Coding

Part:BBa_K5109014:Design

Designed by: Lisa Faccincani   Group: iGEM24_Uni-Padua-IT   (2024-09-26)


LacE


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1579
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 234
    Illegal NgoMIV site found at 455
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1


Design Notes

The enzymatic sequence has two different enzyme restriction sites at the ends: a BsaI restriction site upstream the coding sequence, and a BamHI restriction site downstream the sequence, after the His - Tag. Those sites have been designed in order to extract the Laccase sequence from the surface display system in which it is inserted (BBa_K5109021), in order to exchange it with other enzymes, without modifying the rest of the expression cassette. This part was designed for a multiple cloning experiment which involved it's exchange with three different enzymes, structured in the same way, encoded as part BBa_K5109013, BBa_K5109015, and BBa_K5109016.


Source

The coding sequence of the ECOL laccase was identified inside E. coli sp genome, and on the registry is originally reported as part BBa_K863006

References