Team:ETH Zurich/lab/sequences

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(Sensor Constructs)
 
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[[Team:ETH_Zurich/lab/sequences#BUFFER_Gate_Construct|BUFFER Gate Construct]]
[[Team:ETH_Zurich/lab/sequences#BUFFER_Gate_Construct|BUFFER Gate Construct]]
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[[Team:ETH_Zurich/lab/sequences#Signal_Propagation_Construct|Signal Propagation Construct]]
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[[Team:ETH_Zurich/lab/sequences#Integrase_Reporter_Construct|Integrase Reporter Construct]]
[[Team:ETH_Zurich/lab/sequences#Combined_Sensor_and_Producer_Constructs|Combined Sensor and Producer Constructs]]
[[Team:ETH_Zurich/lab/sequences#Combined_Sensor_and_Producer_Constructs|Combined Sensor and Producer Constructs]]
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[[File:ETH2014_piG0047_Map.png|link=https://static.igem.org/mediawiki/2014/1/1b/ETH2014_piG0047.txt]]
[[File:ETH2014_piG0047_Map.png|link=https://static.igem.org/mediawiki/2014/1/1b/ETH2014_piG0047.txt]]
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==Producer Constructs==
==Producer Constructs==
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[https://static.igem.org/mediawiki/2014/4/43/ETH2014_piG0049.txt piG0049]
[https://static.igem.org/mediawiki/2014/4/43/ETH2014_piG0049.txt piG0049]
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==Sensor Constructs==
==Sensor Constructs==
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[https://static.igem.org/mediawiki/2014/a/a7/ETH2014_piG0058_sensor_plasRstRBS-sfGFP.txt piG0058]
[https://static.igem.org/mediawiki/2014/a/a7/ETH2014_piG0058_sensor_plasRstRBS-sfGFP.txt piG0058]
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[[File:ETH2014 piG0067 logic bxb1 BUFFER-sfGFP Map.png|link=https://static.igem.org/mediawiki/2014/5/59/ETH2014_piG0067_logic_bxb1_BUFFER-sfGFP.txt]]
[[File:ETH2014 piG0067 logic bxb1 BUFFER-sfGFP Map.png|link=https://static.igem.org/mediawiki/2014/5/59/ETH2014_piG0067_logic_bxb1_BUFFER-sfGFP.txt]]
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==Signal Propagation Construct==
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==Integrase Reporter Construct==
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[https://static.igem.org/mediawiki/2014/1/1d/ETH2014_piG0071_sensor_pluxRRR12y_bxb1_mCherry.txt piG0071]
[https://static.igem.org/mediawiki/2014/1/1d/ETH2014_piG0071_sensor_pluxRRR12y_bxb1_mCherry.txt piG0071]
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Expression of the integrase Bxb1 and the fluorophore mCherry is induced when LuxR bound to 3OC6-HSL bind to pLuxR. The cis-repressive element (crR12y) inhibits the translation of Bxb1 and mCherry, since the RBS is blocked by secondary structures of the mRNA. The transcript of the trans-activating element (taR12y) binds to the transcript of the cis-repressive element, hence the RBS is not blocked anymore. The two elements build a [https://2014.igem.org/Team:ETH_Zurich/expresults#Riboregulators riboregulator] that decreases leakiness of pLuxR. The p15A is present at a copy number of approximately 15 to 25<sup>[[Team:ETH_Zurich/project/references#refChang|[35]]]</sup>.
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Expression of the integrase Bxb1 and the fluorophore mCherry is induced when LuxR bound to 3OC6-HSL bind to pLuxR. The cis-repressive element (crR12y) inhibits the translation of Bxb1 and mCherry, since the RBS is blocked by secondary structures of the mRNA. The transcript of the trans-activating element (taR12y) binds to the transcript of the cis-repressive element, hence the RBS is not blocked anymore. The two elements build a [https://2014.igem.org/Team:ETH_Zurich/expresults#Riboregulators riboregulator] that decreases leakiness of pLuxR. A biologically neutral spacer sequence was designed using the web application R2oDNA<sup>[[Team:ETH_Zurich/project/references#refR2oDNA|[34]]]</sup>. The p15A is present at a copy number of approximately 15 to 25<sup>[[Team:ETH_Zurich/project/references#refChang|[35]]]</sup>.
[[File:ETH2014 piG0071 sensor pluxRRR12y bxb1 mCherry Map.png|link=https://static.igem.org/mediawiki/2014/1/1d/ETH2014_piG0071_sensor_pluxRRR12y_bxb1_mCherry.txt]]
[[File:ETH2014 piG0071 sensor pluxRRR12y bxb1 mCherry Map.png|link=https://static.igem.org/mediawiki/2014/1/1d/ETH2014_piG0071_sensor_pluxRRR12y_bxb1_mCherry.txt]]
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==Combined Sensor and Producer Constructs==
==Combined Sensor and Producer Constructs==
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[https://static.igem.org/mediawiki/2014/f/f5/ETH2014_piG0096_pluxRRR12y-sfGFP-maxRBS-luxI.txt piG0096]
