Team:Tsinghua-A/Parts

From 2011.igem.org

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<h1 id="Parts List">Parts List</h1><hr width="100%" size=2 color=gray>
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm; text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Name</SPAN></FONT></FONT></P>
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<H1 id="Parts List"><A NAME="Parts List"></A><FONT COLOR="#006b6b"><FONT FACE="Arial, sans-serif"><FONT SIZE=5 STYLE="font-size: 20pt"><SPAN LANG="en-US"><B>Parts
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm; text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Type</SPAN></FONT></FONT></P>
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List</B></SPAN></FONT></FONT></FONT></H1>
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<HR SIZE=1>
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<TH WIDTH=340 BGCOLOR="#aaaaff">
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<P STYLE="margin-bottom: 0cm; line-height: 0.58cm; widows: 2; orphans: 2">
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm; text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Description</SPAN></FONT></FONT></P>
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</TH>
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<TH WIDTH=130 BGCOLOR="#aaaaff">
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm; text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Length</SPAN></FONT></FONT></P>
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</TH>
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</TR>
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<TR>
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<TD WIDTH=160 BGCOLOR="#ddddff">
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm; text-indent:0em"><FONT FACE="Times New Roman, serif"><SPAN LANG="en-US"><A HREF="http://partsregistry.org/wiki/index.php?title=Part:BBa_K574000"><FONT COLOR="#002bb8"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">BBa_K574000</SPAN></FONT></FONT></SPAN></FONT></A></SPAN></FONT></P>
+
-
</TD>
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<TD WIDTH=130 BGCOLOR="#ddddff">
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm; text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Composite</SPAN></FONT></FONT></P>
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</TD>
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<TD WIDTH=340 BGCOLOR="#ddddff">
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm; text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">pLux + TetR</SPAN></FONT></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=130 BGCOLOR="#ddddff">
+
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm; text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">903</SPAN></FONT></FONT></P>
+
-
</TD>
+
-
</TR>
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-
<TR>
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<TD WIDTH=160 BGCOLOR="#ddddff">
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm; text-indent:0em"><FONT FACE="Times New Roman, serif"><SPAN LANG="en-US"><A HREF="http://partsregistry.org/wiki/index.php?title=Part:BBa_K574001"><FONT COLOR="#002bb8"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">BBa_K574001</SPAN></FONT></FONT></SPAN></FONT></A></SPAN></FONT></P>
+
-
</TD>
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<TD WIDTH=130 BGCOLOR="#ddddff">
+
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm; text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Composite</SPAN></FONT></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=340 BGCOLOR="#ddddff">
+
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm; text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">TetR
+
-
regulated by 3OC6HSL</SPAN></FONT></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=130 BGCOLOR="#ddddff">
+
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm; text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">1890</SPAN></FONT></FONT></P>
+
-
</TD>
+
-
</TR>
+
-
<TR>
+
-
<TD WIDTH=160 BGCOLOR="#ddddff">
+
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm; text-indent:0em"><FONT FACE="Times New Roman, serif"><SPAN LANG="en-US"><A HREF="http://partsregistry.org/wiki/index.php?title=Part:BBa_K574002"><FONT COLOR="#002bb8"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">BBa_K574002</SPAN></FONT></FONT></SPAN></FONT></A></SPAN></FONT></P>
+
-
</TD>
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<TD WIDTH=130 BGCOLOR="#ddddff">
+
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm; text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Composite</SPAN></FONT></FONT></P>
+
-
</TD>
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-
<TD WIDTH=340 BGCOLOR="#ddddff">
+
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm; text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">pluxR
+
-
+ CFP</SPAN></FONT></FONT></P>
+
-
</TD>
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<TD WIDTH=130 BGCOLOR="#ddddff">
+
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm; text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">941</SPAN></FONT></FONT></P>
+
-
</TD>
+
-
</TR>
+
-
<TR>
+
-
<TD WIDTH=160 BGCOLOR="#ddddff">
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm; text-indent:0em"><FONT FACE="Times New Roman, serif"><SPAN LANG="en-US"><A HREF="http://partsregistry.org/wiki/index.php?title=Part:BBa_K574003"><FONT COLOR="#002bb8"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">BBa_K574003</SPAN></FONT></FONT></SPAN></FONT></A></SPAN></FONT></P>
+
-
</TD>
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<TD WIDTH=130 BGCOLOR="#ddddff">
+
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm; text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Device</SPAN></FONT></FONT></P>
+
-
</TD>
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<TD WIDTH=340 BGCOLOR="#ddddff">
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm; text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">3OC6HSL -> TetR && reporter</SPAN></FONT></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=130 BGCOLOR="#ddddff">
+
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm; text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">2839</SPAN></FONT></FONT></P>
+
-
</TD>
+
-
</TR>
+
-
<TR>
+
-
<TD WIDTH=160 BGCOLOR="#ddddff">
+
-
<P ALIGN=CENTER STYLE="border: none; padding: 0cm; text-indent:0em"><FONT FACE="Times New Roman, serif"><SPAN LANG="en-US"><A HREF="http://partsregistry.org/wiki/index.php?title=Part:BBa_K574004"><FONT COLOR="#002bb8"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">BBa_K574004</SPAN></FONT></FONT></SPAN></FONT></A></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=130 BGCOLOR="#ddddff">
+
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm; text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Composite</SPAN></FONT></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=340 BGCOLOR="#ddddff">
+
-
<P ALIGN=CENTER STYLE="border: none; padding: 0cm; text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">3OC12HSL regulated by TetR</SPAN></FONT></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=130 BGCOLOR="#ddddff">
+
-
<P ALIGN=CENTER STYLE="border: none; padding: 0cm; text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">797</SPAN></FONT></FONT></P>
+
-
</TD>
+
-
</TR>
+
-
<TR>
+
-
<TD WIDTH=160 BGCOLOR="#ddddff">
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-
<P ALIGN=CENTER STYLE="border: none; padding: 0cm; text-indent:0em"><FONT FACE="Times New Roman, serif"><SPAN LANG="en-US"><A HREF="http://partsregistry.org/wiki/index.php?title=Part:BBa_K574005"><FONT COLOR="#002bb8"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">BBa_K574005</SPAN></FONT></FONT></SPAN></FONT></A></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=130 BGCOLOR="#ddddff">
+
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm; text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Composite</SPAN></FONT></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=340 BGCOLOR="#ddddff">
+
