Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Dominique Dabija, Christopher Jackson, Debha Amatya
Date created: 2011-12-06 12:00:00
Date modified: 2015-05-08 01:14:04
Spider Silk Actuator
| Types | DnaRegion |
| Roles | Composite engineered_region |
| Sequences | BBa_M36394_sequence (Version 1) |
Description
This is a Spider Silk Generating Device that uses Fos and Jun leucine zipper sequences flanking a spider silk protein hexamer. The leucine zippers are linked to the spider silk hexamer by 8 AA linkers. RBS+Leader+ATG are part of the start of the gene, and the terminator is at the end.Notes
Our overall design had a base pair limit of 3000. This pointed to usage of Fos and Jun leucine zippers to attempt to get the spider silk hexamers to link up head to tail via covalent bonds in the E. coli cells. We were not able to spin the spider silk protein out, isolate it, or analyze its structural properties.Source
RBS Leader and ATG came from BioFAB. Terminator comes from BioFAB. Spider silk monomer sequence was repeated six times, and the monomer AA sequence comes from Xia, X.-X., Z.-G. Qian, C. S. Ki, Y. H. Park, D. L. Kaplan, and S. Y. Lee. "Native-sized Recombinant Spider Silk Protein Produced in Metabolically Engineered Escherichia Coli Results in a Strong Fiber." Proceedings of the National Academy of Sciences 107.32 (2010): 14059-4063. Print. The sequence was codon optimized for E. coli. Fos-Jun leucine zipper sequences were taken from . O'Shea, E., R. Rutkowski, W. Stafford, and P. Kim. "Preferential Heterodimer Formation by Isolated Leucine Zippers from Fos and Jun." Science 245.4918 (1989): 646-48. Print. They were also codon optimized for E.coli.Our system was placed into a Rhamnex 67K Xbrane plasmid and tested using varying concentrations of rhamnose.
| Access | Instance | Definition |
| public public public public public public public | BBa_M36293 BBa_M36501 BBa_K133132 BBa_M36500 BBa_K133132 BBa_M36502 BBa_M36292 | BCRBS Fos linker spilk 7.0. linker Jun BBa_M36292 |
| Sequence Annotation | Location | Component / Role(s) |
| BBa_M36293 BBa_M36501 BBa_K133132 BBa_M36500 BBa_K133132 BBa_M36502 BBa_M36292 | 1,88 97,216 225,248 257,886 895,918 927,1046 1055,1136 | BCRBS Fos linker spilk 7.0. linker Jun BBa_M36292 |