Team:Tianjin/Data

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                 <a href="https://2011.igem.org/Team:Tianjin/Data">
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This page shows a list of all the parts that we have made or used in the project. Click on each link to see more details about that part on the <span  style="color:#7f3f00"><b><a href="http://partsregistry.org/Main_Page">Registry of Standard Biological Parts</a></b></span>.</br></br>
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As our project focus on tor2 gene coming from yeast and contains common restriction sites like EcoRI, NotI and SpeI, the stantard Plasmid Backbone and BioBrick prefix/suffix is no longer appropriate. Furthermore, the size of 7800bp makes it pretty too expensive and inconvenient for chemical synthesis and codon optimization to avoid certain restriction sites. To match with the core gene and protein, a series of parts also give up standard prefix/suffix and backbones. We have used at least 3 kinds of plasmids, including blunt-end vectors to Yeast Surface Display Vector. However, based on the splendid standard plasmid backbone pSB1C3, which has a small size and simple MCS for easy-operation, we modified several vectors, which also render functionality and ease-of-use of promoter parts.
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                According to the reconstruction of TOR (Target of Rapamycin) pathway, our project is aimed at increase the tolerance of Saccharomyces cerevisiae. E. coli (Top 10), Saccharomyces cerevisiae (BY4742), and vectors based on them are frequently used throughout our experiments. Since we are intended to regulate the major signaling transduction network, expressions of hundreds of genes and a certain number of pathways will be affected, which may give rise to anxieties of safety regarding researcher and environment. Thus a whole set of experiment guidelines and safety rules are followed in every aspect of our project to minimize every possible threaten. Inside the lab, protective clothing, gloves and masks are required while performing experiments with latent danger such as gel cutting under UV, using toxic and volatile reagents such as phenol and chloroform in special areas. All researchers are required to clean up their working area and put back the experimental equipments. Furthermore, protocols for preservation and operation of common E. coli and yeast strains as well as the vectors are strictly adhered to, and make sure no pollutants are left unattended or take away from the lab, to prevent any toxicant or gene form spreading to the public or contaminating the environment.
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1.  Inducible pCIT2 – Mut-TOR2 - Terminator, BBa_K563061: Promoter of CIT2 gene would be binding with transcription factor RTG1/3 at the presence of FAP. Then our reconstructed Mut-TOR2 would be expressed and forms in Mut-TORC1 which is not allergic to FAP. The accumulation of Mut-TORC1 will help the intracellular phosphorylation back to normal state.</br></br>
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2. pYE- Reporter, BBa_K563040 ~ BBa_K563046: We have successfully construct vectors, which coding bright fluorescent protein and Luciferase and beta-galactosidase in yeast cell. This series of BioBricks provide a convenient method for the measurement and detection of gene expression and terminate the unsuccessful expression of fluorescent protein in yeast cell.</br></br>
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3.  Modified Plasmid Backbone – pSBY11, BBa_K563051: We reconstruct the Yeast Vector pYD through the combination with pSB1A11. According to specific primers, we amplify the yeast elements (CEN4/ARS6, TRP1 orf and terminator region) by PCR, and then insert the fragment into pSB1A11 by single restriction site PstI. In this way, a modified vector that combines pYD and pSB1A11 together(pSBY1) for large fragment transformation between yeast and E. coli is built.</br></br>
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      None of our new BioBrick would raise safety uneasiness. Following BioBrick requirements, we provide standard biological parts in standard BioBrick shipping plasmid, which can be mainly divided into four types: operators, reporters, target protein mutants and modified vectors. They are primarily involved in the phosphorylation of intracellular protein, thus wouldn’t cause any pollution or contaminant.
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    Yes. Bio-safety group in our chemical engineering institute would operate safety checks of our experiment progress, monitor and evaluate the project periodically, at least once a semester, to make sure our operation won’t bring about safety issues and no pathogen would flow into the public. Besides, all the waste reagents should be specifically collected rather than directly throwing away in trash cans. iGEM laboratory construction of Tianjin University is always keep in paces with policies of state government and WHO, to take responsibility for safety of researchers as well as the public.
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        The most valid way to attract people to focus on dealing with safety issues is to remind them of the significance of maintaining safe experimental and living environment, and to warn them of the potential hazard if biochemical crisis become out of control. As undergraduates, we should spare no effort to broadcast the opinion that bio-safety is not far from daily life, no matter whom you are and where you live. It truly exists in each drop of water we drink, and each breath of air we take in. It will be too late to stop it from spreading once contaminants flow into the public, yet it is never too late to prevent the crisis in advance.
 
