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Parts Bacilus Subtilis

For the expression in B.subtilis, we designed eight different basic parts, six composite parts, and two devices. The plasmid vector that we used was BBa_J428326 a medium copy number plasmid backbone with a resistance gene for Kanamycin.

Table 1: Table of Parameters
Name Type Description Length
BBa_K4998020 Basic PgroES Promoter 49bp
BBa_K4998019 Basic LipA signal peptide >93bp
BBa_K4998018 Basic Laccase gene 1008bp
BBa_K4998026 Basic PgroE promoterr 48bp
BBa_K4998027 Basic Gene that expresses LuxI 582bp
BBa_K4998028 Basic PtrC-2-derived promoter 130bp
BBa_K4998029 Basic Gene that expresses bsrG toxin 291bp
BBa_K4998030 Basic Lambda T0 terminator 60bp
BBa_K4998039 Composite PgroE promoter+ R0 RBS(BBa_K3831018)+ LuxI Gene+ B. subtilis terminator (BBa_K780000) 730bp
BBa_K4998040> Composite Pgrac Promoter (BBa_K1074012)+ R0 RBS (BBa_K3831018)+ LuxI Gene+ B. subtilis terminator (BBa_K780000) 852bp
BBa_K4998041 Composite PtrC-2-derived promoter (BBa_K4998028)+ RBS sequence (BBa_B0034)+ TetR gene (BBa_C0040)+ T1 promoter from E.coli rrnB (BBa_B0010) 929bp
BBa_K4998042 Composite Pgrac promoter (BBa_K1074012)+ RBS sequence (BBa_B0034)+ TetR gene (BBa_C0040)+ T1 promoter from E.coli rrnB (BBa_B0010) 969bp
BBa_K4998043 Composite PLtetO-1 Promoter (BBa_K3332034)+ BsrG toxin gene (BBa_K4998029) + T0 Terminator (BBa_K4998030) 466bp
BBa_K4998036 Composite PgrES Promoter+ LipA Sequence for tat pathway+ lac7 Gene+ Terminator 1224bp
BBa_K4998044 Device This device is a kill-switch cirquit for B. subtilis WB800N strain, that is regulated by the presence of IPTG in the medium. It consists of a lacI composite part (BBa_K1088019), a composite tetR part (BBa_K4998041), and the bsrG toxin transcriptional unit (BBa_K4998043). 5568bp
BBa_K4998045 Device This device is a kill-switch circuit for B. subtilis WB800N strain, regulated by IPTG in the medium. It consists of a lacI composite part (BBa_K1088019), the composite tetR part (BBa_K4998041), and the bsrG toxin transcriptional unit (BBa_K4998043). The device is inserted in the BBa_J428344 plasmid backbone 5568bp

Parts Cyanobacterium

We designed ten basic and four different composite parts that were expressed in our cyanobacterium strain. The plasmid vector that was used for the expression of these parts is BBa_J428326 a medium copy number plasmid backbone with a resistance gene for Kanamycin.




Table 2: Table of Parameters
Name Type Description Length
BBa_K4998021 Basic trc promoter 72bp
BBa_K4998022 Basic Gene that expresses the LuxR protein 756bp
BBa_K4998023 Basic ECK120034435 Terminator 57bp
BBa_K4998031 Basic psbA1 promoter 236bp
BBa_K4998032 Basic Gene that expresses the Hetr protein 900bp
BBa_K4998033 Basic trp terminator 49bp
BBa_K4998034 Basic Promoter for VapB15 gene 93bp
BBa_K4998024 Basic RBS 12bp
BBa_K4998035 Basic Gene that expresses the antitoxin VapB15 243bp
BBa_K4998025 Basic Gene that expresses the toxin VapC15 399bp
BBa_K4998046 Composite Promoter PpsbA1+ RBS+ HetR Gene-+ trp Terminator 1213bp
BBa_K4998037 Composite trc promoter+ RBS + LuxR Gene+ ECK120034435 Terminator 913bp
BBa_K4998038 Composite Promoter PpbsA1 + RBS+ VapC15 Gene+ T2 Terminator 697bp
BBa_K4998047 Composite QS induced Promoter+ RBS+ VapB15 Gene+ T1 Terminator 444bp
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Parts E.coli

We designed one basic part that was expressed in TOP10 E.coli cells.

Table 3: Table of Parameters
Name Type Description Length
BBa_K4998048 Basic Antimicrobial peptide 129

Parts Primers

For our experiments, we also designed the primers for the PCR amplification of our parts.




Table 4: Table of Parameters
Name Type Description Length
BBa_K4998000 Primer LuxI gene left primer 20bp
BBa_K4998001 Primer LuxI gene right primer 20bp
BBa_K4998002 Primer Laccase gene left primer 20bp
BBa_K4998003 Primer Laccase gene right primer 20bp
BBa_K4998004 Primer LuxR gene left primer 20bp
BBa_K4998005 Primer LuxR gene right primer 20bp
BBa_K4998006 Primer HetR gene left primer 20bp
BBa_K4998007 Primer HetR gene right primer 20bp
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