Team:EPF-Lausanne/Parts

From 2008.igem.org

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(Parts submitted to the registry)
 
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===Parts submitted to the registry===
===Parts submitted to the registry===
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[http://partsregistry.org/Part:BBa_K092000 BBa_K092000]: <br>
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*[http://partsregistry.org/Part:BBa_K092000 BBa_K092000]: <br>
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[[Image:K092000.jpg]]<br>
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:[[Image:K092000.jpg]]<br>
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TetR coding sequence with RBS and terminator :  
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:TetR coding sequence with RBS and terminator :  
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Coding sequence of the Tetracycline repressor with the ribosome binding site and a double terminator. The TetR comes from transposon Tn10 and has a LVA tail (SsrA) [cf. C0040 part].  
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::Coding sequence of the Tetracycline repressor with the ribosome binding site and a double terminator. The ''tetR'' comes from transposon ''tn10'' and has a ''LVA'' tail (SsrA) [cf. C0040 part].
 +
::This part has been successfully sequenced.
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This part has been successfully sequenced.
 
 +
*[http://partsregistry.org/Part:BBa_K092100 BBa_K092100] :<br>
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[http://partsregistry.org/Part:BBa_K092100 BBa_K092100] :<br>  
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:[[Image:K092100.jpg]]<br>
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:Constitutive RhlR Receiver
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[[Image:K092100.jpg]]<br>
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::This part has been successfully sequenced.
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Constitutive RhlR Receiver
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This part has been successfully sequenced.
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*[http://partsregistry.org/Part:BBa_K092200 BBa_K092200] :
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[http://partsregistry.org/Part:BBa_K092200 BBa_K092200] :  
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:[[Image:K092200.jpg]] <br>
 +
:RhlI with RBS and terminator
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[[Image:K092200.jpg]] <br>
 
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RhlI with RBS and terminator
 
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*[http://partsregistry.org/Part:BBa_K092300 BBa_K092300] :
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[http://partsregistry.org/Part:BBa_K092300 BBa_K092300] :  
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:[[Image:K092300.jpg]]<br>
 +
:RFP controlled by a TetR promoter :
 +
::Coding sequence of the RFP with a ribosome binding site and p(''tetR'') as promoter. Thus, the transcription of RFP is inhibited when TetR is present.
 +
::This part has been successfully sequenced.
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[[Image:K092300.jpg]]<br>
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*[http://partsregistry.org/Part:BBa_K092400 BBa_K092400] : <br>
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RFP controlled by a TetR promoter :  
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:[[Image:K092400.jpg]]<br>
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Coding sequence of the RFP with a ribosome binding site and pTetR as promoter. Thus, the transcription of RFP is inhibited when TetR and tetracyclin are present.
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:New part created by 2-steps PCR. RBS (B0034) + ''luxI'' + Terminator (B0010+B0012) :
 +
::Coding sequence for the Lux Inducer with a ribosome binding site and terminator. This biobrick is an alternative to the F1610 which didn't work
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This part has been successfully sequenced.
 
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[http://partsregistry.org/Part:BBa_K092400 BBa_K092400] : <br>
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*[http://partsregistry.org/Part:BBa_K092600 BBa_K092600] :<br>
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[[Image:K092400.jpg]]<br>
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:[[Image:K092600.jpg]]<br>
-
New part created by 2-steps PCR. RBS (B0034) + LuxI + Terminator (B0010+B0012) :
+
:TetR cds with RBS and Terminator + p(''tetR''), RFP with RBS
-
Coding sequence for the Lux Inducer with a ribosome binding site and terminator. This biobrick is an alternative to the F1610 which didn't work
+
::The TetR sequence (with a ribosome binding site and terminator) is bound to the part containing the promoter of ''tetR'', a ribosome binding site and the red fluorescent protein (RFP). Thus, the production of RFP directly depends on the production of TetR. The transcription of RFP is inhibited when TetR is present. The production of TetR depends on the promoter you put in front of it.
 +
::This part has been successfully sequenced.
 +
*[http://partsregistry.org/Part:BBa_K092700 BBa_K092700] :
 +
:[[Image:K092700.jpg]]<br>
 +
:TetR cds with RBS and Terminator + p(''tetR'', RFP with RBS + ''luxI'' with RBS and terminator
 +
::The TetR sequence (with a ribosome binding site and terminator) [http://partsregistry.org/Part:BBa_K092000 BBa_K092000] is bound to the part containing the promoter of ''tetR'' and the red fluorescent protein (RFP) with a ribosome binding site. A ''luxI'' part (with RBS and terminator) is added at the end of the cds of the RFP. As no terminator was placed after the RFP, they both depends on the ''tetR'' promoter. The transcription of ''RFP'' and ''luxI'' is inhibited when TetR is present. The production of TetR depends on the promoter you put in front of it.
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[http://partsregistry.org/Part:BBa_K092600 BBa_K092600] :<br>
 
