Coding

Part:BBa_K1336002:Design

Designed by: Yan-Kay Ho   Group: iGEM14_UCL   (2014-10-06)


Spore coat protein, laccase, from Bacillus subtilis


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 867
    Illegal XbaI site found at 1482
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 867
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 867
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 867
    Illegal XbaI site found at 1482
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 867
    Illegal XbaI site found at 1482
    Illegal AgeI site found at 1348
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1509
    Illegal SapI.rc site found at 286


Design Notes

When designing PCR primers to change the CotA gene into the BioBrick format, we also had to design primers for site-directed mutagenesis, as there are 2 illegal restriction sites within the native gene: EcoRI at position 867, and XbaI at position 1482).

Source

From genomic sequence of Bacillus subtilis; CotA gene inserted into a pET-21a (+) expression vector, provided to us by XXX Research Group.

References