BBa_K777104

BBa_K777104 Version 1

Component

Source:
http://parts.igem.org/Part:BBa_K777104
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Team Goettingen
Date created: 2012-09-18 11:00:00
Date modified: 2015-05-08 01:13:16

flhDC operon under the control of constitutive promoter J23106



Types
DnaRegion

Roles
Composite

engineered_region

Sequences BBa_K777104_sequence (Version 1)

Description

[[Image:Table_K777101-K777108_new.jpg|thumb|240px|right|Fig. 1: Overview of BioBricks containing the flhDC operon downstream of Anderson promoters. (Activity according to Berkeley 2006)]]
The flhDC operon is the master regulator of motility and chemotaxis in E. coli. FlhDC forms heterotetramers and activates class II operons in concert with sigma factor 70. Among the gene products of class II operons are several components of the flagellum and the alternative sigma factor FliA which is essential for the transcription of class III genes.

Here we used a set of 8 constitutive Anderson-promoters from the 2006 Berkeley group to test how different levels of constitutive flhDC expression affect the motility of E. coli.

Notes

  • The PstI restriction site in the flhDC sequence was mutated to CTGCGG using the reverse primer
  • flhDC was amplified from genomic DNA of E. coli DH10B via PCR using the following primers:
    • Fwd: 5'-actgaattcgcggccgcttctagatgcatacctccgagttgctg-3'
    • Rev: 5'-tcctgcagcggccgctactagttactgcccgggatggcggttgacataagcCgcaggcaaagctgccaacag-3'
      (the capitalized "C" induces the mutation for removal of the PstI site)


Source

  • The part was amplified from genomic DNA of E. coli str. K-12 substr. DH10B, complete genome (CP000948.1).
  • J23106 information was taken from the partsregistry and physical DNA from the 2012 distribution kit.


igem#experience
None
 
igem#sampleStatus
In stock
igem#status
Available
 
synbiohub#ownedBy
user/james
 
synbiohub#ownedBy
user/myers
 
synbiohub#topLevel
BBa_K777104/1