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<th style="background:#ffbf7f;">&nbsp;</th>
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<th style="background:#ffbf7f;">Name</th>
 +
<th style="background:#ffbf7f;">Type</th>
 +
<th style="background:#ffbf7f;">Description</th>
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<th style="background:#ffbf7f;">Designer</th>
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<th style="background:#ffbf7f;">Length</th>
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</tr>
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<tr>
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<td style="background:#ffdfbf;">1</td>
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<td style="background:#ffdfbf;"><a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K563000">BBa_K563000</a></td>
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<td style="background:#ffdfbf;">Regulatory</td>
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<td style="background:#ffdfbf;">Promotor of HIS4</td>
 +
<td style="background:#ffdfbf;">Boxuan Zeng</td>
 +
<td style="background:#ffdfbf;">508</td>
 +
</tr>
 +
<tr>
 +
<td>2</td>
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<td><a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K563001">BBa_K563001</a></td>
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<td>Regulatory</td>
 +
<td>Promoter of DAL7</td>
 +
<td>Boxuan Zeng</td>
 +
<td>498</td>
 +
</tr>
 +
<tr>
 +
<td style="background:#ffdfbf;">3</td>
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<td style="background:#ffdfbf;"><a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K563002">BBa_K563002</a></td>
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<td style="background:#ffdfbf;">Regulatory</td>
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<td style="background:#ffdfbf;">Promoter of ILV1</td>
 +
<td style="background:#ffdfbf;">Boxuan Zeng</td>
 +
<td style="background:#ffdfbf;">509</td>
 +
</tr>
 +
<tr>
 +
<td>4</td>
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<td><a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K563003">BBa_K563003</a></td>
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<td>Regulatory</td>
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<td>Promoter of CIT2</td>
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<td>Boxuan Zeng</td>
 +
<td>497</td>
 +
</tr>
 +
<tr>
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<td style="background:#ffdfbf;">5</td>
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<td style="background:#ffdfbf;"><a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K563004">BBa_K563004</a></td>
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<td style="background:#ffdfbf;">Regulatory</td>
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<td style="background:#ffdfbf;">Promoter of TEF1</td>
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<td style="background:#ffdfbf;">Boxuan Zeng</td>
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<td style="background:#ffdfbf;">450</td>
 +
</tr>
 +
<tr>
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<td>6</td>
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<td><a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K563010">BBa_K563010</a></td>
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<td>Coding</td>
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<td>Original TOR</td>
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<td>Boxuan Zeng</td>
 +
<td>8118</td>
 +
</tr>
 +
<tr>
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<td style="background:#ffdfbf;">7</td>
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<td style="background:#ffdfbf;"><a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K563011">BBa_K563011</a></td>
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<td style="background:#ffdfbf;">Coding</td>
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<td style="background:#ffdfbf;">TOR (LP Mutation)</td>
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<td style="background:#ffdfbf;">Boxuan Zeng</td>
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<td style="background:#ffdfbf;">8118</td>
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</tr>
 +
<tr>
 +
<td>8</td>
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<td><a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K563012">BBa_K563012</a></td>
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<td>Coding</td>
 +
<td>TOR (ITMutation)</td>
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<td>Boxuan Zeng</td>
 +
<td>8118</td>
 +
</tr>
 +
<tr>
 +
<td style="background:#ffdfbf;">9</td>
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<td style="background:#ffdfbf;"><a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K563013">BBa_K563013</a></td>
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<td style="background:#ffdfbf;">Coding</td>
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<td style="background:#ffdfbf;">TOR (IT/EK Mutation)</td>
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<td style="background:#ffdfbf;">Boxuan Zeng</td>
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<td style="background:#ffdfbf;">8118</td>
 +
</tr>
 +
<tr>
 +
<td>10</td>
 +
<td><a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K563014">BBa_K563014</a></td>
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<td>Coding</td>
 +
<td>TOR (IT/VA Mutation)</td>
 +
<td>Boxuan Zeng</td>
 +
<td>8118</td>
 +
</tr>
 +
<tr>
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<td style="background:#ffdfbf;">11</td>
