Coding

Part:BBa_K4204000:Design

Designed by: Jianfei Song   Group: iGEM22_BNDS_China   (2022-09-20)


Coding sequence of endolysin LysPBC5


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 690
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 742
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 988


Design Notes

It codes for a two-domain protein.The EAD is on the N-terminal(5-198 amino acids, 13-594bp) and the CBD is on the C-terminal (200-399 amino acids, 597-1017bp). Since it codes for a protein that must be extracted to use, the sequence does not contain a start codon and must be cloned to vectors used for protein purification (such as pET-28 in our project) before being used.


Source

Bacteriophage PBC5 genome. Codon was optimized for E.coli

References

[1] Lee, K. O., Kong, M., Kim, I., Bai, J., Cha, S., Kim, B., Ryu, K.-S., Ryu, S., & Suh, J.-Y. (2019). Structural basis for cell-wall recognition by bacteriophage PBC5 endolysin. Structure, 27(9). https://doi.org/10.1016/j.str.2019.07.001