Part:BBa_K1491034

Constitutive T7 RNAP with YFPH

T7 RNA polymerase 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.

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This plasmid was used in a two plasmid system to detect estrogen in water.

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Characterization:

The sensor final sensor system contains a YFP coding region to report the amount of T7 RNA Polymerase that is produced. The following graphs show the fluorescence of the system and its controls before and after the YFP coding region was added in Top10 cells. Two graphs are shown because of the high magnitude of RFU in the RNAP sample after YFP was added. The following graphs show the amount of fluorescence in the red channel, so RFU is indicative of the amount of RFP that was synthesized in the two plasmid system.

This graph shows the difference in red fluorescence from before the T7 RNAP was added (in the sample 115 YFP) and after it was added. We observed a significantly increased signal from RFP (over 400 times that of the RFP signal from the 115 RFP) after the T7 RNAP was added to the plasmid.

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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
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]