Composite

Part:BBa_K5199004:Design

Designed by: Hannah Grover   Group: iGEM24_USAFA   (2024-09-27)


INP Silicatein


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 321
    Illegal XhoI site found at 1246
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 63
    Illegal NgoMIV site found at 396
    Illegal AgeI site found at 498
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 765
    Illegal SapI.rc site found at 229


Design Notes

To make this part, a pET-28a (+) TF-silicatein-α plasmid gifted from Bryan Berger's lab was cut with BamHI and HindIII to remove the TF part. INP (Ice Nucleation Protein) was cloned out of pET-28a INP OspA (USAFA 2023 iGEM Team) with INP specific primers. BamHI and HindIII sites were added to INP via PCR.T4 ligase was used to ligate the INP fragment into the BamHI and HindIII sites of the pET-28a-silicatein-α plasmid.


Source

Previous iGEM plasmids were used to clone out the INP and the silicatein was added behind the INP sequences.

References