Difference between revisions of "Team:CCU Taiwan/Composite Part"

 
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{{Template:CCU_Taiwan/css}}
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{{CCU_Taiwan/css}}
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<html>
 
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<meta name="viewport" content="width=device-width, initial-scale=1">
 
<script>
 
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                 <ul>
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                 <ul class="front">
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                     <div id="home_button" style="cursor:pointer;"  onclick="location.href=
 
                     <div id="home_button" style="cursor:pointer;"  onclick="location.href=
 
'https://2018.igem.org/Team:CCU_Taiwan';"> <img src="https://static.igem.org/mediawiki/2018/0/08/T--CCU_Taiwan--home_button.png"></img></div>
 
'https://2018.igem.org/Team:CCU_Taiwan';"> <img src="https://static.igem.org/mediawiki/2018/0/08/T--CCU_Taiwan--home_button.png"></img></div>
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                     <li class="title" id="Home"><a>Home</a>
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                         <ul class="sub" id="sub_home">
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                     <li class="title" style="cursor:pointer;" id="Home"><img class="img_title" src="https://static.igem.org/mediawiki/2018/2/24/T--CCU_Taiwan--aboutus.png"></img><a>About Us</a>
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                             <li class="list" id="home1"><a href="https://2018.igem.org/Team:CCU_Taiwan/Team">Team</a></li>
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                         <ul class="sub" id="sub_home" style="cursor:default;">
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<li class="list" id="home2"><a href="https://2018.igem.org/Team:CCU_Taiwan/Attributions">Attributions</a></li>
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                             <a href="https://2018.igem.org/Team:CCU_Taiwan/Team"><li class="list" id="home1">Team</li></a>
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<li class="list" id="home3"><a href="https://2018.igem.org/Team:CCU_Taiwan/Judge">For Judge</a></li>
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<a href="https://2018.igem.org/Team:CCU_Taiwan/Attributions"><li class="list" id="home2">Attributions</li></a>
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<a href="https://2018.igem.org/Team:CCU_Taiwan/Medal"><li class="list" id="home3">Medals</li></a>
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<div id="home_img">
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<a href="https://2018.igem.org/Team:CCU_Taiwan/Judge"><li class="list" id="home4">For Judges</li></a>
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<img src="https://static.igem.org/mediawiki/2018/5/57/T--CCU_Taiwan--home.png"></img>
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<a href="https://2018.igem.org/Team:CCU_Taiwan/Achievements"><li class="list" id="home5">Achievements</li></a>
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</div>
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<div id="home_word">
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<p>There's a home for us, always.</p>
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</div>
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                         </ul>
 
                         </ul>
 
                     </li>
 
                     </li>
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                     <li class="title" id="Project"><a href="#">Project</a>
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                     <li class="title" style="cursor:pointer;" id="Project"><img class="img_title" src="https://static.igem.org/mediawiki/2018/6/6f/T--CCU_Taiwan--project.png"></img><a>Project</a>
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<ul class="sub" id="sub_project">
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<ul class="sub" id="sub_project" style="cursor:default;">
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    <li class="list" id="project1"><a href="https://2018.igem.org/Team:CCU_Taiwan/Description">Description</a></li>
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    <a href="https://2018.igem.org/Team:CCU_Taiwan/Description"><li class="list" id="project1">Description</li></a>
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<li class="list" id="project2"><a href="https://2018.igem.org/Team:CCU_Taiwan/Design">Design</a></li>
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<a href="https://2018.igem.org/Team:CCU_Taiwan/Applied_Design"><li class="list" id="project2">Design</li></a>
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<li class="list" id="project3"><a href="https://2018.igem.org/Team:CCU_Taiwan/Results">Results</a></li>
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<a href="https://2018.igem.org/Team:CCU_Taiwan/Results"><li class="list" id="project3">Results</li></a>
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<li class="list" id="project4"><a href="https://2018.igem.org/Team:CCU_Taiwan/Achievements">Achievements</a></li>
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<a href="https://2018.igem.org/Team:CCU_Taiwan/Demonstrate"><li class="list" id="project4">Demonstration</li></a>
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<li class="list" id="project5"><a href="https://2018.igem.org/Team:CCU_Taiwan/InterLab ">InterLab</a></li>
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<a href="https://2018.igem.org/Team:CCU_Taiwan/InterLab"><li class="list" id="project5">InterLab</li></a>
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    <li class="list" id="project6"><a href="https://2018.igem.org/Team:CCU_Taiwan/Demonstrate">Demonstration</a></li>
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</ul>
 
