Reporter

Part:BBa_K2429017:Design

Designed by: Tinna-Solveig F Kosoko-Thoroddsen   Group: iGEM17_MIT   (2017-10-22)
Revision as of 18:41, 22 October 2017 by Tinnasolla (Talk | contribs) (→‎Design Notes)


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 654 from a C to an A because the paper we were referencing had mutated the HBG intron in the same way.

Source

The source of this part

References