Team:ETH Zurich/lab/sequences

From 2014.igem.org

(Difference between revisions)
(Combined Sensor and Producer Constructs)
(Sensor Constructs)
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[http://2014.igem.org/wiki/images/d/da/ETH2014_piG0065_sensor_pluxRRR12y-sfGFP.txt piG0065]
[http://2014.igem.org/wiki/images/d/da/ETH2014_piG0065_sensor_pluxRRR12y-sfGFP.txt piG0065]
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Expression of sfGFP is induced when LuxR bound to 3OC6-HSL bind to pLuxR. The cis-repressive element (crR12y) inhibits the translation of sfGFP, 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 [http://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 sfGFP is induced when LuxR bound to 3OC6-HSL bind to pLuxR. The cis-repressive element (crR12y) inhibits the translation of sfGFP, 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 [http://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 piG0065 sensor pluxRRR12y-sfGFP Map.png|link=http://2014.igem.org/wiki/images/d/da/ETH2014_piG0065_sensor_pluxRRR12y-sfGFP.txt]]
[[File:ETH2014 piG0065 sensor pluxRRR12y-sfGFP Map.png|link=http://2014.igem.org/wiki/images/d/da/ETH2014_piG0065_sensor_pluxRRR12y-sfGFP.txt]]
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[http://2014.igem.org/wiki/images/2/24/ETH2014_piG0066_sensor_prhlRRR12-sfGFP.txt piG0066]
[http://2014.igem.org/wiki/images/2/24/ETH2014_piG0066_sensor_prhlRRR12-sfGFP.txt piG0066]
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Expression of sfGFP is induced when RhlR bound to 4C-HSL bind to pRhlR. The cis-repressive element (crR12) inhibits the translation of sfGFP, since the RBS is blocked by secondary structures of the mRNA. The transcript of the trans-activating element (taR12) binds to the transcript of the cis-repressive element, hence the RBS is not blocked anymore. The two elements build a [http://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 sfGFP is induced when RhlR bound to 4C-HSL bind to pRhlR. The cis-repressive element (crR12) inhibits the translation of sfGFP, since the RBS is blocked by secondary structures of the mRNA. The transcript of the trans-activating element (taR12) binds to the transcript of the cis-repressive element, hence the RBS is not blocked anymore. The two elements build a [http://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 piG0066 sensor prhlRRR12-sfGFP Map.png|link=http://2014.igem.org/wiki/images/2/24/ETH2014_piG0066_sensor_prhlRRR12-sfGFP.txt]]
[[File:ETH2014 piG0066 sensor prhlRRR12-sfGFP Map.png|link=http://2014.igem.org/wiki/images/2/24/ETH2014_piG0066_sensor_prhlRRR12-sfGFP.txt]]
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[http://2014.igem.org/wiki/images/9/95/ETH2014_piG0109_pluxRRR12y-sfGFP_EcoRI-_XbaI-.txt piG0109]
[http://2014.igem.org/wiki/images/9/95/ETH2014_piG0109_pluxRRR12y-sfGFP_EcoRI-_XbaI-.txt piG0109]
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Expression of sfGFP is induced when LuxR bound to 3OC6-HSL bind to pLuxR. The cis-repressive element (crR12y) inhibits the translation of sfGFP, 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 [http://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>. PiG0109 is a derivate of piG0065 where the restriction sites EcoRI and XbaI have been removed. Thus, the two constructs slightly differ in the sequence of the 3'-end of the trans-activating element and in the sequence of the 5'-end of the cis-repressive element.
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Expression of sfGFP is induced when LuxR bound to 3OC6-HSL bind to pLuxR. The cis-repressive element (crR12y) inhibits the translation of sfGFP, 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 [http://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>. PiG0109 is a derivate of piG0065 where the restriction sites EcoRI and XbaI have been removed. Thus, the two constructs slightly differ in the sequence of the 3'-end of the trans-activating element and in the sequence of the 5'-end of the cis-repressive element.
[[File:ETH2014 piG0109 pluxRRR12y-sfGFP EcoRI- XbaI- Map.png|link=http://2014.igem.org/wiki/images/9/95/ETH2014_piG0109_pluxRRR12y-sfGFP_EcoRI-_XbaI-.txt]]
[[File:ETH2014 piG0109 pluxRRR12y-sfGFP EcoRI- XbaI- Map.png|link=http://2014.igem.org/wiki/images/9/95/ETH2014_piG0109_pluxRRR12y-sfGFP_EcoRI-_XbaI-.txt]]
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[http://2014.igem.org/wiki/images/f/f9/ETH2014_piG0110_prhlRRR12-sfGFP_EcoRI-_XbaI-.txt piG0110]
[http://2014.igem.org/wiki/images/f/f9/ETH2014_piG0110_prhlRRR12-sfGFP_EcoRI-_XbaI-.txt piG0110]
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Expression of sfGFP is induced when RhlR bound to 4C-HSL bind to pRhlR. The cis-repressive element (crR12) inhibits the translation of sfGFP, since the RBS is blocked by secondary structures of the mRNA. The transcript of the trans-activating element (taR12) binds to the transcript of the cis-repressive element, hence the RBS is not blocked anymore. The two elements build a [http://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>. PiG0110 is a derivate of piG0066 where the restriction sites EcoRI and XbaI have been removed. Thus, the two constructs slightly differ in the sequence of the 3'-end of the trans-activating element and in the sequence of the 5'-end of the cis-repressive element.
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Expression of sfGFP is induced when RhlR bound to 4C-HSL bind to pRhlR. The cis-repressive element (crR12) inhibits the translation of sfGFP, since the RBS is blocked by secondary structures of the mRNA. The transcript of the trans-activating element (taR12) binds to the transcript of the cis-repressive element, hence the RBS is not blocked anymore. The two elements build a [http://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>. PiG0110 is a derivate of piG0066 where the restriction sites EcoRI and XbaI have been removed. Thus, the two constructs slightly differ in the sequence of the 3'-end of the trans-activating element and in the sequence of the 5'-end of the cis-repressive element.
[[File:ETH2014 piG0110 prhlRRR12-sfGFP EcoRI- XbaI- Map.png|link=http://2014.igem.org/wiki/images/f/f9/ETH2014_piG0110_prhlRRR12-sfGFP_EcoRI-_XbaI-.txt]]
[[File:ETH2014 piG0110 prhlRRR12-sfGFP EcoRI- XbaI- Map.png|link=http://2014.igem.org/wiki/images/f/f9/ETH2014_piG0110_prhlRRR12-sfGFP_EcoRI-_XbaI-.txt]]

Revision as of 21:16, 17 October 2014

iGEM ETH Zurich 2014