Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Bianca Genenncher
Date created: 2012-09-20 11:00:00
Date modified: 2015-05-08 01:13:16
Tar receptor under the control of constitutive promoter J23104
| Types | DnaRegion |
| Roles | engineered_region Composite |
| Sequences | BBa_K777002_sequence (Version 1) |
Description
[[Image:Table_K777001-K777008_new.png|thumb|240px|right|Fig. 1: Overview of BioBricks containing the Tar gene downstream of Anderson promoters. (Activity according to Berkeley 2006)]]The transmembrane chemoreceptor Tar of E. coli mediates chemotaxis towards aspartate and exists as a functional homodimer.
Here we used a set of 8 constitutive Anderson promoters from the 2006 Berkeley group to test how different levels of constitutive Tar expression affect the chemotaxis of E. coli.
Notes
- Genomic sequence was amplified using the following primers.
- Primers PCR: Base pairs in caps represent the respective restriction sites followed by either prefix or suffix. Primers were provided by SIGMA.
- Fwd: Tar for + prefix: 5?? actGAATTCgcggccgctTCTAGAtgattaaccgtatccgcgtagtc 3??
- Rev: Tar rev + suffix: 5?? tcCTGCAGcggccgctACTAGTcaaaatgtttcccagtttggatc 3??
- Primers PCR: Base pairs in caps represent the respective restriction sites followed by either prefix or suffix. Primers were provided by SIGMA.
- One XbaI site was found at position 420. The restriction site was changed from TCTAGt to CCTAGt via QuikChange PCR.
- Primers QuikChange (QC): Primers were provided by SIGMA.
- Fwd: TarQC for: 5?? ACTGATTGATTAcCTAGATTATGGCAATACTGGAG 3??
- Rev: TarQC rev: 5?? TGCCATAATCTAGgTAATCAATCAGTTCAGTTAAC 3??
(the uncapitalized letter induces the mutation for removal of the XbaI site)
- Primers QuikChange (QC): Primers were provided by SIGMA.
Source
*The part was amplified from genomic DNA of E. coli str. K-12 substr. DH10B, complete genome (CP000948.1).*J23100 information was taken from the partsregistry and physical DNA from the 2012 distribution kit.
| Access | Instance | Definition |
| public public | BBa_J23104 BBa_K777000 | BBa_J23104 BBa_K777000 |
| Sequence Annotation | Location | Component / Role(s) |
| BBa_J23104 BBa_K777000 | 1,35 42,1703 | BBa_J23104 BBa_K777000 |