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<html> <head> <meta http-equiv=Content-Type content="text/html; charset=windows-1252"> <meta name=Generator content="Microsoft Word 10 (filtered)"> <title>Part 0: Introduction</title> <style>  </style> </head> <body lang=EN-US link=blue vlink=purple> <div class=Section1> Part 0: Introduction <h4>Last Meeting Summary</h4> - The needed parts have been requested, Randy has said he’ll ship on Nov 11th. Enzymes and buffers have been obtained. - The temperature sensor project name is tentatively the “Cell-see-us” Thermometer, thank Andrew for that one :) - It’s been agreed that both the temperature sensor and etch-a-sketch will be assembled and tested. Note that both can be assembled in a total of 10 addition operations. - It’s been generally decided that the undergrads (Hannah/Emanuel/Joseph) will be working together at Steve’s lab (MSB4232) to combine all the parts. Please let us know if you want in for scheduling purposes… To Do - Assemble, transform E. coli, do multiple trials. Make video (I’m hoping there’s a camcorder available?). - I can make the presentation, everyone please email me any materials/thoughts/suggestions you think may be helpful. Part 1: Overview of Cell-See-Us Thermometer and Bacterial Etch-a-Sketch Tuneable Temperature Sensor v2 (using LacI ts + other parts from Registry 7.05) [BBa_J11021] <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=6155">http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=6155</a> <table class=MsoNormalTable border=0 cellspacing=0 cellpadding=0 width="100%" style='width:100.0%;background:white;border-collapse:collapse'> <tr> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> <table class=MsoNormalTable border=0 cellspacing=0 cellpadding=0 width="100%" style='width:100.0%;background:white;border-collapse:collapse'> <tr> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> pBad/araC I0500 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> B0034 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> lacI ts J06501 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> B0010 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> B0012 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> LacI R0010 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> B0031 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> C0056 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> B0010 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> B0012 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> Prm + I12006 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> B0032 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> E0040 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> B0010 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> B0012 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> LacI R0010 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> B0031 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> mRFP1 E1010 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> B0010 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> B0012 </td> <td width="100%" style='width:100.0%;padding:0in 0in 0in 0in'> </td> </tr> <tr> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=2916"><img border=0 width=49 height=33 src="http://individual.utoronto.ca/nazareth/wiki_files/image001.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=151"><img border=0 width=49 height=33 src="http://individual.utoronto.ca/nazareth/wiki_files/image002.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=5536"><img border=0 width=49 height=33 src="http://individual.utoronto.ca/nazareth/wiki_files/image003.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=603"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image004.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=145"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image004.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=185"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image005.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=148"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image006.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=5244"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image007.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=603"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image004.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=145"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image004.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=4797"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image005.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=149"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image006.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=5309"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image008.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=603"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image004.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=145"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image004.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=185"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image005.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=148"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image006.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=4932"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image008.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=603"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image004.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=145"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image004.