Coding

Part:BBa_K4944000:Design

Designed by: Kasib Mir   Group: iGEM23_Calgary   (2023-08-01)


mShh with a 6x His-tag (mShh-His)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 695
    Illegal NgoMIV site found at 942
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 665


Design Notes

Full coding sequence for mouse Sonic Hedgehog protein (mShh). Produces a full precursor hedgehog protein which will undergo autocatalytic cleavage in the presence of cholesterol, or other molecules that can substitute as a nucleophile (such as dithiothreitol [DTT]) [1]. Polyhistidine tag was added on the N-terminal for purification.

Source

Modified sequence from pBS mShh (CT#258), Cliff Tabin (Addgene plasmid # 13999 ; http://n2t.net/addgene:13999 ; RRID:Addgene_13999)

References

(1) Hall TMT, Porter JA, Young KE, Koonin EV, Beachy PA, Leahy DJ. Crystal Structure of a Hedgehog Autoprocessing Domain: Homology between Hedgehog and Self-Splicing Proteins. Cell 1997;91:85–97. https://doi.org/10.1016/S0092-8674(01)80011-8.