Composite

Part:BBa_K3781205:Design

Designed by: Nicolas Bayer   Group: iGEM21_TU_Kaiserslautern   (2021-10-15)


L1_sAP_RBD_3xHA, MocloMania Composite


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 598
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 598
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 536
    Illegal XhoI site found at 6
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 598
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 598
    Illegal NgoMIV site found at 684
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 708


Design Notes

This composite part was assembled from three basic parts that are part of the MocloMania collection. Due to the highly standardizes set of overhangs that all our basic parts comply with, assembly could be achieved by a simple Moclo ligation. Assembling this part was of great importance to our project's prospect, since our objective was to test our expression system by expressing recombinant SARS-CoV-2 RBD. Having the fusion protein secreted into the cell's exterior, the culture medium, allows for the RBD to undergo glycosylation while passing the secretory pathway. Adding the 3xHA epitope tag allowed for easy detection of the fusion protein in cell culture preparations on western blot.


Source

For information on the origin of the genetic sequences present in this composite part, please consult the individual basic part pages.