Table 1 Strains and plasimds employed in this study
Strain or plasmid | Relevant characteristic(s) | Source and/or reference |
|---|---|---|
Strain | ||
 Escherichia coli DH5α | F-,φ80dlacZ ΔM15,Δ(lacZYA -argF )U169, deoR , recA1 , endA1 ,hsdR17 (rK-, mK+), phoA, supE44 , λ-, thi-1, gyrA96 , relA1 | TransGen, Beijing |
 Escherichia coli BL21(DE3) | fhuA2 [lon] ompT gal (λ DE3) [dcm] ΔhsdS λ DE3 = λ sBamHIo ΔEcoRI-B int::(lacI::PlacUV5::T7 gene) i21 Δnin5 | TransGen, Beijing |
 Escherichia coli BW25113 | F-, DE(araD-araB)567, lacZ4787(del)::rrnB-3, LAM-, rph-1, DE(rhaD-rhaB)568, hsdR514 | Laboratory stock |
 Escherichia coli BW25113-T7 | BW25113 int::(lacI::PlacUV5::T7 gene) ΔybhC | This study |
Plasmids | ||
 pACYCD-Blank | E. coli cloning vector (p15a ori; CmR) | Laboratory stock |
 pACYCD-sYFP | pACYCD-Blank with sYFP gene, LacI gene and T7-LacI promoter | Laboratory stock |
 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 | plasmid for CRISPR (p15a ori; CmR; sgRNA) | Laboratory stock |
 pTarget-ybhC | plasmid for CRISPR (p15a ori; CmR; sgRNA guided to ybhC gene) | This study |
 pACYCD-ybhC | pACYCD-Blank with ybhC gene (Fragment A) | This study |
 pACYCD-Donor DNA | pACYCD-Blank with Donor DNA | This study |
 pACYCD-sYFP-17A (and so on) | pACYCD-sYFP with T7 Promoter Mutation Library (Single-base substitution mutation) | This study |
 pET28b-ALA-LA | plasmid for biosyntheizing ALA (f1 ori; KanR; hemA; hemL; LacI gene and T7-LacI promoter) | Laboratory stock |
 pET28b-ALA-LAR | plasmid for biosyntheizing ALA (f1 ori; KanR; hemA; hemL; RhtA; LacI gene and T7-LacI promoter) | Laboratory stock |