Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Kevin Chen
Date created: 2012-09-30 11:00:00
Date modified: 2015-05-08 01:13:18
[R0010][B0034] - E. coli K12 FliC
| Types | DnaRegion |
| Roles | Composite engineered_region |
| Sequences | BBa_K781000_sequence (Version 1) |
Description
This is the complete flagellin coding sequence with biobrick standard cut sites removed, under the promoter R0010 and RBS B0034. This part can be used to over express the flagellin monomer. Or, this part can be used as a template for PCR overlap extension, which will make an insertion at the annotated site, given that the insert is in the appropriate format. The resulting chimera will express protein immediately after the PCR reaction is transformed.Notes
Note that when expressing this, and resulting overlaps in bacteria. One should be aware that this promoter is sensitive to LacI. Any strains overly expressing LacI will have very little expression of protein.Source
The first constant domains and the D3 domain were amplified out of the separately, incorporating the appropriate silent mutations. The second constant domains were synthesized. The total coding domain was assembled using overlap extension PCR.| Sequence Annotation | Location | Component / Role(s) |
| BBa_R0010 end of LacI coding region (inactive) CAP binding site -35 -10 LacI binding site start conserved BBa_B0034 E. coli Flagellin | 1,200 1,88 89,126 137,142 161,166 166,200 173,173 213,216 209,220 221,1826 | engineered_region feature/BioBrick feature/cds CDS non_covalent_binding_site feature/binding promoter feature/promoter feature/promoter promoter feature/binding non_covalent_binding_site feature/start start_codon feature/conserved conserved_region engineered_region feature/BioBrick CDS feature/protein |