Part:BBa_K4794010:Design
T7-defensin-like peptide 4(DLP4)-intein-PT linker-ELK16
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 62
Illegal XhoI site found at 310 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 778
- 1000COMPATIBLE 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