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PartsOverview.html
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<!DOCTYPE html>
<html lang="en">
<head>
<meta charset="UTF-8">
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<img class="background" src="https://2021.igem.org/wiki/images/c/cf/T--CPU_CHINA--Parts--background.png" alt="">
<h1 id="title">Parts Overview</h1>
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<nav>
<div class="catalogue-title">Parts Overview</div>
<ul class="catalogue">
<li class="num">
<a href="#section1" title="BASIC PARTS">BASIC PARTS</a>
</li>
<li class="num">
<a href="#section2" title="COMPOSITE PARTS">COMPOSITE PARTS</a>
</li>
<li class="num">
<a href="#section3" title="PART COLLECTION">PART COLLECTION</a>
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<img src="https://2021.igem.org/wiki/images/b/b2/T--CPU_CHINA--experimentalize.png" alt="">
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<div id="detail" class="clearfix">
<p><span> </span><span>This year, among the 30 parts designed, 19 of them are basic parts and the rest are
composite parts. All of the parts are designed on our own, and most of them have been tested to
ensure their reliability. </span></p>
<p><span> </span><strong><span><a href="#">Click here</a> </span></strong><span>
to learn more about our project to understand the purpose of each parts! Click the part number to
see detailed information of the parts we design!</span></p>
<div class="section" id="section1">
<h2 id='basic-parts'><span>Basic Parts</span></h2>
<p><span> </span><span>Among the 19 basic parts designed this year by ourselves, <a href="http://parts.igem.org/Part:BBa_K3853008">BBa_K3853008</a> is our
favorite one, which is the coding sequence of manganese peroxidase derived from
</span><em><span>Phanerochaete chrysosporium</span></em><span>. The translated product of this part
enables our multi-enzyme complex to degrade polyethylene efficiently. Besides, many other basic
parts have also been designed to construct our project.</span></p>
<figure>
<table>
<thead>
<tr>
<th><span>Favorite</span></th>
<th><span>Part Name</span></th>
<th><span>Type</span></th>
<th><span>Description</span></th>
<th><span>Designer</span></th>
<th><span>Length</span></th>
</tr>
</thead>
<tbody>
<tr>
<td> </td>
<td><span><a href="http://parts.igem.org/Part:BBa_K3853000">BBa_K3853000</a></span></td>
<td><span>coding</span></td>
<td><span>MnP</span></td>
<td><span>Peng Luo</span></td>
<td><span>1074</span></td>
</tr>
<tr>
<td> </td>
<td><span><a href="http://parts.igem.org/Part:BBa_K3853001">BBa_K3853001</a></span></td>
<td><span>coding</span></td>
<td><span>AAO</span></td>
<td><span>Peng Luo</span></td>
<td><span>1719</span></td>
</tr>
<tr>
<td> </td>
<td><span><a href="http://parts.igem.org/Part:BBa_K3853002">BBa_K3853002</a></span></td>
<td><span>coding</span></td>
<td><span>HFB1</span></td>
<td><span>Peng Luo</span></td>
<td><span>246</span></td>
</tr>
<tr>
<td> </td>
<td><span><a href="http://parts.igem.org/Part:BBa_K3853003">BBa_K3853003</a></span></td>
<td><span>coding</span></td>
<td><span>dCas9</span></td>
<td><span>GongYu Liu</span></td>
<td><span>4104</span></td>
</tr>
<tr>
<td> </td>
<td><span><a href="http://parts.igem.org/Part:BBa_K3853004">BBa_K3853004</a></span></td>
<td><span>coding</span></td>
<td><span>SpyTag</span></td>
<td><span>Peng Luo</span></td>
<td><span>39</span></td>
</tr>
<tr>
<td> </td>
<td><span><a href="http://parts.igem.org/Part:BBa_K3853001">BBa_K3853005</a></span></td>
<td><span>coding</span></td>
<td><span>SpyCatcher</span></td>
