Team:CIDEB-UANL Mexico/project aroma

From 2014hs.igem.org

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<p>Since the beginning of iGEM project, the use of fluorescent reporters has been used in each one of the proposed projects in previous years, trying to test the theoretical presence of other proteins in <i>E. coli</i>. For our iGEM 2014 project, this module proposed to promote the usage of aroma reporters, instead of fluorescent ones.</p>
 
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<td style="padding-left:12px;"><img width=131 height=127 src="https://static.igem.org/mediawiki/2014hs/5/5c/HandsomeAroma.jpg"/>
 
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<p><b>How is the Aroma module composed?</b></p>
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<img width=131 height=127 src="https://static.igem.org/mediawiki/2014hs/5/5c/HandsomeAroma.jpg" align=right hspace=12/>
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<p align="justify">Since the beginning of iGEM project, the use of fluorescent reporters has been used in each one of the proposed projects in previous years, trying to test the theoretical presence of other proteins in <i>E. coli.</i> For our iGEM 2014 project, this module proposed to promote the usage of aroma reporters, instead of fluorescent ones.</p>
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<p>This gene is composed by the following parts (see <b>figure 1A</b>): (1) a constitutive promoter, (2) a RNA thermometer, also called ribo-switch; used to regulate the WinterGreen-odor protein production through temperature, (3) a Wintergreen-odor enzyme generator, used to allow the production of methyl salicylate, induced by salicylic acid, and (4) a terminator. All of these parts are ligated by an 8-bp scar (TACTAGAG).</p><center>
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<p><b><h2>Description</h2></b></p>
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<p>In the module, WinterGreen is the most important part. Normally, it catalyzes the conversion of salicylic acid into methyl salicylate. The protein is excreted by the bacteria and when salicylic acid is added to the medium, a chemical reaction takes place and produces methyl salicylate, which is responsible for the wintergreen odor.</p>
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<p><img width=507 height=221 src="https://static.igem.org/mediawiki/2014hs/5/56/AromaFigure1Aa.png"align=center hspace=12 alt="IMG_0317"></p>
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<p>But in this module, it will be regulated by temperature with the use of the RNA thermometer. When adding salicylic acid to the bacteria in a 32° Celsius environment, the production of the WinterGreen protein will begin.</p><br>
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<p><b>Figure 1.</b> Aroma Module</p></center>
 