</ul>
 
</li>
 
</li>
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                     <li class="title" id="Parts"><a href="#">Parts</a>
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                     <li class="title" style="cursor:pointer;" id="Parts"><img class="img_title" src="https://static.igem.org/mediawiki/2018/1/17/T--CCU_Taiwan--part.png"></img><a>Parts</a>
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                         <ul class="sub" id="sub_parts">
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                         <ul class="sub" id="sub_parts" style="cursor:default;">
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    <li class="list" id="parts1"><a href="https://2018.igem.org/Team:CCU_Taiwan/Parts">Parts</a></li>
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<a href="https://2018.igem.org/Team:CCU_Taiwan/Parts"><li class="list" id="parts1">Overview</li></a>
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<li class="list" id="parts2"><a href="https://2018.igem.org/Team:CCU_Taiwan/Basic_Part">Basic Part</a></li>
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<a href="https://2018.igem.org/Team:CCU_Taiwan/Basic_Part"><li class="list" id="parts1">Basic Part</li></a>
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<li class="list" id="parts3"><a href="https://2018.igem.org/Team:CCU_Taiwan/Composite_Part">Composite Part</a></li>
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<a href="https://2018.igem.org/Team:CCU_Taiwan/Composite_Part"><li class="list" id="parts2">Composite Part</li></a>
−
<li class="list" id="parts4"><a href="https://2018.igem.org/Team:CCU_Taiwan/Improve">Improved Part</a></li>
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                         </ul>
 
                         </ul>
 
                     </li>
 
                     </li>
−
                     <li class="title" id="Modeling"><a href="#">Modeling</a>
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                     <li class="title" style="cursor:pointer;" id="Modeling"><img class="img_title" src="https://static.igem.org/mediawiki/2018/0/09/T--CCU_Taiwan--model.png"></img><a>Modeling</a>
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                         <ul class="sub" id="sub_modeling">
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                         <ul class="sub" id="sub_modeling" style="cursor:default;">
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                            <li class="list" id="model1"><a href="https://2018.igem.org/Team:CCU_Taiwan/Model">Model</a></li>
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                            <a href="https://2018.igem.org/Team:CCU_Taiwan/Model"><li class="list" id="model1">Overview</li></a>
−
                            <li class="list" id="model2"><a href="https://2018.igem.org/Team:CCU_Taiwan/Enzyme">Enzyme Activity</a></li>
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                            <a href="https://2018.igem.org/Team:CCU_Taiwan/Binding"><li class="list" id="model2">Binding Model</li></a>
−
<li class="list" id="model3"><a href="https://2018.igem.org/Team:CCU_Taiwan/Monolignol">Monolignol Binding</a></li>
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<a href="https://2018.igem.org/Team:CCU_Taiwan/Polymer"><li class="list" id="model3">Polymer Model</li></a>
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<li class="list" id="model4"><a href="https://2018.igem.org/Team:CCU_Taiwan/Tools">Tools</a></li>
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                         </ul>
 
