Device

Part:BBa_K1491035

Designed by: Danielle Peters   Group: iGEM14_Carnegie_Mellon   (2014-09-26)

Constitutive ER Intein T7 RNAP with YFPH

T7 RNA polymerase that contains the estrogen receptor intein with a constitutive promoter. YFP is added to the end of the plasmid so that the amount of T7 RNA polymerase that is produced can be quantified through fluorescence analysis of YFP.

Characterization

The fluorescence of the sensor and its controls were measure in the red and yellow channels with and without the presence of beta-estradiol over several time points.

*The magnitude of fluorescence of T7 RNA polymerase between the red and yellow channels was so large that they are presented on two different graphs.

Screen_Shot_2014-10-17_at_8.47.01_PM.png

Screen_Shot_2014-10-17_at_8.47.08_PM.png

TECAN Parameters to Detect RFP Signal from the Sensor:

emission: 585 nm

excitation: 607

bandwidth: 10nm

gain: 175

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2525
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 2950
    Illegal AgeI site found at 2914
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 2425


[edit]
Categories
Parameters
None