Name | Relevant genotype | Source/reference |
---|---|---|
Strains | ||
 B. subtilis 168 | Wild-type strain, trpC2 | BGSCa |
 B. subtilis 168ΔligD | B. subtilis 168ΔligD | This study |
 BS89 | B. subtilis168Δupp ribC*ribO* yhcF*yvrH*ywaA* P43::ribA | [55] |
 BS89ΔligD | BS89ΔligD | This study |
 CY series strains | Modulation library strains generated with CRISPR/Cas9n technique | This study |
 E. coli. DH5α | F−, φ80lacZΔM15, Δ(lacZYA-argF)U169, deoR, recA1, endA1, hsdR17(rk−, mk+), phoA, supE44, λ−, thi-1, gyrA96, relA1 | Lab stock |
Plasmids | ||
 pCas9cur | E. coli cloning vector for the express Cas9 protein, AmpR | Lab stock |
 pBSCas9 | B. subtilis/E. coli shuttle vector, CmR PxylA-Cas9, P43-gRNA | This study |
 pBSCas9n | pBSCas9 harboring Cas9 mutation | This study |
 pDonor | B. subtilis/E. coli shuttle vector, CmR AmpR, Pmanp-rep60gRNA | This study |
 pHG | Derived from pUC18 for multiple gRNA construction | This study |
 pUC18 | E. coli cloning vector, AmpR | Lab stock |
 pEBs-cop1 | B. subtilis/E. coli shuttle vector, EmR, AmpR | Lab stock |
 pAX01 | B. subtilis integration vector, PxylA, EmR, AmpR | Lab stock |
 pHP13 | B. subtilis/E.coli shuttle vector,CmR, EmR | Lab stock |
 pBSCas9-amyEgRNA | Derived from pBSCas9 for targeting amyE | This study |
 pBSCas9n-amyEgRNA | Derived from pBSCas9n for targeting amyE | This study |
 pBSCas9-Ins1 kbamyEgRNA | Derived from pBSCas9 for targeting Δ1kbamyE | This study |
 pBSCas9n-Ins1kbamyEgRNA | Derived from pBSCas9n for targeting Δ1kbamyE | This study |
 pBSCas9-Ins2kbamyEgRNA | Derived from pBSCas9 for targeting Δ2kbamyE | This study |
 pBSCas9n-Ins2kbamyEgRNA | Derived from pBSCas9n for targeting Δ2kbamyE | This study |
 pBSCas9-amyE/uppgRNA | Derived from pBSCas9 for targeting amyE and upp | This study |
 pBSCas9n-amyE/uppgRNA | Derived from pBSCas9n for targeting amyE and upp | This study |
 pBSCas9-amyE/upp/sigEgRNA | Derived from pBSCas9 for targeting amyE,upp and sigE | This study |
 pBSCas9n-amyE/upp/sigEgRNA | Derived from pBSCas9n for targeting amyE,upp and sigE | This study |
 pBSCas9-LGgRNA | Derived from pBSCas9 for targeting large DNA fragment | This study |
 pBSCas9n-LGgRNA | Derived from pBSCas9n for targeting large DNA fragment | This study |
 pDonor-Del1kb | Derived from pDonor for 1 kb deletion | This study |
 pDonor-Del2kb | Derived from pDonor for 2 kb deletion | This study |
 pDonor-Del4kb | Derived from pDonor for 4 kb deletion | This study |
 pDonor-Del6kb | Derived from pDonor for 6 kb deletion | This study |
 pDonor-Del8kb | Derived from pDonor for 8 kb deletion | This study |
 pDonor-Ine1kb | Derived from pDonor for 1 kb insertion | This study |
 pDonor-Ine2kb | Derived from pDonor for 2 kb insertion | This study |
 pDonor-DelLDNA | Derived from pDonor for large DNA fragment deletion | This study |
 pDonor-MuamyE | Derived from pDonor for codon replacement in amyE | This study |
 pDonor-MuamyE/upp | Derived from pDonor for codon replacement in amyE and upp | This study |
 pDonor-MuamyE/upp/sigE | Derived from pDonor for codon replacement in amyE,upp and sigE | This study |
 pBSCas9n-gRNArib | Derived from pBSCas9n for targeting ribB, ribA and ribH | This study |
 pDonor-ribRBSLib | Derived from pDonor for generating a combinatorial library | This study |