TABLE 1.
Strains and plasmids used in this study.
| Name | Characteristics | Source/reference |
| Strain | ||
| DH5α | fhuA2 lacΔU169 phoA glnV44 Φ80′ lacZΔM15 gyrA96 recA1 relA1 endA1 thi-1 hsdR17 | Laboratory stock |
| MG1655 | F– ilvG rfb-50 rph-1 | S. Adhya |
| HK1060 | MG1655, ΔaraBAD:PBAD-dcas9-KmR | This study |
| Plasmid | ||
| pKD46 | pSC101ori ts, araC, λ red genes, AmpR | Datsenko and Wanner (2000) |
| pdCas9 | P15A ori, dcas9 gene, CmR | Addgene #46569 |
| pTargetF | pBR322 ori, crRNA scaffold, SpR | Addgene #62226 |
| pHK459 | PAM(TGG), pBR322 ori, crRNA target 498AGGCTGTAACTGCGGGATCA517 in galK), SpR | This study |
| pBJ003 | PAM(CGG), pBR322 ori, crRNA target 573CGAACAGAAATCACCAATGC592 in galT), SpR | This study |
| pBJ004 | PAM(TGG), pBR322 ori, crRNA target 406TACGTTGAAAGCTTCCCGAC425 in galE), SpR | This study |
| pBJ005 | PAM(TGG), pBR322 ori, crRNA target –30TTCGCATCTTTGTTATGCTA–11 in Pgal), SpR | This study |
| pBJ021 | PAM(CAG), pBR322 ori, crRNA target 503GTAACTGCGGGATCATGGAT522 in galK), SpR | This study |
| pBJ022 | PAM(GCG), pBR322 ori, crRNA target 489CCAGTTTGTAGGCTGTAACT508 in galK), SpR | This study |
| pBJ023 | PAM(ATG), pBR322 ori, crRNA target 497TAGGCTGTAACTGCGGGATC516 in galK), SpR | This study |
| pBJ024 | PAM(GGA), pBR322 ori, crRNA target 499GGCTGTAACTGCGGGATCAT518 in galK), SpR | This study |
| pBJ025 | PAM(AGC), pBR322 ori, crRNA target 504TAACTGCGGGATCATGGATC523 in galK), SpR | This study |
| pBJ026 | PAM(TGT), pBR322 ori, crRNA target 482CAGAAAACCAGTTTGTAGGC501 in galK), SpR | This study |
| pBJ027 | PAM(AAG), pBR322 ori, crRNA target –1GAAATAACCATAGCATAACA–20 in Pgal), SpR | This study |
| pBJ028 | PAM(GCG), pBR322 ori, crRNA target –6AACCATAGCATAACAAAGAT–25 in Pgal), SpR | This study |
| pBJ029 | PAM(ATG), pBR322 ori, crRNA target –31TTTCGCATCTTTGTTATGCT–12 in Pgal), SpR | This study |
| pBJ030 | PAM(AGA), pBR322 ori, crRNA target –2AAATAACCATAGCATAACAA–21 in Pgal), SpR | This study |
| pBJ031 | PAM(TGC), pBR322 ori, crRNA target –5TAACCATAGCATAACAAAGA–24 in Pgal), SpR | This study |
| pBJ032 | PAM(GGT), pBR322 ori, crRNA target –29TCGCATCTTTGTTATGCTAT–10 in Pgal), SpR | This study |