Team:UTK-Knoxville/primers

From 2013.igem.org

(Difference between revisions)
 
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<a href="../../UTK-knoxville/2013.igem.org">2013.igem.org</a></p>
+
<div id="header"><a href="https://2013.igem.org"><img src="https://static.igem.org/mediawiki/2013/thumb/2/20/Header2.jpg/800px-Header2.jpg" alt="Team UTK-Knoxville" width="910" height="234"></a></div>
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  <div id="header"><a href="https://2013.igem.org"><img src="https://lh5.googleusercontent.com/-HN_AgwewsJM/UZW60twqf3I/AAAAAAAAAA8/R7NTt_xL13c/w910-h253-no/header.jpg" alt="Team UTK-Knoxville"></a></div>
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<div class="MenuBar" id="navi">
<div class="MenuBar" id="navi">
<ul>
<ul>
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<li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span><span>Overall Project</span></span></a></li>
<li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span><span>Overall Project</span></span></a></li>
<li><a href="https://2013.igem.org/Team:UTK-Knoxville/methods"><span><span>Material &amp; Methods</span></span></a>
<li><a href="https://2013.igem.org/Team:UTK-Knoxville/methods"><span><span>Material &amp; Methods</span></span></a>
-
</li><li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span><span>SECTION1</span></span></a></li>
+
</li><li><a href="https://2013.igem.org/Team:UTK-Knoxville/partsorigin"><span><span>Parts Origin</span></span></a></li>
-
<li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span><span>SECTION2</span></span></a></li>
+
<li><a href="https://2013.igem.org/Team:UTK-Knoxville/construct"><span><span>Construct</span></span></a></li>
<li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span><span>SECTION3</span></span></a></li>
<li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span><span>SECTION3</span></span></a></li>
<li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span><span>SECTION4</span></span></a></li>
<li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span><span>SECTION4</span></span></a></li>
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  </ul>
  </ul>
  <li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span><span>Characterization</span></span></a></li>
  <li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span><span>Characterization</span></span></a></li>
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  </ul>
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  </ul>
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     </li>
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<ul class="DropDownMenu" id="MB1-DDM4">
<ul class="DropDownMenu" id="MB1-DDM4">
                               <li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span><span>Project Overview</span></span></a></li>
                               <li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span><span>Project Overview</span></span></a></li>
-
      <li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span><span>Section1</span></span></a></li>
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      <li><a href="https://2013.igem.org/Team:UTK-Knoxville/EastmanHITES"><span><span>Eastman HITES</span></span></a></li>
                               <li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span><span>Section2</span></span></a></li>