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<td style="background:#ffdfbf;"><a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K563015">BBa_K563015</a></td>
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<td style="background:#ffdfbf;">Coding</td>
 +
<td style="background:#ffdfbf;">TOR (EK/VA Mutation)</td>
 +
<td style="background:#ffdfbf;">Boxuan Zeng</td>
 +
<td style="background:#ffdfbf;">8118</td>
 +
</tr>
 +
<tr>
 +
<td>12</td>
 +
<td><a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K563016">BBa_K563016</a></td>
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<td>Coding</td>
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<td>TOR (IT/VA/EK Mutation)</td>
 +
<td>Boxuan Zeng</td>
 +
<td>8118</td>
 +
</tr>
 +
<tr>
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<td style="background:#ffdfbf;">13</td>
 +
<td style="background:#ffdfbf;"><a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K563030">BBa_K563030</a></td>
 +
<td style="background:#ffdfbf;">Coding</td>
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<td style="background:#ffdfbf;">pEASY-TOR01</td>
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<td style="background:#ffdfbf;">Wenbin Kuang</td>
 +
<td style="background:#ffdfbf;">3067</td>
 +
</tr>
 +
<tr>
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<td>14</td>
 +
<td><a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K563031">BBa_K563031</a></td>
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<td>Coding</td>
 +
<td>pEASY-TOR11</td>
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<td>Wenbin Kuang</td>
 +
<td>2521</td>
 +
</tr>
 +
<tr>
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<td style="background:#ffdfbf;">15</td>
 +
<td style="background:#ffdfbf;"><a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K563032">BBa_K563032</a></td>
 +
<td style="background:#ffdfbf;">Coding</td>
 +
<td style="background:#ffdfbf;">pEASY-TOR22</td>
 +
<td style="background:#ffdfbf;">Wenbin Kuang</td>
 +
<td style="background:#ffdfbf;">2709</td>
 +
</tr>
 +
<tr>
 +
<td>16</td>
 +
<td><a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K563033">BBa_K563033</a></td>
 +
<td>Coding</td>
 +
<td>pEASY-TOR33</td>
 +
<td>Wenbin Kuang</td>
 +
<td>2499</td>
 +
</tr>
 +
<tr>
 +
<td style="background:#ffdfbf;">17</td>
 +
<td style="background:#ffdfbf;"><a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K563034">BBa_K563034</a></td>
 +
<td style="background:#ffdfbf;">Coding</td>
 +
<td style="background:#ffdfbf;">pEASY-TOR22(LP Mutation)</td>
 +
<td style="background:#ffdfbf;">Mingrui Zhao</td>
 +
<td style="background:#ffdfbf;">2709</td>
 +
</tr>
 +
<tr>
 +
<td>18</td>
 +
<td><a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K563035">BBa_K563035</a></td>
 +
<td>Coding</td>
 +
<td>pEASY-TOR33(IT Mutation)</td>
 +
<td>Mingrui Zhao</td>
 +
<td>2499</td>
 +
</tr>
 +
<tr>
 +
<td style="background:#ffdfbf;">19</td>
 +
<td style="background:#ffdfbf;"><a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K563036">BBa_K563036</a></td>
 +
<td style="background:#ffdfbf;">Coding</td>
 +
<td style="background:#ffdfbf;">pEASY-TOR33(VA Mutation)</td>
 +
<td style="background:#ffdfbf;">Jing Guo</td>
 +
<td style="background:#ffdfbf;">2499</td>
 +
</tr>
 +
<tr>
 +
<td>20</td>
 +
<td><a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K563037">BBa_K563037</a></td>
 +
<td>Coding</td>
 +
<td>pEASY-TOR33(EK Mutation)</td>
 +
<td>Jing Guo</td>
 +
<td>2499</td>
 +
</tr>
 +
<tr>
 +
<td style="background:#ffdfbf;">21</td>
 +
<td style="background:#ffdfbf;"><a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K563040">BBa_K563040</a></td>
 +
<td style="background:#ffdfbf;">Reporter</td>
 +
<td style="background:#ffdfbf;">pYE-EGFP</td>
 +
<td style="background:#ffdfbf;">Bin Jia</td>
 +
<td style="background:#ffdfbf;">739</td>
 +
</tr>
 +
<tr>
 +
<td>22</td>
 +
<td><a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K563041">BBa_K563041</a></td>
 +
<td>Reporter</td>
 +
<td>pYE-ERFP</td>
 +
<td>Bin Jia</td>
 +
<td>744</td>
 +
</tr>
 +
<tr>
 +
<td style="background:#ffdfbf;">23</td>
 +
<td style="background:#ffdfbf;"><a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K563043">BBa_K563043</a></td>
 +
<td style="background:#ffdfbf;">Reporter</td>
 +
<td style="background:#ffdfbf;">pYE-EYFP</td>
 +
<td style="background:#ffdfbf;">Bin Jia</td>
 +
<td style="background:#ffdfbf;">753</td>
 +
</tr>
 +
<tr>
 +
<td>24</td>
 +
<td><a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K563045">BBa_K563045</a></td>
 +
<td>Reporter</td>
 +
<td>pYE-OFL Orange Firefly Luciferase</td>
 +
<td>Bin Jia</td>
 +
<td>1656</td>
 +
</tr>
 +
<tr>
 +
<td style="background:#ffdfbf;">25</td>
 +
<td style="background:#ffdfbf;"><a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K563044">BBa_K563046</a></td>
 +
<td style="background:#ffdfbf;">Reporter</td>
 +
<td style="background:#ffdfbf;">pYE-EPIC Firefly Luciferase</td>
 +
<td style="background:#ffdfbf;">Bin Jia</td>
 +
<td style="background:#ffdfbf;">1656</td>
 +
</tr>
 +
<tr>
 +
<td>26</td>
 +
<td><a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K563042">BBa_K563047</a></td>
 +
<td>Reporter</td>
 +
<td>pYE-EOFP</td>
 +
<td>Bin Jia</td>
 +
<td>744</td>
 +
</tr>
 +
<tr>
 +
<td style="background:#ffdfbf;">27</td>
 +
<td style="background:#ffdfbf;"><a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K563046">BBa_K563048</a></td>
 +
<td style="background:#ffdfbf;">Reporter</td>
 +
<td style="background:#ffdfbf;">pYE-LacZ</td>
 +
<td style="background:#ffdfbf;">Bin Jia</td>
 +
<td style="background:#ffdfbf;">3000</td>
 +
</tr>
 +
 