[https://static.igem.org/mediawiki/2014/f/f5/ETH2014_piG0096_pluxRRR12y-sfGFP-maxRBS-luxI.txt piG0096]
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Expression of sfGFP and LuxI is induced when LuxR bound to 3OC6-HSL bind to pLuxR. The cis-repressive element (crR12y) inhibits the translation of the succeeding gene, since the RBS is blocked by secondary structures of the mRNA. The transcript of the trans-activating element (taR12y) binds to the transcript of the cis-repressive element, hence the RBS is not blocked anymore. The two elements build a [https://2014.igem.org/Team:ETH_Zurich/expresults#Riboregulators riboregulator] that decreases leakiness of pLuxR. The expression level of LuxI is increased by an RBS optimised for LuxI ([https://salis.psu.edu/software/forward RBS calculator]).The p15A is present at a copy number of approximately 15 to 25<sup>[[Team:ETH_Zurich/project/references#refChang|[35]]]</sup>.
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Expression of sfGFP and LuxI is induced when LuxR bound to 3OC6-HSL bind to pLuxR. The cis-repressive element (crR12y) inhibits the translation of the succeeding gene, since the RBS is blocked by secondary structures of the mRNA. The transcript of the trans-activating element (taR12y) binds to the transcript of the cis-repressive element, hence the RBS is not blocked anymore. The two elements build a [https://2014.igem.org/Team:ETH_Zurich/expresults#Riboregulators riboregulator] that decreases leakiness of pLuxR. The expression level of LuxI is increased by an RBS optimised for LuxI ([https://salis.psu.edu/software/forward RBS calculator]). A biologically neutral spacer sequence was designed using the web application R2oDNA<sup>[[Team:ETH_Zurich/project/references#refR2oDNA|[34]]]</sup>. The p15A is present at a copy number of approximately 15 to 25<sup>[[Team:ETH_Zurich/project/references#refChang|[35]]]</sup>.
[[File:ETH2014 piG0096 pluxRRR12y-sfGFP-maxRBS-luxI Map.png|link=https://static.igem.org/mediawiki/2014/f/f5/ETH2014_piG0096_pluxRRR12y-sfGFP-maxRBS-luxI.txt]]
[[File:ETH2014 piG0096 pluxRRR12y-sfGFP-maxRBS-luxI Map.png|link=https://static.igem.org/mediawiki/2014/f/f5/ETH2014_piG0096_pluxRRR12y-sfGFP-maxRBS-luxI.txt]]
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[https://static.igem.org/mediawiki/2014/f/fc/ETH2014_piG0097_pluxRRR12y-sfGFP-stRBS-luxI.txt piG0097]
[https://static.igem.org/mediawiki/2014/f/fc/ETH2014_piG0097_pluxRRR12y-sfGFP-stRBS-luxI.txt piG0097]
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Expression of sfGFP and LuxI is induced when LuxR bound to 3OC6-HSL bind to pLuxR. The cis-repressive element (crR12y) inhibits the translation of the succeeding gene, since the RBS is blocked by secondary structures of the mRNA. The transcript of the trans-activating element (taR12y) binds to the transcript of the cis-repressive element, hence the RBS is not blocked anymore. The two elements build a [https://2014.igem.org/Team:ETH_Zurich/expresults#Riboregulators riboregulator] that decreases leakiness of pLuxR. The expression level of LuxI is influenced by the RBS [http://parts.igem.org/Part:BBa_B0034 BBa_B0034]. The p15A is present at a copy number of approximately 15 to 25<sup>[[Team:ETH_Zurich/project/references#refChang|[35]]]</sup>.
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Expression of sfGFP and LuxI is induced when LuxR bound to 3OC6-HSL bind to pLuxR. The cis-repressive element (crR12y) inhibits the translation of the succeeding gene, since the RBS is blocked by secondary structures of the mRNA. The transcript of the trans-activating element (taR12y) binds to the transcript of the cis-repressive element, hence the RBS is not blocked anymore. The two elements build a [https://2014.igem.org/Team:ETH_Zurich/expresults#Riboregulators riboregulator] that decreases leakiness of pLuxR. The expression level of LuxI is influenced by the RBS [http://parts.igem.org/Part:BBa_B0034 BBa_B0034]. A biologically neutral spacer sequence was designed using the web application R2oDNA<sup>[[Team:ETH_Zurich/project/references#refR2oDNA|[34]]]</sup>. The p15A is present at a copy number of approximately 15 to 25<sup>[[Team:ETH_Zurich/project/references#refChang|[35]]]</sup>.
[[File:ETH2014 piG0097 pluxRRR12y-sfGFP-stRBS-luxI Map.png|link=https://static.igem.org/mediawiki/2014/f/fc/ETH2014_piG0097_pluxRRR12y-sfGFP-stRBS-luxI.txt]]
[[File:ETH2014 piG0097 pluxRRR12y-sfGFP-stRBS-luxI Map.png|link=https://static.igem.org/mediawiki/2014/f/fc/ETH2014_piG0097_pluxRRR12y-sfGFP-stRBS-luxI.txt]]

Latest revision as of 03:28, 18 October 2014

iGEM ETH Zurich 2014