a rpoZ CDS (encoding RNA polymerase omega subunit)
a cMyc linker
the first constant region of a Monobody CDS
different randomized Monobody variable regions
the second constant Region of a Monobody CDS
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 Monobody 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 antibody mimetics, 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 Monobody libraries or specific antibody mimetics. This part (BBa_K2082000) consists of the Monobody constant regions including RFP instead of variable regions, which can easily be replaced with randomized variable regions or a constant Monobody segment for a specific binder. Additionally, we provide a protocol for our collection for using our library and even constructing your own library with our blindfold (BBa_K2082000).
As a positive control for your designed Monobodies for example for colony PCRs or sequencings we supply one single binder of our library with device BBa_K2082009.
To get further insight into the theoretical assembly take a look here.
If you are planning to build your own Monobody library or construct a specific binder, take a look at our hints
Figure 1: Monobody first randomized region of 24 colonies. Top to bottom: Ordered sequence, chromatogram and sequencing result.
Figure 2: Monobody second 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 Nanobody parts for setting up a library.
