Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Mirjam Groenewold
Date created: 2013-10-03 11:00:00
Date modified: 2015-05-08 01:09:05
EstA Strep-tag SpiderSilkSubunitE1
| Types | DnaRegion |
| Roles | CDS Coding |
| Sequences | BBa_K1085035_sequence (Version 1) |
Description
This biobrick contains signal sequence EstA attached to the silk subunit E1 with an N-terminal strep-tag.The Strep-tag, fused at the N-terminal, is used both as a binding tag to use to indicate its production and as a binding component to Streptavidin.
Subunit E1 codes for a spider silk protein that is optimized for maximal expression in Bacillus subtilis 168. The primary goal of the algorithm was to optimize the codon selection based on their availability scores. The secondary goal was to prevent the formation of secondary RNA structures in close proximity to the RBS, and the tertiary goal was to minimize the number of restriction sites.
The amino acid sequence is based on the adapted MaSp2 sequences described in the paper of Brooks et. al. 2008.
Notes
The DNA sequence of subunit E1 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) and are from the MaSp2 of the Argiope aurantia.The DNA sequence was synthesized by Integrated DNA Technologies (IDT).
| Sequence Annotation | Location | Component / Role(s) |
| RBS starting codon (ATG) estA-SRP strep spider silk subunit E1 | 1,6 15,17 18,107 114,137 156,365 | ribosome_entry_site feature/rbs feature/start start_codon CDS feature/cds feature/tag tag CDS feature/cds |