Composite

Part:BBa_K914014

Designed by: Dylan Iverson   Group: iGEM12_Paris_Bettencourt   (2012-10-25)

Pbad/AraC RBS LacI[LVA(-)] Plac GFP

Used to characterize Paris Bettencourt 2012 delay system. When arabinose is no longer present LacI is able to leave by dilution eventually allowing GFP expression.

Characterization:

Paris Bettencourt 2012 Simple delay.png

We characterized the behavior of our transcriptional circuit by first growing cells in the presence of 1% arabinose induction. In this state, LacI expression is highly induced. The cells were then washed, diluted, and characterized in the presence of arabinose, IPTG, or no induction.

The phase between the 'uninduced' and 'IPTG' curves is the delay from dilution of LacI. After removal of arabinose, LacI remains within the cells but soon becomes inefficient at inhibiting GFP expression as the cells divide. The delay is characterized by also adding IPTG to the system, which activates GFP expression despite presence of LacI.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1247
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1187
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 979
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 3099
    Illegal SapI site found at 961


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