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Part:BBa_K2082006

Designed by: Marius Schöller   Group: iGEM16_Bielefeld-CeBiTec   (2016-10-14)


Nanobody library: Fusion proteins of Nanobody and omega subunit of RNA polymerase (rpoZ)

Library Pand-RBS-rpoZ CDS-cMyc linker-Nanobody constant and variable regions-Terminator


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 166
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

This is a part of an example Nanobody library:

  • an Anderson promoter (BBa_J23106)
  • a codon optimized RBS (BBa_B0034)
  • a RNA polymerase omega subunit
  • a cMyc-linker
  • the first Nanobody constant region
  • the Nanobody variable region
  • the second Nanobody constant region
  • a modified thr terminator (BBa_B1006)
    • .

    Libraries are a powerful tool in synthetic biology, but underestimated in iGEM. Unfortunately, the construction of a library is a difficult and time consuming task. To supply this tool for following iGEM teams, we provide this designed Nanobody library to the iGEM community, as implementation of a new category in iGEM parts registry. As a result, this plasmid mix includes not just a few parts, but rather over 100,000 distinct sequences of binding proteins, out of a theoretically size of over one billion (1,073,741,824) possible sequences which is the result of a sophisticated library design.

    On top, we constructed and submitted a fundamental framework to construct further Nanobody libraries or specific Nanobodies. This device (BBa_K2082001) consists of the constant regions only and can be equipped with the missing variable region or a constant Nanobody segment for a specific binder. Read more on how to build your own Nanobody library. As a positive control for your designed Nanobodies for example for colony PCRs or sequencings we supply one single binder of our library with device BBa_K2082010.

    Figure 1: Nanobody randomized region of 24 colonies. Top to bottom: Ordered sequence, chromatogram and sequencing result.


    By submitting this part we want to establish libraries in iGEM and paved the way for every team to access the advantages of large diversity libraries as well as creating and submitting their own. This would push the iGEM parts registry extremely forward. It would be possible for all teams, to submit different plasmid libraries, encoding several binding proteins, enzymes, aptamers, or other useful proteins. So all following iGEM teams can build on it and screen the diversity for their desired protein with special properties.
    Visit our wiki for more information about our library and the opportunity of building your own library.

    If you are interested in other binding proteins, check out our Monobody parts for setting up a library.

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