Supplementary Table 1 Strains and plasmidsused in this work

Name / Relevant genotype / Source
Strains
Escherichia coliJM109 / recA1, endA1, gyrA96, thi-1, hsdR17, supE44, relA1, Δ(lac-proAB)/F’[traD36, proAB+, lacIq, lacZΔM15] / Laboratory stock
BSUΔL / Bacillussubtilis with gene ldh knockout; Kmr / Laboratory stock
BSUΔL01-2 / BSUΔL harbors a constitutive heterologous Ehrlich pathway (P43::kivd-adh2); Emr / Laboratory stock
BSUL04 / BSUΔL integrating P43::kivd-adh2 into the chromosome, and harboring P43::ilvD-ilvC-alsS; Kmr, Spcr, Emr / Laboratory stock
BSUΔL-01 / BSUΔL harbors a Pspac controlled heterologous Ehrlich pathway (Pspac::kivd-adh2, PlacI::lacI); Kmr, Emr / This work
BSUΔL-02 / BSUΔL with a Pspac controlled heterologous Ehrlich pathway (Pspac::kivd-adh2, PlacI::lacI) into the chromosome; Kmr, Spcr / This work
BSUΔL-03 / BSUΔL-02 harbors a PalsSD controlled KIV precursor biosynthetic pathway (PalsSD::ilvD-ilvC-alsS); Kmr, Spcr, Emr / This work
Plasmids
pMUTIN4 / B. subtilisintegration plasmid; Ampr, Emr / BGSCa
pHPL04 / B. subtilis-E. coli shuttle plasmid; P43::kivd-adh2; Emr / Li et al. (2011)
pHPL08 / B. subtilis-E. coli shuttle plasmid; P43::ilvD-ilvC; Emr / Li et al. (2011)
pDGM / B. subtilisintegration plasmid; Ampr, Emr, Spcr / Li et al. (2011)
pHPL10 / B. subtilis-E. coli shuttle plasmid; P43::kivd-adh2, PlacI::lacI; Emr / This work
pHPL11 / B. subtilis-E. coli shuttle plasmid; Pspac::kivd-adh2, PlacI::lacI; Emr / This work
pHPL12 / B. subtilis-E. coli shuttle plasmid; PalsSD::ilvD-ilvC; Emr / This work
pHPL13 / B. subtilis-E. coli shuttle plasmid; PalsSD::ilvD-ilvC-alsS; Emr / This work
pDGMPKAL / B. subtilisintegration plasmid; Pspac::kivd-adh2, PlacI::lacI; Ampr, Emr, Spcr / This work

aBGSC: Bacillus Genetic Stock Center

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Supplementary Table 2 Primers used in this work

Primer name / Sequencea 5'→3' / Source
Pspac-F / CTGGCATGGAGCTCGCTCTAGAATA / This work
Pspac-R / AGGATCTGCAGGTACCATTCCACACA / This work
lacI-F / AATATTCCCGGGTTTCAAAGACTAC / This work
lacI-R / ATAATCGATCGATCCGCTTACAGA / This work
PKAL-F / AGGCAGCATGCTATGACCATGATTACG / This work
PKAL-R / ATAAGTCGACAAGGGCGATCGATCCG / This work
PalsSD-F / CGCGCAAGCTTATGGAGGTCAATTTC / This work
PalsSD-R / GCTCGGTACCGCAATCAACAACAAG / This work
alsS2-F / CATGAAGCGCTAGCATTGCTTAGTGGAG / This work
alsS2-R / AATCTCTCGAGAATCTGGCTCACAGG / This work
RT-ilvC-F / GTCCGCAGAGAAGGCAAAGGAAG / This work
RT-ilvC-R / ATGTATTCGACGGAGTCGGCGAT / This work
RT-ilvD-F / GAATTCAACACCATCGTCGATGA / This work
RT-ilvD-R / ATGTATTCGACGGAGTCGGCGAT / This work
RT-kivd-F / AATGGATTAGCAGGAAGTTACGCCG / This work
RT-kivd-R / CGTCAGCCAGCGTATGATGAACA / This work
RT-adh2-F / GTCGTTGTCGGCATGGGTGAA / This work
RT-adh2-R / GCCATACAAGAACCGTTCAACCA / This work
RT-alsS-F / GGGTGCCTCTAACTTGGC / Li et al. (2011)
RT-alsS-R / TGCACGGATCACGTTTCC / Li et al. (2011)
RT-gapA-F / GGGTGCCTCTAACTTGGC / Li et al. (2011)
RT-gapA-R / TGCACGGATCACGTTTCC / Li et al. (2011)

aUnderline stands for the restriction site

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