                         </ul>
 
                     </li>
 
                     </li>
−
                     <li class="title" id="Drylab"><a href="#">Drylab</a>
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                     <li class="title" style="cursor:pointer;" id="Drylab"><img class="img_title" src="https://static.igem.org/mediawiki/2018/f/fc/T--CCU_Taiwan--Dry_lab.png"></img><a>Product</a>
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                         <ul class="sub" id="sub_drylab">
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                         <ul class="sub" id="sub_drylab" style="cursor:default;">
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                            <li class="list" id="drylab1"><a href="https://2018.igem.org/Team:CCU_Taiwan/Our_Plan">Our Plan</a></li>
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                            <a href="https://2018.igem.org/Team:CCU_Taiwan/Our_Plan"><li class="list" id="drylab1">Analysis</li></a>
−
<li class="list" id="drylab2"><a href="https://2018.igem.org/Team:CCU_Taiwan/Engineering">Engineering</a></li>
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<a href="https://2018.igem.org/Team:CCU_Taiwan/Engineering"><li class="list" id="drylab2">Production Line</li></a>
−
<li class="list" id="drylab3"><a href="https://2018.igem.org/Team:CCU_Taiwan/Safety">Safety</a></li>
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<a href="https://2018.igem.org/Team:CCU_Taiwan/Safety"><li class="list" id="drylab3">Safety</li></a>
 
                         </ul>
 
                         </ul>
 
                     </li>
 
                     </li>
−
<li class="title" id="Human_Practice"><a href="#">Human Practice</a>
+
    <li class="title" style="cursor:pointer;" id="Human_Practice"><img class="img_title" src="https://static.igem.org/mediawiki/2018/9/96/T--CCU_Taiwan--humanpractice.png"></img><a>HP</a>
−
                         <ul class="sub" id="sub_human_practice">
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                         <ul class="sub" id="sub_human_practice" style="cursor:default;">
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                             <li class="list" id="human_practice1"><a href="https://2018.igem.org/Team:CCU_Taiwan/Human_Practices">Human Practice</a></li>
+
                             <a href="https://2018.igem.org/Team:CCU_Taiwan/Human_Practices"><li class="list" id="human_practice1">Human Practice</li></a>
−
<li class="list" id="human_practice2"><a href="https://2018.igem.org/Team:CCU_Taiwan/Engagement">Engagement</a></li>
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<a href="https://2018.igem.org/Team:CCU_Taiwan/Public_Engagement"><li class="list" id="human_practice2">Public Engagement</li></a>
−
<li class="list" id="human_practice3"><a href="https://2018.igem.org/Team:CCU_Taiwan/Entrepreneurship">Entrepreneurship</a></li>
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<a href="https://2018.igem.org/Team:CCU_Taiwan/Entrepreneurship"><li class="list" id="human_practice3">Entrepreneurship</li></a>
−
<li class="list" id="human_practice4"><a href="https://2018.igem.org/Team:CCU_Taiwan/Business_Model">Business Model</a></li>
+
<a href="https://2018.igem.org/Team:CCU_Taiwan/engaging_experts"><li class="list" id="human_practice4">Engaging Experts</li></a>
 +
<a href="https://2018.igem.org/Team:CCU_Taiwan/Integrate"><li class="list" id="human_practice5">Integrated HP</li></a>
 
                         </ul>
 
                         </ul>
 
                     </li>
 
                     </li>
−
<li class="title" id="Notebook"><a href="#">Notebook</a>
+
    <li class="title" style="cursor:pointer;" id="Notebook"><img class="img_title" src="https://static.igem.org/mediawiki/2018/c/c9/T--CCU_Taiwan--notebook.png"></img><a>Notebook</a>
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                         <ul class="sub" id="sub_notebook">
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                         <ul class="sub" id="sub_notebook" style="cursor:default;">
−
                             <li class="list" id="notebook1"><a href="https://2018.igem.org/Team:CCU_Taiwan/Notebook">Notebook</a></li>
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                             <a href="https://2018.igem.org/Team:CCU_Taiwan/Notebook"><li class="list" id="notebook1">Overview</li></a>
−
<li class="list" id="notebook2"><a href="https://2018.igem.org/Team:CCU_Taiwan/Applied_Design">Applied Design</a></li>
+
<a href="https://2018.igem.org/Team:CCU_Taiwan/Collaborations"><li class="list" id="notebook2">Collaborations</li></a>
−
<li class="list" id="notebook3"><a href="https://2018.igem.org/Team:CCU_Taiwan/Experiments">Experiments</a></li>
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<a href="https://2018.igem.org/Team:CCU_Taiwan/Protocols"><li class="list" id="notebook3">Protocols</li></a>
−
<li class="list" id="notebook4"><a href="https://2018.igem.org/Team:CCU_Taiwan/DATABASE">DATABASE</a></li>
+
<a href="https://2018.igem.org/Team:CCU_Taiwan/Experiments"><li class="list" id="notebook4">Experiments</li></a>
 +
<a href="https://2018.igem.org/Team:CCU_Taiwan/Materials"><li class="list" id="notebook5">Materials</li></a>
 
