BBa_K1085012

BBa_K1085012 Version 1

Component

Source:
http://parts.igem.org/Part:BBa_K1085012
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Claudio Tiecher
Date created: 2013-09-09 11:00:00
Date modified: 2016-02-10 01:09:46

RBS Start-codon EstA Strep-tag SpiderSilkSubunitN1



Types
DnaRegion

Roles
Coding

CDS

Sequences BBa_K1085012_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), a coding sequence for the signal protein EstA and a Strep-tag infront of it.
The RBS is there to allow the ribosomes to bind and start to translate the DNA.

The EstA codes for a signal protein that that is there to fasilitate secretion of the protein.

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 MaSp2. The DNA sequence was synthesized by Integrated DNA Technologies (IDT).

Sequence Annotation Location Component / Role(s)
RBS
starting codon (ATG)
estA (Bac. sub.)
strep
spidersilk subunit N1
1,6
14,16
17,649
656,679
698,838
feature/rbs ribosome_entry_site
feature/start start_codon
feature/cds CDS
feature/tag tag
feature/cds CDS
igem#experience
Issues
 
igem#sampleStatus
In stock
igem#status
Available
 
synbiohub#ownedBy
user/james
 
synbiohub#ownedBy
user/myers
 
synbiohub#topLevel
BBa_K1085012/1