Table 1 Plasmids used in this study
Name | Description | Source |
|---|---|---|
pBSMul1 | E. coli-B. subtilis shuttle vector, PHpaII, secretion signal (SPlipA), ColE1, repB, Kmr, Ampr | [31] |
pET22b-sfGFP1-10 | pET22b(+) containing the truncated sfGFP1-10 under control of PT7 | [18] |
pBSMul1lipA_SPBox | Signal peptide (SP) library based on pBSMul3 signal peptide library from [13] containing HindIII–XbaI inserts of the lipase lipA from B. subtilis with all 173 predicted B. subtilis Sec signal peptides | [18] |
pJOE8999.1 | E. coli-B. subtilis shuttle vector, pUCori, rep pE194ts, cas9, KmR, tracrRNA, lacPOZ | [34] |
pJOE-sgRNA:htrA | pJOE8999.1 with inserted sgRNA for htrA locus (5’-TACATCCGTGAGGTCGCTTC-3’) between BsaI restriction sites | This study |
pJOE-sgRNA:htrA_H1H2 | pJOE-sgRNA:htra with 3300 bp SfiI-SfiI fragment harboring the regions upstream and downstream of htrA required for homologous recombination and htrA (5’-proG-htrA-ykcC-3’). | This study |
pJOE-sgRNA:htrA-htrA∷PhtrA-mCherry | pJOE-sgRNA:htrA-H1H2 with mCherry inserted between proG-PhtrA and ykcC using SLIC. | This study |
pJT’Tmcs-mCherry | AmpR, GmR, Ptac, RBS, mCherry | [39] |
pBS-4nt-SPPel-Cut11 | pBSMul1 containing an 827 bp NdeI–XbaI SPpel-cut-11 fragment | [29] |
pBS-4nt-SPEpr-Cut11 | pBSMul1 containing an 845 bp NdeI–XbaI SPepr-cut-11 fragment | [29] |
pBS-4nt-SPBsn-Cut11 | pBSMul1 containing an 848 bp NdeI–XbaI SPbsn-cut-11 fragment | [29] |
pBS-Xnt-SPPel-Cut11 | pBS-4nt-SPPel-Cut11 with a spacer extended by the insertion of 1–8 adenine(s) at the 5’-end from 4 to 12 nt | [29] |
pBS-Xnt-SPEpr-Cut11 | pBS-4nt-SPEpr-Cut11 with a spacer extended by the insertion of 1–8 adenine(s) at the 5’-end from 4 to 12 nt | [29] |
pBS-Xnt-SPBsn-Cut11 | pBS-4nt-SPBsn-Cut11 with a spacer extended by the insertion of 1–8 adenine(s) at the 5’-end from 4 to 12 nt | [29] |
pBSMul1cut11_SPBox | Signal peptide (SP) library based on pBSMul3 signal peptide library from [18] containing HindIII–XbaI inserts of the cutinase cut from F. solani pisi with all 173 predicted B. subtilis Sec signal peptides and the GFP11 tag sequence at the 3’-end of cut | This study |
pBSMul1_SPBox_SPDacB-Cut11 | pBSMul1cut11_SPBox variant with signal sequence from DacB and 657 bp EcoRI-XbaI cut-gfp11 fragment | This study |
pBSMul1_SPBox_SPMpr-Cut11 | pBSMul1cut11_SPBox variant with signal sequence from Mpr and 657 bp EcoRI-XbaI cut-gfp11 fragment | This study |
pBSMul1_SPBox_SPNamZ-Cut11 | pBSMul1cut11_SPBox variant with signal sequence from NamZ and 657 bp EcoRI-XbaI cut-gfp11 fragment | This study |
pBSMul1_SPBox_SPNamZ*-Cut11 | pBSMul1cut11_SPBox variant with signal sequence from NamZ and 657 bp EcoRI-XbaI cut-gfp11 fragment. Amino acid exchange A252S in GFP11-tag. | This study |