Table 3 Bacillus subtilis strains, plasmids, and primers used in this study

Strains

168

trpC2

BGSC

1A474

amyE3 aroI10

BGSC

BSSA/pHCMC04

1A474 carrying pHCMC04

This work

BSSA/pSAAroA

1A474 carrying pSAAroA

This work

BSSA/pSAAroB

1A474 carrying pSAAroB

This work

BSSA/pSAAroC

1A474 carrying pSAAroC

This work

BSSA/pSAAroD

1A474 carrying pSAAroD

This work

BSSA/pHCMC04/ pDG148-stu

1A474 carrying pHCMC04 and pDG148-stu

This work

BSSA/pHCMC04/ pDGSAAroD

1A474 carrying pHCMC04 and pDGSAAroD

This work

BSSA/pSAAroA/ pDGSAAroD

1A474 carrying pSAAroA and pDGSAAroD

This work

BSSA/pSAAroB pDGSAAroD

1A474 carrying pSAAroB and pDGSAAroD

This work

BSSA/pSAAroC pDGSAAroD

1A474 carrying pSAAroC and pDGSAAroD

This work

BSSA/pSAAroD pDGSAAroD

1A474 carrying pSAAroD and pDGSAAroD

This work

BSSA(Ωtkt::pSATkt)

BSSA/pSAAroA/pDGSAAroD harboring Ωtkt::pSATKT

This work

BSSA(Ωpyk::pSAPyk)

BSSA/pSAAroA/pDGSAAroD harboring Ωpyk::pSAPYK

This work

Plasmids

pHCMC04

bla P _{xyl A} xylR cat

BGSC

pDG148-stu

neo ble Pspac lacI bla

BGSC

pMUTIN4

bla Pspac lacZ lacI ermC

BGSC

pSAAroA

pHCMC04 derivative, aroA expressed from P _{xyl A}

This work

pSAAroB

pHCMC04 derivative, aroB expressed from P _{xyl A}

This work

pSAAroC

pHCMC04 derivative, aroC expressed from P _{xyl A}

This work

pSAAroD

pHCMC04 derivative, aroD expressed from P _{xyl A}

This work

pDGSAroD

pDG148-stu derivative, aroD expressed from Pspac

This work

pSATkt

bla Pspac-tkt-lacZ lacI ermC

This work

pSAPyk

bla Pspac-pyk’-lacZ lacI ermC

This work

Primers

aroA-fw-Spe I

TCAACTAGT ATGAGCAACACAGAGTTAG

Cloning of aroA into pHCMC04

aroA-rv-Xma I

TCACCCGGG TTAAGCGTTGACTTTCAC

aroB-fw-Spe I

TCCACTAGT ATGAAGACACTGCATGTTC

Cloning of aroB into pHCMC04

aroB-rv-Xma I

TCAGGATCC TCATGATGTCTCCTCCAATC

aroC-fw-Spe I

TCAACTAGT GTGAACGTGTTAACGATTAAAG

Cloning of aroC into pHCMC04

aroC-rv-Xma I

TCACCCGGG CTATCCCCGTGTGTTTTTATG

aroD-fw-Spe I

TCCACTAGT ATGAAAAAGCTGTACGG

Cloningof aroD into pHCMC04

aroD-rv-Xma I

TCAGGATCC TTAACATTCTGTTCCTCC

aroD-DG-fw

AAGGAGGAAGCAGGTATGAAAAAGCTGTACGG

Cloning of aroD into pDG148-stu

aroD-DG-rv

GACACGCACGAGGTTTAACATTCTGTTCCTCC

tkt-Mu-fw-Xma I

ACTCCCGGG ACATAAGGAAGGGGATTTTTATG

Cloning of tkt into pMUTIN4

tkt-Mu-rv-Bam HI

ACTGGATCC ACTCATCCTCTTTCAAAAGC

tkt-ck-fw

TTTCAGGAATACATAGAG

Confirming the integration of pSATKT

tkt-ck-rv

TTAGACAAAATTTCTTTC

pyk-Mu-fw-Xma I

CCCCCGGG GAAGATTTCAGAAGGAAGTGAACC

Cloning 5’ end of pyk into pMUTIN4

pyk-Mu-rv-Bam HI

CCGGATCC GGAATTTCCACACCTAAGTCTCC

pyk-ck-fw

AACGTACATATGTAATCG

Confirming the integration of pSAPYK

pyk-ck-rv

CCGTTCAGCAAAAATAAAAC