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> </td> </tr> </table> </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> </td> <td width="16%" style='width:16.16%;padding:0in 0in 0in 0in'> </td> </tr> </table> Explanation Summary: Keeping the cells in a L-arabinose solution induces the cells to produce a specific steady state value of lacI monomers. The portion of these monomers that is in dimer form decreases with temperature. As temperature increases, the LacI dimer concentration decreases, resulting in reduced inhibition of the R0010 promoters. Hence RFP concentration increases. At the same time, more cI is produced, which inhibits Prm +, resulting in less GFP at steady state. Explanation Details: 1) Consider a system with just the “LacI” (pLac) promoter and associated coding regions. The cells are in an IPTG solution, which results in the promoters being induced, resulting in 434 cI and RFP being produced at a “maximal” rate. 2) Now consider some LacI ts added to the system, which forms a dimer at some equilibrium concentration. At [LacI dimer] = 0, 434 cI and RFP are produced at maximal rates. At some concentration of LacI dimer, the transcription rate becomes “minimal” since LacI inhibits the “LacI” promoter (pLac). Between the maximal and minimal rates, there is a linear region where transcription rate gradually decreases as a function of LacI concentration. (see Graph 1) 3) The pBad promoter can be induced by keeping the cells in a solution of L-arabinose. When induced, lacI ts is produced. The concentration of L-arabinose can be set so that at 37˚ the [LacI dimer] concentration is centered in the linear region. Hence as temperature deviates from 37˚, R0010 changes accordingly. 4) As the R0010 transcription rate changes, the RFP concentration steady state value changes accordingly. Also, the cI steady state concentration changes. 5) To increase the visual contrast, Prm + driving GFP has been added to the system. For example, if the temperature increases from 37˚ to 42˚, the LacI dimers disassociate more readily, and [LacI dimer] decreases. R0010 transcription rate increases, resulting in higher [RFP] and [434 cI] steady state values. Since 434 cI represses Prm, the Prm transcription rate will decreases, resulting in a lower [GFP]. It is hoped that the decrease in steady state [434 cI] will indeed result in a noticeable change in [GFP], this depends on [434 cI] at 37˚ being in the linear region of a Prm + transcription rate vs. [434 cI] graph. (A way of tuning this will likely be needed – investigating currently) <img border=0 width=566 height=317 src="http://individual.utoronto.ca/nazareth/wiki_files/image009.jpg"> Tuneable Temperature Sensor v3 (Registry 7.05 DNA-1 + Harvard Parts) [BBa_J11022] <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=6160">http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=6160</a> <table class=MsoNormalTable border=0 cellspacing=0 cellpadding=0 width="107%" style='width:107.38%;background:white;border-collapse:collapse'> <tr> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> pBad/araC I0500 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> B0034 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> lacI ts J06501 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> B0010 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> B0012 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> lacI+pL R0011 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> B0031 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> C0056 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> B0010 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> B0012 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> Prm + I12006 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> B0032 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> E0040 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> B0010 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> B0012 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> LacI R0010 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> B0034 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> mRFP1 E1010 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> B0010 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> B0012 </td> <td width="0%" style='width:.14%;padding:0in 0in 0in 0in'> </td> </tr> <tr> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=2916"><img border=0 width=49 height=33 src="http://individual.utoronto.ca/nazareth/wiki_files/image001.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=151"><img border=0 width=49 height=33 src="http://individual.utoronto.ca/nazareth/wiki_files/image002.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=5536"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image007.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=603"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image004.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=145"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image004.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=186"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image005.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=148"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image006.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=5244"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image007.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=603"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image004.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=145"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image004.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=4797"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image005.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=149"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image006.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=5309"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image008.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=603"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image004.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=145"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image004.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=185"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image005.