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<p>Different to fluorescent reporters, this module was made in order to (in the future) perform as an aroma reporter and also to test the correct function of the bacteria, for its future usage as a new reporter and functional part (CDS). It is desired to use this part in the project to replace the red fluorescent protein (RFP) in the <a href="https://2014hs.igem.org/Team:CIDEB-UANL_Mexico/project_capture">Capture module</a>. But it was preferable to test it apart to demonstrate its effectiveness. Similarly, this piece is also helpful for <a href="https://2014hs.igem.org/Team:CIDEB-UANL_Mexico/project_union">Union module</a>, because when performing the filtration by silica, WinterGreen can demonstrate the presence of bacteria in the beads. The team added the RNA thermometer for regulating the production of the aroma in the project. Another reason for selecting the RNA thermometer as a regulator was to continue the <a href="https://2013hs.igem.org/Team:CIDEB-UANL_Mexico">CIDEB UANL 2013 </a> work with it.</p>
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<center><img width=350 height=250 src="https://static.igem.org/mediawiki/2014hs/6/6e/AromaFigure1Bb.png" align=center hspace=12/></center>
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<center><p><b>Figure 1.</b> Production of Wintergreen Odor</p></center>
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<p><b>How does it work?</b></p><center>
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<p><b><h2>Device</h2></b></p>
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<p>Originally, WinterGreen was thought to be the reporter of the Capture module, but in order to prove the function of this gene and also the one in charge of the capture of sodium ions (NhaS), we decided to separate the full device into the actual modules of Capture and Aroma, as seen in the figure below.</p>
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<p><img width=334 height=232 src="https://static.igem.org/mediawiki/2014hs/6/6e/AromaFigure1Bb.png "align=center hspace=12 alt="IMG_0317"></p>
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<p><img width=450 height=250 src="https://static.igem.org/mediawiki/2014hs/3/32/TestCaptureAndAroma.jpg" align=center hspace=12></p>
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<p><b>Figure 2.</b> The device originally thought, and the 2 modules derived from it.</p></center><br>
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<p><b>Figure 2.</b> Production of Wintergreen Odor</p></center>
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<p>This device is composed by the following parts (see <b>figure 3</b>): a constitutive promoter (1), a RNA thermometer (3); used to regulate the WinterGreen-odor protein production through temperature, a Wintergreen-odor enzyme generator, used to allow the production of methyl salicylate, induced by salicylic acid (2); and a terminator (4). All of these parts are ligated by an 8-bp scar (TACTAGAG).</p>
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<p>This module has a constitutive promoter which will be regulated by temperature with the use of the RNA thermometer. When adding salicylic acid to the bacteria in a 32° Celsius environment, the production of the WinterGreen protein will begin.</p>
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<center><img width=400 height=170 src="https://static.igem.org/mediawiki/2014hs/5/56/AromaFigure1Aa.png" align=center hspace=12/>
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<p><b>Figure 3.</b> Aroma Module</p></center><br>
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<p><b>Parts of the module</b></p>
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<p>These 4 genetic parts form the Aroma device of the project. The full device's length is 1,251bp (including restriction sites).</p>
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<p><b><h2>Parts of the module</h2></b></p>
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<table class=MsoTable15Grid7ColorfulAccent3 border=1 cellspacing=0 cellpadding=0 width=625 style='width:468.55pt;border-collapse:collapse; border:none;mso-border-alt:solid #C2D69B .5pt;mso-border-themecolor:accent3; mso-border-themetint:153;mso-yfti-tbllook:1184;mso-padding-alt:0cm 5.4pt 0cm 5.4pt'>  
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<br><center><div><table class=MsoTable15Grid7ColorfulAccent3 border=1 cellspacing=0 cellpadding=0 width=625 style='width:468.55pt;border-collapse:collapse; border:none;mso-border-alt:solid #C2D69B .5pt;mso-border-themecolor:accent3; mso-border-themetint:153;mso-yfti-tbllook:1184;mso-padding-alt:0cm 5.4pt 0cm 5.4pt'> <tr style='mso-yfti-irow:-1;mso-yfti-firstrow:yes;mso-yfti-lastfirstrow:yes; height:15.0pt'> <td width=88 nowrap valign=top style='width:66.1pt;border:none;border-bottom: solid #C2D69B 1.0pt;mso-border-bottom-themecolor:accent3;mso-border-bottom-themetint: 153;mso-border-bottom-alt:solid #C2D69B .5pt;mso-border-bottom-themecolor: accent3;mso-border-bottom-themetint:153;background:white;mso-background-themecolor: background1;padding:0cm 5.4pt 0cm 5.4pt;height:15.0pt'> <p class=MsoNormal align=center style='margin-bottom:0cm;margin-bottom:.0001pt; text-align:center;line-height:normal;mso-yfti-cnfc:517'><b><span lang=ES-MX style='font-family:Oxygen;mso-fareast-font-family:"Times New Roman"; mso-bidi-font-family:"Times New Roman";mso-fareast-language:ES-MX;mso-bidi-font-style: italic'>IMAGE</span></b><span lang=ES-MX style='font-family:Oxygen; mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:"Times New Roman"; mso-fareast-language:ES-MX;mso-bidi-font-style:italic'><o:p></o:p></span></p> </td> <td width=119 valign=top style='width:89.0pt;border:none;border-bottom:solid #C2D69B 1.0pt; mso-border-bottom-themecolor:accent3;mso-border-bottom-themetint:153; mso-border-bottom-alt:solid #C2D69B .5pt;mso-border-bottom-themecolor:accent3; mso-border-bottom-themetint:153;background:white;mso-background-themecolor: background1;padding:0cm 5.4pt 0cm 5.4pt;height:15.0pt'> <p class=MsoNormal align=center style='margin-bottom:0cm;margin-bottom:.0001pt; text-align:center;line-height:normal;mso-yfti-cnfc:1'><b><span lang=ES-MX style='font-family:Oxygen;mso-fareast-font-family:"Times New Roman"; mso-bidi-font-family:"Times New Roman";mso-fareast-language:ES-MX'>CODE</span></b><span lang=ES-MX style='font-family:Oxygen;mso-fareast-font-family:"Times New Roman"; mso-bidi-font-family:"Times New Roman";mso-fareast-language:ES-MX'><o:p></o:p></span></p> </td> <td width=418 nowrap valign=top style='width:313.45pt;border:none;border-bottom: solid #C2D69B 1.0pt;mso-border-bottom-themecolor:accent3;mso-border-bottom-themetint: 153;mso-border-bottom-alt:solid #C2D69B .5pt;mso-border-bottom-themecolor: accent3;mso-border-bottom-themetint:153;background:white;mso-background-themecolor: background1;padding:0cm 5.4pt 0cm 5.4pt;height:15.0pt'> <p class=MsoNormal align=center style='margin-bottom:0cm;margin-bottom:.0001pt; text-align:center;line-height:normal;mso-yfti-cnfc:1'><b><span lang=ES-MX style='font-family:Oxygen;mso-fareast-font-family:"Times New Roman"; mso-bidi-font-family:"Times New Roman";mso-fareast-language:ES-MX'>DESCRIPTION</span></b><span lang=ES-MX style='font-family:Oxygen;mso-fareast-font-family:"Times New Roman"; mso-bidi-font-family:"Times New Roman";mso-fareast-language:ES-MX'><o:p></o:p></span></p> </td> </tr> <tr style='mso-yfti-irow:0;height:83.15pt'> <td width=88 nowrap valign=top style='width:66.1pt;border:none;border-right: solid #C2D69B 1.0pt;mso-border-right-themecolor:accent3;mso-border-right-themetint: 153;mso-border-top-alt:solid #C2D69B .5pt;mso-border-top-themecolor:accent3; mso-border-top-themetint:153;mso-border-top-alt:solid #C2D69B .5pt; mso-border-top-themecolor:accent3;mso-border-top-themetint:153;mso-border-right-alt: solid #C2D69B .5pt;mso-border-right-themecolor:accent3;mso-border-right-themetint: 153;background:white;mso-background-themecolor:background1;padding:0cm 5.4pt 0cm 5.4pt; height:83.15pt'> <p class=MsoNormal align=right style='margin-bottom:0cm;margin-bottom:.0001pt; text-align:right;line-height:normal;mso-yfti-cnfc:68'><!--[if gte vml 1]><v:shape id="_x0000_s1029" type="#_x0000_t75" style='position:absolute;left:0; text-align:left;margin-left:10.95pt;margin-top:3.2pt;width:66pt;height:48pt; z-index:251660800;visibility:visible;mso-wrap-style:square; mso-width-percent:0;mso-height-percent:0;mso-wrap-distance-left:9pt; mso-wrap-distance-top:0;mso-wrap-distance-right:9pt; mso-wrap-distance-bottom:0;mso-position-horizontal:absolute; mso-position-horizontal-relative:text;mso-position-vertical:absolute; mso-position-vertical-relative:text;mso-width-percent:0; mso-height-percent:0;mso-width-relative:page;mso-height-relative:page'> <v:imagedata src="TablasAromaYResistencia_archivos/image007.png" o:title=""/> <w:wrap type="square"/> </v:shape><![endif]--><![if !vml]><img width=88 height=64 src="https://static.igem.org/mediawiki/2014hs/4/43/PConsAroma.png" align=left hspace=12 v:shapes="_x0000_s1029"><![endif]></p> <p class=MsoNormal align=right style='margin-bottom:0cm;margin-bottom:.0001pt; text-align:right;line-height:normal;mso-yfti-cnfc:68'><i><span lang=ES-MX style='font-family:Oxygen;color:#76923C;mso-themecolor:accent3;mso-themeshade: 191;mso-fareast-language:ES-MX;mso-no-proof:yes'><o:p>&nbsp;</o:p></span></i></p> </td> <td width=119 nowrap valign=top style='width:89.0pt;border-top:none; border-left:none;border-bottom:solid #C2D69B 1.0pt;mso-border-bottom-themecolor: accent3;mso-border-bottom-themetint:153;border-right:solid #C2D69B 1.0pt; mso-border-right-themecolor:accent3;mso-border-right-themetint:153; mso-border-top-alt:solid #C2D69B .5pt;mso-border-top-themecolor:accent3; mso-border-top-themetint:153;mso-border-left-alt:solid #C2D69B .5pt; mso-border-left-themecolor:accent3;mso-border-left-themetint:153;mso-border-alt: solid #C2D69B .5pt;mso-border-themecolor:accent3;mso-border-themetint:153; background:#EAF1DD;mso-background-themecolor:accent3;mso-background-themetint: 51;padding:0cm 5.4pt 0cm 5.4pt;height:83.15pt'> <p class=MsoNormal align=center style='margin-bottom:0cm;margin-bottom:.0001pt; text-align:center;line-height:normal;mso-yfti-cnfc:64'><span lang=ES-MX style='font-size:12.0pt;mso-bidi-font-size:11.0pt;font-family:Oxygen; mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:Arial; color:black;mso-fareast-language:ES-MX'><a href="http://parts.igem.org/Part:BBa_J23119">BBa_J23119A</a></span><span lang=ES-MX style='font-family:Oxygen;mso-fareast-font-family:"Times New Roman"; mso-bidi-font-family:Arial;color:black;mso-fareast-language:ES-MX'><o:p></o:p></span></p> <p class=MsoNormal align=center style='margin-bottom:0cm;margin-bottom:.0001pt; text-align:center;line-height:normal;mso-yfti-cnfc:64'>&nbsp;</p> </td> <td width=418 valign=top style='width:313.45pt;border-top:none;border-left: none;border-bottom:solid #C2D69B 1.0pt;mso-border-bottom-themecolor:accent3; mso-border-bottom-themetint:153;border-right:solid #C2D69B 1.0pt;mso-border-right-themecolor: accent3;mso-border-right-themetint:153;mso-border-top-alt:solid #C2D69B .5pt; mso-border-top-themecolor:accent3;mso-border-top-themetint:153;mso-border-left-alt: solid #C2D69B .5pt;mso-border-left-themecolor:accent3;mso-border-left-themetint: 153;mso-border-alt:solid #C2D69B .5pt;mso-border-themecolor:accent3; mso-border-themetint:153;background:#EAF1DD;mso-background-themecolor:accent3; mso-background-themetint:51;padding:0cm 5.4pt 0cm 5.4pt;height:83.15pt'> <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align: justify;line-height:normal;mso-yfti-cnfc:64'><span style='font-size:12.0pt; mso-bidi-font-size:11.0pt;font-family:Oxygen;mso-fareast-font-family:"Times New Roman"; mso-bidi-font-family:Arial;color:black;mso-ansi-language:EN-US;mso-fareast-language: ES-MX'>In the specific case of our aroma module, it will help the bacteria to continuously transcribe the <span class=SpellE>WinterGreen</span> gene in order to allow the bacteria to continuously produce the aroma. </span><span class=SpellE><span lang=ES-MX style='font-size:12.0pt;mso-bidi-font-size: 11.0pt;font-family:Oxygen;mso-fareast-font-family:"Times New Roman"; mso-bidi-font-family:Arial;color:black;mso-fareast-language:ES-MX'>This</span></span><span lang=ES-MX style='font-size:12.0pt;mso-bidi-font-size:11.0pt;font-family: Oxygen;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:Arial; color:black;mso-fareast-language:ES-MX'> <span class=SpellE>promoter</span> has a <span class=SpellE>length</span> of 35bp.<o:p></o:p></span></p> </td> </tr> <tr style='mso-yfti-irow:1;height:77.45pt'> <td width=88 nowrap valign=top style='width:66.1pt;border-top:solid #C2D69B 1.0pt; mso-border-top-themecolor:accent3;mso-border-top-themetint:153;border-left: none;border-bottom:none;border-right:solid #C2D69B 1.0pt;mso-border-right-themecolor: accent3;mso-border-right-themetint:153;mso-border-top-alt:solid #C2D69B .5pt; mso-border-top-themecolor:accent3;mso-border-top-themetint:153;mso-border-right-alt: solid #C2D69B .5pt;mso-border-right-themecolor:accent3;mso-border-right-themetint: 153;background:white;mso-background-themecolor:background1;padding:0cm 5.4pt 0cm 5.4pt; height:77.45pt'> <p class=MsoNormal align=right style='margin-bottom:0cm;margin-bottom:.0001pt; text-align:right;line-height:normal;mso-yfti-cnfc:4'><!--[if gte vml 1]><v:shape id="_x0000_s1028" type="#_x0000_t75" style='position:absolute;left:0; text-align:left;margin-left:16.95pt;margin-top:4.25pt;width:57.75pt; height:44.25pt;z-index:251662848;visibility:visible;mso-wrap-style:square; mso-width-percent:0;mso-height-percent:0;mso-wrap-distance-left:9pt; mso-wrap-distance-top:0;mso-wrap-distance-right:9pt; mso-wrap-distance-bottom:0;mso-position-horizontal:absolute; mso-position-horizontal-relative:text;mso-position-vertical:absolute; mso-position-vertical-relative:text;mso-width-percent:0; mso-height-percent:0;mso-width-relative:page;mso-height-relative:page'> <v:imagedata src="TablasAromaYResistencia_archivos/image009.png" o:title=""/> <w:wrap type="square"/> </v:shape><![endif]--><![if !vml]><img width=77 height=59 src="https://static.igem.org/mediawiki/2014hs/b/b0/RNAthermAroma.png" align=left hspace=12 v:shapes="_x0000_s1028"><![endif]><i><span lang=ES-MX style='font-family: Oxygen;color:#76923C;mso-themecolor:accent3;mso-themeshade:191;mso-fareast-language: ES-MX;mso-no-proof:yes'><o:p></o:p></span></i></p></td> <td width=119 nowrap valign=top style='width:89.0pt;border-top:none; border-left:none;border-bottom:solid #C2D69B 1.0pt;mso-border-bottom-themecolor: accent3;mso-border-bottom-themetint:153;border-right:solid #C2D69B 1.0pt; mso-border-right-themecolor:accent3;mso-border-right-themetint:153; mso-border-top-alt:solid #C2D69B .5pt;mso-border-top-themecolor:accent3; mso-border-top-themetint:153;mso-border-left-alt:solid #C2D69B .5pt; mso-border-left-themecolor:accent3;mso-border-left-themetint:153;mso-border-alt: solid #C2D69B .5pt;mso-border-themecolor:accent3;mso-border-themetint:153; padding:0cm 5.4pt 0cm 5.4pt;height:77.45pt'> <p class=MsoNormal align=center style='margin-bottom:0cm;margin-bottom:.0001pt; text-align:center;line-height:normal'><span lang=ES-MX style='font-size:12.0pt; mso-bidi-font-size:11.0pt;font-family:Oxygen;mso-fareast-font-family:"Times New Roman"; mso-bidi-font-family:Arial;color:black;mso-fareast-language:ES-MX'><a href="http://parts.igem.org/Part:BBa_K115017">BBa_K115017</a><o:p></o:p></span></p></td> <td width=418 valign=top style='width:313.45pt;border-top:none;border-left: none;border-bottom:solid #C2D69B 1.0pt;mso-border-bottom-themecolor:accent3; mso-border-bottom-themetint:153;border-right:solid #C2D69B 1.0pt;mso-border-right-themecolor: accent3;mso-border-right-themetint:153;mso-border-top-alt:solid #C2D69B .5pt; mso-border-top-themecolor:accent3;mso-border-top-themetint:153;mso-border-left-alt: solid #C2D69B .5pt;mso-border-left-themecolor:accent3;mso-border-left-themetint: 153;mso-border-alt:solid #C2D69B .5pt;mso-border-themecolor:accent3; mso-border-themetint:153;padding:0cm 5.4pt 0cm 5.4pt;height:77.45pt'> <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align: justify;line-height:normal'><span style='font-size:12.0pt;mso-bidi-font-size: 11.0pt;font-family:Oxygen;mso-fareast-font-family:"Times New Roman"; mso-bidi-font-family:Arial;color:black;mso-ansi-language:EN-US;mso-fareast-language: ES-MX'>A RNA thermometer, used for temperature post-transcriptional regulation (thermo sensor), and is designed to initiate transcription around 32°C.