 +
<tr>
 +
<td>28</td>
 +
<td><a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K563051">BBa_K563051</a></td>
 +
<td>Coding</td>
 +
<td>TOR in pSBY1</td>
 +
<td>Bin Jia</td>
 +
<td>11914</td>
 +
</tr>
 +
<tr>
 +
<td style="background:#ffdfbf;">29</td>
 +
<td style="background:#ffdfbf;"><a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K563052">BBa_K563052</a></td>
 +
<td style="background:#ffdfbf;">Plasmid</td>
 +
<td style="background:#ffdfbf;">pSBY11(pSB1A11-TRP1 CEN/ARS Yeast Shuttle Vector)</td>
 +
<td style="background:#ffdfbf;">Bin Jia</td>
 +
<td style="background:#ffdfbf;">3830</td>
 +
</tr>
 +
<tr>
 +
<td>30</td>
 +
<td><a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K563053">BBa_K563053</a></td>
 +
<td>Plasmid Backbone</td>
 +
<td>pYE</td>
 +
<td>Bin Jia</td>
 +
<td>5900</td>
 +
</tr>
 +
<tr>
 +
<td style="background:#ffdfbf;">31</td>
 +
<td style="background:#ffdfbf;"><a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K563054">BBa_K563054</a></td>
 +
<td style="background:#ffdfbf;">Plasmid Backbone</td>
 +
<td style="background:#ffdfbf;">pYD</td>
 +
<td style="background:#ffdfbf;">Bin Jia</td>
 +
<td style="background:#ffdfbf;">5090</td>
 +
</tr>
 +
<tr>
 +
<td>32</td>
 +
<td><a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K563050">BBa_K563050</a></td>
 +
<td>Plasmid Backbone</td>
 +
<td>pSB1A11(Modified from pSB1A2)</td>
 +
<td>Bin Jia</td>
 +
<td>2090</td>
 +
</tr>
 +
</table>
 +
</div>
 +
 
