Composite

Part:BBa_K2924054:Design

Designed by: Melanie Sbielut   Group: iGEM19_Duesseldorf   (2019-10-19)
Revision as of 18:58, 21 October 2019 by Mesbi100 (Talk | contribs) (→‎References)

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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.