                               <li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span><span>Section2</span></span></a></li>
                               <li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span><span>Section3</span></span></a></li>
                               <li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span><span>Section3</span></span></a></li>
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<ul class="DropDownMenu" id="MB1-DDM6">
<ul class="DropDownMenu" id="MB1-DDM6">
-
<li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span><span>Overview</span></span></a></li>
+
<li><a href="https://2013.igem.org/Team:UTK-Knoxville/team"><span><span>Overview</span></span></a></li>
-
<li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span><span>Advisors</span></span></a></li>
+
<li><a href="https://2013.igem.org/Team:UTK-Knoxville/advisors"><span><span>Advisors</span></span></a></li>
-
<li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span><span>Undergraduates</span></span></a></li>
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<li><a href="https://2013.igem.org/Team:UTK-Knoxville/undergraduates"><span><span>Undergraduates</span></span></a></li>
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<li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span><span>University</span></span></a></li>
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<li><a href="https://2013.igem.org/Team:UTK-Knoxville/utk"><span><span>University</span></span></a></li>
-
<li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span><span>SECTION4</span></span></a></li>
+
-
<li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span><span>SECTION5</span></span></a></li>
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-
<li><a href="https://2013.igem.org/Team:UTK-Knoxville/comingsoon"><span><span>SECTION6</span></span></a></li>
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</ul>
</ul>
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</li>
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    </li>
<li>
<li>
<a href="https://2013.igem.org/Team:UTK-Knoxville/sponsors" style="color: white">Sponsors</a>
<a href="https://2013.igem.org/Team:UTK-Knoxville/sponsors" style="color: white">Sponsors</a>
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</div>
</div>
-
<h2> <span class="mw-headline" id="Background">Primers</span></h2>
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<h1><span class="mw-headline">Gibson Primers</span></h1>
-
 
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<table cellspacing="0" cellpadding="0">
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<table width="910" border="1">
+
  <col width="120">
 +
  <col width="442">
 +
  <col width="60">
 +
  <col width="766">
   <tr>
   <tr>
-
     <th width="136" scope="col">Name</th>
+
     <td width="120">Name</td>
-
     <th width="497" scope="col">Sequence</th>
+
     <td width="442">Sequence</td>
-
     <th width="69" scope="col">Melting Temp.(&deg;C)</th>
+
     <td width="60">Melting Temperature</td>
-
     <th width="163" scope="col">Function/Comment</th>
+
     <td width="766">Function / Comment</td>
-
    <th width="101" scope="col">Status</th>
+
   </tr>
   </tr>
   <tr>
   <tr>
-
     <td>General (Transmembrane)</td>
+
     <td width="120"></td>
-
     <td>&nbsp;</td>
+
     <td width="442"></td>
-
     <td>&nbsp;</td>
+
     <td width="60"></td>
