Table 3 Strains and plasmids employed in this study

E. coli DH5α

F-,φ80dlacZ ΔM15,Δ(lacZYA -argF)U169, deoR, recA1, endA1,hsdR17 (rK-, mK +), phoA, supE44, λ-, thi-1, gyrA96, relA1

TransGen, Beijing

E. coli BW25113-T7

F-, DE(araD-araB)567, lacZ4787(del)::rrnB-3, LAM-, rph-1, DE(rhaD-rhaB)568, hsdR514, int:: (lacI:: PlacUV5:: T7 gene) ΔybhC

Laboratory stock

E. coli A2

E. coli BW25113-T7 ΔaceB, ΔdppA, ΔhemF, ΔgalR int ppc with J23119, ΔpoxB int glk with J23119)

This study

E. coli 108

E. coli A2 ΔPhemB, Int::J23108

This study

E. coli 111

E. coli A2 ΔPhemB, Int::J23111

This study

E. coli 116

E. coli A2 ΔPhemB, Int::J23116

This study

E. coli BWT7-RSA

E. coli BW25113-T7 harboring pET28b-LAA and pACYCD-hemA-RS

This study

E. coli A2-RSA

E. coli A2 harboring pET28b-LAA and pACYCD-hemA-RS

This study

E. coli BWT7-LAA

E. coli BW25113-T7 harboring pET28b-LAA

This study

E. coli BWT7-LAA(C)

E. coli BW25113-T7 harboring pET28b-LA-eamA(C)

This study

E. coli BWT8-LAA(C1)

E. coli BW25113-T7 harboring pET28b-LA-eamA(C1)

This study

E. coli A2-RSA-C1

E. coli A2 harboring pET28b-LA-eamA(C1) and pACYCD-hemA-RS

This study

E. coli A2-ASK

E. coli A2 harboring pET28b-LA-eamA(C1) and pACYCD-ASK

This study

E. coli 108-ASK

E. coli 108 harboring pET28b-LA-eamA(C1) and pACYCD-ASK

This study

E. coli 111-ASK

E. coli 111 harboring pET28b-LA-eamA(C1) and pACYCD-ASK

This study

E. coli 116-ASK

E. coli 116 harboring pET28b-LA-eamA(C1) and pACYCD-ASK

This study

Plasmids

Relevant characteristic(s)

Source or reference

pUC57

Cloning vector, AmpR, ColE1/pMB1/pBR322/pUC ori

Laboratory stock

pACYCD

Cloning vector, CmR, p15a ori

Laboratory stock

pET-28b

Cloning vector, f1 ori; KanR

Laboratory stock

pUC57-sYFP

pUC57 with sYFP gene and J23107 promoter

Laboratory stock

pUC-sYFP-pHemL

pUC57 with sYFP gene and PhemL (promoter of hemL gene)

This study

pET28b-LAA

f1 ori; KanR; hemA; hemL; eamA; LacI gene and T7-LacI promoter

Laboratory stock

pSC-LA-eamA

temperature sensitive oriR101; KanR; hemA; hemL; eamA; LacI gene and T7-LacI promoter

This study

pET28b-LA-eamA(C)

f1 ori; KanR; hemA; hemL; eamA(C); LacI gene and T7-LacI promoter

This study

pET28b-LA-eamA(C1)

f1 ori; KanR; hemA; hemL; eamA(C1); LacI gene and T7-LacI promoter

This study

pCas

plasmid for CRISPR (temperature sensitive oriR101; KanR; the λ-Red operon under the control of arabinose-inducible promoter; S. pyogenes-derived cas9; sgRNA guided to ori-p15a under the control of lac operator)

Laboratory stock

pTarget-gene

plasmid for CRISPR (p15a ori; CmR; sgRNA guided to targeted gene, such as hemF, aceB, poxB gene and so on)

This study

pACYCD-gene

pACYCD containing targeted gene (such as hemF, aceB, poxB gene and so on)

This study

pACYCD-Donor DNA

pACYCD containing Donor DNA for targeted gene (such as hemF, aceB, poxB gene and so on)

This study

pACYCD-RS-hemA

pACYCD containing hemA from R. sphaeroides (modified) and T7-LacI promoter

This study

pACYCD-ASK

pACYCD containing hemA from R. sphaeroides (modified) and T7-LacI promoter; sodB, katE and J23107 promoter

This study