Difference between revisions of "Part:BBa K3792003"
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<p>
 
<p>
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apFAB40 is a strong promoter taken from the BIOFAB collection. This device uses apFAB40 to express mRFP1, a red fluorescent protein. The apFAB40 test device consists of apFAB40, an RBS, the coding sequence for mRFP1, and a terminator.<br>
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The apFAB40 Measurement Device is composed of a BIOFAB promoter, a ribosome binding site (BBa_B0034), mRFP1 (BBa_E1010), and a terminator (BBa_B0015). The measurement device was inserted into the pSB1K3 backbone using the NEB HiFi DNA Assembly method. The resulting plasmid vectors were used to transform the E. coli NEBExpress chassis. The promoter strengths were measured via the amount of fluorescence detected in a flow cytometer. The levels of fluorescence will be reported in molecules of equivalent PE-Texas Red (MEPTR) units. <strong>The analysis of the flow cytometry data is ongoing and will be shared during the Giant Jamboree. The interactive chart displayed below is a placeholder for the measurements</strong>.
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This test device and two others with BIOFAB promoters of different strengths were each assembled into the pSB1K3 plasmid and then transformed into E. coli NEBExpress.<br>  
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The fluorescence of the cells was measured using flow cytometry. By comparing the relative fluorescence, we can determine which promoters caused higher levels of expression of mRFP1.<br>
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       <caption>Promoter Strength Measurement</caption>
 
       <caption>Promoter Strength Measurement</caption>
 
       <tr>
 
       <tr>
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         <th scope="row">apFAB40<br>BBa_K3792003</th>
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         <th scope="row">Negative<br>Control</th>
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         <td style="--size: calc(2.477/2.5)"><span class="tooltip">Supernatant of the Chitin Secretion Cells</span><span class="data">2.477</span></td>
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         <td style="--size: calc(1000/2000)"><span class="tooltip">Negative Control</span><span class="data">1000</span></td>
 
       </tr>
 
       </tr>
 
       <tr>
 
       <tr>
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       <th scope="row">LB and Assay</th>
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       <th scope="row">apFAB40<br>BBa_K3792003</th>
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       <td style="--size: calc(1.515/2.5)"><span class="tooltip">LB Growth Medium with the Chitosan Detection Assay</span><span class="data">1.515</span></td>
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       <td style="--size: calc(1700/2000)"><span class="tooltip">apFAB40<br>BBa_K3792003</span><span class="data">1700</span></td>
 
       </tr>
 
       </tr>
 
       <tr>
 
       <tr>
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         <th scope="row">DI Water and Assay</th>
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         <th scope="row">apFAB98<br>BBa_K3792004</th>
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         <td style="--size: calc(0.084/2.5)"><span class="tooltip">Deionized Water with the Chitosan Assay</span><span class="data">0.084</span></td>  
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         <td style="--size: calc(1700/2000)"><span class="tooltip">apFAB98<br>BBa_K3792004</span><span class="data">1700</span></td>  
 
       </tr>
 
       </tr>
 
       <tr>
 
       <tr>
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         <th scope="row">LB and No Assay</th>       
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         <th scope="row">apFAB51<br>BBa_K3792005</th>       
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         <td style="--size: calc(0.08/2.5)"><span class="tooltip">LB Growth Medium with No Chitosan Assay</span><span class="data">0.08</span></td>
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         <td style="--size: calc(1700/2000)"><span class="tooltip">apFAB51<br>BBa_K3792005</span><span class="data">1700</span></td>
 
     </tr>
 
     </tr>
 
   </table>
 
   </table>
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   <div class="data-axis">Absorbance of 540 nm Light</div>
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   <div class="data-axis">molecules of equivalent PE-Texas Red (MEPTR)</div>
 
</div>
 
</div>
  

Revision as of 03:05, 22 October 2021


BIOFAB apFAB40 Measurement Device

The apFAB40 Measurement Device is composed of a BIOFAB promoter, a ribosome binding site (BBa_B0034), mRFP1 (BBa_E1010), and a terminator (BBa_B0015). The measurement device was inserted into the pSB1K3 backbone using the NEB HiFi DNA Assembly method. The resulting plasmid vectors were used to transform the E. coli NEBExpress chassis. The promoter strengths were measured via the amount of fluorescence detected in a flow cytometer. The levels of fluorescence will be reported in molecules of equivalent PE-Texas Red (MEPTR) units. The analysis of the flow cytometry data is ongoing and will be shared during the Giant Jamboree. The interactive chart displayed below is a placeholder for the measurements.

Promoter Strength Measurement
Negative
Control		 Negative Control1000
apFAB40
BBa_K3792003		 apFAB40
BBa_K37920031700
apFAB98
BBa_K3792004		 apFAB98
BBa_K37920041700
apFAB51
BBa_K3792005		 apFAB51
BBa_K37920051700
molecules of equivalent PE-Texas Red (MEPTR)

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 614
    Illegal AgeI site found at 726
  • 1000
    COMPATIBLE WITH RFC[1000]