Difference between revisions of "Part:BBa K921002"
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<partinfo>BBa_K921002 short</partinfo>
 
<partinfo>BBa_K921002 short</partinfo>
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<br>
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This is a mutant T7 promoter that includes a lacO sequence. This is repressible by the LacI protein and is induced in the presence of IPTG. The promoter sits in the 5' region of the gene of interest and initiates transcription when the cell expresses T7 RNA polymerase. Be sure you are using a cell strain that is compatible with T7 expression vectors. <br>
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The following parameters were calculated and compared against the "wild-type"</html> [https://parts.igem.org/part:BBa_K613007] <html> The following parameters were calculated:
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<table border="1">
  
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This is a mutant T7 promoter that includes a lacO sequence. This is repressible by the LacI protein and is induced in the presence of IPTG. The promoter sits in the 5' region of the gene of interest and initiates transcription when the cell expresses T7 RNA polymerase. Be sure you are using a cell strain that includes this gene (DE3).
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<tr>
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<th>Promoter</th>
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<td> BBa K921000 </td>
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<td> <i>BBa K921001</i> </td>
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<td> BBa K921002 </td>
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</tr>
  
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<tr>
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<th>Transcription Strength</th>
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<td> 97% </td>
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<td> <i>72%</i> </td>
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<td> 127% </td>
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</tr>
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<tr>
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<th>Translational Efficiency</th>
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<td> 6%</td>
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<td> <i>6%</i></td>
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<td> 94%</td>
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</tr>
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</table>
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<p><img src="https://static.igem.org/mediawiki/igem.org/d/d0/CMU_FAP-MG2.jpg", width="689", height="384"><br>
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Fluorogen activating proteins were used as protein reporters in our characterization experiment as described <a href="http://2012.igem.org/Team:Carnegie_Mellon/Met-Overview">here</a>. Here, a saturating amount of malachite green was added to each well of a 96 well plate and fluorescence values were recorded over time. Our <a href="http://2012.igem.org/Team:Carnegie_Mellon/Mod-Overview">model</a> was used to describe the behavior of our time lapse data using differential equations and fitting algorithms.
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<br><br>
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<img src="https://static.igem.org/mediawiki/igem.org/5/5c/CMU_Spin-DFHBI2.jpg", width="689", height="384"><br>
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The Spinach aptamer was used as an RNA reporter in our characterization experiment as described <a href="http://2012.igem.org/Team:Carnegie_Mellon/Met-Overview">here</a>. Here, a saturating amount of DFHBI was added to each well of a 96 well plate and fluorescence values were recorded over time. Our <a href="http://2012.igem.org/Team:Carnegie_Mellon/Mod-Overview">model</a> was used to describe the behavior of our time lapse data using differential equations and fitting algorithms.<br>
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<br>
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Note: Not included here are the properties of our characterization method. To learn more about how we characterized this set of promoters, <a href="http://2012.igem.org/Team:Carnegie_Mellon/Met-Overview">click here</a>.
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</p>
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</html>
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here
 
===Usage and Biology===
 
===Usage and Biology===

Revision as of 02:45, 2 October 2012

T7 RNAP + IPTG->PoPs (Mutant III)
This is a mutant T7 promoter that includes a lacO sequence. This is repressible by the LacI protein and is induced in the presence of IPTG. The promoter sits in the 5' region of the gene of interest and initiates transcription when the cell expresses T7 RNA polymerase. Be sure you are using a cell strain that is compatible with T7 expression vectors.
The following parameters were calculated and compared against the "wild-type" [1] The following parameters were calculated:

Promoter BBa K921000 BBa K921001 BBa K921002
Transcription Strength 97% 72% 127%
Translational Efficiency 6% 6% 94%


Fluorogen activating proteins were used as protein reporters in our characterization experiment as described here. Here, a saturating amount of malachite green was added to each well of a 96 well plate and fluorescence values were recorded over time. Our model was used to describe the behavior of our time lapse data using differential equations and fitting algorithms.


The Spinach aptamer was used as an RNA reporter in our characterization experiment as described here. Here, a saturating amount of DFHBI was added to each well of a 96 well plate and fluorescence values were recorded over time. Our model was used to describe the behavior of our time lapse data using differential equations and fitting algorithms.

Note: Not included here are the properties of our characterization method. To learn more about how we characterized this set of promoters, click here.

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


Functional Parameters

directionForward
efficiency160%
negative_regulatorsLacI (Hypothesized LacIQ compatible)