-
     <td>&nbsp;</td>
+
     <td width="766"></td>
-
    <td>&nbsp;</td>
+
   </tr>
   </tr>
   <tr>
   <tr>
-
     <td>EnvZ_r</td>
+
     <td width="120">Transmembrane    Construct</td>
-
     <td>aaaaaactcgagttaggatccTTACCCTTCTTTTGTCGTGCC</td>
+
     <td width="442"></td>
-
     <td>57</td>
+
     <td width="60"></td>
-
     <td>Lowercase part has restriction sites: XhoI-ctcgag BamHI-ggatcc</td>
+
     <td width="766"></td>
-
    <td>In</td>
+
   </tr>
   </tr>
   <tr>
   <tr>
-
     <td>OmpR_r</td>
+
     <td width="120">EnvZ_F</td>
-
     <td>AAAAAACTCGAGAGGGGATCtcatgctttagagccgtccgg</td>
+
     <td width="442">gtaagtcacgacttgcgcacgccgctgac</td>
-
     <td>60</td>
+
     <td width="60">70</td>
-
    <td>Restriction sites: XhoI-ctcgag BamHI-ggatcc</td>
+
     <td width="766">Amplifies EnvZ does not include    any overlap of the sensor part.</td>
-
     <td>In</td>
+
   </tr>
   </tr>
   <tr>
   <tr>
-
     <td>OmpR_f</td>
+
     <td width="120">Envz_R</td>
-
     <td>aaaaaaCATATGCAAGAGACCTACAAGATTCTG</td>
+
     <td width="442">ggcggccgctctattagtgatggtggtgatgatgttacccttcttttgtcgtgc</td>
-
     <td>53</td>
+
     <td width="60">75</td>
-
     <td>Restriction site: NdeI-catatg</td>
+
     <td width="766">Amplifies EnvZ (20 bp) and    includes overlap (34 bp) of the free space between it and the terminator.</td>
-
    <td>In</td>
+
   </tr>
   </tr>
   <tr>
   <tr>
-
     <td>&nbsp;</td>
+
     <td width="120">OmpR_F</td>
-
     <td>&nbsp;</td>
+
     <td width="442">agaaataattttgtttaactataagaaggagatatacatatgcaagagaactacaagatt</td>
-
     <td>&nbsp;</td>
+
     <td width="60">64</td>
-
    <td>&nbsp;</td>
+
     <td width="766">Amplifies OmpR and overlaps with    the RBS and T7 promoter.&nbsp; Note the    backbone needs to have the T7 promoter and RBS already.</td>
-
     <td>&nbsp;</td>
+
   </tr>
   </tr>
   <tr>
   <tr>
-
     <td>Aer</td>
+
     <td width="120">OmpR_R</td>
-
     <td>&nbsp;</td>
+
     <td width="442">tcatgctttagagccgtccggtacaaagac</td>
-
     <td>&nbsp;</td>
+
     <td width="60">65</td>
-
     <td>&nbsp;</td>
+
     <td width="766">Amplifies OmpR and does not    include the second RBS to avoid homology.</td>
-
    <td>&nbsp;</td>
+
   </tr>
   </tr>
   <tr>
   <tr>
-
     <td>Aer_f</td>
+
     <td width="120"></td>
-
     <td>aaaaaagaattctaaagatctaaggagatataATGTCTTCTCATCCGTATGTC</td>
+
     <td width="442"></td>
-
     <td>53</td>
+
     <td width="60"></td>
-
     <td>Lowercase part has restriction sites: EcoRI-gaattc BglII-agatct</td>
+
     <td width="766">&nbsp;</td>
-
    <td>In 5/30</td>
+
   </tr>
   </tr>
   <tr>
   <tr>
-
     <td>Aer-EnvZ_1r</td>
+
     <td width="120">Aer</td>
-
     <td>GCGCAAGTCGTGACTTACCCatttctgacactggacacctgg</td>
+
     <td width="442"></td>
-
     <td>58</td>
+
     <td width="60"></td>
-
     <td>Overlapping PCR primers</td>
+
     <td width="766">&nbsp;</td>
-
    <td>In 5/30</td>
+
   </tr>
   </tr>
   <tr>
   <tr>
-
     <td>Aer-EnvZ_1f</td>
+
     <td width="120">Aer_f</td>
-
     <td>ccaggtgtccagtgtcagaaatGGGTAAGTCACGACTTGCGC</td>
+
     <td width="442">tctttgtaccggacggctctaaagcatgaaaggagatataatgtcttctcatccgtatgt</td>
-
     <td>59</td>
+
     <td width="60">71</td>
-
     <td>Overlapping PCR primers</td>
+
     <td width="766">Includes overlap with OmpR (29    bp), the RBS (11 bp) and overlap with Aer (20 bp)</td>
-
    <td>In 5/30</td>
+
   </tr>
   </tr>
   <tr>
   <tr>
-
     <td>&nbsp;</td>
+
