Composite

Part:BBa_K1716002:Design

Designed by: Pernille Neve Myers   Group: iGEM15_DTU-Denmark   (2015-09-16)


NahR Biosensor for detection of acetylsalicylic acid/aspirin with blue chromoprotein reporter


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 786
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 77
    Illegal NgoMIV site found at 618
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The construct was originally assembled into pDG268neo using Gibson assembly. pDG268neo contains amyE sites for integration into Bacillus subtilis. The expression cassette with promoter, RBS, CDS, and terminator was amplified by PCR. The PCR product was inserted into pSB1C3 using Gibson assembly. The plasmid was transformed into E. coli and subsequently purified and sequenced. The plasmid was linearised before transformation into B. subtilis.


Source

Bacillus subtilis phage SSP1 gene product 35 Homoloogus regions upstream and downstream from must (genomic DNA template) Promoter: Terminator: Neomycin resistance marker from Bacillus subtilis plasmid pDG268neo

References