<o:p></o:p></span></p> </td> </tr> <tr style='mso-yfti-irow:2;height:190.6pt'> <td width=88 nowrap valign=top style='width:66.1pt;border-top:solid #C2D69B 1.0pt; mso-border-top-themecolor:accent3;mso-border-top-themetint:153;border-left: none;border-bottom:none;border-right:solid #C2D69B 1.0pt;mso-border-right-themecolor: accent3;mso-border-right-themetint:153;mso-border-top-alt:solid #C2D69B .5pt; mso-border-top-themecolor:accent3;mso-border-top-themetint:153;mso-border-right-alt: solid #C2D69B .5pt;mso-border-right-themecolor:accent3;mso-border-right-themetint: 153;background:white;mso-background-themecolor:background1;padding:0cm 5.4pt 0cm 5.4pt; height:190.6pt'> <p class=MsoNormal align=right style='margin-bottom:0cm;margin-bottom:.0001pt; text-align:right;line-height:normal;mso-yfti-cnfc:68'><!--[if gte vml 1]><v:shape id="_x0000_s1027" type="#_x0000_t75" style='position:absolute;left:0; text-align:left;margin-left:13.95pt;margin-top:3.2pt;width:63pt;height:51.75pt; z-index:251664896;visibility:visible;mso-wrap-style:square; mso-width-percent:0;mso-height-percent:0;mso-wrap-distance-left:9pt; mso-wrap-distance-top:0;mso-wrap-distance-right:9pt; mso-wrap-distance-bottom:0;mso-position-horizontal:absolute; mso-position-horizontal-relative:text;mso-position-vertical:absolute; mso-position-vertical-relative:text;mso-width-percent:0; mso-height-percent:0;mso-width-relative:page;mso-height-relative:page'> <v:imagedata src="TablasAromaYResistencia_archivos/image011.png" o:title=""/> <w:wrap type="square"/> </v:shape><![endif]--><![if !vml]><img width=84 height=69 src="https://static.igem.org/mediawiki/2014hs/b/b1/WinterGreenAroma.png" align=left hspace=12 v:shapes="_x0000_s1027"><![endif]><i><span style='font-size:3.0pt;font-family: Oxygen;color:#76923C;mso-themecolor:accent3;mso-themeshade:191;mso-ansi-language: EN-US;mso-fareast-language:ES-MX;mso-no-proof:yes'><o:p></o:p></span></i></p></td> <td width=119 nowrap valign=top style='width:89.0pt;border-top:none; border-left:none;border-bottom:solid #C2D69B 1.0pt;mso-border-bottom-themecolor: accent3;mso-border-bottom-themetint:153;border-right:solid #C2D69B 1.0pt; mso-border-right-themecolor:accent3;mso-border-right-themetint:153; mso-border-top-alt:solid #C2D69B .5pt;mso-border-top-themecolor:accent3; mso-border-top-themetint:153;mso-border-left-alt:solid #C2D69B .5pt; mso-border-left-themecolor:accent3;mso-border-left-themetint:153;mso-border-alt: solid #C2D69B .5pt;mso-border-themecolor:accent3;mso-border-themetint:153; background:#EAF1DD;mso-background-themecolor:accent3;mso-background-themetint: 51;padding:0cm 5.4pt 0cm 5.4pt;height:190.6pt'> <p class=MsoNormal align=center style='margin-bottom:0cm;margin-bottom:.0001pt; text-align:center;line-height:normal;mso-yfti-cnfc:64'><span style='font-size:12.0pt;mso-bidi-font-size:11.0pt;font-family:Oxygen; mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:Arial; mso-ansi-language:EN-US;mso-fareast-language:ES-MX'><a href="http://parts.igem.org/Part:BBa_K1255001:Design">BBa_K1255001</a><o:p></o:p></span></p> <p class=MsoNormal align=center style='margin-bottom:0cm;margin-bottom:.0001pt; text-align:center;line-height:normal;mso-yfti-cnfc:64'>&nbsp;</p> </td> <td width=418 valign=top style='width:313.45pt;border-top:none;border-left: none;border-bottom:solid #C2D69B 1.0pt;mso-border-bottom-themecolor:accent3; mso-border-bottom-themetint:153;border-right:solid #C2D69B 1.0pt;mso-border-right-themecolor: accent3;mso-border-right-themetint:153;mso-border-top-alt:solid #C2D69B .5pt; mso-border-top-themecolor:accent3;mso-border-top-themetint:153;mso-border-left-alt: solid #C2D69B .5pt;mso-border-left-themecolor:accent3;mso-border-left-themetint: 153;mso-border-alt:solid #C2D69B .5pt;mso-border-themecolor:accent3; mso-border-themetint:153;background:#EAF1DD;mso-background-themecolor:accent3; mso-background-themetint:51;padding:0cm 5.4pt 0cm 5.4pt;height:190.6pt'> <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align: justify;line-height:normal;mso-yfti-cnfc:64'><span style='font-size:12.0pt; mso-bidi-font-size:11.0pt;font-family:Oxygen;mso-fareast-font-family:"Times New Roman"; mso-bidi-font-family:Arial;color:black;mso-ansi-language:EN-US;mso-fareast-language: ES-MX'>Produces a <span class=SpellE>transferase</span> to convert salicylic acid into methyl salicylate (<span class=SpellE>WinterGreen</span> odor). The wintergreen odor generator requires of 2mM of salicylic acid to produce methyl salicylate. BSMT1 (<a href="http://parts.igem.org/Part:BBa_J45004">BBa_J45004</a>), <span class=SpellE>WinterGreen</span> Odor Generator original name, was created by <a href="https://2006.igem.org/wiki/index.php/MIT_2006">MIT 2006</a>. This year, the team optimized the sequence for <i>Escherichia coli</i>. </span><span class=SpellE><span lang=ES-MX style='font-size:12.0pt; mso-bidi-font-size:11.0pt;font-family:Oxygen;mso-fareast-font-family:"Times New Roman"; mso-bidi-font-family:Arial;color:black;mso-fareast-language:ES-MX'>The</span></span><span lang=ES-MX style='font-size:12.0pt;mso-bidi-font-size:11.0pt;font-family: Oxygen;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:Arial; color:black;mso-fareast-language:ES-MX'> new <span class=SpellE>biobrick</span> has a <span class=SpellE>length</span> of 1,074bp.<o:p></o:p></span></p> </td> </tr> <tr style='mso-yfti-irow:3;mso-yfti-lastrow:yes;height:70.7pt'> <td width=88 nowrap valign=top style='width:66.1pt;border-top:solid #C2D69B 1.0pt; mso-border-top-themecolor:accent3;mso-border-top-themetint:153;border-left: none;border-bottom:none;border-right:solid #C2D69B 1.0pt;mso-border-right-themecolor: accent3;mso-border-right-themetint:153;mso-border-top-alt:solid #C2D69B .5pt; mso-border-top-themecolor:accent3;mso-border-top-themetint:153;mso-border-right-alt: solid #C2D69B .5pt;mso-border-right-themecolor:accent3;mso-border-right-themetint: 153;background:white;mso-background-themecolor:background1;padding:0cm 5.4pt 0cm 5.4pt; height:70.7pt'> <p class=MsoNormal align=right style='margin-bottom:0cm;margin-bottom:.0001pt; text-align:right;line-height:normal;mso-yfti-cnfc:4'><!--[if gte vml 1]><v:shape id="_x0000_s1026" type="#_x0000_t75" style='position:absolute;left:0; text-align:left;margin-left:18pt;margin-top:1.85pt;width:59.1pt;height:49.4pt; z-index:251666944;visibility:visible;mso-wrap-style:square; mso-width-percent:0;mso-height-percent:0;mso-wrap-distance-left:9pt; mso-wrap-distance-top:0;mso-wrap-distance-right:9pt; mso-wrap-distance-bottom:0;mso-position-horizontal:absolute; mso-position-horizontal-relative:text;mso-position-vertical:absolute; mso-position-vertical-relative:text;mso-width-percent:0; mso-height-percent:0;mso-width-relative:page;mso-height-relative:page'> <v:imagedata src="TablasAromaYResistencia_archivos/image013.png" o:title=""/> <w:wrap type="square"/> </v:shape><![endif]--><![if !vml]><img width=79 height=66 src="https://static.igem.org/mediawiki/2014hs/6/62/TerminatorAroma.png" align=left hspace=12 v:shapes="_x0000_s1026"><![endif]></p> </td> <td width=119 nowrap valign=top style='width:89.0pt;border-top:none; border-left:none;border-bottom:solid #C2D69B 1.0pt;mso-border-bottom-themecolor: accent3;mso-border-bottom-themetint:153;border-right:solid #C2D69B 1.0pt; mso-border-right-themecolor:accent3;mso-border-right-themetint:153; mso-border-top-alt:solid #C2D69B .5pt;mso-border-top-themecolor:accent3; mso-border-top-themetint:153;mso-border-left-alt:solid #C2D69B .5pt; mso-border-left-themecolor:accent3;mso-border-left-themetint:153;mso-border-alt: solid #C2D69B .5pt;mso-border-themecolor:accent3;mso-border-themetint:153; padding:0cm 5.4pt 0cm 5.4pt;height:70.7pt'> <p class=MsoNormal align=center style='margin-bottom:0cm;margin-bottom:.0001pt; text-align:center;line-height:normal'><span lang=ES-MX style='font-size:12.0pt; mso-bidi-font-size:11.0pt;font-family:Oxygen;mso-fareast-font-family:"Times New Roman"; mso-bidi-font-family:Arial;mso-fareast-language:ES-MX'><a href="http://parts.igem.org/Part:BBa_B1002">BBa_B1002</a><o:p></o:p></span></p> <p class=MsoNormal align=center style='margin-bottom:0cm;margin-bottom:.0001pt; text-align:center;line-height:normal'>&nbsp;</p> </td> <td width=418 valign=top style='width:313.45pt;border-top:none;border-left: none;border-bottom:solid #C2D69B 1.0pt;mso-border-bottom-themecolor:accent3; mso-border-bottom-themetint:153;border-right:solid #C2D69B 1.0pt;mso-border-right-themecolor: accent3;mso-border-right-themetint:153;mso-border-top-alt:solid #C2D69B .5pt; mso-border-top-themecolor:accent3;mso-border-top-themetint:153;mso-border-left-alt: solid #C2D69B .5pt;mso-border-left-themecolor:accent3;mso-border-left-themetint: 153;mso-border-alt:solid #C2D69B .5pt;mso-border-themecolor:accent3; mso-border-themetint:153;padding:0cm 5.4pt 0cm 5.4pt;height:70.7pt'> <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align: justify;line-height:normal'><span style='font-size:12.0pt;mso-bidi-font-size: 11.0pt;font-family:Oxygen;mso-fareast-font-family:"Times New Roman"; mso-bidi-font-family:Arial;color:black;mso-ansi-language:EN-US;mso-fareast-language: ES-MX'>Part made of 6bp, responsible for transcription stop. The terminator stops the production of methyl salicylate. <o:p></o:p></span></p> </td> </tr></table></div></center><br>
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<tr style='mso-yfti-irow:-1;mso-yfti-firstrow:yes;mso-yfti-lastfirstrow:yes; height:15.0pt'>  
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<td width=88 nowrap valign=top style='width:66.1pt;border:none;border-bottom: solid #C2D69B 1.0pt;mso-border-bottom-themecolor:accent3;mso-border-bottom-themetint: 153;mso-border-bottom-alt:solid #C2D69B .5pt;mso-border-bottom-themecolor: accent3;mso-border-bottom-themetint:153;background:white;mso-background-themecolor: background1;padding:0cm 5.4pt 0cm 5.4pt;height:15.0pt'>  
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<p class=MsoNormal align=center style='margin-bottom:0cm;margin-bottom:.0001pt; text-align:center;line-height:normal;mso-yfti-cnfc:517'><b>
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<p><b><h2>Justification</h2></b></p>
 +
<p>Different to fluorescent reporters, this module was made in order to (in the future) perform as an aroma reporter and also to test the correct function of the bacteria, for its future usage as a new reporter and functional part (CDS). It is desired to use this part in the project to replace the red fluorescent protein (RFP) in the Capture module. But it was preferable to test it apart to demonstrate its effectiveness. Similarly, this piece is also helpful when the biofilter is assembled, because when performing the filtration by silica, WinterGreen can demonstrate the presence of bacteria in the beads. </p>
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<span lang=ES-MX style='font-family:Oxygen;mso-fareast-font-family:"Times New Roman"; mso-bidi-font-family:"Times New Roman";mso-fareast-language:ES-MX;mso-bidi-font-style: italic'>IMAGE
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<p>The team added the RNA thermometer in the device for regulating the production of the aroma. Another reason for selecting the RNA thermometer as a regulator was to continue the CIDEB UANL 2013 work with it.</p>
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</span></b>
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<span lang=ES-MX style='font-family:Oxygen; mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:"Times New Roman"; mso-fareast-language:ES-MX;mso-bidi-font-style:italic'><o:p></o:p>
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<p><b><h2>Other teams that used RNA thermometer and BSMT1</h2></b></p>
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</span></p>  
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</td>
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<td width=119 valign=top style='width:89.0pt;border:none;border-bottom:solid #C2D69B 1.0pt; mso-border-bottom-themecolor:accent3;mso-border-bottom-themetint:153; mso-border-bottom-alt:solid #C2D69B .5pt;mso-border-bottom-themecolor:accent3; mso-border-bottom-themetint:153;background:white;mso-background-themecolor: background1;padding:0cm 5.4pt 0cm 5.4pt;height:15.0pt'>  
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<p class=MsoNormal align=center style='margin-bottom:0cm;margin-bottom:.0001pt; text-align:center;line-height:normal;mso-yfti-cnfc:1'><b>
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<div align=center width="80%" >
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<table border=1 cellspacing=0 cellpadding=0 style='border-collapse:collapse;border:none;mso-border-alt:solid #C2D69B .5pt; mso-yfti-tbllook:1184;mso-padding-alt:0cm 5.4pt 0cm 5.4pt;mso-border-insideh: .5pt solid #C2D69B;mso-border-insidev:.5pt solid #C2D69B'>
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<tr style='mso-yfti-irow:0;mso-yfti-firstrow:yes'>  
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<td width=170 valign=top style='width:127.35pt;border:solid #9BBB59 1.0pt; border-right:none;mso-border-top-alt:solid #9BBB59 .5pt;mso-border-left-alt: solid #9BBB59 .5pt;mso-border-bottom-alt:solid #9BBB59 .5pt;background:#9BBB59; padding:0cm 5.4pt 0cm 5.4pt'>  
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<span lang=ES-MX style='font-family:Oxygen;mso-fareast-font-family:"Times New Roman"; mso-bidi-font-family:"Times New Roman";mso-fareast-language:ES-MX'>CODE
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<p class=MsoNormal align=center style='margin-bottom:0cm;margin-bottom:.0001pt; text-align:center;line-height:normal'><b>
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</span></b>
