Table 1.

Bacterial strains, plasmids and oligonucleotides used in this study

Bacterial strains	Genotype	Source/reference
E. coli OmniMAX	F′ {proAB lacIq lacZΔM15 Tn10(TetR) Δ(ccdAB)} mcrA Δ(mrr hsdRMS-mcrBC) Φ 80(lacZ)ΔM15 Δ(lacZYA-argF)U169 endA1 recA1 supE44 thi-1 gyrA96 relA1 tonA panD; used for cloning	Invitrogen
B. subtilis 1012	leuA8 metB5 trpC2 hsrM1	[18]

Plasmids	Description	Source/references
pHCMC02-bgaB	PlepA-bgaB, inducer-free expression plasmid	[12]
pNDH33-bgaB	Pgrac01-bgaB, inducible	[3]
pLacI	Expression of LacI, used with E. coli cloning strain	Novagen
pHT01	Pgrac01, inducible	[9]
pHT01-bgaB	Pgrac01-bgaB, inducible	[9]
pHT24	Pgrac01, Strep-tag II, inducible	[9]
pHT10-gfp+	Pgrac01-gfp+, inducible	[9]
pHT100	Pgrac100-bgaB, inducible	[4]
pHT255	Pgrac100, inducible	[4]
pHT1168	Pgrac100-gfp+, inducible	[4]
pHT1650	Pgrac01-gfp+, ΔlacI, lacO1-lacO3 548 bp	This study
pHT1651	Pgrac01-gfp+, ΔlacI, lacO1-lacO3 787 bp	This study
pHT1653	Pgrac01-bgaB, ΔlacI, lacO1-lacO3 787 bp	This study
pHT1654	Pgrac100-bgaB, ΔlacI, lacO1-lacO3 732 bp	This study
pHT1655	Pgrac01-bgaB, ΔlacI, lacO1-lacO3 143 bp	This study
pHT1656	Pgrac100-bgaB, ΔlacI, lacO1-lacO3 88 bp	This study
pHT1657	Basic plasmid, used to construct pHT1655	This study
pHT1658	Basic plasmid, used to construct pHT1656, pHT1681, pHT1682	This stuy
pHT1660	Pgrac01-bgaB, ΔlacI, lacO1-lacO3 548 bp	This study
pHT1661	Basic plasmid, used to construct pHT1660	This study
pHT1674	Pgrac100-bgaB, ΔlacI, lacO1-lacO3 493 bp	This study
pHT1695	Pgrac100-gfp+, ΔlacI, lacO1-lacO3 493 bp	This study
pHT1696	Pgrac100-gfp+, ΔlacI, lacO1-lacO3 732 bp	This study
pHT2071	Pgrac01-bgaB, ΔlacI, lacO1-lacO3 280 bp	This study
pHT2079	Pgrac100-bgaB, ΔlacI, lacO1-lacO3 255 bp	This study

Oligonucleotide	Sequence 5′→3′	Used for
ON224	CCGGATGACGTCGAATTCTAAACCTTCCCGGCTTCATCATG	pHT1655, pHT1660, pHT1656
ON925	GAATTAGCTTGGTACCAAAGGAGGTAAGGATCACTAG	To amplify promoter Pgrac100 to construct pHT1674
ON926B	GACGTCGACTCTAGACATGGATCCTTCCTCCTTTAATTGG	To amplify promoter Pgrac100 to construct pHT1674
ON941	AAAGGAGGAAGGATCCATGAATGTGTTATC	pHT1655, pHT1660, pHT1656
ON985	CAATTGCGTTGCGCTCACTGCCGGTAC	pHT1657, pHT1658
ON986	CGGCAGTGAGCGCAACGCAATTGAGCT	pHT1657
ON1975	CAATTGCGTTGCGCTCACTGCCAGCGCT	To create lacO3 in pHT2071
ON1976	AGCGCTGGCAGTGAGCGCAACGCAATTGAGCT

Pgrac01 (another name is Pgrac) and Pgrac100 are the name of two different promoters