     <td width="120">Aer_R</td>
-
     <td>&nbsp;</td>
+
     <td width="442">TACGCgtcagcggcgtgcgcaagtcgtgacttacatttctgacactggacacctg</td>
-
     <td>&nbsp;</td>
+
     <td width="60">77</td>
-
    <td>&nbsp;</td>
+
     <td width="766">Include overlap with EnvZ (34    bp) and Aer (21 bp).&nbsp; The temperature    differance is quite extereme.</td>
-
     <td>&nbsp;</td>
+
   </tr>
   </tr>
   <tr>
   <tr>
-
     <td>NarX</td>
+
     <td width="120"></td>
-
     <td>&nbsp;</td>
+
     <td width="442"></td>
-
     <td>&nbsp;</td>
+
     <td width="60"></td>
-
     <td>&nbsp;</td>
+
     <td width="766"></td>
-
    <td>&nbsp;</td>
+
   </tr>
   </tr>
   <tr>
   <tr>
-
     <td>NarX_F</td>
+
     <td width="120">NarX</td>
-
     <td>&nbsp;</td>
+
     <td width="442">&nbsp;</td>
-
     <td>&nbsp;</td>
+
     <td width="60"></td>
-
    <td>&nbsp;</td>
+
     <td width="766"></td>
-
     <td>We can order both sets of primers (transmembrane region from either protein) </td>
+
   </tr>
   </tr>
   <tr>
   <tr>
-
     <td>NarX_R</td>
+
     <td width="120">NarX_F</td>
-
     <td>&nbsp;</td>
+
     <td width="442">tctttgtaccggacggctctaaagcatgaAAGGAGATATAatgcttaaacgttgtctctc</td>
-
     <td>&nbsp;</td>
+
     <td width="60">71</td>
-
     <td>&nbsp;</td>
+
     <td width="766">Includes overlap with OmpR (29    bp), the RBS (11 bp) and overlap with NarX (20 bp)</td>
-
    <td>&nbsp;</td>
+
   </tr>
   </tr>
   <tr>
   <tr>
-
     <td>Nar-EnvZ_R</td>
+
     <td width="120">NarX_R</td>
-
     <td>&nbsp;</td>
+
     <td width="442">tacgcgtcagcggcgtgcgcaagtcgtgacttacatttttatgctccagcccggc</td>
-
     <td>&nbsp;</td>
+
     <td width="60">79</td>
-
    <td>&nbsp;</td>
+
     <td width="766">Include overlap with EnvZ (34    bp) and NarX (21 bp).&nbsp; The temperature    differance is quite extereme.</td>
-
     <td>&nbsp;</td>
+
   </tr>
   </tr>
   <tr>
   <tr>
-
     <td>&nbsp;</td>
+
     <td width="120"></td>
-
     <td>&nbsp;</td>
+
     <td width="442"></td>
-
     <td>&nbsp;</td>
+
     <td width="60"></td>
-
     <td>&nbsp;</td>
+
     <td width="766"></td>
-
    <td>&nbsp;</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td>HemAT</td>
+
-
    <td>&nbsp;</td>
+
-
    <td>&nbsp;</td>
+
-
    <td>&nbsp;</td>
+
-
    <td>&nbsp;</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td>HemAT_F</td>
+
-
    <td>aaaaaagaattctaaagatctaaggagatataTTGTTATTTAAAAAAGACAGAAAACAAG</td>
+
-
    <td>52</td>
+
-
    <td>Lowercase part has restriction sites: EcoRI-gaattc BglII-agatct</td>
+
-
    <td>Ordered 6/12</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td>HemAT_R</td>
+
-
    <td>ggcgctggttatccatcggactggttgtactcgctttgaaacg</td>
+
-
    <td>58</td>
+
-
    <td>Overlapping PCR primers</td>
+
-
    <td>Ordered 6/12</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td>HemAT_NtrB_F</td>
+
-
    <td>cgtttcaaagcgagtacaaccagtccgatggataaccagcgcc</td>
+
-
    <td>61</td>
+
-
    <td>Overlapping PCR primers</td>
+
-
    <td>Ordered 6/12</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td>&nbsp;</td>
+
-
    <td>&nbsp;</td>
+
-
    <td>&nbsp;</td>
+
-
    <td>&nbsp;</td>
+
-
    <td>&nbsp;</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td>TodS</td>
+
-
    <td>&nbsp;</td>
+
-
    <td>&nbsp;</td>
+
-
    <td>&nbsp;</td>
+
-
    <td>&nbsp;</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td>&nbsp;</td>
+
-
    <td>&nbsp;</td>
+
-
    <td>&nbsp;</td>
+
-
    <td>&nbsp;</td>
+
-
