Coding

Part:BBa_K3793005:Design

Designed by: Rupesh Balaji Jayachandran, Enrique de Dios Mateos   Group: iGEM21_Aalto-Helsinki   (2021-10-21)


CDS - Human AhR Nuclear Translocator (Truncated)


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 104
    Illegal EcoRI site found at 1210
    Illegal SpeI site found at 1054
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 104
    Illegal EcoRI site found at 1210
    Illegal SpeI site found at 1054
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 104
    Illegal EcoRI site found at 1210
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 104
    Illegal EcoRI site found at 1210
    Illegal SpeI site found at 1054
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 104
    Illegal EcoRI site found at 1210
    Illegal SpeI site found at 1054
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The residues comprising of the different domains were to be identified for creating a truncated version. The restriction sites of type IIS enzymes like BsaI and BbsI were to removed from the CDS. An N-Terminal 6x HIS tag has been added to facilitate downstream purification.


Source

The sequence is modified from the NCBI entry of AhR (NM_001668.4). The residues encoding for the basic helix-loop-helix, PAS-A and PAS-B domains were obtained from Schulte et al., 2017.


References

Schulte KW, Green E, Wilz A, Platten M, Daumke O. Structural Basis for Aryl Hydrocarbon Receptor-Mediated Gene Activation. Structure. 2017 Jul 5;25(7):1025-1033.e3. doi: 10.1016/j.str.2017.05.008. Epub 2017 Jun 9. PMID: 28602820.