Composite

Part:BBa_K3781217:Design

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


L1_sAP_3xFLAG_RBD_Strep8His, MocloMania Composite


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 709
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 709
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 647
    Illegal XhoI site found at 6
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 709
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 709
    Illegal NgoMIV site found at 795
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 819


Design Notes

This composite part was assembled from four 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, secreting and purifying recombinant SARS-CoV2-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.

Since facilitating the production of complexly glycosylated proteins is a core motivation in our project, generating and testing L1 constructs like this is very beneficial to achieving our research goal. Furthermore, assembly of this part allowed us to verify the correct construction of its sub-parts.


Source

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