Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Claudio Tiecher
Date created: 2013-09-09 11:00:00
Date modified: 2015-06-02 12:19:48
RBS Start-codon SpiderSilkSubunitN1 Strep-tag
| Types | DnaRegion |
| Roles | CDS Coding |
| Sequences | BBa_K1085004_sequence (Version 1) |
Description
This BioBrick contains the coding sequence for part of the spider silk protein (SubunitN1) with a Bacillus Subtilis ribosome binding site (RBS) and the Strep-tag infront of it.The RBS is there to allow the ribosomes to bind and start to translate the DNA.
The Strep-tag, fused at the N terminal, is used both as a binding tag to use to inidicate its production and as a binding component to biotin.
SubunitN1 codes for a spider silk protein that is optimized for maximal expression in Bacillus subtillus 168. The primary goal of the algorithm was to optimize the codon selection based on their availbility scores. The secondary goal was to prevent the formation of secondary RNA structures in close proximity to the RBS, and the tetrary goal was to minimize the number of restriction sites.
Notes
The DNA sequence was codon-optimized for Bacillus Subtilis. Moreover the nucleotide sequence was optimized in order to accomplish the production standard of IDT. A couple of optimized codons were affected by this.Source
The amino acids sequence of the silk protein was obtained from the paper by Brooks et al. (2008). The DNA sequence was synthesized by Integrated DNA Technologies (IDT).| Sequence Annotation | Location | Component / Role(s) |
| RBS starting codon (ATG) strep spidersilk subunit N1 | 1,6 14,16 17,40 59,199 | ribosome_entry_site feature/rbs start_codon feature/start tag feature/tag CDS feature/cds |