<td><span>Peng Luo</span></td>
<td><span>351</span></td>
</tr>
<tr>
<td> </td>
<td><span><a href="http://parts.igem.org/Part:BBa_K3853006">BBa_K3853006</a></span></td>
<td><span>DNA</span></td>
<td><span>dsDNA</span></td>
<td><span>GongYu Liu</span></td>
<td><span>189</span></td>
</tr>
<tr>
<td> </td>
<td><span><a href="http://parts.igem.org/Part:BBa_K3853007">BBa_K3853007</a></span></td>
<td><span>DNA</span></td>
<td><span>gRNA scaffold</span></td>
<td><span>GongYu Liu</span></td>
<td><span>80</span></td>
</tr>
<tr>
<td><span>♥</span></td>
<td><span><a href="http://parts.igem.org/Part:BBa_K3853008">BBa_K3853008</a></span></td>
<td><span>coding</span></td>
<td><span>his-tag-SpyTag-MnP</span></td>
<td><span>Peng Luo</span></td>
<td><span>1248</span></td>
</tr>
<tr>
<td> </td>
<td><span><a href="http://parts.igem.org/Part:BBa_K3853009">BBa_K3853009</a></span></td>
<td><span>coding</span></td>
<td><span>his-tag-SpyTag-AAO</span></td>
<td><span>Peng Luo</span></td>
<td><span>1893</span></td>
</tr>
<tr>
<td> </td>
<td><span><a href="http://parts.igem.org/Part:BBa_K3853010">BBa_K3853010</a></span></td>
<td><span>coding</span></td>
<td><span>his-tag-SpyTag-HFB1</span></td>
<td><span>Peng Luo</span></td>
<td><span>420</span></td>
</tr>
<tr>
<td> </td>
<td><span><a href="http://parts.igem.org/Part:BBa_K3853011">BBa_K3853011</a></span></td>
<td><span>coding</span></td>
<td><span>dCas9-SpyCatcher</span></td>
<td><span>GongYu Liu</span></td>
<td><span>4530</span></td>
</tr>
<tr>
<td> </td>
<td><span><a href="http://parts.igem.org/Part:BBa_K3853012">BBa_K3853012</a></span></td>
<td><span>coding</span></td>
<td><span>sfGFP</span></td>
<td><span>Peng Luo</span></td>
<td><span>720</span></td>
</tr>
<tr>
<td> </td>
<td><span><a href="http://parts.igem.org/Part:BBa_K3853013">BBa_K3853013</a></span></td>
<td><span>coding</span></td>
<td><span>MnP(E74P)</span></td>
<td><span>RuYi Ma</span></td>
<td><span>1074</span></td>
</tr>
<tr>
<td> </td>
<td><span><a href="http://parts.igem.org/Part:BBa_K3853014">BBa_K3853014</a></span></td>
<td><span>coding</span></td>
<td><span>MnP(E74M)</span></td>
<td><span>RuYi Ma</span></td>
<td><span>1074</span></td>
</tr>
<tr>
<td> </td>
<td><span><a href="http://parts.igem.org/Part:BBa_K3853015">BBa_K3853015</a></span></td>
<td><span>coding</span></td>
<td><span>MnP(D182T)</span></td>
<td><span>RuYi Ma</span></td>
<td><span>1074</span></td>
</tr>
<tr>
<td> </td>
<td><span><a href="http://parts.igem.org/Part:BBa_K3853016">BBa_K3853016</a></span></td>
<td><span>coding</span></td>
<td><span>MnP(S232P)</span></td>
<td><span>RuYi Ma</span></td>
<td><span>1074</span></td>
</tr>
<tr>
<td> </td>
<td><span><a href="http://parts.igem.org/Part:BBa_K3853017">BBa_K3853017</a></span></td>
<td><span>coding</span></td>
<td><span>MnP(E74L)</span></td>
<td><span>RuYi Ma</span></td>
<td><span>1074</span></td>
</tr>
<tr>
<td> </td>
<td><span><a href="http://parts.igem.org/Part:BBa_K3853018">BBa_K3853018</a></span></td>
<td><span>coding</span></td>
<td><span>MnP(S78P)</span></td>
<td><span>RuYi Ma</span></td>
<td><span>1074</span></td>
</tr>
<tr>
<td> </td>
<td><span><a href="http://parts.igem.org/Part:BBa_K3853019">BBa_K3853019</a></span></td>
<td><span>coding</span></td>
<td><span>his-tag</span></td>
<td><span>Peng Luo</span></td>
<td><span>1074</span></td>
</tr>
</tbody>
</table>
</figure>
<p> </p>
</div>
<div class="section" id="section2">
<h2 id='composite-parts'><span>Composite Parts</span></h2>
<p><span> </span><span>A series of composite parts have been designed by us this year, and most of them
are
used for the assembly of the PE-degrading complex in our project throughout the season. </span>
