Translational_Unit

Part:BBa_K4429015:Design

Designed by: Anish Mulchandani   Group: iGEM22_IISER-Pune2_India   (2022-10-08)


Full Length aMBP IgG-IgY Chimera


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 693
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 693
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 693
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Construct intended to be produced in a pET-21b vector. Codes for 2 cistrons, for heavy and light chains, each cistron having its own ribosome binding site and both under a single T7 promoter. [1]


Source

IgY heavy and light chain cistrons were assembled as per BBa_K4429000 and BBa_K4429001. They were substituted onto existing DNA fragment of a full length IgG translational unit as per Robinson et al. 2015 [2]. -

References

[1] Eaglesham, J. B., Garcia, A., & Berkmen, M. (2021). Production of antibodies in SHuffle Escherichia coli strains. Methods in enzymology, 659, 105–144. https://doi.org/10.1016/bs.mie.2021.06.040

[2] Robinson, MP., Ke, N., Lobstein, J. et al. Efficient expression of full-length antibodies in the cytoplasm of engineered bacteria. Nat Commun 6, 8072 (2015). https://doi.org/10.1038/ncomms9072