Composite
Blue Light

Part:BBa_K892800

Designed by: Rada Jumroon   Group: iGEM12_Washington   (2012-10-03)
Revision as of 03:41, 13 October 2012 by Felixekn (Talk | contribs)

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Blue Light Sensor, TetR Inverter and sfGFP Output

pLac I - LovTAP - Term - pTrp - TetR - Term - pTetO - sfGFP - Term

Part BBa_892800 is an improvement on part BBa_K191003. It fixes a frameshift deletion in LovTAP and includes the inverter and sfGFP readout on a single plasmid. The hybrid protein LovTAP has a light-sensitive domain (Lov: Light, oxygen, voltage) and a regulatory domain (part of the E. coli Trp repressor). The fused protein allows transmission of the conformational change induced by light (on the light-sensitive domain) to the DNA-binding domain, therefore resulting in an increase or decrease of the regulatory domain's affinity for its DNA binding site. The activation of the LovTAP protein (binding to DNA) is achieved by illumination @ 470 nm.

Washington BlueLightSensor.png

For more details, please consult the section of LovTAP system on team [http://2009.igem.org/Team:EPF-Lausanne/LOVTAP/ EPF-Lausanne wiki], or the [http://2012.igem.org/Team:Washington/Optogenetics Optogenetics page] on the 2012 University of Washington wiki.

Usage and Biology

In order to be able to test the efficiency of the fused protein LovTAP, its expression is induced by IPTG (Due to the LacI-sensitive promoter). The readout of the system is also included on the plasmid. The TetR driven by the TrpR promoter acts as an inverter, while the super-folder GFP (sfGFP) driven by the TetO promoter is the ultimate readout. Also available in the registry are the two parts: BBa_K191004 and BBa_K191005.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal SpeI site found at 1104
    Illegal SpeI site found at 1112
    Illegal PstI site found at 370
    Illegal PstI site found at 627
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal SpeI site found at 1104
    Illegal SpeI site found at 1112
    Illegal PstI site found at 370
    Illegal PstI site found at 627
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal SpeI site found at 1104
    Illegal SpeI site found at 1112
    Illegal PstI site found at 370
    Illegal PstI site found at 627
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal SpeI site found at 1104
    Illegal SpeI site found at 1112
    Illegal PstI site found at 370
    Illegal PstI site found at 627
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 764
    Illegal SapI.rc site found at 2074


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