    <td>&nbsp;</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td>&nbsp;</td>
+
-
    <td>&nbsp;</td>
+
-
    <td>&nbsp;</td>
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-
    <td>&nbsp;</td>
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-
    <td>&nbsp;</td>
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-
  </tr>
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-
  <tr>
+
-
    <td>&nbsp;</td>
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-
    <td>&nbsp;</td>
+
-
    <td>&nbsp;</td>
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-
    <td>&nbsp;</td>
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-
    <td>&nbsp;</td>
+
-
  </tr>
+
-
  <tr>
+
-
    <td>&nbsp;</td>
+
-
    <td>&nbsp;</td>
+
-
    <td>&nbsp;</td>
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    <td>&nbsp;</td>
+
-
    <td>&nbsp;</td>
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-
  </tr>
+
-
  <tr>
+
-
 
+
-
    <td>RcoM</td>
+
-
    <td>&nbsp;</td>
+
-
    <td>&nbsp;</td>
+
-
    <td>&nbsp;</td>
+
-
    <td>&nbsp;</td>
+
   </tr>
   </tr>
   <tr>
   <tr>
-
     <td>RcoM_F</td>
+
     <td width="120">Cytoplasmic    Construct</td>
-
     <td>aaaaaagaattctaaagatctaaggagatataTTAGTTTTTACTTGCTTCAGTTGG</td>
+
     <td width="442"></td>
-
     <td>54</td>
+
     <td width="60"></td>
-
     <td>Lowercase part has restriction sites: EcoRI-gaattc BglII-agatct</td>
+
     <td width="766"></td>
-
    <td>Ordered 6/12</td>
+
   </tr>
   </tr>
   <tr>
   <tr>
-
     <td>RcoM_R</td>
+
     <td width="120">NtrC_F</td>
-
     <td>ggcgctggttatccatcggaatgtttaaatattttgcaattgcttc</td>
+
     <td width="442">TAGAAATAATTTTGTTTAACTataagaaggagatatacatatgcaacgagggatagtctg</td>
-
     <td>52</td>
+
     <td width="60">65</td>
-
     <td>Overlapping PCR primers</td>
+
     <td width="766">Amplifies NtrC and overlaps with    the RBS and T7 promoter.&nbsp; Note the    backbone needs to have the T7 promoter and RBS already.</td>
-
    <td>Ordered 6/12</td>
+
   </tr>
   </tr>
   <tr>
   <tr>
-
     <td>RcoM_NtrB_F</td>
+
     <td width="120">NtrC_R</td>
-
     <td>gaagcaattgcaaaatatttaaacattccgatggataaccagcgcc</td>
+
     <td width="442">gagctggggatggagtgaTTTTAACTTACGC</td>
-
     <td>61</td>
+
     <td width="60">64</td>
-
     <td>Overlapping PCR primers</td>
+
     <td width="766">Amplifies NtrC and does not    include the second RBS to avoid homology.</td>
-
    <td>Ordered 6/12</td>
+
   </tr>
   </tr>
   <tr>
   <tr>
-
     <td>&nbsp;</td>
+
     <td width="120">NtrB_F</td>
-
     <td>&nbsp;</td>
+
     <td width="442">ccgatggataaccagcgccgcTTAAGTCAGG</td>
-
     <td>&nbsp;</td>
+
     <td width="60">69</td>
-
     <td>&nbsp;</td>
+
     <td width="766">Amplifies NtrB does not include    any overlap of the sensor part.</td>
-
    <td>&nbsp;</td>
+
   </tr>
   </tr>
   <tr>
   <tr>
-
     <td>LasR</td>
+
     <td width="120">NtrB_R</td>
-
     <td>&nbsp;</td>
+
     <td width="442">GGCGGCCGCTCTATTAGTGATGGTGGTGATGATGttatttcctgataggcaggt</td>
-
     <td>&nbsp;</td>
+
     <td width="60">74</td>
-
     <td>&nbsp;</td>
+
     <td width="766">Amplifies NtrB (20 bp) and    includes overlap (34 bp) of the free space between it and the terminator.</td>
-
    <td>&nbsp;</td>
+
   </tr>
   </tr>
   <tr>
   <tr>
-
     <td>&nbsp;</td>
+
     <td width="120"></td>
-
     <td>&nbsp;</td>
+
     <td width="442"></td>
-
     <td>&nbsp;</td>
+
     <td width="60"></td>
-
