Table 2 Strains, plasmids, and primers used in this study

RL3000Δferm

E. coli MG1655 ilvG⁺ rph⁺ rfb-50 ybhJ(L54→I) yebN(G25→D) ycfK::97 bp. ΔinsB-5 ΔinsA-5 ΔinsAB-5 ΔackA ΔfrdA ΔldhA ΔadhE ΔilvC ΔlacY

[19]

LJ110

E. coli W3110 Fnr⁺

[57]

LK3S

LJ110 ΔldhA ΔpflB

Lab collection

SB001

LJ110 ΔldhA ΔpflB ΔfrdA

This study

RL3000Δferm-pIBA4

RL3000Δferm transformed with plasmid pIBA4

[19]

SB001-pIBA4

SB001 transformed with plasmid pIBA4

This study

pIBA4

Expression plasmid encoding the iBuOH pathway

[pBBRori-aadA1(Sp^R)-T_L-lacI-P_lacIq] (814bb)-P_T7A1-O34-RBS_AS7-alsS_Bsu-RBS_AC7-ilvC_Eco-6E6-RBS_AD5-ilvD_Eco-RBS_AK7-kivd_Lla-RBS_AA2-adhA_Lla-M5-gfp-T_P22

[19]

pKD4

Template plasmid for generation of deletion cassette

[56]

Primer name

Sequence (5ʹ-3ʹ)

 P1_frdA_fw

CAAACCTTTCAAGCCGATCTTGCCATTGTAGGCGCCGTGTAGGCTGGAGCTGCTTCG

 P2_frdA_rv

CGCCTTCTCCTTCTTATTGGCTGCTTCCGCCTTATCCATATGAATATCCTCCTTA