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Table 1 Bacterial strains and plasmids used in this study

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