Difference between revisions of "Part:BBa K2100004"
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K2100004 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K2100004 SequenceAndFeatures</partinfo>
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We characterized our TRE promoter induced with a fine sweep of Doxycycline from a concentration of 0 uM to 2000 uM. We transfected HEK293 cells with 250ng of TRE-mKate and 250 ng hEF1a-eYFP. We analyzed fluorescent output in the PE Texas Red channel to measure the activity of the TRE promoter under different DOX concentrations. We used 0 uM of DOX as a control.
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https://static.igem.org/mediawiki/parts/9/96/T--MIT--TRE.png
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The results indicate the optimal DOX concentration was 500 uM for maximum TRE activity. Beyond that point, adding a higher concentration of DOX does not significantly increase fluorescent output from the TRE-mKate construct.
  
  

Latest revision as of 23:06, 28 October 2016


pENTR pTRE

This construct is an entry vector containing the TRE promoter. The promoter is structured as six tetO sites upstream of a minimal CMV promoter. The induction of doxycycline activates a tetracycline transactivator protein which in turn binds to the tetO sites to initiate transcription of the gene.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 6
    Illegal EcoRI site found at 322
    Illegal XbaI site found at 30
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 6
    Illegal EcoRI site found at 322
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 6
    Illegal EcoRI site found at 322
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 6
    Illegal EcoRI site found at 322
    Illegal XbaI site found at 30
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 6
    Illegal EcoRI site found at 322
    Illegal XbaI site found at 30
  • 1000
    COMPATIBLE WITH RFC[1000]

We characterized our TRE promoter induced with a fine sweep of Doxycycline from a concentration of 0 uM to 2000 uM. We transfected HEK293 cells with 250ng of TRE-mKate and 250 ng hEF1a-eYFP. We analyzed fluorescent output in the PE Texas Red channel to measure the activity of the TRE promoter under different DOX concentrations. We used 0 uM of DOX as a control.

T--MIT--TRE.png

The results indicate the optimal DOX concentration was 500 uM for maximum TRE activity. Beyond that point, adding a higher concentration of DOX does not significantly increase fluorescent output from the TRE-mKate construct.