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm; text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">3OC12HSL and YFP regulated by pBad</SPAN></FONT></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=130 BGCOLOR="#ddddff">
+
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm; text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">1712</SPAN></FONT></FONT></P>
+
-
</TD>
+
-
</TR>
+
-
<TR>
+
-
<TD WIDTH=160 BGCOLOR="#ddddff">
+
-
<P ALIGN=CENTER STYLE="border: none; padding: 0cm; text-indent:0em"><FONT FACE="Times New Roman, serif"><SPAN LANG="en-US"><A HREF="http://partsregistry.org/wiki/index.php?title=Part:BBa_K574006"><FONT COLOR="#002bb8"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">BBa_K574006</SPAN></FONT></FONT></SPAN></FONT></A></SPAN></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=130 BGCOLOR="#ddddff">
+
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm; text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Device</SPAN></FONT></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=340 BGCOLOR="#ddddff">
+
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm; text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">3OC12HSL regulated by pTet and pBad</SPAN></FONT></FONT></P>
+
-
</TD>
+
-
<TD WIDTH=130 BGCOLOR="#ddddff">
+
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm; text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">2517</SPAN></FONT></FONT></P>
+
-
</TD>
+
-
</TR>
+
-
<TR>
+
-
<TD WIDTH=160 BGCOLOR="#ddddff">
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm; text-indent:0em"><FONT FACE="Times New Roman, serif"><SPAN LANG="en-US"><A HREF="http://partsregistry.org/wiki/index.php?title=Part:BBa_K574007"><FONT COLOR="#002bb8"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">BBa_K574007</SPAN></FONT></FONT></SPAN></FONT></A></SPAN></FONT></P>
+
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</TD>
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<TD WIDTH=130 BGCOLOR="#ddddff">
+
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm; text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Composite</SPAN></FONT></FONT></P>
+
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</TD>
+
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<TD WIDTH=340 BGCOLOR="#ddddff">
+
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm; text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">lasR + TT</SPAN></FONT></FONT></P>
+
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</TD>
+
-
<TD WIDTH=130 BGCOLOR="#ddddff">
+
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm; text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">918</SPAN></FONT></FONT></P>
+
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</TD>
+
-
</TR>
+
-
<TR>
+
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<TD WIDTH=160 BGCOLOR="#ddddff">
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm; text-indent:0em"><FONT FACE="Times New Roman, serif"><SPAN LANG="en-US"><A HREF="http://partsregistry.org/wiki/index.php?title=Part:BBa_K574008"><FONT COLOR="#002bb8"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">BBa_K574008</SPAN></FONT></FONT></SPAN></FONT></A></SPAN></FONT></P>
+
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</TD>
+
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<TD WIDTH=130 BGCOLOR="#ddddff">
+
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm; text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Composite</SPAN></FONT></FONT></P>
+
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</TD>
+
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<TD WIDTH=340 BGCOLOR="#ddddff">
+
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm; text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">a constitutive LasR producer</SPAN></FONT></FONT></P>
+
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</TD>
+
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<TD WIDTH=130 BGCOLOR="#ddddff">
+
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm; text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">982</SPAN></FONT></FONT></P>
+
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</TD>
+
-
</TR>
+
-
<TR>
+
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<TD WIDTH=160 BGCOLOR="#ddddff">
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm; text-indent:0em"><FONT FACE="Times New Roman, serif"><SPAN LANG="en-US"><A HREF="http://partsregistry.org/wiki/index.php?title=Part:BBa_K574009"><FONT COLOR="#002bb8"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">BBa_K574009</SPAN></FONT></FONT></SPAN></FONT></A></SPAN></FONT></P>
+
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</TD>
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<TD WIDTH=130 BGCOLOR="#ddddff">
+
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm; text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Composite</SPAN></FONT></FONT></P>
+
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</TD>
+
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<TD WIDTH=340 BGCOLOR="#ddddff">
+
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm; text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">3OC12HSL -> PoPS Receiver</SPAN></FONT></FONT></P>
+
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</TD>
+
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<TD WIDTH=130 BGCOLOR="#ddddff">
+
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm; text-indent:0em"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">1147</SPAN></FONT></FONT></P>
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</TD>
+
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</TR>
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</TABLE>
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<BR>
<BR>
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<h1 id="Descriptions">Descriptions</h1><hr width="100%" size=2 color=gray>
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</P>
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<br>
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<CENTER>
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<p>We assembled <A HREF="http://partsregistry.org/wiki/index.php/Part:BBa_E0840"><FONT COLOR="#002bb8">BBa_E0840</FONT></A>
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<TABLE WIDTH=760 BORDER=4 CELLPADDING=0 CELLSPACING=0 BGCOLOR="#ffffff">
-
under the pLas promoter  
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<COL WIDTH=160>
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<A HREF="http://partsregistry.org/wiki/index.php/Part:BBa_R0079"><FONT COLOR="#002bb8">BBa_R0079</FONT></A>
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<COL WIDTH=130>
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that was contained into K574009. We kept pSB1A2 as the scaffold vector.</p>
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<COL WIDTH=340>
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<br>
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<COL WIDTH=130>
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<h1 id="Methods">Methods</h1><hr width="100%" size=2 color=gray>
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<TR>
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<br>
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<TH WIDTH=160 BGCOLOR="#aaaaff">
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<p>1. Ligate
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Name</SPAN></FONT></FONT></FONT></P>
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<A HREF="http://partsregistry.org/wiki/index.php/Part:BBa_E0840"><FONT COLOR="#002bb8">BBa_E0840</FONT></A>
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</TH>
-
downstream of
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<TH WIDTH=130 BGCOLOR="#aaaaff">
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<A HREF="http://partsregistry.org/wiki/index.php/Part:BBa_K574009"><FONT COLOR="#002bb8">BBa_K574009</FONT></A>
+
<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Type</SPAN></FONT></FONT></FONT></P>
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, which acts as a reporter.</p>
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</TH>
-
 