                         </ul>
 
                         </ul>
 
                     </li>
 
                     </li>
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             </div>
 
             </div>
 
         </nav>
 
         </nav>
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     </header>
 
     </header>
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</body>
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<div class="indicator">
  
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<div class="column full_size judges-will-not-evaluate">
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<div class="pointerProject" id="2"><a href="#ca1">laccase 1</a></div>
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<h3>★  ALERT! </h3>
+
<div class="pointerProject" id="3"><a href="#ca2">peroxidase 18</a></div>
−
<p>This page is used by the judges to evaluate your team for the <a href="https://2018.igem.org/Judging/Medals">medal criterion</a> or <a href="https://2018.igem.org/Judging/Awards"> award listed below</a>. </p>
+
<div class="pointerProject" id="4"><a href="#ca3">peroxidase 16</a></div>
−
<p> Delete this box in order to be evaluated for this medal criterion and/or award. See more information at <a href="https://2018.igem.org/Judging/Pages_for_Awards"> Instructions for Pages for awards</a>.</p>
+
</div>
−
</div>
+
 +
<div class="backgroundParts">
 +
<div class="photoComposite"><h1 class="bigtitle">COMPOSITE PART<h1></div>
 +
<div class="content">
 +
<p class="second" id="ca1"><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2809011">BBa_K2809011</a><br>
 +
laccase 1
 +
</p>
 +
<p class="description">&emsp;&emsp;We use pGAPZ A vector from invitrogen and insert this part to construct our vector. We constructed this part by combining BBa_K2809001, BBa_K2809002 and BBa_K2809010.<br>
 +
  &emsp;&emsp;This part contain a Kozak sequence for initiating and enhancing translation, Prepro-alpha-factor as a signal peptide for secretion, codon-optimized version of laccase gene sequence, HA tag for Western Blot protein detection and 6xHis-tag for protein purification.
 +
</p><br>
  
 +
        <div id="Composite1" class="polaroid">
 +
                  <img src="https://static.igem.org/mediawiki/parts/5/59/T--CCU_Taiwan--111.png" style="width:100%">
 +
                  <div class="container">
 +
                    <p>Figure 1: The schematic structure of gene design- Lac1.</p>
 +
                  </div>
 +
                </div><br><br>
 +
<p class="description">&emsp;&emsp;It will produce the mature laccase 1 without Prepro-alpha-factor, and the molecular weight of enzyme will be 63.65 kilodaltons.
 +
</p><br>
 +
        <div id="Composite1-2" class="polaroid">
 +
                  <img src="https://static.igem.org/mediawiki/parts/2/2f/T--CCU_Taiwan--_lac1_western_blot.jpeg" style="width:100%">
 +
                  <div class="container">
 +
                    <p>Figure 2: The result of western blot— laccase 1.</p>
 +
                  </div>
 +
                </div>
 +
<br><br><br>
  
