Composite

Part:BBa_K2474000:Design

Designed by: J. Baggman, J. Bergqvist, H. Karlsson, L. Karlsson, J. Larsson, M. Lindberg, M. Nilsson, M. Peterson, O. Reinhed Gustafsson, S. Stridh Karppinen & J. Ybrahim   Group: iGEM17_Linkoping_Sweden   (2017-10-27)


Amyloid-B mNeonGreen


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1205
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1267
    Illegal BamHI site found at 1144
    Illegal BamHI site found at 2190
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 979
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 961


Design Notes

We designed this biobrick with amyloid-beta bound C-terminally to the fluorescent protein mNeonGreen. For this plasmid we used the arabinose induced promotor BBa_I0500 which includes the regulatory protein araC which binds to the promoter-region pBAD in the absence of arabinose. To connect the fusion proteins we created a GS-linker between Amyloid-Beta and mNeonGreen to make sure it was motile. The fusion protein carries a histidine tag (N-terminally) to enable IMAC purification.

Source

mNeonGreen and Amyloid-Beta was synthetically made by the company IDT. The vector psb1C3 came from iGEM and pBAD is a BioBrick (BBa_10500).

References