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<span lang=ES-MX style='font-family:Oxygen;mso-fareast-font-family:"Times New Roman"; mso-bidi-font-family:"Times New Roman";mso-fareast-language:ES-MX'><o:p></o:p>
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<span style='font-size:12.0pt; color:white;mso-ansi-language:EN-US'>Team<o:p></o:p>
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</span></p>  
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</span></b></p>  
</td>  
</td>  
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<td width=418 nowrap valign=top style='width:313.45pt;border:none;border-bottom: solid #C2D69B 1.0pt;mso-border-bottom-themecolor:accent3;mso-border-bottom-themetint: 153;mso-border-bottom-alt:solid #C2D69B .5pt;mso-border-bottom-themecolor: accent3;mso-border-bottom-themetint:153;background:white;mso-background-themecolor: background1;padding:0cm 5.4pt 0cm 5.4pt;height:15.0pt'>  
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<td width=170 valign=top style='width:127.35pt;border:solid #9BBB59 1.0pt; border-left:none;mso-border-top-alt:solid #9BBB59 .5pt;mso-border-bottom-alt: solid #9BBB59 .5pt;mso-border-right-alt:solid #9BBB59 .5pt;background:#9BBB59; padding:0cm 5.4pt 0cm 5.4pt'>  
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<p class=MsoNormal align=center style='margin-bottom:0cm;margin-bottom:.0001pt; text-align:center;line-height:normal;mso-yfti-cnfc:1'><b>
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<p class=MsoNormal align=center style='margin-bottom:0cm;margin-bottom:.0001pt; text-align:center;line-height:normal'><b>
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<span lang=ES-MX style='font-family:Oxygen;mso-fareast-font-family:"Times New Roman"; mso-bidi-font-family:"Times New Roman";mso-fareast-language:ES-MX'>DESCRIPTION
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<span style='font-size:12.0pt; color:white;mso-ansi-language:EN-US'>Part Used<o:p></o:p>
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</span></b>
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</span></b></p>  
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<span lang=ES-MX style='font-family:Oxygen;mso-fareast-font-family:"Times New Roman"; mso-bidi-font-family:"Times New Roman";mso-fareast-language:ES-MX'><o:p></o:p>
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</span></p>  
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</td>  
</td>  
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<td width=387 valign=top style='width:290.6pt;border:solid #9BBB59 1.0pt; border-left:none;mso-border-top-alt:solid #9BBB59 .5pt;mso-border-bottom-alt: solid #9BBB59 .5pt;mso-border-right-alt:solid #9BBB59 .5pt;background:#9BBB59; padding:0cm 5.4pt 0cm 5.4pt'>  
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<tr style='mso-yfti-irow:0;height:83.15pt'>
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<td width=88 nowrap valign=top style='width:66.1pt;border:none;border-right: solid #C2D69B 1.0pt;mso-border-right-themecolor:accent3;mso-border-right-themetint: 153;mso-border-top-alt:solid #C2D69B .5pt;mso-border-top-themecolor:accent3; mso-border-top-themetint:153;mso-border-top-alt:solid #C2D69B .5pt; mso-border-top-themecolor:accent3;mso-border-top-themetint:153;mso-border-right-alt: solid #C2D69B .5pt;mso-border-right-themecolor:accent3;mso-border-right-themetint: 153;background:white;mso-background-themecolor:background1;padding:0cm 5.4pt 0cm 5.4pt; height:83.15pt'>  
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<p class=MsoNormal align=right style='margin-bottom:0cm;margin-bottom:.0001pt; text-align:right;line-height:normal;mso-yfti-cnfc:68'>
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<p class=MsoNormal align=center style='margin-bottom:0cm;margin-bottom:.0001pt; text-align:center;line-height:normal'><b>
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<!--[if gte vml 1]><v:shape id="_x0000_s1029" type="#_x0000_t75" style='position:absolute;left:0; text-align:left;margin-left:10.95pt;margin-top:3.2pt;width:66pt;height:48pt; z-index:251660800;visibility:visible;mso-wrap-style:square; mso-width-percent:0;mso-height-percent:0;mso-wrap-distance-left:9pt; mso-wrap-distance-top:0;mso-wrap-distance-right:9pt; mso-wrap-distance-bottom:0;mso-position-horizontal:absolute; mso-position-horizontal-relative:text;mso-position-vertical:absolute; mso-position-vertical-relative:text;mso-width-percent:0; mso-height-percent:0;mso-width-relative:page;mso-height-relative:page'> <v:imagedata src="TablasAromaYResistencia_archivos/image007.png" o:title=""/> <w:wrap type="square"/> </v:shape><![endif]--><![if !vml]><img width=88 height=64 src="https://static.igem.org/mediawiki/2014hs/4/43/PConsAroma.png" align=left hspace=12 v:shapes="_x0000_s1029"><![endif]></p>  
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<p class=MsoNormal align=right style='margin-bottom:0cm;margin-bottom:.0001pt; text-align:right;line-height:normal;mso-yfti-cnfc:68'><i>
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<span style='font-size:12.0pt; color:white;mso-ansi-language:EN-US'>Use<o:p></o:p>
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</span></b></p>  
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<span lang=ES-MX style='font-family:Oxygen;color:#76923C;mso-themecolor:accent3;mso-themeshade: 191;mso-fareast-language:ES-MX;mso-no-proof:yes'><o:p>&nbsp;</o:p>
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</span></i></p>  
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</td>  
</td>  
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<td width=119 nowrap valign=top style='width:89.0pt;border-top:none; border-left:none;border-bottom:solid #C2D69B 1.0pt;mso-border-bottom-themecolor: accent3;mso-border-bottom-themetint:153;border-right:solid #C2D69B 1.0pt; mso-border-right-themecolor:accent3;mso-border-right-themetint:153; mso-border-top-alt:solid #C2D69B .5pt;mso-border-top-themecolor:accent3; mso-border-top-themetint:153;mso-border-left-alt:solid #C2D69B .5pt; mso-border-left-themecolor:accent3;mso-border-left-themetint:153;mso-border-alt: solid #C2D69B .5pt;mso-border-themecolor:accent3;mso-border-themetint:153; background:#EAF1DD;mso-background-themecolor:accent3;mso-background-themetint: 51;padding:0cm 5.4pt 0cm 5.4pt;height:83.15pt'>  
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</tr>
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<tr style='mso-yfti-irow:1;height:33.0pt'>
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<td width=170 valign=top style='width:127.35pt;border:solid #C2D69B 1.0pt; border-top:none;mso-border-top-alt:solid #C2D69B .5pt;mso-border-alt:solid #C2D69B .5pt; background:#EAF1DD;padding:0cm 5.4pt 0cm 5.4pt;height:33.0pt'>  
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<p class=MsoNormal align=center style='margin-bottom:0cm;margin-bottom:.0001pt; text-align:center;line-height:normal;mso-yfti-cnfc:64'>
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<p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;line-height: normal'><b><u>
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<span lang=ES-MX style='font-size:12.0pt;mso-bidi-font-size:11.0pt;font-family:Oxygen; mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:Arial; color:black;mso-fareast-language:ES-MX'><a href="http://parts.igem.org/Part:BBa_J23119">BBa_J23119A</a>
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<span lang=FR style='font-size:12.0pt;color:#4F81BD;mso-ansi-language: FR'><a href="https://2008.igem.org/Team:TUDelft" target="_blank">TuDelft 2008</a>
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</span>
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</span></u></b>
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<span lang=ES-MX style='font-family:Oxygen;mso-fareast-font-family:"Times New Roman"; mso-bidi-font-family:Arial;color:black;mso-fareast-language:ES-MX'><o:p></o:p>
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<span lang=FR style='font-size:6.0pt;mso-bidi-font-size:12.0pt;mso-ansi-language:FR; mso-bidi-font-weight:bold'>:
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</span></p>  
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</span><b>
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<p class=MsoNormal align=center style='margin-bottom:0cm;margin-bottom:.0001pt; text-align:center;line-height:normal;mso-yfti-cnfc:64'>&nbsp;</p>  
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<span lang=FR style='font-size:12.0pt; mso-ansi-language:FR'><o:p></o:p>
 +
</span></b></p>  
</td>  
</td>  
-
<td width=418 valign=top style='width:313.45pt;border-top:none;border-left: none;border-bottom:solid #C2D69B 1.0pt;mso-border-bottom-themecolor:accent3; mso-border-bottom-themetint:153;border-right:solid #C2D69B 1.0pt;mso-border-right-themecolor: accent3;mso-border-right-themetint:153;mso-border-top-alt:solid #C2D69B .5pt; mso-border-top-themecolor:accent3;mso-border-top-themetint:153;mso-border-left-alt: solid #C2D69B .5pt;mso-border-left-themecolor:accent3;mso-border-left-themetint: 153;mso-border-alt:solid #C2D69B .5pt;mso-border-themecolor:accent3; mso-border-themetint:153;background:#EAF1DD;mso-background-themecolor:accent3; mso-background-themetint:51;padding:0cm 5.4pt 0cm 5.4pt;height:83.15pt'>  
+
<td width=170 valign=top style='width:127.35pt;border-top:none;border-left: none;border-bottom:solid #C2D69B 1.0pt;border-right:solid #C2D69B 1.0pt; mso-border-top-alt:solid #C2D69B .5pt;mso-border-left-alt:solid #C2D69B .5pt; mso-border-alt:solid #C2D69B .5pt;background:#EAF1DD;padding:0cm 5.4pt 0cm 5.4pt; height:33.0pt'>  
-
<p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align: justify;line-height:normal;mso-yfti-cnfc:64'>
+
<p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;line-height: normal'>
-
<span style='font-size:12.0pt; mso-bidi-font-size:11.0pt;font-family:Oxygen;mso-fareast-font-family:"Times New Roman"; mso-bidi-font-family:Arial;color:black;mso-ansi-language:EN-US;mso-fareast-language: ES-MX'>In the specific case of our aroma module, it will help the bacteria to continuously transcribe the
+
<span style='font-size:12.0pt;mso-ansi-language:EN-US'>RNA themometer<o:p></o:p>
 +
</span></p>  
-
<span class=SpellE>WinterGreen
+
<p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;line-height: normal'>
-
</span> gene in order to allow the bacteria to continuously produce the aroma.  
+
-
</span>
+
-
<span class=SpellE>
+
<span style='font-size:12.0pt;mso-ansi-language:EN-US'><o:p>&nbsp;</o:p>
 +
</span></p>
 +
</td>
 +
<td width=387 valign=top style='width:290.6pt;border-top:none;border-left: none;border-bottom:solid #C2D69B 1.0pt;border-right:solid #C2D69B 1.0pt; mso-border-top-alt:solid #C2D69B .5pt;mso-border-left-alt:solid #C2D69B .5pt; mso-border-alt:solid #C2D69B .5pt;background:#EAF1DD;padding:0cm 5.4pt 0cm 5.4pt; height:33.0pt'>  
-
<span lang=ES-MX style='font-size:12.0pt;mso-bidi-font-size: 11.0pt;font-family:Oxygen;mso-fareast-font-family:"Times New Roman"; mso-bidi-font-family:Arial;color:black;mso-fareast-language:ES-MX'>This
+
<p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;line-height: normal'>
-
</span>
+
-
</span>
+
-
<span lang=ES-MX style='font-size:12.0pt;mso-bidi-font-size:11.0pt;font-family: Oxygen;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:Arial; color:black;mso-fareast-language:ES-MX'>  
+
<span style='font-size:12.0pt;mso-ansi-language:EN-US'>Temperature-sensing bacteria that changes color at different temperatures; as a temperature reporter system inlarge-scale fermentations, or as a temperature-inducible protein production system.<o:p></o:p>
 +
</span></p>  
-
<span class=SpellE>promoter
+
<p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;line-height: normal'>
-
</span> has a
+
-
<span class=SpellE>length
+
<span style='font-size:12.0pt;mso-ansi-language:EN-US'><o:p>&nbsp;</o:p>
-
</span> of 35bp.<o:p></o:p>
+
</span></p>  
</span></p>  
</td>  
</td>  
</tr>  
</tr>  
-
<tr style='mso-yfti-irow:1;height:77.45pt'>  
+
<tr style='mso-yfti-irow:2'>  
-
<td width=88 nowrap valign=top style='width:66.1pt;border-top:solid #C2D69B 1.0pt; mso-border-top-themecolor:accent3;mso-border-top-themetint:153;border-left: none;border-bottom:none;border-right:solid #C2D69B 1.0pt;mso-border-right-themecolor: accent3;mso-border-right-themetint:153;mso-border-top-alt:solid #C2D69B .5pt; mso-border-top-themecolor:accent3;mso-border-top-themetint:153;mso-border-right-alt: solid #C2D69B .5pt;mso-border-right-themecolor:accent3;mso-border-right-themetint: 153;background:white;mso-background-themecolor:background1;padding:0cm 5.4pt 0cm 5.4pt; height:77.45pt'>  
+
<td width=170 valign=top style='width:127.35pt;border:solid #C2D69B 1.0pt; border-top:none;mso-border-top-alt:solid #C2D69B .5pt;mso-border-alt:solid #C2D69B .5pt; padding:0cm 5.4pt 0cm 5.4pt'>  
-
<p class=MsoNormal align=right style='margin-bottom:0cm;margin-bottom:.0001pt; text-align:right;line-height:normal;mso-yfti-cnfc:4'>
+
<p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;line-height: normal'><b><u>
-
<!--[if gte vml 1]><v:shape id="_x0000_s1028" type="#_x0000_t75" style='position:absolute;left:0; text-align:left;margin-left:16.95pt;margin-top:4.25pt;width:57.75pt; height:44.25pt;z-index:251662848;visibility:visible;mso-wrap-style:square; mso-width-percent:0;mso-height-percent:0;mso-wrap-distance-left:9pt; mso-wrap-distance-top:0;mso-wrap-distance-right:9pt; mso-wrap-distance-bottom:0;mso-position-horizontal:absolute; mso-position-horizontal-relative:text;mso-position-vertical:absolute; mso-position-vertical-relative:text;mso-width-percent:0; mso-height-percent:0;mso-width-relative:page;mso-height-relative:page'> <v:imagedata src="TablasAromaYResistencia_archivos/image009.png" o:title=""/> <w:wrap type="square"/> </v:shape><![endif]--><![if !vml]><img width=77 height=59 src="https://static.igem.org/mediawiki/2014hs/b/b0/RNAthermAroma.png" align=left hspace=12 v:shapes="_x0000_s1028"><![endif]><i>
+
-
<span lang=ES-MX style='font-family: Oxygen;color:#76923C;mso-themecolor:accent3;mso-themeshade:191;mso-fareast-language: ES-MX;mso-no-proof:yes'><o:p></o:p>
+
<span style='font-size:12.0pt;color:#4F81BD;mso-ansi-language: EN-US'><a href="https://2009.igem.org/Team:VictoriaBC" target="_blank">Victoria BC 2009</a>
-
</span></i></p>
+
</span></u></b>
-
</td>
+
-
<td width=119 nowrap valign=top style='width:89.0pt;border-top:none; border-left:none;border-bottom:solid #C2D69B 1.0pt;mso-border-bottom-themecolor: accent3;mso-border-bottom-themetint:153;border-right:solid #C2D69B 1.0pt; mso-border-right-themecolor:accent3;mso-border-right-themetint:153; mso-border-top-alt:solid #C2D69B .5pt;mso-border-top-themecolor:accent3; mso-border-top-themetint:153;mso-border-left-alt:solid #C2D69B .5pt; mso-border-left-themecolor:accent3;mso-border-left-themetint:153;mso-border-alt: solid #C2D69B .5pt;mso-border-themecolor:accent3;mso-border-themetint:153; padding:0cm 5.4pt 0cm 5.4pt;height:77.45pt'>  
+
-
<p class=MsoNormal align=center style='margin-bottom:0cm;margin-bottom:.0001pt; text-align:center;line-height:normal'>
+
<span style='font-size:6.0pt;mso-bidi-font-size:12.0pt;mso-ansi-language:EN-US; mso-bidi-font-weight:bold'>:<b> <o:p></o:p></b>
-
 