+
<TH WIDTH=340 BGCOLOR="#aaaaff">
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<p>2. Culture the positive colony at 37&deg;C,
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Description</SPAN></FONT></FONT></FONT></P>
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220 rpm for 12 hours, as well as the DH5alpha.</p>
+
</TH>
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+
<TH WIDTH=130 BGCOLOR="#aaaaff">
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<p>3. Dilute 1:25 the former in 15 falcon tubes (4 ml cultures),  
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Length</SPAN></FONT></FONT></FONT></P>
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each 3 tubes as a group. The later is also diluted in 3 tubes as  
+
</TH>
-
the negative control.</p>
+
</TR>
-
 
+
<TR>
-
<p>4. After culturing for another 2 hours, as their OD600 reach 0.2, induce
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<TD WIDTH=160 BGCOLOR="#ddddff">
-
with 3OC12HSL 0, 1e-7 M, 1e-6 M, 1e-5 M and 1e-4 M respectively.</p>
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT FACE="Times New Roman, serif"><SPAN LANG="en-US"><A HREF="http://partsregistry.org/wiki/index.php?title=Part:BBa_K574000"><FONT COLOR="#002bb8"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">BBa_K574000</SPAN></FONT></FONT></SPAN></FONT></A></SPAN></FONT></P>
-
 
+
</TD>
-
<p>5. Take samples after 1 hour, 4 hours, 8 hours and 24 hours, that is,
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<TD WIDTH=130 BGCOLOR="#ddddff">
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centrifugalizate 0.5 mL of each culture and suspend them by 1 mL of PBS
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Composite</SPAN></FONT></FONT></FONT></P>
-
(phosphate-buffered saline).</p>
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</TD>
-
 
