Coding

Part:BBa_K4623003:Design

Designed by: Ruixiao Tan   Group: iGEM23_BNU-China   (2023-10-09)


TrwC, a protein linker to covalently link ssDNA 5' end


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 286
    Illegal PstI site found at 604
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 286
    Illegal PstI site found at 604
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 286
    Illegal PstI site found at 604
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 286
    Illegal PstI site found at 604
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 409
    Illegal SapI.rc site found at 124


Design Notes

The sequences were codon-optimized for E.coli and a stop codon was added at the C-terminus.

Source

Amino acid series were obtained from ref and transformed into corresponding DNA sequences, and codon optimization was performed against E.coli.

References

[1]Lucas M, González-Pérez B, Cabezas M, Moncalian G, Rivas G, de la Cruz F. Relaxase DNA binding and cleavage are two distinguishable steps in conjugative DNA processing that involve different sequence elements of the nic site. J Biol Chem. 2010 Mar 19;285(12):8918-26. doi: 10.1074/jbc.M109.057539. Epub 2010 Jan 8. PMID: 20061574; PMCID: PMC2838313.

[2]Sagredo, Sandra et al. “Orthogonal Protein Assembly on DNA Nanostructures Using Relaxases.” *Angewandte Chemie (International ed. in English)* vol. 55,13 (2016): 4348-52. doi:10.1002/anie.201510313.