Reporter

Part:BBa_K2429034:Design

Designed by: Nia Myrie   Group: iGEM17_MIT   (2017-10-24)
Revision as of 23:38, 24 October 2017 by Nmyrie (Talk | contribs) (→‎Design Notes)

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phEF1a 2 Exon mKate-HBG Reporter


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 2764
    Illegal PstI site found at 315
    Illegal PstI site found at 820
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 2764
    Illegal PstI site found at 315
    Illegal PstI site found at 820
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 2764
    Illegal BglII site found at 569
    Illegal BamHI site found at 1183
    Illegal XhoI site found at 968
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 2764
    Illegal PstI site found at 315
    Illegal PstI site found at 820
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 2764
    Illegal PstI site found at 315
    Illegal PstI site found at 820
    Illegal NgoMIV site found at 703
    Illegal AgeI site found at 81
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2669
    Illegal SapI.rc site found at 1237


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