Encapsulin protein with HexaHistidine insert & sfGFP under native promoter included between BsaI cut
Composite part designed to serve as a rapid platform for fusing antigen coding sequences to the C-terminus of the encapsulin protein.
Usage and Biology
This Encapsulin protein derived from K2686002 has a hexahistidine insert to increase heat stability, and endow better hydrodynamic properties (Moon et al., 2014). After the encapsulin site on the plasmid (After the C-terminus), is a site for sfGFP under native promoter included within BsaI cut sites. These BsaI cut sites would allow for the rapid, scarless introduction of antigen peptides which will be expressed on the surface of the Encapsulin subunit (After the C-terminus). The insert in between the two BsaI cut sites, consisting of sfGFP with a native promoter and terminator, allows for checking the success of the insertion of the antigen (green colonies do not contain the desired peptide insert).
In our case this part was used in order to build K2686000 using a OT1 peptide coding sequence (Choi et al., 2016).
Improvement upon BBa_K192000
Expression of BBa_K2686005 precursors compared to BBa_K192000
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.
Sequence and Features
- 10COMPATIBLE WITH RFC
- 12COMPATIBLE WITH RFC
- 21Illegal BglII site found at 77
Illegal BglII site found at 492
- 23COMPATIBLE WITH RFC
- 25COMPATIBLE WITH RFC
- 1000Illegal SapI.rc site found at 426
Illegal SapI.rc site found at 457
Illegal SapI.rc site found at 1031