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=151"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image006.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=4932"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image008.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=603"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image004.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=145"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image004.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> </td> </tr> </table> Explanation Summary: The same as BBa_J11022, however the parts have been moved around so that currently available parts can be used to reduce the overall number of parts (no change in functionality). Also, some minor substitutes have been made (ex. R0010->R0011) to further reduce the number of parts. For synthesis, only 8 parts have to be assembled together. <table class=MsoNormalTable border=0 cellspacing=0 cellpadding=0 width="100%" style='width:100.0%;background:white;border-collapse:collapse'> <tr> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> pBad/araC I0500 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> J06801 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> B0031 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> C0056 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> B0015 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> Prm + I12006 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> E0240 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> J04450 </td> <td width="100%" style='width:100.0%;padding:0in 0in 0in 0in'> </td> </tr> <tr> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=2916"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image005.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=5534"><img border=0 width=49 height=33 src="http://individual.utoronto.ca/nazareth/wiki_files/image010.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=148"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image006.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=5244"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image007.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=202"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image004.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=4797"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image005.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=5339"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image008.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=5499"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image008.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> </td> </tr> </table> Bacterial Whiteboard by Hannah (w/ Minimal Available Parts) [BBa_J11031] <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=6154">http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=6154</a> <table class=MsoNormalTable border=0 cellspacing=0 cellpadding=0 width="100%" style='width:100.0%;background:white;border-collapse:collapse'> <tr> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> tetR R0040 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> B0034 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> lacZ a E0033 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> B0010 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> B0012 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> lacI+pL R0011 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> B0034 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> ECFP E0022 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> B0010 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> B0012 </td> <td width="50%" style='width:50.08%;padding:0in 0in 0in 0in'> </td> </tr> <tr> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=187"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image005.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=151"><img border=0 width=49 height=33 src="http://individual.utoronto.ca/nazareth/wiki_files/image002.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=3911"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image008.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=603"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image004.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=145"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image004.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=186"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image005.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=151"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image006.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=162"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image008.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=603"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image004.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=145"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image004.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> </td> </tr> </table> Explanation Summary: When lactose is added to the system, bind to lac repressor (endogenous) therefore CFP (fast degrading) produced - glow cyan. When Tet added to the system, drive transcription of lacZ beta galactosidase which hydrolyses latose and lac repressor active. Stop Glowing cyan. ***NOTE*** This system will ONLY WORK IN LAC Z- bugs!!!!! For synthesis, only 4 parts have to be assembled together: <table class=MsoNormalTable border=0 cellspacing=0 cellpadding=0 width="100%" style='width:100.0%;background:white;border-collapse:collapse'> <tr> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> tetR R0040 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> E0433 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> lacI+pL R0011 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> E0422 </td> <td width="100%" style='width:100.0%;padding:0in 0in 0in 0in'> </td> </tr> <tr> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=187"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image005.