Table 1.
Strains and plasmids used in this study.
| Strain or Plasmid | Relevant properties | Sources |
|---|---|---|
| Strain | ||
| E. coli DH10B | F- endA1 deoR+ recA1 galE15 galK16 nupG rpsL Δ(lac)X74 φ80lacZΔM15 araD139 Δ(ara,leu)7697 mcrA Δ(mrr-hsdRMS-mcrBC) StrR λ- | Invitrogen |
| E. coli MG1655 | F- lambda- ilvG- rfb-50 rph-1 | Lab stock |
| MG1655ΔlacZ::bla | MG1655 with lacZ inserted with bla gene, Ampr | This study |
| MG1655-lacZ(TAA) | MG1655 with the initial codon of lacZ mutated to ‘TAA,’ plasmids cured | This study |
| MG1655-lacZ(His) | MG1655 with the His-tag DNA sequence inserted after the initial codon of lacZ, plasmids cured | This study |
| MG1655-lacZ(amilGFP) | MG1655 with the amilGFP CDS fused to the 5′-end of lacZ, plasmids cured | This study |
| MG1655Δfrr::bla | MG1655 with the bla gene and RBS inserted to the front of frr gene, Ampr | This study |
| MG1655-frr(His) | MG1655 with the His-tag DNA sequence inserted after the initial codon of frr, plasmids cured | This study |
| Plasmid | ||
| pUC18 | pUC, bla | Lab stock |
| pSY1071 | pSY1069 carrying amilGFP (BBa_K592010) | Li et al., 2016 |
| pTarget | pMB1, aadA, sgRNA-fts | Jiang et al., 2015 |
| pCas | repA101(Ts), kanr, Pcas-cas9, ParaB-Red, lacIq, Ptrc-sgRNA-pMB1; temperature-sensitive replication vector | Jiang et al., 2015 |
| pCasM | A derivative plasmid from pCas, harboring a mutant repA101 (RepAA56V) | This study |
| psgRNA_bla | A derivative plasmid from pTarget, expressing an sgRNA targeting bla gene | This study |