Other
Part:BBa_K2082004
Designed by: Mikail Sahin Group: iGEM16_Bielefeld-CeBiTec (2016-10-14)
Monobody: Fusion protein of Monobody constant regions with RFP (exhange) and rpoZ
Pand-RBS-rpoZ CDS-cMyc linker-Monobody constant regions-RFP-Terminator
With this fundamental Monobody framework, consisting just the constant regions of a Monobody, we provide a construct to build Monobody libraries or specific antibody mimetics.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30
Illegal NheI site found at 481 - 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 166
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1067
Illegal AgeI site found at 1179 - 1000COMPATIBLE WITH RFC[1000]
This device consists of:
For constructing the Monobody rpoZ fusion library we decided to set the design up in a way that one can exchange RFP for the fragment containing the variable (randomized) regions.
Figure 1: Monobody rpoZ fusion with RFP. Construct of the Monobody rpoZ fusion with RFP insert exchangable for variable regions.
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
By using the RFP CDS the red color of colonies is a great visual control when cloning a library (figure 2). Colonies that are red represent starting plasmids and not parts of the library.
Figure 2: Colonies of dH5alpha with plasmid BBa_K2082000 shows slightly red color.
This device is a component of our part collection, which also includes our designed Monobody library (BBa_K2082005) and also a single example monobody ( BBa_K2082009) as a positive control for your designed Monobodies for colony PCRs, sequencings and more.
Visit our wiki for more information about our library and the opportunity of building your own library and further about our part collection. With this collection we provide every iGEM team the unprecedented chance for easily building their own Monobodies!
If you are interested in other binding proteins, check out our Nanobody parts for setting up a library.
If you are planning to build your own Monobody library or construct a specific binder, take a look at our hints
By using the RFP CDS the red color of colonies is a great visual control when cloning a library (figure 2). Colonies that are red represent starting plasmids and not parts of the library.
Figure 2: Colonies of dH5alpha with plasmid BBa_K2082000 shows slightly red color.
This device is a component of our part collection, which also includes our designed Monobody library (BBa_K2082005) and also a single example monobody ( BBa_K2082009) as a positive control for your designed Monobodies for colony PCRs, sequencings and more.
Visit our wiki for more information about our library and the opportunity of building your own library and further about our part collection. With this collection we provide every iGEM team the unprecedented chance for easily building their own Monobodies!
If you are interested in other binding proteins, check out our Nanobody parts for setting up a library.
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