+
-
<span lang=ES-MX style='font-size:12.0pt; mso-bidi-font-size:11.0pt;font-family:Oxygen;mso-fareast-font-family:"Times New Roman"; mso-bidi-font-family:Arial;color:black;mso-fareast-language:ES-MX'><a href="http://parts.igem.org/Part:BBa_K115017">BBa_K115017</a><o:p></o:p>
+
-
</span></p>
+
-
</td>
+
-
<td width=418 valign=top style='width:313.45pt;border-top:none;border-left: none;border-bottom:solid #C2D69B 1.0pt;mso-border-bottom-themecolor:accent3; mso-border-bottom-themetint:153;border-right:solid #C2D69B 1.0pt;mso-border-right-themecolor: accent3;mso-border-right-themetint:153;mso-border-top-alt:solid #C2D69B .5pt; mso-border-top-themecolor:accent3;mso-border-top-themetint:153;mso-border-left-alt: solid #C2D69B .5pt;mso-border-left-themecolor:accent3;mso-border-left-themetint: 153;mso-border-alt:solid #C2D69B .5pt;mso-border-themecolor:accent3; mso-border-themetint:153;padding:0cm 5.4pt 0cm 5.4pt;height:77.45pt'>
+
-
 
+
-
<p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align: justify;line-height:normal'>
+
-
 
