Composite

Part:BBa_K3889122

Designed by: Prameya Garge, Ashwin Sharma   Group: iGEM21_IISER-Tirupati_India   (2021-10-14)


Bacillus subtilis mf-Lon cassette

This gene cassette has been designed to produce an anti-toxin mf-Lon continuously. The modified mf-lon gene has been placed downstream to a constitutive Synthetic promoter (SP146) of Bacillus subtilis and an RBS of Bacillus subtilis (BBa_K1351030). The gene producing mf-Lon is upstream to the porcine teschovirus-1 P2A Peptide Linker (BBa_K3889069) and the reporter gene sfGFP (BBa_K3889002). This reporter will help in determining the working of the cassette in Bacillus subtilis. Finally, the cassette is completed by adding a double terminator of Bacillus subtilis (BBa_B0015) downstream to the reporter gene.

This cassette has been developed to be a part of a light-inducible kill switch in Bacillus subtilis. The constitutive production of the anti-toxin mf-Lon will aid in controlling any leaky expression of the bpDNase1 toxin by forming a complex with it. This will help in avoiding the accidental death of bacteria due to leaky expression of the toxin.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 2503


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