Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Morgan Litschko, Malak Elbatarny, Joanna Semrau, Justine Ring
Date created: 2015-09-10 11:00:00
Date modified: 2015-09-26 08:08:01
T3-10 Self-Assembling Scaffold with K-coil & His tag
| Types | DnaRegion |
| Roles | CDS Coding |
| Sequences | BBa_K1831002_sequence (Version 1) |
Description
A single subunit of the T3-10 self-assembling protein scaffold (Baker 2008) is fused in-frame to the K=coil from Team Calgary 2013???s parallel coiled-coil proteins (BBa_K1189010). The ???K??? denotes the charged lysine residues in the ???e??? and ???g??? positions of the helical wheel representation of the coils. A His-tag fused to the C-terminus of the coils allows quick purification. The flexible linker Gly-Ser between the K-coil and the His-tag prevents potential electrostatic interference between the two domains.This fusion of the K-coil to each individual subunit means that once assembled into a multimer, multiple proteins with a complementary E-coil domain (such as our Type III AFPs: BBa_K1831001, BBa_K1831003) can be non-covalently attached to the scaffold.
Notes
The K-coil was created as an in-frame fusion protein, added to the C-terminus of the T3-10 subunit. By adding a flexible linker of Gly-Ser, between the scaffold and the coil, we incorporated a unique BamHI restriction site into the DNA sequence. This acts as a diagnostic marker for simplified cloning of our BioBrick into the expression vector of your choosing.Source
This part contains the sequence for the T3-10 subunit (PDB file 4EGG) which was computationally designed by Neil P. King of the Todd O. Yeates & David Baker lab (www.sciencemag.org/cgi/content/full/336/6085/1171/DC1)| Sequence Annotation | Location | Component / Role(s) |
| T7 promoter + RBS NdeI site T3-10 subunit K-coil BamHI Site 734 stop x2 XhoI site | 1,61 62,67 68,664 665,727 728,733 734,748 749,754 755,760 | promoter feature/promoter sequence_feature feature/misc feature/protein CDS feature/misc sequence_feature feature/misc sequence_feature feature/tag tag feature/stop stop_codon sequence_feature feature/misc |