+
-
<span style='font-size:12.0pt;mso-bidi-font-size: 11.0pt;font-family:Oxygen;mso-fareast-font-family:"Times New Roman"; mso-bidi-font-family:Arial;color:black;mso-ansi-language:EN-US;mso-fareast-language: ES-MX'>A RNA thermometer, used for temperature post-transcriptional regulation (thermo sensor), and is designed to initiate transcription around 32°C.<o:p></o:p>
+
</span></p>  
</span></p>  
-
</td>
 
-
</tr>
 
-
<tr style='mso-yfti-irow:2;height:190.6pt'>
 
-
<td width=88 nowrap valign=top style='width:66.1pt;border-top:solid #C2D69B 1.0pt; mso-border-top-themecolor:accent3;mso-border-top-themetint:153;border-left: none;border-bottom:none;border-right:solid #C2D69B 1.0pt;mso-border-right-themecolor: accent3;mso-border-right-themetint:153;mso-border-top-alt:solid #C2D69B .5pt; mso-border-top-themecolor:accent3;mso-border-top-themetint:153;mso-border-right-alt: solid #C2D69B .5pt;mso-border-right-themecolor:accent3;mso-border-right-themetint: 153;background:white;mso-background-themecolor:background1;padding:0cm 5.4pt 0cm 5.4pt; height:190.6pt'>
 
-
<p class=MsoNormal align=right style='margin-bottom:0cm;margin-bottom:.0001pt; text-align:right;line-height:normal;mso-yfti-cnfc:68'>
+
<p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;line-height: normal'><b>
-
<!--[if gte vml 1]><v:shape id="_x0000_s1027" type="#_x0000_t75" style='position:absolute;left:0; text-align:left;margin-left:13.95pt;margin-top:3.2pt;width:63pt;height:51.75pt; z-index:251664896;visibility:visible;mso-wrap-style:square; mso-width-percent:0;mso-height-percent:0;mso-wrap-distance-left:9pt; mso-wrap-distance-top:0;mso-wrap-distance-right:9pt; mso-wrap-distance-bottom:0;mso-position-horizontal:absolute; mso-position-horizontal-relative:text;mso-position-vertical:absolute; mso-position-vertical-relative:text;mso-width-percent:0; mso-height-percent:0;mso-width-relative:page;mso-height-relative:page'> <v:imagedata src="TablasAromaYResistencia_archivos/image011.png" o:title=""/> <w:wrap type="square"/> </v:shape><![endif]--><![if !vml]><img width=84 height=69 src="https://static.igem.org/mediawiki/2014hs/b/b1/WinterGreenAroma.png" align=left hspace=12 v:shapes="_x0000_s1027"><![endif]><i>
+
-
<span style='font-size:3.0pt;font-family: Oxygen;color:#76923C;mso-themecolor:accent3;mso-themeshade:191;mso-ansi-language: EN-US;mso-fareast-language:ES-MX;mso-no-proof:yes'><o:p></o:p>
+
<span style='font-size:12.0pt;mso-ansi-language:EN-US'><o:p>&nbsp;</o:p>
-
</span></i></p>
+
</span></b></p>  
</td>  
</td>  
-
<td width=119 nowrap valign=top style='width:89.0pt;border-top:none; border-left:none;border-bottom:solid #C2D69B 1.0pt;mso-border-bottom-themecolor: accent3;mso-border-bottom-themetint:153;border-right:solid #C2D69B 1.0pt; mso-border-right-themecolor:accent3;mso-border-right-themetint:153; mso-border-top-alt:solid #C2D69B .5pt;mso-border-top-themecolor:accent3; mso-border-top-themetint:153;mso-border-left-alt:solid #C2D69B .5pt; mso-border-left-themecolor:accent3;mso-border-left-themetint:153;mso-border-alt: solid #C2D69B .5pt;mso-border-themecolor:accent3;mso-border-themetint:153; background:#EAF1DD;mso-background-themecolor:accent3;mso-background-themetint: 51;padding:0cm 5.4pt 0cm 5.4pt;height:190.6pt'>  
+
<td width=170 valign=top style='width:127.35pt;border-top:none;border-left: none;border-bottom:solid #C2D69B 1.0pt;border-right:solid #C2D69B 1.0pt; mso-border-top-alt:solid #C2D69B .5pt;mso-border-left-alt:solid #C2D69B .5pt; mso-border-alt:solid #C2D69B .5pt;padding:0cm 5.4pt 0cm 5.4pt'>  
-
<p class=MsoNormal align=center style='margin-bottom:0cm;margin-bottom:.0001pt; text-align:center;line-height:normal;mso-yfti-cnfc:64'>
+
<p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;line-height: normal'>
-
<span style='font-size:12.0pt;mso-bidi-font-size:11.0pt;font-family:Oxygen; mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:Arial; mso-ansi-language:EN-US;mso-fareast-language:ES-MX'><a href="http://parts.igem.org/Part:BBa_K1255001:Design">BBa_K1255001</a><o:p></o:p>
+
<span style='font-size:12.0pt;mso-ansi-language:EN-US'>RNA themometer<o:p></o:p>
</span></p>  
</span></p>  
-
<p class=MsoNormal align=center style='margin-bottom:0cm;margin-bottom:.0001pt; text-align:center;line-height:normal;mso-yfti-cnfc:64'>&nbsp;</p>
+
<p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;line-height: normal'>
-
</td>
+
-
<td width=418 valign=top style='width:313.45pt;border-top:none;border-left: none;border-bottom:solid #C2D69B 1.0pt;mso-border-bottom-themecolor:accent3; mso-border-bottom-themetint:153;border-right:solid #C2D69B 1.0pt;mso-border-right-themecolor: accent3;mso-border-right-themetint:153;mso-border-top-alt:solid #C2D69B .5pt; mso-border-top-themecolor:accent3;mso-border-top-themetint:153;mso-border-left-alt: solid #C2D69B .5pt;mso-border-left-themecolor:accent3;mso-border-left-themetint: 153;mso-border-alt:solid #C2D69B .5pt;mso-border-themecolor:accent3; mso-border-themetint:153;background:#EAF1DD;mso-background-themecolor:accent3; mso-background-themetint:51;padding:0cm 5.4pt 0cm 5.4pt;height:190.6pt'>  
+
-
<p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align: justify;line-height:normal;mso-yfti-cnfc:64'>
+
<span style='font-size:12.0pt;mso-ansi-language:EN-US'><o:p>&nbsp;</o:p>
-
 
+
-
<span style='font-size:12.0pt; mso-bidi-font-size:11.0pt;font-family:Oxygen;mso-fareast-font-family:"Times New Roman"; mso-bidi-font-family:Arial;color:black;mso-ansi-language:EN-US;mso-fareast-language: ES-MX'>Produces a
+
-
 
+
-
<span class=SpellE>transferase
+
-
</span> to convert salicylic acid into methyl salicylate (
+
-
 