−
<div class="clear"></div>
+
<p class="second" id="ca2"><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2809021">BBa_K2809021</a><br>
 +
peroxidase 18
 +
</p>
 +
<p class="description">&emsp;&emsp;We use pGAPZ A vector from invitrogen and insert this part to construct our vector. We constructed this part by combining BBa_K2809001, BBa_K2809002 and BBa_K2809020.<br>
 +
  This part contain a Kozak sequence for initiating and enhancing translation, Prepro-alpha-factor as a signal peptide for secretion, codon-optimized version of peroxidase 18 gene sequence, V5-tag for Western Blot protein detection and 6xHis-tag for protein purification.
 +
 +
</p><br><br>
 +
        <div id="Composite2" class="polaroid">
 +
                  <img src="https://static.igem.org/mediawiki/parts/9/9e/T--CCU_Taiwan--114.png" style="width:100%">
 +
                  <div class="container">
 +
                    <p>Figure 3: The schematic structure of gene design- Px18.</p>
 +
                  </div>
 +
                </div><br><br>
 +
<p class="description">&emsp;&emsp;It will produce the mature peroxidase 16 without Prepro-alpha-factor, and the molecular weight of enzyme should be 35.29 kilodaltons. However, because of protein N-linked glycosylation, the molecular weight of enzyme will be about 37~45 kilodaltons according to the Western blot result.
 +
</p><br>
 +
        <div id="Composite2-2" class="polaroid">
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                  <img src="https://static.igem.org/mediawiki/parts/6/62/T--CCU_Taiwan--_px16_western_blot-p.png" style="width:100%">
 +
                  <div class="container">
 +
                    <p>Figure 4: The result of Western blot— peroxidase 16.</p>
 +
                  </div>
 +
                </div><br><br
 +
<br>
 +
<p class="second" id="ca3"><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2809031">BBa_K2809031</a><br>
 +
peroxidase 16
 +
</p>
 +
<p class="description">&emsp;&emsp;We use pGAPZ A vector from invitrogen and insert this part to construct our vector. We constructed this part by combining BBa_K2809001, BBa_K2809002 and BBa_K2809030.<br>
 +
  This part contain a Kozak sequence for initiating and enhancing translation, Prepro-alpha-factor as a signal peptide for secretion, codon-optimized version of peroxidase 16 gene sequence, FLAG-tag for Western Blot protein detection and 6xHis-tag for protein purification.
  
 +
</p><br><br>         <div id="Composite3" class="polaroid">
 +
                  <img src="https://static.igem.org/mediawiki/parts/1/1e/T--CCU_Taiwan--117.png" style="width:100%">
 +
                  <div class="container">
 +
                    <p>Figure 5: The schematic structure of gene design- Px16.</p>
 +
                  </div>
 +
                </div><br><br>
 +
<p class="description">&emsp;&emsp;We use pGAPZ A vector from invitrogen and insert this part to construct our vector. We constructed this part by combining BBa_K2809001, BBa_K2809002 and BBa_K2809030.<br>
 +
  This part contain a Kozak sequence for initiating and enhancing translation, Prepro-alpha-factor as a signal peptide for secretion, codon-optimized version of peroxidase 16 gene sequence, FLAG-tag for Western Blot protein detection and 6xHis-tag for protein purification.
  
 +
</p><br><br>
 +
        <div id="Composite3-2" class="polaroid">
 +
                  <img src="https://static.igem.org/mediawiki/parts/4/4b/T--CCU_Taiwan--_px18_western_blot-s.png" style="width:100%">
 +
                  <div class="container">
 +
                    <p>Figure 6: The result of Western blot— peroxidase 18(soup).</p>
 +
                  </div>
 +
                </div>
 +
<br>
 +
<br>
 +
        <div id="Composite3-3" class="polaroid">
 +
                  <img src="https://static.igem.org/mediawiki/parts/3/37/T--CCU_Taiwan--_px18_western_blot-p.png" style="width:100%">
 +
                  <div class="container">
 +
                    <p>Figure 7: The result of Western blot— peroxidase 18(protein extraction from pellet).</p>
 +
                  </div>
 +
                </div>
  
  
  
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<div class="column full_size">
 
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<h1>Composite Parts</h1>
 
  
  
