Coding

Part:BBa_K2429050:Design

Designed by: Nia Myrie   Group: iGEM17_MIT   (2017-10-24)


pTRE 2 Exon mKate-HBG Reporter


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 6
    Illegal EcoRI site found at 322
    Illegal EcoRI site found at 1920
    Illegal XbaI site found at 30
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 6
    Illegal EcoRI site found at 322
    Illegal EcoRI site found at 1920
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 6
    Illegal EcoRI site found at 322
    Illegal EcoRI site found at 1920
    Illegal BamHI site found at 339
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 6
    Illegal EcoRI site found at 322
    Illegal EcoRI site found at 1920
    Illegal XbaI site found at 30
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 6
    Illegal EcoRI site found at 322
    Illegal EcoRI site found at 1920
    Illegal XbaI site found at 30
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1825
    Illegal SapI.rc site found at 393


Design Notes

This part is human-codon optimized. We found a BsaI site in the middle of mKate Exon 2, so we use site-directed mutagenesis to change the base pair at position 1450 from a G to an A. We also mutated the HBG Intron 2 at position 888 from a C to an A because the paper we were referencing had mutated the HBG intron in the same way.



Source

The HBG intron came from HEK genome, and mKate from jellyfish.


References