimReg1 disrupted simReg1 gene  
S. antibioticus ΔsimReg1 × pSSimR1-1 ΔsimReg1 strain carrying plasmid with the intact simReg1 gene under its own promoter, used for complementation studies This work
S. lividans 1326 Wild type Hopwood et al. 1985
S. lividans × pSimD4script Derivative of S. lividans 1326 carrying plasmid with gusA gene under the control of the putative promoter of the simD4 gene This work
S. lividans × pSimD4script/pUWLsimReg1 Derivative of S. lividans 1326 carrying plasmid with gusA gene under the control of putative promoter of the simD4 gene and second plasmid with simReg1 gene under the control of P ermE This work
S. lividans ×pGUS Derivative of S. lividans 1326 carrying plasmid with promoterless reporter gene gusA This work
S. lividans × pGUS/pUWLsimReg1 Derivative of S. lividans 1326 carrying plasmid with promoterless reporter gene gusA and plasmid with simReg1 gene under the control of the P ermE promoter This work
pBluescriptIIKS + General purpose cloning vector; Apr MBI Fermentas
pUC19 General purpose cloning vector; Apr MBI Fermentas
pSET152 E. coli/Streptomyces shuttle vector with φC31 integration system for streptomycetes; Amr Bierman et al. 1992
pKC1218E pKC1218 derivative expression vector with P ermE promoter and SCP2* replicon; Amr Ostash et al. 2004
pHYG1 pLitmus38 containing hygromycin resistance cassette hyg C. Olano Univ. de Oviedo, Spain
pKC1139 E. coli/Streptomyces shuttle vector with temperature sensitive replicon pSG5, Amr Muth et al. 1989
pUWL-oriT pUWL-KS derivative harboring oriT from pSET152 Zelyas et al. 2009
pET21d Vector for His-tagged protein expression Novagen
5JH10 pUC plus simB3-D4 segment Trefzer et al. 2002
pUCsimR1 pUC19 derivative containing simReg1 gene This work
pUCsimR1-hyg pUCsimR1 derivative with hyg cassette cloned into the simReg1 coding region This work
pKCsimR1-hyg pKC1139 derivative with cloned simReg1::hyg construction used for simReg1 gene inactivation This work
pKCEsimR1 pKCE1218 derivative containing simReg1 gene under the control of P ermE Rebets et al. 2008
pSSimR1 pSET152 plus 2.3 kb simD4-X1 segment This work
pSSimR1-1 pSET152 derivative containing simReg1 gene under the control of its own promoter This work
pMA-simR1 plasmid containing synthetic codon-optimized simReg1 gene Mr. Gene, Heidelberg
pETSR1c-15 pET21d derivative containing synthetic codon-optimized simReg1 gene This work
pGUS pSET152 derivative containing promoterless reporter gene gusA Myronovskyi et al.2011
pSimD4script derivative of pGUS harboring gusA reporter gene under the promoter of the simD4 gene This work
pUWLsimReg1 derivative of pUWL containing gene simReg1 under the control of P ermE This work