Table 1 Strains and plasmids

Strains

 E. coli

  JM83

F’, ara, Δ(lac-pro AB), rpsL, (Str^r), Φ80, lacZΔM15

[27]

  ET12567/pUZ8002

dam-13::Tn9 dcm-6 hsdM; containing the non-transmissible RP4 derivative plasmid pUZ8002

[27]

 S17-1

recA, pro, hsdR, RP4-2-Tc::Mu-Km::Tn7

[37]

 S. lincolnensis

  NRRL 2936

Wild-type (WT), lincomycin producer

NRRL, USA

  ΔlmbU

NRRL 2936 with in-frame deletion of lmbU

This study

  WT/p0469TE

NRRL 2936 attBφC31::p0469TE

This study

  ΔlmbU/p0469TE

ΔlmbU attBφC31::p0469TE

This study

  WT/p1037TE

NRRL 2936 attBφC31::p1037TE

This study

  ΔlmbU/p1037TE

ΔlmbU attBφC31::p1037TE

This study

  WT/p8097TE

NRRL 2936 attBφC31::p8097TE

This study

  ΔlmbU/p8097TE

ΔlmbU attBφC31::p8097TE

This study

  ΔSLINC_0469

NRRL 2936 with in-frame deletion of SLINC_0469

This study

  ΔSLINC_1037

NRRL 2936 with in-frame deletion of SLINC_1037

This study

ΔSLINC_8097

NRRL 2936 with in-frame deletion of SLINC_8097

This study

  CSLINC_0469

SLINC_0469 complementation strain

This study

  CSLINC_1037

SLINC_1037 complementation strain

This study

  CSLINC_8097

SLINC_8097 complementation strain

This study

Plasmids

 pLU-1

lmbU cloned in NdeI/EcoRI sites of pET-28a ( +), for LmbU expression

[27]

 pKCcas9dO

actII-orf4-specific guide-RNA, homologous arms flanking actII-orf4, aac(3)IV, pSG5

[41]

 pKCcas9dlmbU

lmbU-specific guide-RNA, homologous arms flanking lmbU, aac(3)IV, pSG5

This study

 pIB139

Integrative vector based on φC31 integrase, ermE*p

[27]

 pPU139

pIB139 carrying lmbU gene under its own promoter

This study

 pATE152

pSET152 derivative carrying xylTE gene under lmbAp promoter

[42]

 pSET152

Integrative vector based on φC31 integrase

[43]

 p0469TE

pSET152 carrying xylTE gene under the control of SLINC_0469p

This study

 p1037TE

pSET152 carrying xylTE gene under the control of SLINC_1037p

This study

 p8097TE

pSET152 carrying xylTE gene under the control of SLINC_8097p

This study

 pKCcas9d0469

SLINC_0469-specific guide-RNA, homologous arms flanking SLINC_0469, aac(3)IV, pSG5

This study

 pKCcas9d1037

SLINC_1037-specific guide-RNA, homologous arms flanking SLINC_1037, aac(3)IV, pSG5

This study

 pKCcas9d8097

SLINC_8097-specific guide-RNA, homologous arms flanking SLINC_8097, aac(3)IV, pSG5

This study