Table 3 Strains and plasmids used in this study

Strains

 E. coli Nova blue

endA1 hsdR17(r_K12⁻m_K12⁺) supE44 thi-I gyrA96 relA1 lac recA1/F’[proAB + lacIq ZΔM15::Tn10(Tetr)]

Novagen

 B. subtilis W168

trpC2

JMC^a

 B. subtilis W168∆2

trpC2∆ackA∆pta

This study

 B. subtilis W168∆3

trpC2∆ackA∆pta∆lctE

This study

 B. subtilis W168∆4

trpC2∆ackA∆pta∆lctE∆mmgA

This study

Plasmids

 pJOE8999

Kan^R, P_manPA-cas9, P_vanP*, lacPOZ’-gRNA, oop ter, T7P, repE194^ts, pUCori

Altenbuchner [30]

 pJOE_ackA

pJOE8999 derivative, containing 20-nt spacer targeting ackA and fused homologous arms of ackA

This study

 pJOE_pta

pJOE8999 derivative, containing 20-nt spacer targeting pta and fused homologous arms of pta

This study

 pJOE_lctE

pJOE8999 derivative, containing 20-nt spacer targeting lctE and fused homologous arms of lctE

This study

 pJOE_mmgA

pJOE8999 derivative, containing 20-nt spacer targeting mmgA and fused homologous arms of mmgA

This study