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[[Image:K092600.jpg]]<br>
 
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TetR cds with RBS and Terminator + pTetR, RFP with RBS
+
*[http://partsregistry.org/Part:BBa_K092800 BBa_K092800] :<br>
 +
:[[Image:K092800bis.jpg]]
 +
:Sequence that codes the same amino acid sequence than LacI but coded with a codon optimized sequence. It will thus have a different rate of translation than LacI.
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The TetR sequence (with a ribosome binding site and terminator) is bound to the part containing the promoter of TetR, a ribosome binding site and the red fluorescent protein (RFP). Thus, the production of RFP directly depends on the production of TetR. The transcription of RFP is inhibited when TetR is present. The production of TetR depends on the promoter you put in front of it.
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::Source of LacI<sub>m</sub> : Basu et al."A synthetic multicelluar system for programmed pattern formation", Nature. 2005 Apr 28;434(7037):1130-4
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This part has been successfully sequenced.
 
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[http://partsregistry.org/Part:BBa_K092700 BBa_K092700] :
 
 +
*[http://partsregistry.org/Part:BBa_K092900 BBa_K092900] :
 +
:[[Image:K092900.jpg]]<br>
 +
:''rhlR'' controlled by a constitutive promoter, and ''lacI<sub>m</sub>'' with a RhlR promoter.
-
[[Image:K092700.jpg]]<br>
 
-
TetR cds with RBS and Terminator + pTetR, RFP with RBS + LuxI with RBS and terminator
 
-
The TetR sequence (with a ribosome binding site and terminator) [http://partsregistry.org/Part:BBa_K092000 BBa_K092000] is bound to the part containing the promoter of TetR and the red fluorescent protein (RFP) with a ribosome binding site. A luxI part (with RBS and terminator) is added at the end of the cds of the RFP. As no terminator was placed after the RFP, they both depends on the TetR promoter. The transcription of RFP and LuxI is inhibited when TetR is present. The production of TetR depends on the promoter you put in front of it.
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*[http://partsregistry.org/Part:BBa_K092901 BBa_K092901]: Project Part
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:[[Image:K092001.jpg]]<br>
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:Sequence containing : - ''rhlR'' controlled by a constitutive promoter - ''lacI<sub>m</sub>'' modified with a RhlR promoter - ''tetR'' controlled by a Lac Promoter - ''RFP'' controlled by a TetR promoter - ''luxI'' is added at the end of the cds of the ''RFP''. As no terminator was placed after the ''RFP'', they both depends on the TetR promoter. The transcription of ''RFP'' and ''luxI'' is inhibited when TetR is present. The production of TetR depends on the promoter you put in front of it.
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[http://partsregistry.org/Part:BBa_K092800 BBa_K092800] :<br>
+
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[[Image:K092800bis.jpg]]<br>
+
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Coding sequence for LacI modified with a different rate of translation than LacI, RBS.
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-
 
+
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Source of LacIm : Basu et al."A synthetic multicelluar system for programmed pattern formation", Nature. 2005 Apr 28;434(7037):1130-4
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+
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[http://partsregistry.org/Part:BBa_K092900 BBa_K092900] :
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+
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+
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[[Image:K092900.jpg]]<br>
+
-
 
