Difference between revisions of "Team:Lambert GA/Composite Part"

 
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       <a href="https://2017.igem.org/Team:Lambert_GA/Part Collection" class="dropbtn">Parts</a>
 
       <a href="https://2017.igem.org/Team:Lambert_GA/Part Collection" class="dropbtn">Parts</a>
 
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       <a class="drplink" style="transition: color 0.5s ease-in-out;" href="https://2017.igem.org/Team:Lambert_GA/Part Collection">Part_Collection</a>
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       <a class="drplink" style="transition: color 0.5s ease-in-out;" href="https://2017.igem.org/Team:Lambert_GA/Part Collection">Part Collection</a>
 
       <a class="drplink" style="transition: color 0.5s ease-in-out;"  
 
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href="https://2017.igem.org/Team:Lambert_GA/Updated_Part">Updated Parts</a>
 
href="https://2017.igem.org/Team:Lambert_GA/Updated_Part">Updated Parts</a>
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--><a  href="https://2017.igem.org/Team:Lambert_GA/Safety"class="dropbtn">Safety</a><!--
 
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       <a class="drplink" style="transition: color 0.5s ease-in-out;" href="https://2017.igem.org/Team:Lambert_GA/HP/Silver">Silver HP</a>
 
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       <a class="drplink" style="transition: color 0.5s ease-in-out;" href="https://2017.igem.org/Team:Lambert_GA/HP/Gold_Integrated">Integrated and Gold</a>
 
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       <a class="drplink" style="transition: color 0.5s ease-in-out;" href="https://2017.igem.org/Team:Lambert_GA/Applied_Design">Applied Design</a>
 
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<a class="drplink" style="transition: color 0.5s ease-in-out;" href="https://2017.igem.org/Team:Lambert_GA/Hardware">Hardware</a>
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      <a class="drplink" style="transition: color 0.5s ease-in-out;" href="https://2017.igem.org/Team:Lambert_GA/Measurement">Measurement</a>
 
 
       <a class="drplink" style="transition: color 0.5s ease-in-out;" href="https://2017.igem.org/Team:Lambert_GA/Model">Model</a>
 
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<div class="col-md-12 col-sm-10">
 
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<center><font color= "#D49AE6">BBa_K2353000: tsPurple </font>
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<center><font color= "#D49AE6"><a href="http://parts.igem.org/Part:BBa_K2353000" style="text-decoration: none;"> BBa_K2353000: tsPurple</a> </font>
 
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<br>
 
<img src="https://static.igem.org/mediawiki/2017/4/4f/TsPurple.png"><br></center>
 
<img src="https://static.igem.org/mediawiki/2017/4/4f/TsPurple.png"><br></center>
 
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<center><font color= "#D49AE6">BBa_K2353001: tsPurpleDAS </font>
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<center><font color= "#D49AE6"><a href="http://parts.igem.org/Part:BBa_K2353001" style="text-decoration: none;">BBa_K2353001: tsPurpleDAS</a> </font>
 
<br><img src="https://static.igem.org/mediawiki/2017/a/a7/TsPurpleDAS.png"><br></center>
 
<br><img src="https://static.igem.org/mediawiki/2017/a/a7/TsPurpleDAS.png"><br></center>
 
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<center><font color= "#D49AE6">BBa_K2353002: tsPurpleLAA </font>
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<center><font color= "#D49AE6"><a href="http://parts.igem.org/Part:BBa_K2353002" style="text-decoration: none;">BBa_K2353002: tsPurpleLAA</a> </font>
 
<br><img src="https://static.igem.org/mediawiki/2017/a/a2/TsPurpleLAA.png"><br></center>
 
<br><img src="https://static.igem.org/mediawiki/2017/a/a2/TsPurpleLAA.png"><br></center>
 