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<TD WIDTH=340 BGCOLOR="#ddddff">
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<P>6. Measure the fluorescence of the samples with the flow cytometer.</P>
+
<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">pLux
-
<br>
+
+ TetR</SPAN></FONT></FONT></FONT></P>
-
<h1 id="Results">Results</h1><hr width="100%" size=2 color=gray>
+
</TD>
-
<br>
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<TD WIDTH=130 BGCOLOR="#ddddff">
-
<p style="text-indent:0em"align="CENTER"><img src="https://static.igem.org/mediawiki/2011/c/ca/K574009_chart.png" NAME="K574009_chart" /></p>
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<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">903</SPAN></FONT></FONT></FONT></P>
-
<P>From
+
</TD>
 +
</TR>
 +
<TR>
 +
<TD WIDTH=160 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT FACE="Times New Roman, serif"><SPAN LANG="en-US"><A HREF="http://partsregistry.org/wiki/index.php?title=Part:BBa_K574001"><FONT COLOR="#002bb8"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">BBa_K574001</SPAN></FONT></FONT></SPAN></FONT></A></SPAN></FONT></P>
 +
</TD>
 +
<TD WIDTH=130 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Composite</SPAN></FONT></FONT></FONT></P>
 +
</TD>
 +
<TD WIDTH=340 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">TetR
 +
regulated by 3OC6HSL</SPAN></FONT></FONT></FONT></P>
 +
</TD>
 +
<TD WIDTH=130 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">1890</SPAN></FONT></FONT></FONT></P>
 +
</TD>
 +
</TR>
 +
<TR>
 +
<TD WIDTH=160 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT FACE="Times New Roman, serif"><SPAN LANG="en-US"><A HREF="http://partsregistry.org/wiki/index.php?title=Part:BBa_K574002"><FONT COLOR="#002bb8"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">BBa_K574002</SPAN></FONT></FONT></SPAN></FONT></A></SPAN></FONT></P>
 +
</TD>
 +
<TD WIDTH=130 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Composite</SPAN></FONT></FONT></FONT></P>
 +
</TD>
 +
<TD WIDTH=340 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">pluxR
 +
+ CFP</SPAN></FONT></FONT></FONT></P>
 +
</TD>
 +
<TD WIDTH=130 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">941</SPAN></FONT></FONT></FONT></P>
 +
</TD>
 +
</TR>
 +
<TR>
 +
<TD WIDTH=160 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT FACE="Times New Roman, serif"><SPAN LANG="en-US"><A HREF="http://partsregistry.org/wiki/index.php?title=Part:BBa_K574003"><FONT COLOR="#002bb8"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">BBa_K574003</SPAN></FONT></FONT></SPAN></FONT></A></SPAN></FONT></P>
 +
</TD>
 +
<TD WIDTH=130 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Device</SPAN></FONT></FONT></FONT></P>
 +
</TD>
 +
<TD WIDTH=340 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">3OC6HSL
 +
-&gt; TetR &amp;&amp; reporter</SPAN></FONT></FONT></FONT></P>
 +
</TD>
 +
<TD WIDTH=130 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">2839</SPAN></FONT></FONT></FONT></P>
 +
</TD>
 +
</TR>
 +
<TR>
 +
<TD WIDTH=160 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT FACE="Times New Roman, serif"><SPAN LANG="en-US"><A HREF="http://partsregistry.org/wiki/index.php?title=Part:BBa_K574004"><FONT COLOR="#002bb8"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">BBa_K574004</SPAN></FONT></FONT></SPAN></FONT></A></SPAN></FONT></P>
 +
</TD>
 +
<TD WIDTH=130 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Composite</SPAN></FONT></FONT></FONT></P>
 +
</TD>
 +
<TD WIDTH=340 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">3OC12HSL
 +
regulated by TetR</SPAN></FONT></FONT></FONT></P>
 +
</TD>
 +