     <td>&nbsp;</td>
+
     <td width="766"></td>
-
    <td>&nbsp;</td>
+
   </tr>
   </tr>
   <tr>
   <tr>
-
     <td>&nbsp;</td>
+
     <td width="120">HemAT</td>
-
     <td>&nbsp;</td>
+
     <td width="442"></td>
-
     <td>&nbsp;</td>
+
     <td width="60"></td>
-
     <td>&nbsp;</td>
+
     <td width="766"></td>
-
    <td>&nbsp;</td>
+
   </tr>
   </tr>
   <tr>
   <tr>
-
     <td>&nbsp;</td>
+
     <td width="120">HemAT_F</td>
-
     <td>&nbsp;</td>
+
     <td width="442">gtaagttaaaatcactccatccccagctcAAGGAGATATAttgttatttaaaaaagacag</td>
-
     <td>&nbsp;</td>
+
     <td width="60">67</td>
-
     <td>&nbsp;</td>
+
     <td width="766">Includes overlap with NtrC (29    bp), the RBS (11 bp) and overlap with HemAT (20 bp)</td>
-
    <td>&nbsp;</td>
+
   </tr>
   </tr>
   <tr>
   <tr>
-
     <td>&nbsp;</td>
+
     <td width="120">HemAT_R</td>
-
     <td>&nbsp;</td>
+
     <td width="442">CTGTTCCTGACTTAAGCggcgctggttatccatcggctggttgtactcgctttgaaac</td>
-
     <td>&nbsp;</td>
+
     <td width="60">75</td>
-
    <td>&nbsp;</td>
+
     <td width="766">Include overlap with NtrB (34    bp) and HemAT (21 bp).&nbsp; The temperature    differance is quite extereme.</td>
-
     <td>&nbsp;</td>
+
   </tr>
   </tr>
   <tr>
   <tr>
-
     <td>Cytoplasmic Construct</td>
+
     <td width="120"></td>
-
     <td>&nbsp;</td>
+
     <td width="442"></td>
-
     <td>&nbsp;</td>
+
     <td width="60"></td>
-
     <td>&nbsp;</td>
+
     <td width="766"></td>
-
    <td>&nbsp;</td>
+
   </tr>
   </tr>
   <tr>
   <tr>
-
     <td>NtrC_F</td>
+
     <td width="120">RcoM</td>
-
     <td>aaaaaacatatgatgcaacgagggatagtctgg</td>
+
     <td width="442"></td>
-
     <td>58</td>
+
     <td width="60"></td>
-
     <td>Restriction site: NdeI-catatg</td>
+
     <td width="766"></td>
-
    <td>Ordered 6/12</td>
+
   </tr>
   </tr>
   <tr>
   <tr>
-
     <td>NtrC_R</td>
+
     <td width="120">RcoM_F</td>
-
     <td>agatccgagctggggatggagtgactcgagaaaaaa</td>
+
     <td width="442">gtaagttaaaatcactccatccccagctcAAGGAGATATAatgtttaaatattttgcaatt</td>
-
     <td>57</td>
+
     <td width="60">67</td>
-
     <td>XhoI-ctcgag</td>
+
     <td width="766">Includes overlap with NtrC (29    bp), the RBS (11 bp) and overlap with RcoM (20 bp)</td>
-
    <td>Ordered 6/12</td>
+
   </tr>
   </tr>
   <tr>
   <tr>
-
     <td>NtrB_R</td>
+
     <td width="120">RcoM_R</td>
-
     <td>aaaaaactcgagttaggatccTTATTTCCTGATAGGCAGGTAAAC</td>
+
     <td width="442">gttcctgacttaagcggcgctggttatccatcggcttatggacggttacactacc</td>
-
     <td>54</td>
+
     <td width="60">75</td>
-
     <td>Lowercase part has restriction sites: XhoI-ctcgag BamHI-ggatcc</td>
+
     <td width="766">Include overlap with NtrB (34    bp) and RcoM (21 bp).&nbsp; The temperature    differance is quite extereme.</td>
-
    <td>Ordered 6/12</td>
+
   </tr>
   </tr>
   <tr>
   <tr>
-
     <td>&nbsp;</td>
+
     <td width="120"></td>
-
     <td>&nbsp;</td>
+
     <td width="442"></td>
-
     <td>&nbsp;</td>
+
     <td width="60"></td>
-
     <td>&nbsp;</td>
+
     <td width="766"></td>
-
    <td>&nbsp;</td>
+
   </tr>
   </tr>
   <tr>
   <tr>
-
     <td>&nbsp;</td>
+
     <td width="120">TodS</td>
-
     <td>&nbsp;</td>
+
     <td width="442"></td>
-
     <td>&nbsp;</td>
+
     <td width="60"></td>
-
     <td>&nbsp;</td>
+
     <td width="766"></td>
-
    <td>&nbsp;</td>
+
   </tr>
   </tr>
   <tr>
   <tr>
-
     <td>&nbsp;</td>
+
     <td width="120">TodS_F</td>
-
     <td>&nbsp;</td>
