Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Daniela Geist
Date created: 2014-09-30 11:00:00
Date modified: 2015-05-08 01:09:54
StrepDARPidin
| Types | DnaRegion |
| Roles | engineered_region Device |
| Sequences | BBa_K1329000_sequence (Version 1) |
Description
StrepDARPidin is a fusion protein combining the monomeric streptavidin from Streptomyces avidinii and the DARPin (Designed Ankrin Repeat Protein) EC1 for targeting a tumor surface marker.Streptavidin monomers build up tetrameric constructs and have high and specific affinity to biotin or Strep-Tags.
The DARPin EC1 was designed for an specific binding of EpCAM which is a surface marker of tumor cells derived from epithelial tissue.
Our idea was to create a fusion protein which can be used for targeting appropiate cells with increased affinity by creating induced proximity using tetrameric structures. Additionally these can be used for linking biotinylated or Strep-tagged constructs at the same time.
Notes
We designed the construct with an N-terminal His-Tag for purification and Antibody targeting. GS-linkers were used to give the construct flexibility to provide steric tensions.Source
The sequence for streptavidin was found in the Streptomyces avidinii genome (http://blast.st-va.ncbi.nlm.nih.gov/Blast.cgi#alnHdr_134951). The sequence was synthesized and optimized for E. Coli codon usage.The aminoacid sequence of DARPin Ec1 was found in the supplementary materials of the publication "DARPins Recognizing the Tuor-Associated Antigen EpCAM Selected by Phage and Ribosome Display and Engineered for Multivalency". The sequence was synthesized and optimized for E. Coli codon usage.