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=4644"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image011.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=186"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image005.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=2678"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image008.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> </td> </tr> </table> Part 2: Temperature Sensor Modeling <img border=0 width=939 height=333 src="http://individual.utoronto.ca/nazareth/wiki_files/image012.jpg"> Note: “In the presence of lactose or IPTG, an analog of lactose, LacI is unable to correctly bind and inhibit transcription.” [http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=185] Part 3: Assembly Schedule <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0 style='border-collapse:collapse;border:none'> <tr> <td valign=top style='border:solid windowtext 1.0pt;padding:0in 5.4pt 0in 5.4pt'> Part </td> <td valign=top style='border:solid windowtext 1.0pt;border-left:none; padding:0in 5.4pt 0in 5.4pt'> Day 1 </td> <td valign=top style='border:solid windowtext 1.0pt;border-left:none; padding:0in 5.4pt 0in 5.4pt'> Day 2 </td> <td width=375 valign=top style='width:281.2pt;border:solid windowtext 1.0pt; border-left:none;padding:0in 5.4pt 0in 5.4pt'> Day 3 </td> </tr> <tr> <td valign=top style='border:solid windowtext 1.0pt;border-top:none; padding:0in 5.4pt 0in 5.4pt'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=2916"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image005.jpg"></a> I0500 </td> <td rowspan=2 valign=top style='border-top:none;border-left:none;border-bottom: solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:0in 5.4pt 0in 5.4pt'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=2916"><img border=0 width=48 height=33 src="http://individual.utoronto.ca/nazareth/wiki_files/image013.jpg"></a><a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=5534"><img border=0 width=49 height=33 src="http://individual.utoronto.ca/nazareth/wiki_files/image010.jpg"></a> I0500+ J06801 </td> <td rowspan=4 valign=top style='border-top:none;border-left:none;border-bottom: solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:0in 5.4pt 0in 5.4pt'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=2916"><img border=0 width=48 height=33 src="http://individual.utoronto.ca/nazareth/wiki_files/image013.jpg"></a><a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=5534"><img border=0 width=49 height=33 src="http://individual.utoronto.ca/nazareth/wiki_files/image010.jpg"></a><a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=148"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image006.jpg"></a><a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=5244"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image007.jpg"></a> (I0500+ J06801)+( B0031+ C0056) </td> <td width=375 rowspan=8 valign=top style='width:281.2pt;border-top:none; border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; padding:0in 5.4pt 0in 5.4pt'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=2916"><img border=0 width=48 height=33 src="http://individual.utoronto.ca/nazareth/wiki_files/image013.jpg"></a><a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=5534"><img border=0 width=49 height=33 src="http://individual.utoronto.ca/nazareth/wiki_files/image010.jpg"></a><a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=148"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image006.jpg"></a><a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=5244"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image007.jpg"></a><a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=202"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image004.jpg"></a><a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=4797"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image005.jpg"></a><a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=5339"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image008.jpg"></a><a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=5499"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image008.jpg"></a> {(I0500+ J06801)+( B0031+ C0056)}+{ (B0015+ I12006)+( E0240+ J04450)} </td> </tr> <tr> <td style='border:solid windowtext 1.0pt;border-top:none;padding:0in 5.4pt 0in 5.4pt'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=5534"><img border=0 width=49 height=33 src="http://individual.utoronto.ca/nazareth/wiki_files/image010.jpg"></a>J06801 </td> </tr> <tr> <td valign=top style='border:solid windowtext 1.0pt;border-top:none; padding:0in 5.4pt 0in 5.4pt'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=148"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image006.jpg"></a> B0031 </td> <td rowspan=2 valign=top style='border-top:none;border-left:none;border-bottom: solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:0in 5.4pt 0in 5.4pt'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=148"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image006.jpg"></a><a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=5244"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image007.jpg"></a> B0031+ C0056 </td> </tr> <tr> <td valign=top style='border:solid windowtext 1.0pt;border-top:none; padding:0in 5.4pt 0in 5.4pt'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=5244"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image007.jpg"></a> C0056 </td> </tr> <tr> <td valign=top style='border:solid windowtext 1.0pt;border-top:none; padding:0in 5.4pt 0in 5.4pt'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=202"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image004.jpg"></a> B0015 </td> <td rowspan=2 valign=top style='border-top:none;border-left:none;border-bottom: solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:0in 