+
-
<span class=SpellE>WinterGreen
+
-
</span> odor). The wintergreen odor generator requires of 2mM of salicylic acid to produce methyl salicylate. BSMT1 (<a href="http://parts.igem.org/Part:BBa_J45004">BBa_J45004</a>),
+
-
 
+
-
<span class=SpellE>WinterGreen
+
-
</span> Odor Generator original name, was created by <a href="https://2006.igem.org/wiki/index.php/MIT_2006">MIT 2006</a>. This year, the team optimized the sequence for <i>Escherichia coli</i>.
+
-
</span>
+
-
 
+
-
<span class=SpellE>
+
-
 
+
-
<span lang=ES-MX style='font-size:12.0pt; mso-bidi-font-size:11.0pt;font-family:Oxygen;mso-fareast-font-family:"Times New Roman"; mso-bidi-font-family:Arial;color:black;mso-fareast-language:ES-MX'>The
+
-
</span>
+
-
</span>
+
-
 
+
-
<span lang=ES-MX style='font-size:12.0pt;mso-bidi-font-size:11.0pt;font-family: Oxygen;mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:Arial; color:black;mso-fareast-language:ES-MX'> new
+
-
 
+
-
<span class=SpellE>biobrick
+
-
</span> has a
+
-
 
+
-
<span class=SpellE>length
+
-
</span> of 1,074bp.<o:p></o:p>
+
</span></p>  
</span></p>  
</td>  
</td>  
-
</tr>
+
<td width=387 valign=top style='width:290.6pt;border-top:none;border-left: none;border-bottom:solid #C2D69B 1.0pt;border-right:solid #C2D69B 1.0pt; mso-border-top-alt:solid #C2D69B .5pt;mso-border-left-alt:solid #C2D69B .5pt; mso-border-alt:solid #C2D69B .5pt;padding:0cm 5.4pt 0cm 5.4pt'>  
-
<tr style='mso-yfti-irow:3;mso-yfti-lastrow:yes;height:70.7pt'>
+
-
<td width=88 nowrap valign=top style='width:66.1pt;border-top:solid #C2D69B 1.0pt; mso-border-top-themecolor:accent3;mso-border-top-themetint:153;border-left: none;border-bottom:none;border-right:solid #C2D69B 1.0pt;mso-border-right-themecolor: accent3;mso-border-right-themetint:153;mso-border-top-alt:solid #C2D69B .5pt; mso-border-top-themecolor:accent3;mso-border-top-themetint:153;mso-border-right-alt: solid #C2D69B .5pt;mso-border-right-themecolor:accent3;mso-border-right-themetint: 153;background:white;mso-background-themecolor:background1;padding:0cm 5.4pt 0cm 5.4pt; height:70.7pt'>  
+
-
<p class=MsoNormal align=right style='margin-bottom:0cm;margin-bottom:.0001pt; text-align:right;line-height:normal;mso-yfti-cnfc:4'>
+
<p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;line-height: normal'>
-
<!--[if gte vml 1]><v:shape id="_x0000_s1026" type="#_x0000_t75" style='position:absolute;left:0; text-align:left;margin-left:18pt;margin-top:1.85pt;width:59.1pt;height:49.4pt; z-index:251666944;visibility:visible;mso-wrap-style:square; mso-width-percent:0;mso-height-percent:0;mso-wrap-distance-left:9pt; mso-wrap-distance-top:0;mso-wrap-distance-right:9pt; mso-wrap-distance-bottom:0;mso-position-horizontal:absolute; mso-position-horizontal-relative:text;mso-position-vertical:absolute; mso-position-vertical-relative:text;mso-width-percent:0; mso-height-percent:0;mso-width-relative:page;mso-height-relative:page'> <v:imagedata src="TablasAromaYResistencia_archivos/image013.png" o:title=""/> <w:wrap type="square"/> </v:shape><![endif]--><![if !vml]><img width=79 height=66 src="https://static.igem.org/mediawiki/2014hs/6/62/TerminatorAroma.png" align=left hspace=12 v:shapes="_x0000_s1026"><![endif]></p>
+
-
</td>
+
-
<td width=119 nowrap valign=top style='width:89.0pt;border-top:none; border-left:none;border-bottom:solid #C2D69B 1.0pt;mso-border-bottom-themecolor: accent3;mso-border-bottom-themetint:153;border-right:solid #C2D69B 1.0pt; mso-border-right-themecolor:accent3;mso-border-right-themetint:153; mso-border-top-alt:solid #C2D69B .5pt;mso-border-top-themecolor:accent3; mso-border-top-themetint:153;mso-border-left-alt:solid #C2D69B .5pt; mso-border-left-themecolor:accent3;mso-border-left-themetint:153;mso-border-alt: solid #C2D69B .5pt;mso-border-themecolor:accent3;mso-border-themetint:153; padding:0cm 5.4pt 0cm 5.4pt;height:70.7pt'>  
+
-
<p class=MsoNormal align=center style='margin-bottom:0cm;margin-bottom:.0001pt; text-align:center;line-height:normal'>
+
<span style='font-size:12.0pt;mso-ansi-language:EN-US'>NAND logic gate using the ribo-key/ribo-lock system designed by <a href="https://2006.igem.org/wiki/index.php/Berkeley">Berkeley 2006</a> team , producing RFP except when the cells are grown in the presence of both arabinose and IPTG, also coupling fluorescent outputs with the ribo-thermometers made by TUDelft 2008 team.<o:p></o:p>
-
 
+
-
<span lang=ES-MX style='font-size:12.0pt; mso-bidi-font-size:11.0pt;font-family:Oxygen;mso-fareast-font-family:"Times New Roman"; mso-bidi-font-family:Arial;mso-fareast-language:ES-MX'><a href="http://parts.igem.org/Part:BBa_B1002">BBa_B1002</a><o:p></o:p>
+
</span></p>  
</span></p>  
-
<p class=MsoNormal align=center style='margin-bottom:0cm;margin-bottom:.0001pt; text-align:center;line-height:normal'>&nbsp;</p>
+
<p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;line-height: normal'>
-
</td>
+
-
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<p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align: justify;line-height:normal'>
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<span style='font-size:12.0pt;mso-bidi-font-size: 11.0pt;font-family:Oxygen;mso-fareast-font-family:"Times New Roman"; mso-bidi-font-family:Arial;color:black;mso-ansi-language:EN-US;mso-fareast-language: ES-MX'>Part made of 6bp, responsible for transcription stop. The terminator stops the production of methyl salicylate. <o:p></o:p>
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<span style='font-size:12.0pt;mso-ansi-language:EN-US'><o:p>&nbsp;</o:p>
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<br>
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<p><b>Full Device:</b></p>
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<p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;line-height: normal'><b><u>
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<p>These 4 genetic parts form the Aroma device of the project (see <b>Figure 3</b>). The full device's length is 1,251bp (including restriction sites).</p><center>
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<span style='font-size:12.0pt;color:#4F81BD;mso-ansi-language: EN-US'><a href="https://2013hs.igem.org/Team:CIDEB-UANL_Mexico" target="_blank">CIDEB UANL 2013</a>
 +
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<p><img width=629 height=106 src="https://static.igem.org/mediawiki/2014hs/3/3a/AromaFigure4.png"align=center hspace=12 alt="IMG_0317"></p>
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<span style='font-size:6.0pt;mso-bidi-font-size:12.0pt;mso-ansi-language:EN-US; mso-bidi-font-weight:bold'>:<b> <o:p></o:p></b>
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<p><b>Figure 3.</b> Aroma Device</p></center>
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<p><b>Other teams that use RNA thermometer and WinterGreen (BSMT1)</b></p>
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<span style='font-size:12.0pt;mso-ansi-language:EN-US'><o:p>&nbsp;</o:p>
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<p><i>RNA thermometer</i></p>
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<span style='font-size:12.0pt;mso-ansi-language:EN-US'>RNA themometer<o:p></o:p>
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<span class=SpellE><b style='mso-bidi-font-weight:normal'>
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<span style='font-size:12.0pt;mso-ansi-language:EN-US'>Production of Vip3ca3, which acts as a pesticide protein, regulated by specific temperatures in order to avoid overproduction and it will show activity against target organisms Coleoptera and Lepidoptera..<o:p></o:p>
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</span></p>  
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<span><a href="https://2008.igem.org/Team:TUDelft">TUDelft
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<span> 2008:</a>
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<span style='font-size:12.0pt;mso-ansi-language:EN-US'><o:p>&nbsp;</o:p>
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<td width=170 valign=top style='width:127.35pt;border:solid #C2D69B 1.0pt; border-top:none;mso-border-top-alt:solid #C2D69B .5pt;mso-border-alt:solid #C2D69B .5pt; background:#EAF1DD;padding:0cm 5.4pt 0cm 5.4pt'>  
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<span> Temperature-sensing bacteria that changescolor at different temperatures; as a temperature reporter system inlarge-scale fermentations, or as a temperature-inducible protein productionsystem. <o:p></o:p>
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<p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;line-height: normal'><b><u>
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</span></p><center>
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<p><img width=571 height=129 src="https://static.igem.org/mediawiki/2014hs/e/e0/AromaFigure5.png"align=center hspace=12 alt="IMG_0317"></p>
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<span lang=ES-MX style='font-size:12.0pt;color:#4F81BD'><a href="https://2006.igem.org/wiki/index.php/MIT_2006" target="_blank">MIT 2006</a>
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</span></u></b>
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<p><b>Figure 4.</b> RNA Thermometer circuit, excerpted from <a href="https://2008.igem.org/Team:TUDelft">TUDelft 2008</a> team</p></center>
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<span lang=ES-MX style='font-size:7.0pt;mso-bidi-font-size:12.0pt;mso-bidi-font-weight: bold'>:<b> <o:p></o:p></b>
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<td width=170 valign=top style='width:127.35pt;border-top:none;border-left: none;border-bottom:solid #C2D69B 1.0pt;border-right:solid #C2D69B 1.0pt; mso-border-top-alt:solid #C2D69B .5pt;mso-border-left-alt:solid #C2D69B .5pt; mso-border-alt:solid #C2D69B .5pt;background:#EAF1DD;padding:0cm 5.4pt 0cm 5.4pt'>  
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<span style='font-size:12.0pt;mso-ansi-language:EN-US'>BSMT1<o:p></o:p>
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<td width=387 valign=top style='width:290.6pt;border-top:none;border-left: none;border-bottom:solid #C2D69B 1.0pt;border-right:solid #C2D69B 1.0pt; mso-border-top-alt:solid #C2D69B .5pt;mso-border-left-alt:solid #C2D69B .5pt; mso-border-alt:solid #C2D69B .5pt;background:#EAF1DD;padding:0cm 5.4pt 0cm 5.4pt'>  
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<span> <a href="https://2009.igem.org/Team:VictoriaBC">VictoriaBC
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<span style='font-size:12.0pt;mso-ansi-language:EN-US'>This device produces methyl salicylate in the presence of salicylic acid. Methyl salicylate smells strongly of mint (wintergreen). Production of methyl salicylate was verified both by scent and by gas chromatography: E. coli with no WGD did not produce methyl salicylate when SA was added to the medium, while E. coli with the WGD did produce methyl salicylate when SA was added to the medium..<o:p></o:p>
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<span> 2009: </a>
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<span>NAND logic gate using the ribo-key/ribo-lock system designed by <a href="https://2006.igem.org/wiki/index.php/Berkeley">Berkeley 2006</a> team , producing RFP except when the cells are grown in the presence of both arabinose and IPTG, also coupling fluorescent outputs with the ribo-thermometers made by <a href="https://2008.igem.org/Team:TUDelft">TUDelft 2008</a> team. <o:p></o:p>
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<span><a href="https://2013hs.igem.org/Team:CIDEB-UANL_Mexico">iGEM_CIDEB
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<span> Production of Vip3ca3, which acts as a pesticide protein, regulated by specific temperatures in order to avoid overproduction and it will show activity against target organisms Coleoptera and Lepidoptera. <o:p></o:p>
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</span></p><center>
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<p><img width=474 height=146 src="https://static.igem.org/mediawiki/2014hs/3/3f/AromaFigure6.jpg "align=center hspace=12 alt="IMG_0317"></p>
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<p><b>Figure 5.</b> Circuit from <a href="https://2013hs.igem.org/Team:CIDEB-UANL_Mexico">iGEM CIDEB UANL 2013</a> team</p></center>
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<p><i>WinterGreen (BSMT1)</i></p>
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<span> This device produces methyl salicylate in the presence of salicylic acid. Methyl salicylate smells strongly of mint (wintergreen). Production of methyl salicylate was verified both by scent and by gas chromatography: E. coli with no WGD did not produce methyl salicylate when SA was added to the medium, while E. coli with the WGD did produce methyl salicylate when SA was added to the medium. <o:p></o:p>
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<p><b>Figure 6.</b> Wintergreen odor enzyme (BSMT1) generator circuit by <a href="https://2006.igem.org/wiki/index.php/MIT_2006">MIT 2006</a></p></center>
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<p><b><h2>Project Zoom in</h2></b></p>
<p><center><iframe width="600" height="500" src="//www.youtube.com/embed/fd9EQNCOr2k" frameborder="0" allowfullscreen></iframe></center></p>
<p><center><iframe width="600" height="500" src="//www.youtube.com/embed/fd9EQNCOr2k" frameborder="0" allowfullscreen></iframe></center></p>
<br>
<br>
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<p><b>Bibliography/References</b></p><font size="2">
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<p><b><h2>Bibliography/References</h2></b></p><font size="2">
<p>● Huang, H. (2006, August 30). <i>Part:BBa_B1002. </i> Retrieved August 30, 2014, from <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_B1002"> http://parts.igem.org/wiki/index.php?title=Part:BBa_B1002</a>.</p>
<p>● Huang, H. (2006, August 30). <i>Part:BBa_B1002. </i> Retrieved August 30, 2014, from <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_B1002"> http://parts.igem.org/wiki/index.php?title=Part:BBa_B1002</a>.</p>