</p>
<p><span> </span><span>To achieve this assembly, we designed fusion enzymes and proteins containing
SpyTag
and fusion deactivated CRISPR-associated protein 9 (dCas9) containing SpyCatcher, enabling the
incubation-induced connection of multi-protein complexes via covalent bond formation mediated by
the
two Spy domains. Subsequently, these multi-protein complexes could anchor to designated
positions on
the designed double-stranded DNA (dsDNA) template through the guidance of dCas9-combined
sgRNAs.</span></p>
<p><span> </span><span>Our favorite composite part is <a href="http://parts.igem.org/Part:BBa_K3853051">BBa_K3853051</a> , which contains the sequence of
manganese peroxidase (MnP) for polyethylene degradation, and the sequence of SpyTag for
multi-domain
complex formation, as well as sequences of AOX1 promoter and AOX1 terminator, enabling its
efficient
inducible expression in </span><em><span>Pichia pastoris</span></em><span>.</span></p>
<figure>
<table>
<thead>
<tr>
<th><span>Favorite</span></th>
<th><span>Part Name</span></th>
<th><span>Type</span></th>
<th><span>Description</span></th>
<th><span>Designer</span></th>
<th><span>Length</span></th>
</tr>
</thead>
<tbody>
<tr>
<td> </td>
<td><span><a href="http://parts.igem.org/Part:BBa_K3853050">BBa_K3853050</a></span></td>
<td><span>composite</span></td>
<td><span>P</span><sub><span>AOX1</span></sub><span>-α-factor-sfGFP-AOX1
Terminator</span>
</td>
<td><span>Peng Luo</span></td>
<td><span>2136</span></td>
</tr>
<tr>
<td><span>♥</span></td>
<td><span><a href="http://parts.igem.org/Part:BBa_K3853051">BBa_K3853051</a></span></td>
<td><span>composite</span></td>
<td><span>P</span><sub><span>AOX1</span></sub><span>-α-factor-his-tag-SpyTag-MnP-AOX1
Terminator</span></td>
<td><span>Peng Luo</span></td>
<td><span>2664</span></td>
</tr>
<tr>
<td> </td>
<td><span><a href="http://parts.igem.org/Part:BBa_K3853053">BBa_K3853053</a></span></td>
<td><span>composite</span></td>
<td><span>P</span><sub><span>AOX1</span></sub><span>-α-factor-his-tag-SpyTag-AAO-AOX1
Terminator</span></td>
<td><span>Peng Luo</span></td>
<td><span>3309</span></td>
</tr>
<tr>
<td> </td>
<td><span><a href="http://parts.igem.org/Part:BBa_K3853054">BBa_K3853054</a></span></td>
<td><span>composite</span></td>
<td><span>P</span><sub><span>AOX1</span></sub><span>-α-factor-his-tag-SpyTag-HFB1-AOX1
Terminator</span></td>
<td><span>Peng Luo</span></td>
<td><span>1836</span></td>
</tr>
<tr>
<td> </td>
<td><span><a href="http://parts.igem.org/Part:BBa_K3853055">BBa_K3853055</a></span></td>
<td><span>composite</span></td>
<td><span>R0085-C0012-B2002-his-tag-dCas9-SpyCatcher-B0012</span></td>
<td><span>Peng Luo</span></td>
<td><span>5758</span></td>
</tr>
<tr>
<td> </td>
<td><span><a href="http://parts.igem.org/Part:BBa_K3853056">BBa_K3853056</a></span></td>
<td><span>composite</span></td>
<td><span>P</span><sub><span>AOX1</span></sub><span>-α-factor-his-tag-MnP(E74P)-AOX1
Terminator</span></td>
<td><span>Peng Luo</span></td>
<td><span>2508</span></td>
</tr>
<tr>
<td> </td>
<td><span><a href="http://parts.igem.org/Part:BBa_K3853057">BBa_K3853057</a></span></td>
<td><span>composite</span></td>
<td><span>P</span><sub><span>AOX1</span></sub><span>-α-factor-his-tag-MnP(E74M)-AOX1
Terminator</span></td>
<td><span>Peng Luo</span></td>
<td><span>2508</span></td>
</tr>
<tr>
<td> </td>
<td><span><a href="http://parts.igem.org/Part:BBa_K3853058">BBa_K3853058</a></span></td>
<td><span>composite</span></td>
<td><span>P</span><sub><span>AOX1</span></sub><span>-α-factor-his-tag-MnP(D182T)-AOX1
Terminator</span></td>