5.4pt 0in 5.4pt'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=202"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image004.jpg"></a><a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=4797"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image005.jpg"></a> B0015+ I12006 </td> <td rowspan=4 valign=top style='border-top:none;border-left:none;border-bottom: solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:0in 5.4pt 0in 5.4pt'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=202"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image004.jpg"></a><a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=4797"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image005.jpg"></a><a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=5339"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image008.jpg"></a><a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=5499"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image008.jpg"></a> (B0015+ I12006)+( E0240+ J04450) </td> </tr> <tr> <td valign=top style='border:solid windowtext 1.0pt;border-top:none; padding:0in 5.4pt 0in 5.4pt'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=4797"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image005.jpg"></a> I12006 </td> </tr> <tr> <td valign=top style='border:solid windowtext 1.0pt;border-top:none; padding:0in 5.4pt 0in 5.4pt'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=5339"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image008.jpg"></a> E0240 </td> <td rowspan=2 valign=top style='border-top:none;border-left:none;border-bottom: solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:0in 5.4pt 0in 5.4pt'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=5339"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image008.jpg"></a><a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=5499"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image008.jpg"></a> E0240+ J04450 </td> </tr> <tr> <td valign=top style='border:solid windowtext 1.0pt;border-top:none; padding:0in 5.4pt 0in 5.4pt'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=5499"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image008.jpg"></a> J04450 </td> </tr> <tr> <td valign=top style='border:solid windowtext 1.0pt;border-top:none; padding:0in 5.4pt 0in 5.4pt'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=187"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image005.jpg"></a> R0040 </td> <td rowspan=2 valign=top style='border-top:none;border-left:none;border-bottom: solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:0in 5.4pt 0in 5.4pt'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=187"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image005.jpg"></a><a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=4644"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image011.jpg"></a> R0040+ E0433 </td> <td rowspan=4 valign=top style='border-top:none;border-left:none;border-bottom: solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:0in 5.4pt 0in 5.4pt'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=187"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image005.jpg"></a><a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=4644"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image011.jpg"></a><a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=186"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image005.jpg"></a><a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=2678"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image008.jpg"></a> (R0040+ E0433)+( R0011+ E0422) </td> <td width=375 valign=top style='width:281.2pt;border-top:none;border-left: none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; padding:0in 5.4pt 0in 5.4pt'> </td> </tr> <tr> <td valign=top style='border:solid windowtext 1.0pt;border-top:none; padding:0in 5.4pt 0in 5.4pt'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=4644"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image011.jpg"></a> E0433 </td> <td width=375 valign=top style='width:281.2pt;border-top:none;border-left: none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; padding:0in 5.4pt 0in 5.4pt'> </td> </tr> <tr> <td valign=top style='border:solid windowtext 1.0pt;border-top:none; padding:0in 5.4pt 0in 5.4pt'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=186"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image005.jpg"></a> R0011 </td> <td rowspan=2 valign=top style='border-top:none;border-left:none;border-bottom: solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:0in 5.4pt 0in 5.4pt'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=186"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image005.jpg"></a><a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=2678"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image008.jpg"></a> R0011+ E0422 </td> <td width=375 valign=top style='width:281.2pt;border-top:none;border-left: none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; padding:0in 5.4pt 0in 5.4pt'> </td> </tr> <tr> <td valign=top style='border:solid windowtext 1.0pt;border-top:none; padding:0in 5.4pt 0in 5.4pt'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=2678"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image008.jpg"></a> E0422 </td> <td width=375 valign=top style='width:281.2pt;border-top:none;border-left: none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; padding:0in 5.4pt 0in 5.4pt'> </td> </tr> </table> BBa_J11022 <table class=MsoNormalTable border=0 cellspacing=0 cellpadding=0 width="100%" style='width:100.0%;background:white;border-collapse:collapse'> <tr> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> pBad/araC I0500 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> J06801 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> B0031 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> C0056 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> B0015 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> Prm + I12006 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> E0240 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> J04450 </td> <td width="100%" style='width:100.0%;padding:0in 0in 0in 0in'> </td> </tr> <tr> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=2916"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image005.