<TD WIDTH=130 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">797</SPAN></FONT></FONT></FONT></P>
 +
</TD>
 +
</TR>
 +
<TR>
 +
<TD WIDTH=160 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT FACE="Times New Roman, serif"><SPAN LANG="en-US"><A HREF="http://partsregistry.org/wiki/index.php?title=Part:BBa_K574005"><FONT COLOR="#002bb8"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">BBa_K574005</SPAN></FONT></FONT></SPAN></FONT></A></SPAN></FONT></P>
 +
</TD>
 +
<TD WIDTH=130 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Composite</SPAN></FONT></FONT></FONT></P>
 +
</TD>
 +
<TD WIDTH=340 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">3OC12HSL
 +
and YFP regulated by pBad</SPAN></FONT></FONT></FONT></P>
 +
</TD>
 +
<TD WIDTH=130 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">1712</SPAN></FONT></FONT></FONT></P>
 +
</TD>
 +
</TR>
 +
<TR>
 +
<TD WIDTH=160 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT FACE="Times New Roman, serif"><SPAN LANG="en-US"><A HREF="http://partsregistry.org/wiki/index.php?title=Part:BBa_K574006"><FONT COLOR="#002bb8"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">BBa_K574006</SPAN></FONT></FONT></SPAN></FONT></A></SPAN></FONT></P>
 +
</TD>
 +
<TD WIDTH=130 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Device</SPAN></FONT></FONT></FONT></P>
 +
</TD>
 +
<TD WIDTH=340 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">3OC12HSL
 +
regulated by pTet and pBad</SPAN></FONT></FONT></FONT></P>
 +
</TD>
 +
<TD WIDTH=130 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">2517</SPAN></FONT></FONT></FONT></P>
 +
</TD>
 +
</TR>
 +
<TR>
 +
<TD WIDTH=160 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT FACE="Times New Roman, serif"><SPAN LANG="en-US"><A HREF="http://partsregistry.org/wiki/index.php?title=Part:BBa_K574007"><FONT COLOR="#002bb8"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">BBa_K574007</SPAN></FONT></FONT></SPAN></FONT></A></SPAN></FONT></P>
 +
</TD>
 +
<TD WIDTH=130 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Composite</SPAN></FONT></FONT></FONT></P>
 +
</TD>
 +
<TD WIDTH=340 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">lasR
 +
+ TT</SPAN></FONT></FONT></FONT></P>
 +
</TD>
 +
<TD WIDTH=130 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">918</SPAN></FONT></FONT></FONT></P>
 +
</TD>
 +
</TR>
 +
<TR>
 +
<TD WIDTH=160 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT FACE="Times New Roman, serif"><SPAN LANG="en-US"><A HREF="http://partsregistry.org/wiki/index.php?title=Part:BBa_K574008"><FONT COLOR="#002bb8"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">BBa_K574008</SPAN></FONT></FONT></SPAN></FONT></A></SPAN></FONT></P>
 +
</TD>
 +
<TD WIDTH=130 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Composite</SPAN></FONT></FONT></FONT></P>
 +
</TD>
 +
<TD WIDTH=340 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">a
 +
constitutive LasR producer</SPAN></FONT></FONT></FONT></P>
 +
</TD>
 +
<TD WIDTH=130 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">982</SPAN></FONT></FONT></FONT></P>
 +
</TD>
 +
</TR>
 +
<TR>
 +
<TD WIDTH=160 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT FACE="Times New Roman, serif"><SPAN LANG="en-US"><A HREF="http://partsregistry.org/wiki/index.php?title=Part:BBa_K574009"><FONT COLOR="#002bb8"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">BBa_K574009</SPAN></FONT></FONT></SPAN></FONT></A></SPAN></FONT></P>
 +
</TD>
 +
<TD WIDTH=130 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">Composite</SPAN></FONT></FONT></FONT></P>
 +
</TD>
 +
<TD WIDTH=340 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">3OC12HSL
 +
-&gt; PoPS Receiver</SPAN></FONT></FONT></FONT></P>
 +
</TD>
 +
<TD WIDTH=130 BGCOLOR="#ddddff">
 +
