Part:BBa_K4998036:Design

Composite SilA gene

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 480
  Illegal XhoI site found at 353
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 284
  Illegal NgoMIV site found at 946
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 230
  

Design Notes

For the constant expression of our gene, we had to use a constitutive promoter. We wanted our laccase to be active extracellularly so we needed to add a signal peptide either from the tat or sec pathway. We chose the tat pathway because we preferred our protein to exit the cell fully folded.

Source

The PgroES promoter and the LipA signal peptide originate from B.subtilis. The silA gene originates from Streptomyces ipomonae.

References

[1] Song, Y., Nikoloff, J. M., Fu, G., Chen, J., Li, Q., Xie, N., Zheng, P., Sun, J., & Zhang, D. (2016). Promoter screening from bacillus subtilis in various conditions hunting for synthetic biology and Industrial Applications. PLOS ONE, 11(7). https://doi.org/10.1371/journal.pone.0158447

[2]Zhang, W., Yang, M., Yang, Y., Zhan, J., Zhou, Y., & Zhao, X. (2016). Optimal secretion of alkali-tolerant xylanase in bacillus subtilis by signal peptide screening. Applied Microbiology and Biotechnology, 100(20), 8745–8756. https://doi.org/10.1007/s00253-016-7615-4

[3] Janusz, G., Pawlik, A., Świderska-Burek, U., Polak, J., Sulej, J., Jarosz-Wilkołazka, A., & Paszczyński, A. (2020). Laccase properties, physiological functions, and evolution. International Journal of Molecular Sciences, 21(3), 966. https://doi.org/10.3390/ijms21030966