Supplementary material

Table S1 Strains and plasmids

Strains or plasmids	Description	Resource
Strains
E. coli DH5α	F- φ80dlacZΔM15 Δ(lacZYAargF)U169 deoR recA1 endA1 hsdR17(rk-mk+) phoA supE44 λ-, thi-1gyrA96relA1	Lab stock
E. coli BL21(DE3)	F- ompT gal dcm lon hsdSB(rB- mB-) λ(DE3 [lacI lacUV5-T7 gene 1 ind1 sam7 nin5])	Lab stock
E. coli BL21(DE3)/pCDFDuet-1-PalsI-sfGFP	E. coli BL21(DE3), pCDFDuet-1-PalsI-sfGFP	This study
E. coli BL21(DE3)/pCDFDuet-1-PalsI-sfGFP/pETDuet-1-nativeP-alsR	E. coli BL21(DE3), pCDFDuet-1-PalsI-sfGFP, pETDuet-1-nativeP-alsR	This study
E. coli BL21(DE3)/pCDFDuet-1-alsR-P-sfGFP	E. coli BL21(DE3), pCDFDuet-1-alsR-P-sfGFP	This study
E. coli BL21(DE3)/pCDFDuet-1-alsR*-P-sfGFP	E. coli BL21(DE3), pCDFDuet-1-alsR*-P-sfGFP	This study
E. coli BL21(DE3)/pET28a-rpiB	E. coli BL21(DE3), pET28a-rpiB	This study
E. coli BL21(DE3)/pET28a-rpiB/ pCDFDuet-1-alsR*-P-sfGFP	E. coli BL21(DE3), pET28a-rpiB, pCDFDuet-1-alsR*-P-sfGFP	This study
Plasmids
pET28a	pMB1, Kanr	Lab stock
pETDuet1	pMB1, Ampr	Lab stock
pCDFDuet1	CloDF13, Specr	Lab stock
pET28a-sfGFP	pET28a derivate, containing sfGFP gene	Lab stock
pCDFDuet-1-PalsI-sfGFP	pCDFDuet1 derivate, containing bifunctional promoter between alsR gene and rpiB gene in E. coli BL21(DE3) and sfGFP gene	This work
pETDuet-1-nativeP-alsR	pETDuet1 derivate, containing bifunctional promoter between alsR gene and rpiB gene in E. coli BL21(DE3) and alsR gene in E. coli BL21(DE3)	This work
pCDFDuet-1-alsR-P-sfGFP	pCDFDuet1 derivate, containing alsR gene in E. coli BL21(DE3), bifunctional promoter between alsR gene and rpiB gene in E. coli BL21(DE3) and sfGFP gene	This work
pCDFDuet-1-alsR*-P-sfGFP	pCDFDuet1 derivate, containing mutant alsR gene of M2 from directed evolution, bifunctional promoter between alsR gene and rpiB gene in E. coli BL21(DE3) and sfGFP gene	This work
pET28a-rpiB	pET28a derivate, containing rpiB gene from Clostridium thermocellum	This work

Table S2 Primers used in this study

Application	Primer name	sequence (5’-3’)
pCDFDuet-1-	pCDF-gj-For	ctgtacaaataactactagcgcagcttaattaac
PalsI-sfGFP	pCDF-gj-Rev	aaacccggtaacgcctctccccgcgcgttggc
	PalsI-For	gcggggagaggcgttaccgggtttgagtaacc
	PalsI-Rev	ctttacgcatatcacagccaaatgcaatct
	sfGFP-For	tttggctgtgatatgcgtaaaggtgaagaact
	sfGFP-Rev	gcgctagtagttatttgtacagttcgtccatac
:--	:--	:--
pETDuet-1-	pETDuet1-gj-For	gcgctgaaataactactagcgcagcttaattaac
nativeP-alsR	pETDuet1-gj-Rev	cggggaagatgacgcctctccccgcgcgttggc
	PalsR-For	cggggagaggcgtcatcttccccgtccgtgcaaaac
	alsR-Rev	tgcgctagtagttatttcagcgcgcctggtgag
:--	:--	:--
pCDFDuet-1-	pCDF-gj-For	ctgtacaaataactactagcgcagcttaattaac
alsR-P-sfGFP	pCDF-gj-Rev	caggcgcgctgaaataagcctctccccgcgcgttggc
	PalsR-For	ctttacgcatatcacagccaaatgcaatct
	alsR-Rev	gcgcggggagaggcttatttcagcgcgcctggtgag
	sfGFP-For	tttggctgtgatatgcgtaaaggtgaagaact
	sfGFP-Rev	gcgctagtagttatttgtacagttcgtccatac
:--	:--	:--
pCDFDuet-1-	pCDF-gj-For	ctgtacaaataactactagcgcagcttaattaac
alsR*-P-sfGFP	pCDF-gj-Rev	caggcgcgctgaaataagcctctccccgcgcgttggc
	PalsR-For	ctttacgcatatcacagccaaatgcaatct
	alsR-Rev	gcgcggggagaggcttatttcagcgcgcctggtgag
	sfGFP-For	tttggctgtgatatgcgtaaaggtgaagaact
	sfGFP-Rev	gcgctagtagttatttgtacagttcgtccatac
:--	:--	:--
pET28a-rpiB	pET28a-For	gaaatactccaagcaccaccaccaccaccactgag
	pET28a-Rev	accgatcttcatgcgacccatttgctgtccaccag
	rpiB-For	aaatgggtcgcatgaagatcggtatcggtagc
	rpiB-Rev	tggtggtggtgcttggagtatttcttctcgatttc
:--	:--	:--
Error-Prone PCR of alsR	alsR-EP-For	gcgatggcggaattacctttttctta
	alsR-EP-Rev	cttcaattcaaggacaagagaacgtgatg
	pCDF-gj-For	ctgactggctcatcacgttctcttgtccttgaattg
	pCDF-gj-Rev	ttccgccatcgcattcattaatgcagctggcac

1.pCDFDuet-1-Palsl-sfGFP Plasmid map

2.pETDuet-1-nativeP-alsR Plasmid map

3.pCDFDuet-1-alsR-P-sfGFP Plasmid map