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The middle part of the full three part construct is one of three tsPurple sequences. Each sequence is composed of BBa_B0034 (RBS) and BBa_K1033906 (tsPurple- a purple chromoprotein also known as TinselPurple). When assembled with p-lambda-r-LacI and pLac-ClpXP-CI, the tsPurple constructs are expressed by p-lambda-r-LacI. The terminator for tsPurple is located at the beginning of the pLac-ClpXP-CI sequence, before the promoter for ClpXP-CI. When the construct is fully assembled (and induced with IPTG) ClpXP is expressed, which degrades tsPurple when there is a degradation tag attached at the end of the tsPurple construct. BBa_K2353000, the tsPurple construct without any degradation tag, shows the baseline expression of tsPurple as a basis for comparison. BBa_K2353001 has degradation tag DAS (BBa_M0052) which should cause moderately fast degradation of tsPurple, while BBa_K2353002 has the degradation tag LAA (BBa_M0050), which should cause high rates of degradation due to the high affinity of ClpXP for LAA.
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<p style="font-size:20px; text-indent: 50px; color: white;">The middle part of the full three part construct is one of three tsPurple sequences. Each sequence is composed of BBa_B0034 (RBS) and BBa_K1033906 (tsPurple- a purple chromoprotein also known as TinselPurple). When assembled with p-lambda-r LacI and pLac-ClpXP-CI, the tsPurple constructs are expressed by p-lambda-r LacI. The terminator for tsPurple is located at the beginning of the pLac-ClpXP-CI sequence, before the promoter for ClpXP-CI. When the construct is fully assembled and induced with IPTG, ClpXP is expressed and causes degradation when there is a degradation tag attached at the end of the tsPurple construct. BBa_K2353000, the tsPurple construct without any degradation tag, shows the baseline expression of tsPurple as a basis for comparison. BBa_K2353001 has degradation tag DAS (BBa_M0052) which should cause moderately fast degradation of tsPurple, while BBa_K2353002 has the degradation tag LAA (BBa_M0050), which should cause high rates of degradation due to the high affinity of ClpXP for LAA.
  
 
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<img style="width:600px; align:center" src="https://static.igem.org/mediawiki/2017/5/58/Tspurples_gel.jpeg">
 
<img style="width:600px; align:center" src="https://static.igem.org/mediawiki/2017/5/58/Tspurples_gel.jpeg">
 
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<i style="font-size: 14px;">gel results of digest of all three tsPurple parts. from left to right: das, laa, no deg tag</i>
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<i style="font-size: 14px;">Gel results from a digest of all three tsPurple parts. From top to bottom: BBa_K2353000, BBa_K2353002, BBa_K2353001</i>
 
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Latest revision as of 03:40, 2 November 2017


Composite Parts



BBa_K2353000: tsPurple



BBa_K2353001: tsPurpleDAS



BBa_K2353002: tsPurpleLAA


The middle part of the full three part construct is one of three tsPurple sequences. Each sequence is composed of BBa_B0034 (RBS) and BBa_K1033906 (tsPurple- a purple chromoprotein also known as TinselPurple). When assembled with p-lambda-r LacI and pLac-ClpXP-CI, the tsPurple constructs are expressed by p-lambda-r LacI. The terminator for tsPurple is located at the beginning of the pLac-ClpXP-CI sequence, before the promoter for ClpXP-CI. When the construct is fully assembled and induced with IPTG, ClpXP is expressed and causes degradation when there is a degradation tag attached at the end of the tsPurple construct. BBa_K2353000, the tsPurple construct without any degradation tag, shows the baseline expression of tsPurple as a basis for comparison. BBa_K2353001 has degradation tag DAS (BBa_M0052) which should cause moderately fast degradation of tsPurple, while BBa_K2353002 has the degradation tag LAA (BBa_M0050), which should cause high rates of degradation due to the high affinity of ClpXP for LAA.



Gel results from a digest of all three tsPurple parts. From top to bottom: BBa_K2353000, BBa_K2353002, BBa_K2353001