Coding

Part:BBa_K4701303:Design

Designed by: Henri Sundquist   Group: iGEM23_Aalto-Helsinki   (2023-07-18)


SpyTag-MHETase


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
    COMPATIBLE WITH RFC[1000]


Design Notes

The sequence for MHETase is presented here. The first 75 bases were removed as in this part we didn't require the signal sequence. Furthermore ColabFold results indicated, that the N-terminus of MHETase contains some unstructured sequence, which was removed. Figure 1 shows ColabFold results before and after the trimming of the N-terminus. Lastly, a His-tag was added for protein purification purposes.

SpyCatcher/SpyTag fusion of FAST-PETase and MHETase, before and after trimmed MHETase N-terminus
Figure 1: ColabFold prediction of the SpyCatcher/SpyTag fusion of FAST-PETase and MHETase. Left: MHETase with unstructured N-terminus. Right: unstructured MHETase N-terminus removed. (cyan: MHETase, magenta: linkers, yellow: SpyCatcher002, red: SpyTag, orange: FAST-PETase, gray: His-tags)