<P ALIGN=CENTER STYLE="border: none; padding: 0cm"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=4><SPAN LANG="en-US">1147</SPAN></FONT></FONT></FONT></P>
 +
</TD>
 +
</TR>
 +
</TABLE>
 +
</CENTER>
 +
<P STYLE="line-height: 0.58cm; widows: 2; orphans: 2"><BR><BR>
 +
</P>
 +
<H1 id="Descriptions" STYLE="page-break-before: always; page-break-after: avoid"><A NAME="Descriptions"></A>
 +
<FONT COLOR="#006b6b"><FONT FACE="Arial, sans-serif"><FONT SIZE=5 STYLE="font-size: 20pt"><SPAN LANG="en-US">Descriptions</SPAN></FONT></FONT></FONT></H1>
 +
<HR SIZE=1>
 +
<P STYLE="line-height: 0.58cm; widows: 2; orphans: 2"><BR><BR>
 +
</P>
 +
<P STYLE="margin-left: 1.13cm; line-height: 200%; widows: 2; orphans: 2">
 +
<FONT FACE="Times New Roman, serif"><SPAN LANG="en-US"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=3 STYLE="font-size: 12pt"><SPAN STYLE="font-style: normal"><SPAN STYLE="font-weight: normal">We
 +
assembled&nbsp;</SPAN></SPAN></FONT></FONT></FONT><A HREF="http://partsregistry.org/wiki/index.php/Part:BBa_E0840"><FONT COLOR="#002bb8"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial, sans-serif"><FONT SIZE=3 STYLE="font-size: 12pt"><SPAN STYLE="font-style: normal"><SPAN STYLE="font-weight: normal">BBa_E0840</SPAN></SPAN></FONT></FONT></SPAN></FONT></A><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=3 STYLE="font-size: 12pt"><SPAN STYLE="font-style: normal"><SPAN STYLE="font-weight: normal">&nbsp;under
 +
the pLas promoter&nbsp;</SPAN></SPAN></FONT></FONT></FONT><A HREF="http://partsregistry.org/wiki/index.php/Part:BBa_R0079"><FONT COLOR="#002bb8"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial, sans-serif"><FONT SIZE=3 STYLE="font-size: 12pt"><SPAN STYLE="font-style: normal"><SPAN STYLE="font-weight: normal">BBa_R0079</SPAN></SPAN></FONT></FONT></SPAN></FONT></A><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=3 STYLE="font-size: 12pt"><SPAN STYLE="font-style: normal"><SPAN STYLE="font-weight: normal">&nbsp;that
 +
was contained into K574009. We kept pSB1A2 as the scaffold vector.</SPAN></SPAN></FONT></FONT></FONT></SPAN></FONT></P>
 +
<P STYLE="line-height: 0.58cm; widows: 2; orphans: 2"><BR><BR>
 +
</P>
 +
<H1 id="Methods" STYLE="page-break-before: always; page-break-after: avoid"><A NAME="Methods"></A>
 +
<FONT COLOR="#006b6b"><FONT FACE="Arial, sans-serif"><FONT SIZE=5 STYLE="font-size: 20pt"><SPAN LANG="en-US">Methods</SPAN></FONT></FONT></FONT></H1>
 +
<HR SIZE=1>
 +
<P STYLE="line-height: 0.58cm; widows: 2; orphans: 2"><BR><BR>
 +
</P>
 +
<P STYLE="margin-left: 1.16cm; line-height: 200%; widows: 2; orphans: 2">
 +
<FONT FACE="Times New Roman, serif"><SPAN LANG="en-US"><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=3 STYLE="font-size: 12pt"><SPAN STYLE="font-style: normal"><SPAN STYLE="font-weight: normal">1.
 +
Ligate&nbsp;</SPAN></SPAN></FONT></FONT></FONT><A HREF="http://partsregistry.org/wiki/index.php/Part:BBa_E0840"><FONT COLOR="#002bb8"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial, sans-serif"><FONT SIZE=3 STYLE="font-size: 12pt"><SPAN STYLE="font-style: normal"><SPAN STYLE="font-weight: normal">BBa_E0840</SPAN></SPAN></FONT></FONT></SPAN></FONT></A><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=3 STYLE="font-size: 12pt"><SPAN STYLE="font-style: normal"><SPAN STYLE="font-weight: normal">&nbsp;downstream
 +
of&nbsp;</SPAN></SPAN></FONT></FONT></FONT><A HREF="http://partsregistry.org/wiki/index.php/Part:BBa_K574009"><FONT COLOR="#002bb8"><SPAN STYLE="text-decoration: none"><FONT FACE="Arial, sans-serif"><FONT SIZE=3 STYLE="font-size: 12pt"><SPAN STYLE="font-style: normal"><SPAN STYLE="font-weight: normal">BBa_K574009</SPAN></SPAN></FONT></FONT></SPAN></FONT></A><FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=3 STYLE="font-size: 12pt"><SPAN STYLE="font-style: normal"><SPAN STYLE="font-weight: normal">&nbsp;,