+
     <td width="442">gtaagttaaaatcactccatccccagctcAAGGAGATATAatgagctccttggatagaaa</td>
-
     <td>&nbsp;</td>
+
     <td width="60">70</td>
-
     <td>&nbsp;</td>
+
     <td width="766">Includes overlap with NtrC (29    bp), the RBS (11 bp) and overlap with TodS (20 bp)</td>
-
    <td>&nbsp;</td>
+
   </tr>
   </tr>
   <tr>
   <tr>
-
     <td>&nbsp;</td>
+
     <td width="120">TodS_R</td>
-
     <td>&nbsp;</td>
+
     <td width="442">GTTCCTGACTTAAgcggcgctggttatccatcggatcggtaatattgcgcccttc</td>
-
     <td>&nbsp;</td>
+
     <td width="60">75</td>
-
     <td>&nbsp;</td>
+
     <td width="766">Include overlap with NtrB (34    bp) and TodS (21 bp)</td>
-
    <td>&nbsp;</td>
+
   </tr>
   </tr>
   <tr>
   <tr>
-
     <td>&nbsp;</td>
+
     <td width="120"></td>
-
     <td>&nbsp;</td>
+
     <td width="442"></td>
-
     <td>&nbsp;</td>
+
     <td width="60"></td>
-
     <td>&nbsp;</td>
+
     <td width="766"></td>
-
    <td>&nbsp;</td>
+
   </tr>
   </tr>
   <tr>
   <tr>
-
     <td>&nbsp;</td>
+
     <td width="120">LasR</td>
-
    <td>&nbsp;</td>
+
     <td width="442">&nbsp;</td>
-
     <td>&nbsp;</td>
+
     <td width="60">&nbsp;</td>
-
     <td>&nbsp;</td>
+
     <td width="766">&nbsp;</td>
-
     <td>&nbsp;</td>
+
   </tr>
   </tr>
   <tr>
   <tr>
-
     <td>&nbsp;</td>
+
     <td width="120">LasR_F</td>
-
    <td>&nbsp;</td>
+
     <td width="442">&nbsp;</td>
-
     <td>&nbsp;</td>
+
     <td width="60">&nbsp;</td>
-
     <td>&nbsp;</td>
+
     <td width="766">&nbsp;</td>
-
     <td>&nbsp;</td>
+
   </tr>
   </tr>
   <tr>
   <tr>
-
     <td>&nbsp;</td>
+
     <td width="120">LasR_R</td>
-
    <td>&nbsp;</td>
+
     <td width="442">&nbsp;</td>
-
     <td>&nbsp;</td>
+
     <td width="60">&nbsp;</td>
-
     <td>&nbsp;</td>
+
     <td width="766">&nbsp;</td>
-
     <td>&nbsp;</td>
+
   </tr>
   </tr>
   <tr>
   <tr>
-
     <td>The backbone will be cut with NedI on the OmpR and NtrC side and BamHI on the EnvZ and NtrB side</td>
+
     <td width="120">&nbsp;</td>
-
    <td>&nbsp;</td>
+
     <td width="442">&nbsp;</td>
-
     <td>&nbsp;</td>
+
     <td width="60">&nbsp;</td>
-
     <td>&nbsp;</td>
+
     <td width="766">Note: APE and IDT primer Tm    differ</td>
-
     <td>&nbsp;</td>
+
   </tr>
   </tr>
</table>
</table>
-
<p><br>
+
<p> [<a href="https://2013.igem.org/Team:UTK-Knoxville" class="external text" rel="nofollow">back</a>]</p>
-
[<a href="https://2013.igem.org/Team:UTK-Knoxville" class="external text" rel="nofollow">back</a>]
+
<p><a href="http://www.engr.utk.edu/cbe/"><img src="https://static.igem.org/mediawiki/2013/2/22/Footerutk.jpg" width="910" height="250" border="0" usemap="#Map">
-
</p>
+
    <map name="Map">
 +
      <area shape="poly" coords="6,2,6,5" href="#">
 +
      <area shape="poly" coords="6,134,7,131,11,7,404,5,402,134" href="http://utk.edu" alt="UTK.edu">
 +
      <area shape="poly" coords="6,140" alt="UTK CBE">
 +
      <area shape="poly" coords="5,142,906,140,902,236,10,240" href="http://www.engr.utk.edu/cbe/" alt="UTK CBE">
 +
      <area shape="poly" coords="409,6,904,5,904,132,409,132" href="http://web.utk.edu/~ctrinh/" alt="Trinh Lab">
 +
    </map>
 +
</a></p>

Latest revision as of 03:40, 30 July 2013

Team:UK-Knoxville - 2013.igem.org

Gibson Primers

Name Sequence Melting Temperature Function / Comment
Transmembrane Construct
EnvZ_F gtaagtcacgacttgcgcacgccgctgac 70 Amplifies EnvZ does not include any overlap of the sensor part.