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=5534"><img border=0 width=49 height=33 src="http://individual.utoronto.ca/nazareth/wiki_files/image010.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=148"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image006.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=5244"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image007.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=202"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image004.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=4797"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image005.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=5339"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image008.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=5499"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image008.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> </td> </tr> </table> BBa_J11031 <table class=MsoNormalTable border=0 cellspacing=0 cellpadding=0 width="100%" style='width:100.0%;background:white;border-collapse:collapse'> <tr> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> tetR R0040 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> E0433 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> lacI+pL R0011 </td> <td valign=top style='padding:0in 1.5pt 0in 1.5pt'> E0422 </td> <td width="100%" style='width:100.0%;padding:0in 0in 0in 0in'> </td> </tr> <tr> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=187"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image005.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=4644"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image011.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=186"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image005.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> <a href="http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=2678"><img border=0 width=37 height=25 src="http://individual.utoronto.ca/nazareth/wiki_files/image008.jpg"></a> </td> <td style='padding:0in 0in 0in 0in'> </td> </tr> </table> Part 4: Part Locations <table class=MsoTableGrid border=1 cellspacing=0 cellpadding=0 style='border-collapse:collapse;border:none'> <tr> <td width=67 valign=top style='width:.7in;border:solid windowtext 1.0pt; padding:0in 5.4pt 0in 5.4pt'> Part </td> <td width=672 valign=top style='width:7.0in;border:solid windowtext 1.0pt; border-left:none;padding:0in 5.4pt 0in 5.4pt'> Registry 7.05 DNA-1 Well </td> </tr> <tr> <td width=67 valign=top style='width:.7in;border:solid windowtext 1.0pt; border-top:none;padding:0in 5.4pt 0in 5.4pt'> I0500 </td> <td width=672 valign=top style='width:7.0in;border-top:none;border-left:none; border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; padding:0in 5.4pt 0in 5.4pt'> <table class=MsoNormalTable border=0 cellspacing=0 cellpadding=0 style='background:white'> <tr> <td valign=bottom style='padding:3.75pt 30.0pt 3.75pt 3.75pt'> Registry 7.05 </td> <td valign=bottom style='padding:3.75pt 30.0pt 3.75pt 3.75pt'> 9I </td> <td valign=bottom style='padding:3.75pt 30.0pt 3.75pt 3.75pt'> Registry 7.05 DNA-2 </td> <td valign=bottom style='padding:3.75pt 30.0pt 3.75pt 3.75pt'> pSB2K3 </td> <td valign=bottom style='padding:3.75pt 30.0pt 3.75pt 3.75pt'> V1003 </td> <td valign=bottom style='padding:3.75pt 3.75pt 3.75pt 3.75pt'> </td> </tr> <tr> <td colspan=2 style='padding:3.75pt 3.75pt 3.75pt 12.0pt'> () </td> <td style='padding:3.75pt 3.75pt 3.75pt 3.75pt'> </td> <td style='padding:3.75pt 3.75pt 3.75pt 3.75pt'> </td> <td style='padding:3.75pt 3.75pt 3.75pt 3.75pt'> </td> <td style='padding:3.75pt 3.75pt 3.75pt 3.75pt'> </td> </tr> </table> </td> </tr> <tr> <td width=67 valign=top style='width:.7in;border:solid windowtext 1.0pt; border-top:none;padding:0in 5.4pt 0in 5.4pt'> J06801 </td> <td width=672 valign=top style='width:7.0in;border-top:none;border-left:none; border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; padding:0in 5.4pt 0in 5.4pt'> <table class=MsoNormalTable border=0 cellspacing=0 cellpadding=0 style='background:white'> <tr> <td valign=bottom style='padding:3.75pt 30.0pt 3.75pt 3.75pt'> Registry Update </td> <td valign=bottom style='padding:3.75pt 30.0pt 3.75pt 3.75pt'> 17 </td> <td valign=bottom style='padding:3.75pt 30.0pt 3.75pt 3.75pt'> Harvard iGEM2005 - 1 </td> <td valign=bottom style='padding:3.75pt 30.0pt 3.75pt 3.75pt'> pSB2K3 </td> <td valign=bottom style='padding:3.75pt 30.0pt 3.75pt 3.75pt'> V1009 </td> </tr> <tr> <td colspan=2 style='padding:3.75pt 3.75pt 3.75pt 12.0pt'> (TK Freezer) </td> <td style='padding:3.75pt 3.75pt 3.75pt 3.75pt'> </td> <td style='padding:3.75pt 3.75pt 3.75pt 3.75pt'> </td> <td style='padding:3.75pt 3.75pt 3.75pt 3.75pt'> </td> </tr> </table> </td> </tr> <tr> <td width=67 valign=top style='width:.7in;border:solid windowtext 1.0pt; border-top:none;padding:0in 5.4pt 0in 5.4pt'> B0031 </td> <td width=672 valign=top style='width:7.0in;border-top:none;border-left:none; border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; padding:0in 5.4pt 0in 5.4pt'> 3I </td> </tr> <tr> <td width=67 valign=top style='width:.7in;border:solid windowtext 1.0pt; border-top:none;padding:0in 5.4pt 0in 5.4pt'> C0056 </td> <td width=672 valign=top style='width:7.0in;border-top:none;border-left:none; border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; padding:0in 5.4pt 0in 5.4pt'> 18L </td> </tr> <tr> <td width=67 valign=top style='width:.7in;border:solid windowtext 1.0pt; border-top:none;padding:0in 5.4pt 0in 5.4pt'> B0015 </td> <td width=672 valign=top