Part:BBa_K5427003:Design
MaSp1
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
The MaSp1 sequence was optimized for E. coli expression by performing codon optimization and manually removing repetitive sequences to maintain genetic stability. We reviewed and adjusted the sequence to reduce complexity, repeating the process until complexity was minimized and no major issues remained. The GC content was manually adjusted (to 49.9%) to avoid high values, ensuring codons were spaced to prevent hairpin formations. Additionally, illegal restriction sites were removed to adhere to Biobrick standards. A His-tag was added to the N-terminus of the sequence based on the source information, facilitating easy protein purification. It is noted that in the future a plasmid containing tRNA for alanine and glycine would be necessary in order to optimize spider silk production, but we did not use that in our design.
Source
Arndt, T., Greco, G. H., Schmuck, B., Bunz, J., Shilkova, O., Francis, J., Pugno, N., Kristaps Jaudzems, Barth, A., Johansson, J., & Rising, A. (2022). Engineered Spider Silk Proteins for Biomimetic Spinning of Fibers with Toughness Equal to Dragline Silks. Advanced Functional Materials, 32(23), 2200986–2200986. https://doi.org/10.1002/adfm.202200986