Latest revision as of 03:27, 21 June 2014

iGEM CIDEB 2014 - Project

Aroma Module


Since the beginning of iGEM project, the use of fluorescent reporters has been used in each one of the proposed projects in previous years, trying to test the theoretical presence of other proteins in E. coli. For our iGEM 2014 project, this module proposed to promote the usage of aroma reporters, instead of fluorescent ones.


Description

In the module, WinterGreen is the most important part. Normally, it catalyzes the conversion of salicylic acid into methyl salicylate. The protein is excreted by the bacteria and when salicylic acid is added to the medium, a chemical reaction takes place and produces methyl salicylate, which is responsible for the wintergreen odor.

But in this module, it will be regulated by temperature with the use of the RNA thermometer. When adding salicylic acid to the bacteria in a 32° Celsius environment, the production of the WinterGreen protein will begin.


Figure 1. Production of Wintergreen Odor

Device

Originally, WinterGreen was thought to be the reporter of the Capture module, but in order to prove the function of this gene and also the one in charge of the capture of sodium ions (NhaS), we decided to separate the full device into the actual modules of Capture and Aroma, as seen in the figure below.

Figure 2. The device originally thought, and the 2 modules derived from it.


This device is composed by the following parts (see figure 3): a constitutive promoter (1), a RNA thermometer (3); used to regulate the WinterGreen-odor protein production through temperature, a Wintergreen-odor enzyme generator, used to allow the production of methyl salicylate, induced by salicylic acid (2); and a terminator (4). All of these parts are ligated by an 8-bp scar (TACTAGAG).

Figure 3. Aroma Module


These 4 genetic parts form the Aroma device of the project. The full device's length is 1,251bp (including restriction sites).

Parts of the module


IMAGE

CODE

DESCRIPTION

 

BBa_J23119A

 

In the specific case of our aroma module, it will help the bacteria to continuously transcribe the WinterGreen gene in order to allow the bacteria to continuously produce the aroma. This promoter has a length of 35bp.

BBa_K115017

A RNA thermometer, used for temperature post-transcriptional regulation (thermo sensor), and is designed to initiate transcription around 32°C.

BBa_K1255001

 

Produces a transferase to convert salicylic acid into methyl salicylate (WinterGreen odor). The wintergreen odor generator requires of 2mM of salicylic acid to produce methyl salicylate. BSMT1 (BBa_J45004), WinterGreen Odor Generator original name, was created by MIT 2006. This year, the team optimized the sequence for Escherichia coli. The new biobrick has a length of 1,074bp.

BBa_B1002

 

Part made of 6bp, responsible for transcription stop. The terminator stops the production of methyl salicylate.


Justification

Different to fluorescent reporters, this module was made in order to (in the future) perform as an aroma reporter and also to test the correct function of the bacteria, for its future usage as a new reporter and functional part (CDS). It is desired to use this part in the project to replace the red fluorescent protein (RFP) in the Capture module. But it was preferable to test it apart to demonstrate its effectiveness. Similarly, this piece is also helpful when the biofilter is assembled, because when performing the filtration by silica, WinterGreen can demonstrate the presence of bacteria in the beads.

The team added the RNA thermometer in the device for regulating the production of the aroma. Another reason for selecting the RNA thermometer as a regulator was to continue the CIDEB UANL 2013 work with it.

Other teams that used RNA thermometer and BSMT1

Team

Part Used

Use

TuDelft 2008 :

RNA themometer

 

Temperature-sensing bacteria that changes color at different temperatures; as a temperature reporter system inlarge-scale fermentations, or as a temperature-inducible protein production system.

 

Victoria BC 2009 :

 

RNA themometer

 

NAND logic gate using the ribo-key/ribo-lock system designed by Berkeley 2006 team , producing RFP except when the cells are grown in the presence of both arabinose and IPTG, also coupling fluorescent outputs with the ribo-thermometers made by TUDelft 2008 team.

 

CIDEB UANL 2013 :

 

RNA themometer

 

Production of Vip3ca3, which acts as a pesticide protein, regulated by specific temperatures in order to avoid overproduction and it will show activity against target organisms Coleoptera and Lepidoptera..

 

MIT 2006 :

 

BSMT1

This device produces methyl salicylate in the presence of salicylic acid. Methyl salicylate smells strongly of mint (wintergreen). Production of methyl salicylate was verified both by scent and by gas chromatography: E. coli with no WGD did not produce methyl salicylate when SA was added to the medium, while E. coli with the WGD did produce methyl salicylate when SA was added to the medium..

Project Zoom in


Bibliography/References

● Huang, H. (2006, August 30). Part:BBa_B1002. Retrieved August 30, 2014, from http://parts.igem.org/wiki/index.php?title=Part:BBa_B1002.

● iGEM2006_Berkeley. (2006). Part:BBa_J23100. Retrieved August 30, 2014, from http://parts.igem.org/Part:BBa_J23100.

● iGEM2006_MIT. (2006). Part:BBa_J45004. Retrieved August 30, 2014, from http://parts.igem.org/Part:BBa_J45004.

● iGEM CIDEB Team. (2013). iGEM CIDEB UANL 2013. Retrieved on March 31th, 2014. https://2013hs.igem.org/Team:CIDEB-UANL_Mexico/Project.

● iGEM08_TUDelft. (2008). Part:BBa_K115017. Retrieved August 30, 2014, from http://parts.igem.org/Part:BBa_K115017.

● MIT IGEM Team. (2006). MIT 2006. Retrieved on March 31th, 2014, from: https://2006.igem.org/wiki/index.php/MIT_2006.

● TUDelft iGEM Team. (2008). TUDelft 2008. Retrieved on March 31th, 2014, from: https://2008.igem.org/Team:TUDelft.

● VictoriaBC. (2009). VictoriaBC 2009. Retrieved on March 31th, 2014, from: https://2009.igem.org/Team:VictoriaBC.

● Zubieta, Chole et al. (2003). Structural Basis for Substrate Recognition in the Salicylic Acid Carboxyl Methyltransferase Family. Manuscript submitted for publication. Retrieved from www.plantcell.org.


iGEM CIDEB 2014 - Footer