Reporter

Part:BBa_K2429017:Design

Designed by: Tinna-Solveig F Kosoko-Thoroddsen   Group: iGEM17_MIT   (2017-10-22)


2 Exon mKate-HBG Reporter


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1582
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1582
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1582
    Illegal BamHI site found at 1
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1582
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1582
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1487
    Illegal SapI.rc site found at 55


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.

This basic part entry vector is flanked by L1 and L2 sites, which are used to denote a gene.

Source

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

References