Composite

Part:BBa_K2257000:Design

Designed by: Ruoyun Wang   Group: iGEM17_Nanjing-China   (2017-10-22)


hydrogen sensor sequence, HoxA-HoxB-HoxC-HoxJ-terminator-HoxP-EGFP


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 5912
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 6989
    Illegal BamHI site found at 1
    Illegal XhoI site found at 409
    Illegal XhoI site found at 2562
    Illegal XhoI site found at 2574
    Illegal XhoI site found at 3102
    Illegal XhoI site found at 3129
    Illegal XhoI site found at 3876
    Illegal XhoI site found at 4152
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 816
    Illegal NgoMIV site found at 1767
    Illegal NgoMIV site found at 1959
    Illegal NgoMIV site found at 2074
    Illegal NgoMIV site found at 2343
    Illegal NgoMIV site found at 2751
    Illegal NgoMIV site found at 3501
    Illegal AgeI site found at 891
    Illegal AgeI site found at 1476
    Illegal AgeI site found at 3519
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 150
    Illegal BsaI.rc site found at 3199
    Illegal BsaI.rc site found at 3516
    Illegal BsaI.rc site found at 4347
    Illegal SapI.rc site found at 2304
    Illegal SapI.rc site found at 4991


Design Notes

We synthesized the genes to remove any illegal restriction sites. Protein hoxB and hoxC form a Heterodimeric core they acts as an hydrogen sensor in concert with the histidine protein kinase HoxJ . This RH-HoxJ complex detects the presence of hydrogen in the environment and transmits the signal to a response regulator HoxA. The response regulator, HoxA, is active in its non-phosphorylated form and inactivated by HoxJ-mediated phosphorylation. In the hydrogen atmosphere, HoxA is unphosphorylated and interacts with a promotor Phox, and it is capable of activating downstream gene transcription. Now these genes are ligated one after another by isocaudomers. And we change its downstream gene to GFP, so when the hydrogen comes , our E. colis turn to green.


Source

The sequence of HoxABCJP comes from Ralstonia eutropha H16 megaplasmid pHG1 in the NCBI data base.

References