Composite

Part:BBa_K808030:Design

Designed by: Marie Burghard, Henrik Cordes, Jascha Diemer, Sven Jager, Philipp Rottmann, Rene Sahm, Arne Wehling   Group: iGEM12_TU_Darmstadt   (2012-09-02)

RBS-PhoA-His6tag-pNBEst13-Myctag-EstA


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1174
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1700
    Illegal NgoMIV site found at 1838
    Illegal NgoMIV site found at 2378
    Illegal NgoMIV site found at 2747
    Illegal AgeI site found at 1245
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

We got it synthesized by GENEART but added the RBS via PCR.



Source

This is no natural or even native construct. We got it synthesized by GENEART. but added the RBS via PCR.


References

  • Fakult, M., & Enzymtechnologie, M. (2006). biochemische Charakterisierung Dialkylphthalat spaltender Esterasen.
  • Becker, S., Theile, S., Heppeler, N., Michalczyk, A., Wentzel, A., Wilhelm, S., Jaeger, K.-E., et al. (2005). A generic system for the Escherichia coli cell-surface display of lipolytic enzymes. FEBS letters, 579(5), 1177–82. doi:10.1016/j.febslet.2004.12.087
  • Yang, T. H., Pan, J. G., Seo, Y. S., & Rhee, J. S. (2004). Use of Pseudomonas putida EstA as an Anchoring Motif for Display of a Periplasmic Enzyme on the Surface of Escherichia coli, 70(12), 6968–6976. doi:10.1128/AEM.70.12.6968