Composite
1Gate-NpuD

Part:BBa_K1362100:Design

Designed by: Constantin Ahlmann-Eltze, Charlotte Bunne, Magdalena Buescher, Jan Gleixner, Max Horn, Anna Huhn, Nils Klughammer, Jakob Kreft, Elisabeth Schaefer, Carolin Schmelas, Silvan Schmitz, Max Waldhauer   Group: iGEM14_Heidelberg   (2014-10-05)

RBS + NpuDnaE N-intein RFC[105] assembly construct (with His6)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 826
    Illegal AgeI site found at 938
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1115
    Illegal BsaI.rc site found at 40


Design Notes

The part is designed in compliance with RFC[105] [1]. For an exact description of how to use it or how to design other intein constructs accordingly, please read the iGEM team Heidelberg 2014's wikipage or the RFC[???].

Source

The part is assembled from different sources. The intein sequences come addgene or were kindly provided by different working groups. BBa_J04450 was used as mRFP selection marker. The scars and overhangs are part of the RFC[105] intein cloning strategy and are further explained on our wiki and in RFC[105] [1]. Please see the corresponding basic parts for the individual detailed information.

References

[1] [link RFC[105]]

[2] Zettler, J., Schütz, V. & Mootz, H. D. The naturally split Npu DnaE intein exhibits an extraordinarily high rate in the protein trans-splicing reaction. FEBS Lett. 583, 909–14 (2009). http://www.ncbi.nlm.nih.gov/pubmed/19302791