Composite

Part:BBa_K914004

Designed by: Dylan Iverson   Group: iGEM12_Paris_Bettencourt   (2012-09-20)

Pbad/AraC RBS LacI

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

Used as part of the simple delay mechanism for bWARE suicide mechanism. When arabinose is no longer present, LacI will no longer be present to repress another plasmid that expresses a restriction enzyme under Plac.

Characterization of K914014 the complete delay system.

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
    COMPATIBLE WITH RFC[1000]


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