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<p>
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<br><br><br>
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A composite part is a functional unit of DNA consisting of two or more basic parts assembled together. <a href="http://parts.igem.org/wiki/index.php/Part:BBa_I13507">BBa_I13507</a> is an example of a composite part, consisting of an RBS, a protein coding region for a red fluorescent protein, and a terminator.
+
              <p class="second">Part construction in pGAPZ A:</p><br>
 +
<div class="row">
 +
        <div id="Composite4" class="polaroid">
 +
                  <img src="https://static.igem.org/mediawiki/parts/f/f2/T--CCU_Taiwan--_pGAPZ_A_Lac1%2C_pGAPZ_A_Px16%2C_pGAPZ_A_Px18_Colony_PCR.jpeg" style="width:100%">
 +
                  <div class="container">
 +
                    <p>Figure 4: PCR result.<br> Well1: Marker/ Well2, 3, 4: Lac1 Colony 1, 2, 3/ Well5, 6, 7: Px16 Colony 1, 2, 3/ Well8, 9, 10: Px18 Colony 1, 2, 3/ Well11: pGAPZ A Colony
 +
<br>
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Insert long after PCR — pGAPZ A_Px16 :1377 bp/ pGAPZ A_Px18 :1359 bp/ pGAPZ A_Lac1 :2124bp
 
</p>
 
</p>
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                  </div>
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<p>New composite BioBrick devices can be made by combining existing BioBrick Parts (like Inverters, Amplifiers, Smell Generators, Protein Balloon Generators, Senders, Receivers, Actuators, and so on).</p>
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                </div>
 +
        <div id="Composite5" class="polaroid">
 +
                  <img src="https://static.igem.org/mediawiki/parts/a/ae/T--CCU_Taiwan--_pSB1C3_Lac1%2C_pSB1C3_Px16%2C_pSB1C3_Px18_Miniprep_and_digest_with_EcoRI%2CAgeI.jpeg" style="width:100%">
 +
                  <div class="container">
 +
                    <p>Figure 5: Miniprep plasmid digest by EcoRI, PstI.<br>Well 1: Marker/ Well2, 3, 4: Plasmid of Lac1 colony1, 2, 3 after EcoRI, AgeI digest/ Well5, 6, 7: Plasmid of Px16 colony1, 2, 3 after EcoRI, AgeI digest/ Well8, 9, 10: Plasmid of Px18 colony1, 2, 3 after EcoRI, AgeI digest
 +
<br>
 +
Vector long after digest — pGAPZ A: 2884bp
 +
<br>
 +
Insert long after digest — Px16: 1295bp/ Px18: 1277bp/ Lac1: 2042bp
 +
</p>
 +
                  </div>
 +
                </div>
 
</div>
 
</div>
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<br>
 +
<br>
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<br>
  
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<div class="column full_size">
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<div class="highlight decoration_background">
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</div>
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<h3>Note</h3>
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</body>
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<p>This page should list all the composite parts your team has made during your project. You must add all characterization information for your parts on the Registry. You should not put characterization information on this page. Remember judges will only look at the first part in the list for the Best Composite Part award, so put your best part first!</p>
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<footer class="footer">
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</div>
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            <a href="http://huitong.com.tw">
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</div>
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              <img  id="BSlogo" src="https://static.igem.org/mediawiki/2018/0/07/T--CCU_Taiwan--CCUP1.png" align="bottom">
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            </a>
  
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            <a href="https://www.facebook.com/ccuigemteam/?ref=br_rs">
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            <img  id="CCUTEAMlogo" src="https://static.igem.org/mediawiki/2018/e/e3/T--CCU_Taiwan--CCUP2.png" align="bottom">
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            </a>
  
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            <a href="http://ewww.ccu.edu.tw/">
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<div class="column full_size">
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            <img  id="CCUlogo" src="https://static.igem.org/mediawiki/2018/8/89/T--CCU_Taiwan--CCUP3.png" align="bottom">
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<h3>Best Composite Part Special Prize</h3>
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            </a>
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</footer>
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<p>To be eligible for this award, this part must adhere to <a href="http://parts.igem.org/DNA_Submission">Registry sample submission guidelines</a> and have been sent to the Registry of Standard Biological Parts. If you have a part you wish to nominate your team for this <a href="https://2018.igem.org/Judging/Awards">special prize</a>, make sure you add your part number to your <a href="https://2018.igem.org/Judging/Judging_Form">judging form</a> and delete the box at the top of this page.
+
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<br><br>
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<b>Please note:</b> Judges will only look at the first part number you list, so please only enter ONE (1) part number in the judging form for this prize. </p>
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</div>
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</html>
 
