Part:BBa_K3843000:Design
PEA-binding salmon trypsin (1UTM)
- 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 GenBank sequence coded for a propeptide that also contained a signal sequence. We first codon optimized the sequence for Escherichia coli K12 and simultaneously removed illegal restriction sites for RFC[25]. Next, we identified the signal peptide using the bioinformatics server SignalP 5.0, then we removed the signal peptide. We also removed the last 142 bp from the sequence, as these were annotated to not be present in the mature peptide by the authors of the GenBank submission (Male et al., 2005). The resulting sequence gives rise to functional salmon trypsin, appropriate for use in a fusion protein.
Source
The sequence of this part was obtained from GenBank: https://www.ncbi.nlm.nih.gov/nuccore/X70071.
References
Leiros, H. K. S., Brandsdal, B. O., Andersen, O. A., Os, V., Leiros, I., Helland, R., Otlewski, J., Willassen, N. P., & Smalås, A. O. (2009, January 1). Trypsin specificity as elucidated by lie calculations, x‐ray structures, and association constant measurements. Protein Science, 13(4), 1056-1070. https://onlinelibrary.wiley.com/doi/full/10.1110/ps.03498604.
Male, R., Lorens, J. B., Smalås, A. O., & Torrissen, K. R. (2005, March 3). Molecular cloning and characterization of anionic and cationic variants of trypsin from Atlantic Salmon. European Journal of Biochemistry, 232(2), 677-685. https://febs.onlinelibrary.wiley.com/doi/full/10.1111/j.1432-1033.1995.677zz.x?sid=nlm%3Apubmed.