Composite

Part:BBa_K5403015:Design

Designed by: Lizette van der Ziel   Group: iGEM24_TU-Eindhoven   (2024-10-02)


Fusion protein for membrane anchoring of GFP based on OmpAtb and OmpA


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 1696
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 30
    Illegal AgeI site found at 169
    Illegal AgeI site found at 628
    Illegal AgeI site found at 782
    Illegal AgeI site found at 807
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 16
    Illegal BsaI.rc site found at 744
    Illegal SapI.rc site found at 1043


Design Notes

A flexible linker with the sequence GSGSGSGSAS was used to fuse the membrane anchoring domain and GFP. The flexibility prevents the individual domains from misfolding as a result of being fused to one another.

The sequence was codon-optimized for expression in M. smegmatis.

Several modelling tools were used to aid in the design of this fusion protein (DeepTMHMM, Alphafold).


Source

E. coli and M. smegmatis

References