DNA

Part:BBa_K4794010:Design

Designed by: wong yik teng   Group: iGEM23_HK-Joint-School   (2023-10-07)


T7-defensin-like peptide 4(DLP4)-intein-PT linker-ELK16


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 62
    Illegal XhoI site found at 310
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 778
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

(1) Intein is added to ensure continuous expression of AMP. (2) Sal1 and Kpn1 are added on both ends of intein-DLP4 to allow the assembly of intein-DLP4 to other construct or expression system. (3) SacI is located between intein and DLP4. It can be used with KpnI for the change of another AMP easily. (4) It is codon optimised to lower the complexity of the sequence.


Source

https://parts.igem.org/Part:BBa_I719005, https://parts.igem.org/Part:BBa_K3815000 and https://www.ncbi.nlm.nih.gov/nuccore/LC741404.1?from=1&to=246

References

Xia J, Ge C, Yao H. Antimicrobial Peptides from Black Soldier Fly (Hermetia illucens) as Potential Antimicrobial Factors Representing an Alternative to Antibiotics in Livestock Farming. Animals (Basel). 2021;11(7):1937. Published 2021 Jun 29. doi:10.3390/ani11071937

Wu C-L, Chih Y-H, Hsieh H-Y, Peng K-L, Lee Y-Z, Yip B-S, Sue S-C, Cheng J-W. High Level Expression and Purification of Cecropin-like Antimicrobial Peptides in Escherichia coli. Biomedicines. 2022; 10(6):1351. https://doi.org/10.3390/biomedicines10061351

Wang, M., Zheng, K., Lin, J., Huang, M., Ma, Y., Li, S., Luo, X., and Wang, J. (2018). Rapid and efficient production of cecropin A antibacterial peptide in Escherichia coli by fusion with a self-aggregating protein. BMC Biotechnol. 18, 62