Coding

Part:BBa_K4212003:Design

Designed by: Shirin Bamezai   Group: iGEM22_Imperial_College_London   (2022-09-29)


CotG_AHx3_ChiS


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 897
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 897
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 897
    Illegal BamHI site found at 882
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 897
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 897
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

AHx3 - rigid linker


Source

The CotG CDS comes from the genome of B. subtilis, and was produced through genome amplification with appropriately designed primers. The reverse primer included the sequence for the synthetic rigid linker. The chitinase protein sequence comes from the B. pumilus, however this was codon-optimised for B. subtilis and synthesized as a gBlock (IDT). The individual CDSs were also either ordered or amplified with Type IIS restriction sites on either end, to enable scarless fusion as a single CDS in a level 0 plasmid within a Golden Gate system.

References