Envz_R ggcggccgctctattagtgatggtggtgatgatgttacccttcttttgtcgtgc 75 Amplifies EnvZ (20 bp) and includes overlap (34 bp) of the free space between it and the terminator.
OmpR_F agaaataattttgtttaactataagaaggagatatacatatgcaagagaactacaagatt 64 Amplifies OmpR and overlaps with the RBS and T7 promoter.  Note the backbone needs to have the T7 promoter and RBS already.
OmpR_R tcatgctttagagccgtccggtacaaagac 65 Amplifies OmpR and does not include the second RBS to avoid homology.
 
Aer  
Aer_f tctttgtaccggacggctctaaagcatgaaaggagatataatgtcttctcatccgtatgt 71 Includes overlap with OmpR (29 bp), the RBS (11 bp) and overlap with Aer (20 bp)
Aer_R TACGCgtcagcggcgtgcgcaagtcgtgacttacatttctgacactggacacctg 77 Include overlap with EnvZ (34 bp) and Aer (21 bp).  The temperature differance is quite extereme.
NarX  
NarX_F tctttgtaccggacggctctaaagcatgaAAGGAGATATAatgcttaaacgttgtctctc 71 Includes overlap with OmpR (29 bp), the RBS (11 bp) and overlap with NarX (20 bp)
NarX_R tacgcgtcagcggcgtgcgcaagtcgtgacttacatttttatgctccagcccggc 79 Include overlap with EnvZ (34 bp) and NarX (21 bp).  The temperature differance is quite extereme.
Cytoplasmic Construct
NtrC_F TAGAAATAATTTTGTTTAACTataagaaggagatatacatatgcaacgagggatagtctg 65 Amplifies NtrC and overlaps with the RBS and T7 promoter.  Note the backbone needs to have the T7 promoter and RBS already.
NtrC_R gagctggggatggagtgaTTTTAACTTACGC 64 Amplifies NtrC and does not include the second RBS to avoid homology.
NtrB_F ccgatggataaccagcgccgcTTAAGTCAGG 69 Amplifies NtrB does not include any overlap of the sensor part.
NtrB_R GGCGGCCGCTCTATTAGTGATGGTGGTGATGATGttatttcctgataggcaggt 74 Amplifies NtrB (20 bp) and includes overlap (34 bp) of the free space between it and the terminator.
HemAT
HemAT_F gtaagttaaaatcactccatccccagctcAAGGAGATATAttgttatttaaaaaagacag 67 Includes overlap with NtrC (29 bp), the RBS (11 bp) and overlap with HemAT (20 bp)
HemAT_R CTGTTCCTGACTTAAGCggcgctggttatccatcggctggttgtactcgctttgaaac 75 Include overlap with NtrB (34 bp) and HemAT (21 bp).  The temperature differance is quite extereme.
RcoM
RcoM_F gtaagttaaaatcactccatccccagctcAAGGAGATATAatgtttaaatattttgcaatt 67 Includes overlap with NtrC (29 bp), the RBS (11 bp) and overlap with RcoM (20 bp)
RcoM_R gttcctgacttaagcggcgctggttatccatcggcttatggacggttacactacc 75 Include overlap with NtrB (34 bp) and RcoM (21 bp).  The temperature differance is quite extereme.
TodS
TodS_F gtaagttaaaatcactccatccccagctcAAGGAGATATAatgagctccttggatagaaa 70 Includes overlap with NtrC (29 bp), the RBS (11 bp) and overlap with TodS (20 bp)
TodS_R GTTCCTGACTTAAgcggcgctggttatccatcggatcggtaatattgcgcccttc 75 Include overlap with NtrB (34 bp) and TodS (21 bp)
LasR      
LasR_F      
LasR_R      
      Note: APE and IDT primer Tm differ

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