Part:BBa_K2924054:Design

Hpall + SPYurl + alpha s2 + fd terminator

Assembly Compatibility:

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

  Illegal BamHI site found at 1127
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 557
  

Design Notes

Expression system for the expression and secretion of the protein in B. subtilis

Source

PBSMul1-SPYurl plasmid was kindly provided by Dr. Andreas Knapp from the research center Jülich. The DNA sequence was acquired by reverse-translating the protein sequence of a-s2-casein. a-s2-casein was commercial synthesized by GenScript.

References

Brockmeier, U., Caspers, M., Freudl, R., Jockwer, A., Noll, T., & Eggert, T. (2006). Systematic screening of all signal peptides from Bacillus subtilis: a powerful strategy in optimizing heterologous protein secretion in Gram-positive bacteria. Journal of molecular biology, 362(3), 393-402.

Knapp, A., Ripphahn, M., Volkenborn, K., Skoczinski, P., & Jaeger, K. E. (2017). Activity-independent screening of secreted proteins using split GFP. Journal of biotechnology, 258, 110-116.