Coding

Part:BBa_K4429001:Design

Designed by: Anish Mulchandani   Group: iGEM22_IISER-Pune2_India   (2022-06-10)


Anti-MBP Recombinant IgY Heavy Chain


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 870
    Illegal SapI.rc site found at 1450


Design Notes

Assembled from Variable Heavy regions against Maltose Binding Protein, and Gallus gallus IgY constant region taken from IMGT [2].

Consideration was taken to ensure the correct V-C region interface between a mouse IgG variable region (aMBP) and Chicken IgY constant region.

Codon optimisation performed for E. coli K12, removing biobrick illegal restriction sites. Adding polyhistidene tag.

Source

anti-MBP variable region sequence comes from the coding sequence of anti-MBP IgG expressed in SHuffle E. coli in this Robinson et al. 2015 [1].

References

[1] 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

[2] https://www.imgt.org/ligmdb/view?id=X07174