Coding

Part:BBa_K4011011:Design

Designed by: Meilun Wu   Group: iGEM21_LINKS_China   (2021-10-21)


Ma-CBM3-NT2RepCT-CBM3


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 576
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 948
    Illegal AgeI site found at 2359
    Illegal AgeI site found at 2403
    Illegal AgeI site found at 2575
    Illegal AgeI site found at 2641
    Illegal AgeI site found at 3739
    Illegal AgeI site found at 3783
    Illegal AgeI site found at 3955
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 424


Design Notes

1. The following mutations are added in Mα_Mut: A9D, A20T, Q32H, F48S, and G62R.

2. All codons were optimized for S. cerevisiae based on S. cerevisiae codon bias.

3. Linker EAEAFGS used between Ma and first CBM3, flexible linker GSGGGS used between first CBM3 and NT2RepCT, flexible linker GGGGS used between NT2RepCT and second CBM3.


Source

Ma comes from S. cerevisiae, NT2RepCT is an artificial spidroin sequence, CBM3 comes from Ruminiclostridium thermocellum.

References