 +
which acts as a reporter.</SPAN></SPAN></FONT></FONT></FONT></SPAN></FONT></P>
 +
<P STYLE="margin-left: 1.16cm; line-height: 200%; widows: 2; orphans: 2">
 +
<FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=3 STYLE="font-size: 12pt"><SPAN LANG="en-US">2.
 +
Culture the positive colony at 37&deg;C, 220 rpm for 12 hours, as
 +
well as the DH5alpha.</SPAN></FONT></FONT></FONT></P>
 +
<P STYLE="margin-left: 1.16cm; line-height: 200%; widows: 2; orphans: 2">
 +
<FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=3 STYLE="font-size: 12pt"><SPAN LANG="en-US">3.
 +
Dilute 1:25 the former in 15 falcon tubes (4 ml cultures), each 3
 +
tubes as a group. The later is also diluted in 3 tubes as the
 +
negative control.</SPAN></FONT></FONT></FONT></P>
 +
<P STYLE="margin-left: 1.16cm; line-height: 200%; widows: 2; orphans: 2">
 +
<FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=3 STYLE="font-size: 12pt"><SPAN LANG="en-US">4.
 +
After culturing for another 2 hours, as their OD600 reach 0.2, induce
 +
with 3OC12HSL 0, 1e-7 M, 1e-6 M, 1e-5 M and 1e-4 M respectively.</SPAN></FONT></FONT></FONT></P>
 +
<P STYLE="margin-left: 1.16cm; line-height: 200%; widows: 2; orphans: 2">
 +
<FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=3 STYLE="font-size: 12pt"><SPAN LANG="en-US">5.
 +
Take samples after 1 hour, 4 hours, 8 hours and 24 hours, that is,
 +
centrifugalizate 0.5 mL of each culture and suspend them by 1 mL of
 +
PBS (phosphate-buffered saline).</SPAN></FONT></FONT></FONT></P>
 +
<P STYLE="margin-left: 1.16cm; line-height: 200%; widows: 2; orphans: 2">
 +
<FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=3 STYLE="font-size: 12pt"><SPAN LANG="en-US">6.
 +
Measure the fluorescence of the samples with the flow cytometer.</SPAN></FONT></FONT></FONT></P>
 +
<P STYLE="line-height: 0.58cm; widows: 2; orphans: 2"><BR><BR>
 +
</P>
 +
<H1 id="Results" STYLE="page-break-before: always; page-break-after: avoid"><A NAME="Results"></A>
 +
<FONT COLOR="#006b6b"><FONT FACE="Arial, sans-serif"><FONT SIZE=5 STYLE="font-size: 20pt"><SPAN LANG="en-US">Results</SPAN></FONT></FONT></FONT></H1>
 +
<HR SIZE=1>
 +
<P STYLE="line-height: 0.58cm; widows: 2; orphans: 2"><BR><BR>
 +
</P>
 +
<P ALIGN=CENTER STYLE="line-height: 200%; widows: 2; orphans: 2"><FONT COLOR="#000000"><IMG SRC="https://static.igem.org/mediawiki/2011/c/ca/K574009_chart.png" NAME="K574009_chart" ALIGN=BOTTOM WIDTH=770 HEIGHT=490 BORDER=0></FONT></P>
 +
<P ALIGN=CENTER STYLE="line-height: 200%; widows: 2; orphans: 2"><BR><BR>
 +
</P>
 +
<P STYLE="margin-left: 1.16cm; line-height: 200%; widows: 2; orphans: 2">
 +
<FONT COLOR="#000000"><FONT FACE="Arial, sans-serif"><FONT SIZE=3 STYLE="font-size: 12pt"><SPAN LANG="en-US">From
the chart, we can see that cells were hardly induced in the control
the chart, we can see that cells were hardly induced in the control
group, and with the concentration of inducer growing, the intensity
group, and with the concentration of inducer growing, the intensity
of GFP increased by groups. The most efficient concentration of
of GFP increased by groups. The most efficient concentration of
-
inducer was around 10<sup>-5</sup>M, and higher concentration may lead to the
+
inducer was around 10<SUP>-5</SUP>M, and higher concentration may
-
expression of GFP decreasing. Additionally, to most groups, the intensity of GFP
+
lead to the expression of GFP decreasing. Additionally, to most
-
reached its maxium after 4 hours.</P>
+
groups, the intensity of GFP reached its maximum after 4 hours.</SPAN></FONT></FONT></FONT></P>
-
 