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Latest revision as of 00:50, 6 October 2011

Template:Https://2011.igem.org/Team:Peking S/bannerhidden Template:Https://2011.igem.org/Team:Peking S/back2 Untitled

This page shows a list of all the parts that we have made or used in the project. Click on each link to see more details about that part on the Registry of Standard Biological Parts.

As our project focus on tor2 gene coming from yeast and contains common restriction sites like EcoRI, NotI and SpeI, the stantard Plasmid Backbone and BioBrick prefix/suffix is no longer appropriate. Furthermore, the size of 7800bp makes it pretty too expensive and inconvenient for chemical synthesis and codon optimization to avoid certain restriction sites. To match with the core gene and protein, a series of parts also give up standard prefix/suffix and backbones. We have used at least 3 kinds of plasmids, including blunt-end vectors to Yeast Surface Display Vector. However, based on the splendid standard plasmid backbone pSB1C3, which has a small size and simple MCS for easy-operation, we modified several vectors, which also render functionality and ease-of-use of promoter parts.

1. Inducible pCIT2 – Mut-TOR2 - Terminator, BBa_K563061: Promoter of CIT2 gene would be binding with transcription factor RTG1/3 at the presence of FAP. Then our reconstructed Mut-TOR2 would be expressed and forms in Mut-TORC1 which is not allergic to FAP. The accumulation of Mut-TORC1 will help the intracellular phosphorylation back to normal state.

2. pYE- Reporter, BBa_K563040 ~ BBa_K563046: We have successfully construct vectors, which coding bright fluorescent protein and Luciferase and beta-galactosidase in yeast cell. This series of BioBricks provide a convenient method for the measurement and detection of gene expression and terminate the unsuccessful expression of fluorescent protein in yeast cell.

3. Modified Plasmid Backbone – pSBY11, BBa_K563051: We reconstruct the Yeast Vector pYD through the combination with pSB1A11. According to specific primers, we amplify the yeast elements (CEN4/ARS6, TRP1 orf and terminator region) by PCR, and then insert the fragment into pSB1A11 by single restriction site PstI. In this way, a modified vector that combines pYD and pSB1A11 together(pSBY1) for large fragment transformation between yeast and E. coli is built.

  Name Type Description Designer Length
1 BBa_K563000 Regulatory Promotor of HIS4 Boxuan Zeng 508
2 BBa_K563001 Regulatory Promoter of DAL7 Boxuan Zeng 498
3 BBa_K563002 Regulatory Promoter of ILV1 Boxuan Zeng 509
4 BBa_K563003 Regulatory Promoter of CIT2 Boxuan Zeng 497
5 BBa_K563004 Regulatory Promoter of TEF1 Boxuan Zeng 450
6 BBa_K563010 Coding Original TOR Boxuan Zeng 8118
7 BBa_K563011 Coding TOR (LP Mutation) Boxuan Zeng 8118
8 BBa_K563012 Coding TOR (ITMutation) Boxuan Zeng 8118
9 BBa_K563013 Coding TOR (IT/EK Mutation) Boxuan Zeng 8118
10 BBa_K563014 Coding TOR (IT/VA Mutation) Boxuan Zeng 8118
11 BBa_K563015 Coding TOR (EK/VA Mutation) Boxuan Zeng 8118
12 BBa_K563016 Coding TOR (IT/VA/EK Mutation) Boxuan Zeng 8118
13 BBa_K563030 Coding pEASY-TOR01 Wenbin Kuang 3067
14 BBa_K563031 Coding pEASY-TOR11 Wenbin Kuang 2521
15 BBa_K563032 Coding pEASY-TOR22 Wenbin Kuang 2709
16 BBa_K563033 Coding pEASY-TOR33 Wenbin Kuang 2499
17 BBa_K563034 Coding pEASY-TOR22(LP Mutation) Mingrui Zhao 2709
18 BBa_K563035 Coding pEASY-TOR33(IT Mutation) Mingrui Zhao 2499
19 BBa_K563036 Coding pEASY-TOR33(VA Mutation) Jing Guo 2499
20 BBa_K563037 Coding pEASY-TOR33(EK Mutation) Jing Guo 2499
21 BBa_K563040 Reporter pYE-EGFP Bin Jia 739
22 BBa_K563041 Reporter pYE-ERFP Bin Jia 744
23 BBa_K563043 Reporter pYE-EYFP Bin Jia 753
24 BBa_K563045 Reporter pYE-OFL Orange Firefly Luciferase Bin Jia 1656
25 BBa_K563046 Reporter pYE-EPIC Firefly Luciferase Bin Jia 1656
26 BBa_K563047 Reporter pYE-EOFP Bin Jia 744
27 BBa_K563048 Reporter pYE-LacZ Bin Jia 3000
28 BBa_K563051 Coding TOR in pSBY1 Bin Jia 11914
29 BBa_K563052 Plasmid pSBY11(pSB1A11-TRP1 CEN/ARS Yeast Shuttle Vector) Bin Jia 3830
30 BBa_K563053 Plasmid Backbone pYE Bin Jia 5900
31 BBa_K563054 Plasmid Backbone pYD Bin Jia 5090
32 BBa_K563050 Plasmid Backbone pSB1A11(Modified from pSB1A2) Bin Jia 2090