Composite

Part:BBa_K2686005:Design

Designed by: Thomas Jordan   Group: iGEM18_EPFL   (2018-10-07)


Encapsulin protein with HexaHistidine insert & sfGFP under native promoter included between BsaI cut


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 77
    Illegal BglII site found at 492
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 426
    Illegal SapI.rc site found at 457
    Illegal SapI.rc site found at 1031


Design Notes

Two amino acids were removed at the C terminus, in that way the future antigen sequence is expressed in the same fashion as specified in the literature (Choi et al., 2016), and the BsaI cut sites are on an insert coding for sfGFP. A HexaHistidine insert is present for better heat resistance and hydrodynamic properties (Moon et al., 2014).

Source

Derived from BBa_K2686002 and a sfGFP coding construct from our host laboratory.

References

Choi, B., Moon, H., Hong, S., Shin, C., Do, Y., Ryu, S. and Kang, S. (2016). Effective Delivery of Antigen–Encapsulin Nanoparticle Fusions to Dendritic Cells Leads to Antigen-Specific Cytotoxic T Cell Activation and Tumor Rejection. ACS Nano, 10(8), pp.7339-7350.

Moon, H., Lee, J., Min, J. and Kang, S. (2014). Developing Genetically Engineered Encapsulin Protein Cage Nanoparticles as a Targeted Delivery Nanoplatform. Biomacromolecules, 15(10), pp.3794-3801.