+
<P><FONT FACE="Arial, sans-serif"><SPAN LANG="en-US"><BR></SPAN></FONT><BR><BR>
-
 
+
</P>
-
<br><br><br>
+
-
<p align="CENTER" STYLE="text-indent:0em"><a href="https://2011.igem.org/Team:Tsinghua-A"><img src="https://static.igem.org/mediawiki/2011/9/92/Killbanner_header.jpg" alt="" width="960"/><a href="https://2011.igem.org"><img src="https://static.igem.org/mediawiki/igem.org/2/29/Killbanner_header2.jpg" alt="" width="960"/></p>
+
</BODY>
</BODY>
</HTML>
</HTML>

Latest revision as of 02:28, 28 October 2011

JavaScript Slideshow - TinySlideshow



Contents

Parts List


Name

Type

Description

Length

BBa_K574000

Composite

pLux + TetR

903

BBa_K574001

Composite

TetR regulated by 3OC6HSL

1890

BBa_K574002

Composite

pluxR + CFP

941

BBa_K574003

Device

3OC6HSL -> TetR && reporter

2839

BBa_K574004

Composite

3OC12HSL regulated by TetR

797

BBa_K574005

Composite

3OC12HSL and YFP regulated by pBad

1712

BBa_K574006

Device

3OC12HSL regulated by pTet and pBad

2517

BBa_K574007

Composite

lasR + TT

918

BBa_K574008

Composite

a constitutive LasR producer

982

BBa_K574009

Composite

3OC12HSL -> PoPS Receiver

1147



Descriptions



We assembled BBa_E0840 under the pLas promoter BBa_R0079 that was contained into K574009. We kept pSB1A2 as the scaffold vector.



Methods



1. Ligate BBa_E0840 downstream of BBa_K574009 , which acts as a reporter.

2. Culture the positive colony at 37°C, 220 rpm for 12 hours, as well as the DH5alpha.

3. Dilute 1:25 the former in 15 falcon tubes (4 ml cultures), each 3 tubes as a group. The later is also diluted in 3 tubes as the negative control.

4. After culturing for another 2 hours, as their OD600 reach 0.2, induce with 3OC12HSL 0, 1e-7 M, 1e-6 M, 1e-5 M and 1e-4 M respectively.

5. Take samples after 1 hour, 4 hours, 8 hours and 24 hours, that is, centrifugalizate 0.5 mL of each culture and suspend them by 1 mL of PBS (phosphate-buffered saline).

6. Measure the fluorescence of the samples with the flow cytometer.



Results





From the chart, we can see that cells were hardly induced in the control group, and with the concentration of inducer growing, the intensity of GFP increased by groups. The most efficient concentration of inducer was around 10-5M, and higher concentration may lead to the expression of GFP decreasing. Additionally, to most groups, the intensity of GFP reached its maximum after 4 hours.




Retrieved from "http://2011.igem.org/Team:Tsinghua-A/Parts"