style='width:7.0in;border-top:none;border-left:none; border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; padding:0in 5.4pt 0in 5.4pt'> 1I </td> </tr> <tr> <td width=67 valign=top style='width:.7in;border:solid windowtext 1.0pt; border-top:none;padding:0in 5.4pt 0in 5.4pt'> I12006 </td> <td width=672 valign=top style='width:7.0in;border-top:none;border-left:none; border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; padding:0in 5.4pt 0in 5.4pt'> <table class=MsoNormalTable border=0 cellspacing=0 cellpadding=0 style='background:white'> <tr> <td valign=bottom style='padding:3.75pt 30.0pt 3.75pt 3.75pt'> Registry Update </td> <td valign=bottom style='padding:3.75pt 30.0pt 3.75pt 3.75pt'> 24 </td> <td valign=bottom style='padding:3.75pt 30.0pt 3.75pt 3.75pt'> Box 5 </td> <td valign=bottom style='padding:3.75pt 30.0pt 3.75pt 3.75pt'> pSB2K3 </td> <td valign=bottom style='padding:3.75pt 30.0pt 3.75pt 3.75pt'> V1006 </td> </tr> <tr> <td colspan=2 style='padding:3.75pt 3.75pt 3.75pt 12.0pt'> (TK -80 (1D5C)) </td> <td colspan=4 style='padding:3.75pt 3.75pt 3.75pt 3.0pt'> CE100-pSB2K3-I12006 </td> </tr> <tr> <td valign=bottom style='padding:3.75pt 30.0pt 3.75pt 3.75pt'> Registry 7.05 </td> <td valign=bottom style='padding:3.75pt 30.0pt 3.75pt 3.75pt'> 8E </td> <td valign=bottom style='padding:3.75pt 30.0pt 3.75pt 3.75pt'> Source Plate 5 </td> <td valign=bottom style='padding:3.75pt 30.0pt 3.75pt 3.75pt'> pSB2K3 </td> <td valign=bottom style='padding:3.75pt 30.0pt 3.75pt 3.75pt'> V1006 </td> <td valign=bottom style='padding:3.75pt 3.75pt 3.75pt 3.75pt'> </td> </tr> <tr> <td colspan=2 style='padding:3.75pt 3.75pt 3.75pt 12.0pt'> (TK -80) </td> <td style='padding:3.75pt 3.75pt 3.75pt 3.75pt'> </td> <td style='padding:3.75pt 3.75pt 3.75pt 3.75pt'> </td> <td style='padding:3.75pt 3.75pt 3.75pt 3.75pt'> </td> <td style='padding:3.75pt 3.75pt 3.75pt 3.75pt'> </td> </tr> <tr> <td valign=bottom style='padding:3.75pt 30.0pt 3.75pt 3.75pt'> Registry 7.05 </td> <td valign=bottom style='padding:3.75pt 30.0pt 3.75pt 3.75pt'> 15I </td> <td valign=bottom style='padding:3.75pt 30.0pt 3.75pt 3.75pt'> Registry 7.05 DNA-2 </td> <td valign=bottom style='padding:3.75pt 30.0pt 3.75pt 3.75pt'> pSB2K3 </td> <td valign=bottom style='padding:3.75pt 30.0pt 3.75pt 3.75pt'> V1006 </td> <td valign=bottom style='padding:3.75pt 3.75pt 3.75pt 3.75pt'> </td> </tr> <tr> <td colspan=2 style='padding:3.75pt 3.75pt 3.75pt 12.0pt'> () </td> <td style='padding:3.75pt 3.75pt 3.75pt 3.75pt'> </td> <td style='padding:3.75pt 3.75pt 3.75pt 3.75pt'> </td> <td style='padding:3.75pt 3.75pt 3.75pt 3.75pt'> </td> <td style='padding:3.75pt 3.75pt 3.75pt 3.75pt'> </td> </tr> </table> </td> </tr> <tr> <td width=67 valign=top style='width:.7in;border:solid windowtext 1.0pt; border-top:none;padding:0in 5.4pt 0in 5.4pt'> E0240 </td> <td width=672 valign=top style='width:7.0in;border-top:none;border-left:none; border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; padding:0in 5.4pt 0in 5.4pt'> 16A </td> </tr> <tr> <td width=67 valign=top style='width:.7in;border:solid windowtext 1.0pt; border-top:none;padding:0in 5.4pt 0in 5.4pt'> J04450 </td> <td width=672 valign=top style='width:7.0in;border-top:none;border-left:none; border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; padding:0in 5.4pt 0in 5.4pt'> <table class=MsoNormalTable border=0 cellspacing=0 cellpadding=0 style='background:white'> <tr> <td valign=bottom style='padding:3.75pt 30.0pt 3.75pt 3.75pt'> Registry Update </td> <td valign=bottom style='padding:3.75pt 30.0pt 3.75pt 3.75pt'> 81 </td> <td valign=bottom style='padding:3.75pt 30.0pt 3.75pt 3.75pt'> Box 5 </td> <td valign=bottom style='padding:3.75pt 30.0pt 3.75pt 3.75pt'> pSB1A2 </td> <td valign=bottom style='padding:3.75pt 30.0pt 3.75pt 3.75pt'> V1010 </td> </tr> <tr> <td colspan=2 style='padding:3.75pt 3.75pt 3.75pt 12.0pt'> (TK -80 (1D5C)) </td> <td style='padding:3.75pt 3.75pt 3.75pt 3.75pt'> </td> <td style='padding:3.75pt 3.75pt 3.75pt 3.75pt'> </td> <td style='padding:3.75pt 3.75pt 3.75pt 3.75pt'> </td> </tr> <tr> <td valign=bottom style='padding:3.75pt 30.0pt 3.75pt 3.75pt'> Registry Update </td> <td valign=bottom style='padding:3.75pt 30.0pt 3.75pt 3.75pt'> 91 </td> <td valign=bottom style='padding:3.75pt 30.0pt 3.75pt 3.75pt'> Box 5 </td> <td valign=bottom style='padding:3.75pt 30.0pt 3.75pt 3.75pt'> pSB1A2 </td> <td valign=bottom style='padding:3.75pt 30.0pt 3.75pt 3.75pt'> V1010 </td> </tr> <tr> <td colspan=2 style='padding:3.75pt 3.75pt 3.75pt 12.0pt'> (TK -80 (1D5C)) </td> <td style='padding:3.75pt 3.75pt 3.75pt 3.75pt'> </td> <td style='padding:3.75pt 3.75pt 3.75pt 3.75pt'> </td> <td style='padding:3.75pt 3.75pt 3.75pt 3.75pt'> </td> </tr> </table> </td> </tr> <tr> <td width=67 valign=top style='width:.7in;border:solid windowtext 1.0pt; border-top:none;padding:0in 5.4pt 0in 5.4pt'> R0040 </td> <td width=672 valign=top style='width:7.0in;border-top:none;border-left:none; border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; padding:0in 5.4pt 0in 5.4pt'> 7O </td> </tr> <tr> <td width=67 valign=top style='width:.7in;border:solid windowtext 1.0pt; border-top:none;padding:0in 5.4pt 0in 5.4pt'> E0433 </td> <td width=672 valign=top style='width:7.0in;border-top:none;border-left:none; border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; padding:0in 5.4pt 0in 5.4pt'> 14K </td> </tr> <tr> <td width=67 valign=top style='width:.7in;border:solid windowtext 1.0pt; border-top:none;padding:0in 5.4pt 0in 5.4pt'> R0011 </td> <td width=672 valign=top style='width:7.0in;border-top:none;border-left:none; border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; padding:0in 5.4pt 0in 5.4pt'> 7M </td> </tr> <tr> <td width=67 valign=top style='width:.7in;border:solid windowtext 1.0pt; border-top:none;padding:0in 5.4pt 0in 5.4pt'> E0422 </td> <td width=672 valign=top style='width:7.0in;border-top:none;border-left:none; border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt; padding:0in 5.4pt 0in 5.4pt'> 11G </td> </tr> </table> </div> </body> </html>

