Composite

Part:BBa_K389015

Designed by: Jonas Aretz   Group: iGEM10_Bielefeld-Germany   (2010-09-28)
Revision as of 15:16, 26 October 2010 by Jaretz (Talk | contribs)

VirA/G reporter device luc

This BioBrick contains a complete VirA/G receptor system with a firefly luciferase (from Promega's pGL4.10[luc2] vector) under the control of a vir promoter as reporter gene.

Input: acetosyringone

Output: expression of luciferase


Usage and Biology

This BioBrick is for testing, characterizing and measuring the VirA/G signaling system. This system contains an unmutated virA, so it is possible to measure how the natural VirA/G system works and reacts. It is also possible to determine the basal transcription of the vir promoter and its activity after inducing the system with acetosyringone.


Important parameters

Experiment Characteristic Value
Transfer Function Maximum induction level 2.2 fold
Maximum induction level reached 200 µM acetosyringone
Hill coefficient 1.09
Switch Point 31.6 µM acetosyringone
Doubling time / h without plasmid 1.98
carrying K389015 2.24
carrying K389015 with 400 µM acetosyringone 2.67
Response time Induction: exponential phase >1 h
Induction: begin of cultivation max. induction at OD600 = 1 +/- 0.5

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal NheI site found at 647
    Illegal NheI site found at 2581
    Illegal NheI site found at 2604
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1632
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 3769
    Illegal NgoMIV site found at 5113
    Illegal NgoMIV site found at 5134
    Illegal AgeI site found at 4837
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 1768
    Illegal SapI.rc site found at 5019


[edit]
Categories
//cds/membrane/receptor
//chassis/prokaryote/ecoli
//function/reporter/light
Parameters
None