</html>

Latest revision as of 08:47, 1 December 2018

COMPOSITE PART

BBa_K2809011
laccase 1

  We use pGAPZ A vector from invitrogen and insert this part to construct our vector. We constructed this part by combining BBa_K2809001, BBa_K2809002 and BBa_K2809010.
  This part contain a Kozak sequence for initiating and enhancing translation, Prepro-alpha-factor as a signal peptide for secretion, codon-optimized version of laccase gene sequence, HA tag for Western Blot protein detection and 6xHis-tag for protein purification.


Figure 1: The schematic structure of gene design- Lac1.



  It will produce the mature laccase 1 without Prepro-alpha-factor, and the molecular weight of enzyme will be 63.65 kilodaltons.


Figure 2: The result of western blot— laccase 1.




BBa_K2809021
peroxidase 18

  We use pGAPZ A vector from invitrogen and insert this part to construct our vector. We constructed this part by combining BBa_K2809001, BBa_K2809002 and BBa_K2809020.
This part contain a Kozak sequence for initiating and enhancing translation, Prepro-alpha-factor as a signal peptide for secretion, codon-optimized version of peroxidase 18 gene sequence, V5-tag for Western Blot protein detection and 6xHis-tag for protein purification.



Figure 3: The schematic structure of gene design- Px18.



  It will produce the mature peroxidase 16 without Prepro-alpha-factor, and the molecular weight of enzyme should be 35.29 kilodaltons. However, because of protein N-linked glycosylation, the molecular weight of enzyme will be about 37~45 kilodaltons according to the Western blot result.


Figure 4: The result of Western blot— peroxidase 16.



BBa_K2809031
peroxidase 16

  We use pGAPZ A vector from invitrogen and insert this part to construct our vector. We constructed this part by combining BBa_K2809001, BBa_K2809002 and BBa_K2809030.
This part contain a Kozak sequence for initiating and enhancing translation, Prepro-alpha-factor as a signal peptide for secretion, codon-optimized version of peroxidase 16 gene sequence, FLAG-tag for Western Blot protein detection and 6xHis-tag for protein purification.



Figure 5: The schematic structure of gene design- Px16.



  We use pGAPZ A vector from invitrogen and insert this part to construct our vector. We constructed this part by combining BBa_K2809001, BBa_K2809002 and BBa_K2809030.
This part contain a Kozak sequence for initiating and enhancing translation, Prepro-alpha-factor as a signal peptide for secretion, codon-optimized version of peroxidase 16 gene sequence, FLAG-tag for Western Blot protein detection and 6xHis-tag for protein purification.



Figure 6: The result of Western blot— peroxidase 18(soup).



Figure 7: The result of Western blot— peroxidase 18(protein extraction from pellet).




Part construction in pGAPZ A:


Figure 4: PCR result.
Well1: Marker/ Well2, 3, 4: Lac1 Colony 1, 2, 3/ Well5, 6, 7: Px16 Colony 1, 2, 3/ Well8, 9, 10: Px18 Colony 1, 2, 3/ Well11: pGAPZ A Colony
Insert long after PCR — pGAPZ A_Px16 :1377 bp/ pGAPZ A_Px18 :1359 bp/ pGAPZ A_Lac1 :2124bp

Figure 5: Miniprep plasmid digest by EcoRI, PstI.
Well 1: Marker/ Well2, 3, 4: Plasmid of Lac1 colony1, 2, 3 after EcoRI, AgeI digest/ Well5, 6, 7: Plasmid of Px16 colony1, 2, 3 after EcoRI, AgeI digest/ Well8, 9, 10: Plasmid of Px18 colony1, 2, 3 after EcoRI, AgeI digest
Vector long after digest — pGAPZ A: 2884bp
Insert long after digest — Px16: 1295bp/ Px18: 1277bp/ Lac1: 2042bp