<td><span>Peng Luo</span></td>
<td><span>2508</span></td>
</tr>
<tr>
<td> </td>
<td><span><a href="http://parts.igem.org/Part:BBa_K3853059">BBa_K3853059</a></span></td>
<td><span>composite</span></td>
<td><span>P</span><sub><span>AOX1</span></sub><span>-α-factor-his-tag-MnP(S232P)-AOX1
Terminator</span></td>
<td><span>Peng Luo</span></td>
<td><span>2508</span></td>
</tr>
<tr>
<td> </td>
<td><span><a href="http://parts.igem.org/Part:BBa_K3853060">BBa_K3853060</a></span></td>
<td><span>composite</span></td>
<td><span>P</span><sub><span>AOX1</span></sub><span>-α-factor-his-tag-MnP(E74L)-AOX1
Terminator</span></td>
<td><span>Peng Luo</span></td>
<td><span>2508</span></td>
</tr>
<tr>
<td> </td>
<td><span><a href="http://parts.igem.org/Part:BBa_K3853061">BBa_K3853061</a></span></td>
<td><span>composite</span></td>
<td><span>P</span><sub><span>AOX1</span></sub><span>-α-factor-his-tag-MnP(S78P)-AOX1
Terminator</span></td>
<td><span>Peng Luo</span></td>
<td><span>2508</span></td>
</tr>
</tbody>
</table>
</figure>
<p> </p>
</div>
<div class="section" id="section3">
<h2 id='part-collection'><strong><span>Part Collection</span></strong></h2>
<p><span> </span><span>This year, we developed a part collection related to the CRSIPR/dCas9 anchoring
system. By introducing SpyTag into the N-terminus of different proteins to develop fusion
proteins,
their connection to dCas9-SpyCatcher and the following sgRNA-guided anchoring on dsDNA can be
realized. The type, quantity and proportion of anchoring components will remarkably affect their
individual function, as well as the overall performance of the complex. It depends on how the
system
designer would like to arrange the dCas9-binding sequence on dsDNA, and how the assembly process
is
conducted through specific order for acquiring expected component array of the complex.</span>
</p>
<p><span> </span><span> For our part collection, we characterized three different fusion proteins, i.e.
SpyTag-MnP, SpyTag-AAO, SpyTag-HFB1, and uploaded their complete and detailed characterization
data
to the iGEM part registry. We hope these organized information can be instructive for the design
and
charaterization of upcoming teams who wish to adopt these protein-nucleic-acid complex.</span>
</p>
<figure>
<table>
<thead>
<tr>
<th><span>Peptide segment</span></th>
<th><span>Fusion</span></th>
<th><span>Part Name</span></th>
<th><span>Type</span></th>
<th><span>Description</span></th>
<th><span>Designer</span></th>
<th><span>Length</span></th>
</tr>
</thead>
<tbody>
<tr>
<td rowspan="3"><span>SpyTag</span></td>
<td rowspan="3"><span>N-terminal</span></td>
<td><span><a href="http://parts.igem.org/Part:BBa_K3853051">BBa_K3853051</a></span></td>
<td><span>composite</span></td>
<td><span>P</span><sub><span>AOX1</span></sub><span>-α-factor-his-tag-SpyTag-MnP-AOX1
Terminator</span></td>
<td><span>Peng Luo</span></td>
<td><span>2664</span></td>
</tr>
<tr>
<td><span><a href="http://parts.igem.org/Part:BBa_K3853053">BBa_K3853053</a></span></td>
<td><span>composite</span></td>
<td><span>P</span><sub><span>AOX1</span></sub><span>-α-factor-his-tag-SpyTag-AAO-AOX1
Terminator</span></td>
<td><span>Peng Luo</span></td>
<td><span>3309</span></td>
</tr>
<tr>
<td><span><a href="http://parts.igem.org/Part:BBa_K3853054">BBa_K3853054</a></span></td>
<td><span>composite</span></td>
<td><span>P</span><sub><span>AOX1</span></sub><span>-α-factor-his-tag-SpyTag-HFB1-AOX1
Terminator</span></td>
<td><span>Peng Luo</span></td>
<td><span>1836</span></td>
</tr>
</tbody>
</table>
</figure>
</div>
</div>
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