Difference between revisions of "Part:BBa K5317011"

 
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<!-- Add more about the biology of this part here
 
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===Usage and Biology===
 
===Usage and Biology===
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=Cloning=
  
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===Theoretical Part Design===
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Placing the MRE containing promoter upstream of the reporter gene EGFP allows the visualization of primarily metal-dependent activation of MTF-1.
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===Sequence and Features===
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<partinfo>BBa_K5317011 SequenceAndFeatures</partinfo>
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===Cloning===
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To test the MREdada promoter functionality the reporter gene EGFP (<span class="plainlinks">[https://parts.igem.org/Part:BBa_K3338006 K3338006]</span>) was cloned downstream of the promoter by inserting the MREdada promoter into the AseI- and NheI-digested EGFP-C2 backbone (<span class="plainlinks">[https://parts.igem.org/Part:BBa_K3338020 K3338020]</span>) using NEB Hifi Assembly.
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<html>
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  <head>
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      <title>HTML Table Caption</title>
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  </head>
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<body>
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<caption>Table1: Primers used to create matching overhangs on promoter amplicon to digested pEGFP-C2 backbone</caption>
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<table style="width:70%">
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  <tr>
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    <th>Primer name</th>
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    <th>Sequence</th>
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  </tr>
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  <tr>
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    <td>MREdada_fw</td>
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    <td>CCGCCATGCATTAGTTATGCACACTGGCGCT</td>
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  </tr>
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  <tr>
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    <td>MREdada_rev</td>
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    <td>TGGCGACCGGTAGCGGACGCTTAGAGGACAGC</td>
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  </tr>
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</table>
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</body>
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</html>
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The vector map of the assembled construct is shown in figure 1.
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>

Revision as of 19:05, 14 September 2024


MREdada-EGFP

The MRE-sites containing promoter enables the metal-dependent expression of a downstream positioned reporter gene via the metal ion-dependent transcription factor MTF-1 for cell-based metal detection.

In order to integrate the findings of Searle and colleagues (1985) and Wang and colleagues (2004) regarding the metal inducibility of a promoter with two MREa sites and a high affinity between MREd and MTF-1, we designed a synthetic promoter with two MREa and two MREd sites that alternate. The aim is to enhance the sensitivity and efficiency of the metal-dependent promoter.

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]