From: Construction of an easy-to-use CRISPR-Cas9 system by patching a newly designed EXIT circuit
Strain or plasmid | Relevant characteristic(s) | References |
---|---|---|
Strains | ||
  Escherichia coli NEB10β | Δ(ara-leu) 7697 araD139 fhuA ΔlacX74 galK16 galE15 e14- ϕ80dlacZΔM15 recA1 relA1 endA1 nupG rpsL (StrR) rph spoT1 Δ(mrr-hsdRMS-mcrBC) | NEB |
  E. coli MG1655 | K-12 F− λ− ilvG − rfb-50 rph-1 | Lab stock |
Plasmids | ||
 pACYC184 | p15A, CmR | [42] |
 pCP202 | Broad host vector pBBR1-MCS2 derivative, TET, PCN-GFPmut3b-T1T2 | Lab stock |
 pKD46 | Exo, bet, gam, rep and arabinose operon | [55] |
 pEC100 | p15A, CmR, TET-araC-PBAD-I-SceI-T1T2 | This study |
 pEC001 | p15A, CmR, araC-PBAD-I-SceI-GFPmut3b fusion | This study |
 pEC101 | p15A, CmR, the control module composed of TET-araC-PBAD-I-SceI-T1T2, bordered by two I-SceI cleavage sites | This study |
 pP103 | p15A, CmR, PJ23119-GFPmut3b | Lab stock |
 pEC201 | p15A, CmR, the control module, PJ23119-GFPmut3b | This study |
 pET-19(b) | pBR322, AmpR | Lab stock |
 pEC102 | pBR322, AmpR, the control module | This study |
 pIC202 | colE1, CmR | Lab stock |
 pEC103 | colE1, CmR, the control module | This study |
 pUC-EM | pMB1, EmR | Lab stock |
 pEC104 | pMB1, EmR, the control module | This study |
 pbeloBac11 | repE, CmR, sopA, sopB, sopC, cos | Lab stock |
 pEC105 | pBeloBAC11 derivative, CmR, the control module | This study |
 pEM106 | pBR322, the exit module of AmpR | This study |
 pCAS9 | Cas9 | [8] |
 pICK1 | p15A, KanR, lacI-Plac-GFP | Lab stock |
 pICK2 | p15A, lacI-Plac-GFP, the exit module of KanR | This study |
 pCAS92 | p15A, lacI-Plac-Cas9, the exit module of KanR | This study |
 pGRNA2 | colE, CmR, sgRNA scaffold, the control module | This study |
 pGRNA2-1 | pGRNA2 derivative, sgRNA with the N20 sequence targeting region between ompW and yciE | This study |
 pCM1001 | incP broad host vector, PTEF1-lacZ-T1T2 | This study |
 PCT100 | Broad host vector pBBR1-MCS2 derivative, PCN23-atzA-T1T2 | This study |