Good news, there is an E coli strain availabe that contains Tn10 inserted into LacZ, meaning it is lacZ- and Tetracycline sensitive.

about the Tet operon and Tn10 http://www.biomedcentral.com/1471-2180/5/1 http://www.biomedcentral.com/1471-2199/3/13

-Hannah

--

UPdate ~ 12 Oct 2005

--

15:54, 3 Oct 2005 (EDT)15:54, 3 Oct 2005 (EDT)~~ Hi everyone, I made a biobrick version of my system, http://parts2.mit.edu/r/parts/partsdb/view.cgi?part_id=6153 It is part # BBa_J11030 I hope all the parts are available... Also very important NOTE that this system MUST MUST MUST be in LacZ- bugs! So lets try and get a hold of those if we want to make my system

Hannah

[http://pubs.acs.org/cgi-bin/archive.cgi/chreay/2000/100/i07/pdf/cr990115p.pdf Genetically engineered whole-cell sensing systems: coupling biological recognition with reporter genes.]

Chem Rev. 2000 Jul 12;100(7):2705-38. Daunert S, Barrett G, Feliciano JS, Shetty RS, Shrestha S, Smith-Spencer W.

Discusses important biosensing systems, mainly in E. coli.

On another note, currently if anyone wants to post files (Powerpoint presentations, papers, etc.) the file must be available online somewhere and linked to from this Wiki. If you need a file to be posted online, feel free to email it to me at emanuelnazareth@hotmail.com and I'll get back to you with how to link to it.

~ Emanuel

Toronto

From 2006.igem.org

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