+
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RhlR controlled by a constitutive promoter, and LacI modified with a Rhl R promoter.
+
-
 
+
-
 
+
-
 
+
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[http://partsregistry.org/Part:BBa_K092901 BBa_K092901]: Project Part
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-
 
+
-
 
+
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[[Image:K092001.jpg]]<br>
+
-
 
+
-
Sequence containing : - RhlR controlled by a constitutive promoter - LacI modified with a RhlR promoter - TetR controlled by a Lac Promoter - RFP controlled by a TetR promoter - LuxI is added at the end of the cds of the RFP. As no terminator was placed after the RFP, they both depends on the TetR promoter. The transcription of RFP and LuxI is inhibited when TetR is present. The production of TetR depends on the promoter you put in front of it.
+
Unfortunately, as we are finishing our wiki, the last ligation is running. Thus, we can not submit our project part.
Unfortunately, as we are finishing our wiki, the last ligation is running. Thus, we can not submit our project part.

Latest revision as of 18:38, 29 October 2008


Cloning Scheme


First plasmid : the basic network components

Plasmid1.jpg

Second plasmid : the testing component

Plasmid2.jpg


Parts submitted to the registry


K092000.jpg
TetR coding sequence with RBS and terminator :
Coding sequence of the Tetracycline repressor with the ribosome binding site and a double terminator. The tetR comes from transposon tn10 and has a LVA tail (SsrA) [cf. C0040 part].
This part has been successfully sequenced.


K092100.jpg
Constitutive RhlR Receiver
This part has been successfully sequenced.
K092200.jpg
RhlI with RBS and terminator


K092300.jpg
RFP controlled by a TetR promoter :
Coding sequence of the RFP with a ribosome binding site and p(tetR) as promoter. Thus, the transcription of RFP is inhibited when TetR is present.
This part has been successfully sequenced.
K092400.jpg
New part created by 2-steps PCR. RBS (B0034) + luxI + Terminator (B0010+B0012) :
Coding sequence for the Lux Inducer with a ribosome binding site and terminator. This biobrick is an alternative to the F1610 which didn't work


K092600.jpg
TetR cds with RBS and Terminator + p(tetR), RFP with RBS
The TetR sequence (with a ribosome binding site and terminator) is bound to the part containing the promoter of tetR, a ribosome binding site and the red fluorescent protein (RFP). Thus, the production of RFP directly depends on the production of TetR. The transcription of RFP is inhibited when TetR is present. The production of TetR depends on the promoter you put in front of it.
This part has been successfully sequenced.
K092700.jpg
TetR cds with RBS and Terminator + p(tetR, RFP with RBS + luxI with RBS and terminator
The TetR sequence (with a ribosome binding site and terminator) BBa_K092000 is bound to the part containing the promoter of tetR and the red fluorescent protein (RFP) with a ribosome binding site. A luxI part (with RBS and terminator) is added at the end of the cds of the RFP. As no terminator was placed after the RFP, they both depends on the tetR promoter. The transcription of RFP and luxI is inhibited when TetR is present. The production of TetR depends on the promoter you put in front of it.


K092800bis.jpg
Sequence that codes the same amino acid sequence than LacI but coded with a codon optimized sequence. It will thus have a different rate of translation than LacI.
Source of LacIm : Basu et al."A synthetic multicelluar system for programmed pattern formation", Nature. 2005 Apr 28;434(7037):1130-4


K092900.jpg
rhlR controlled by a constitutive promoter, and lacIm with a RhlR promoter.


K092001.jpg
Sequence containing : - rhlR controlled by a constitutive promoter - lacIm modified with a RhlR promoter - tetR controlled by a Lac Promoter - RFP controlled by a TetR promoter - luxI is added at the end of the cds of the RFP. As no terminator was placed after the RFP, they both depends on the TetR promoter. The transcription of RFP and luxI is inhibited when TetR is present. The production of TetR depends on the promoter you put in front of it.

Unfortunately, as we are finishing our wiki, the last ligation is running. Thus, we can not submit our project part.