Composite

Part:BBa_K1341000

Designed by: Run-ze Zhang   Group: iGEM14_AHUT_China   (2014-09-27)

switch device(LTT gfp)for our bio-compass

Ahutk0006.png



We use this device as a part of a switch device designed for our bio-compass.It can help us screen out the possible path way which include the start point,terminal point, and the particular intermediate nodes(such as the museum(No.5 node)in our story).It has a LTT Promoter,with elements of the tet, lac, they will compose as a device and is responsive to the commonly used inducers IPTG and aTc, producing GFP as an output signal.If this device and tetR(BBa_K1341003)&lacI(BBa_K1341002)successful inserted in the path way by using the typically enzyme site. It will drive expression of green fluorescent protein (GFP). This characteristic will help us to screen out the possible way we want to find.PS: We build a measurement system to test the screen functional about this device in our project(BBa_K1341001).





model

Ahutk0002.png


The x and y axes form a grid of 36 pairs of inducer concentrations. The concentrations tested were 0, 1, 10, 50,100, and 200 ng/mL for aTC and 0, 0.001, 0.002, 0.005,0.010, 0.100, and 1.000mM for IPTG behavior is depicted 6 h after induction. The plotted model values are the means of 1000 independent stochastic kinetic simulations, the source and the reference are in the design page.









characterization

Ahut k10.jpg




Conditions are ±100 ng/mL aTc and ±1mM IPTG, This panel depicts an LTT promoter design, where the single lac operator site is upstream of the −35 sequence. In this case, LacI is no longer able to entirely suppress transcription and an intermediate level of induction (leakiness) occurs at conditions of high aTc but no IPTG. The fluorescence level still has an apparent change when we add aTc firstly, so the figure can explain why we first use IPTG for testing firstly.(All data is obtained 6 h after induction.)








Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 75
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 732
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