Composite

Part:BBa_K2686006:Design

Designed by: Thomas Jordan   Group: iGEM18_EPFL   (2018-10-08)


Encapsulin protein with HexaHistidine insert, OT1 peptide at C terminus and a sfGFP-tag protein.


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 902
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 77
    Illegal BglII site found at 492
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 902
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 902
  • 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 955


Design Notes

Two amino acids were removed at the C terminus, a HexaHistidine insert was added for better heat resistance, a OT1 peptide was fused to the C terminus of the protein (Choi et al., 2016). The sequence is followed by a T7 RBS and a sfGFP coding protein with a 15 amino acid tag binding Encapsulin's interior surface (Cassidy-Armstutz et al., 2016, Addgene plasmid #86405).

Source

Derived from BBa_K2686005.

References

Cassidy-Amstutz, C., Oltrogge, L., Going, C., Lee, A., Teng, P., Quintanilla, D., East-Seletsky, A., Williams, E. and Savage, D. (2016). Identification of a Minimal Peptide Tag for in Vivo and in Vitro Loading of Encapsulin. Biochemistry, 55(24), pp.3461-3468.

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.