Team:ETH Zurich/lab/sequences

From 2014.igem.org

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(Combined Sensor and Producer 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=http://2014.igem.org/wiki/images/1/1b/ETH2014_piG0047.txt]]
[[File:ETH2014_piG0047_Map.png|link=http://2014.igem.org/wiki/images/1/1b/ETH2014_piG0047.txt]]
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==Producer Constructs==
==Producer Constructs==
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[http://2014.igem.org/wiki/images/4/43/ETH2014_piG0049.txt piG0049]
[http://2014.igem.org/wiki/images/4/43/ETH2014_piG0049.txt piG0049]
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==Sensor Constructs==
==Sensor Constructs==
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[http://2014.igem.org/wiki/images/a/a7/ETH2014_piG0058_sensor_plasRstRBS-sfGFP.txt piG0058]
[http://2014.igem.org/wiki/images/a/a7/ETH2014_piG0058_sensor_plasRstRBS-sfGFP.txt piG0058]
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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>.
+
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>.
+
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.
+
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]]
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[[File:ETH2014 piG0067 logic bxb1 BUFFER-sfGFP Map.png|link=http://2014.igem.org/wiki/images/5/59/ETH2014_piG0067_logic_bxb1_BUFFER-sfGFP.txt]]
[[File:ETH2014 piG0067 logic bxb1 BUFFER-sfGFP Map.png|link=http://2014.igem.org/wiki/images/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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[http://2014.igem.org/wiki/images/1/1d/ETH2014_piG0071_sensor_pluxRRR12y_bxb1_mCherry.txt piG0071]
[http://2014.igem.org/wiki/images/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 [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 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 [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 piG0071 sensor pluxRRR12y bxb1 mCherry Map.png|link=http://2014.igem.org/wiki/images/1/1d/ETH2014_piG0071_sensor_pluxRRR12y_bxb1_mCherry.txt]]
[[File:ETH2014 piG0071 sensor pluxRRR12y bxb1 mCherry Map.png|link=http://2014.igem.org/wiki/images/1/1d/ETH2014_piG0071_sensor_pluxRRR12y_bxb1_mCherry.txt]]
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 +
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==Combined Sensor and Producer Constructs==
==Combined Sensor and Producer Constructs==
 +
[http://2014.igem.org/wiki/images/f/f5/ETH2014_piG0096_pluxRRR12y-sfGFP-maxRBS-luxI.txt piG0096]
[http://2014.igem.org/wiki/images/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 [http://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 [http://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=http://2014.igem.org/wiki/images/f/f5/ETH2014_piG0096_pluxRRR12y-sfGFP-maxRBS-luxI.txt]]
[[File:ETH2014 piG0096 pluxRRR12y-sfGFP-maxRBS-luxI Map.png|link=http://2014.igem.org/wiki/images/f/f5/ETH2014_piG0096_pluxRRR12y-sfGFP-maxRBS-luxI.txt]]
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[http://2014.igem.org/wiki/images/f/fc/ETH2014_piG0097_pluxRRR12y-sfGFP-stRBS-luxI.txt piG0097]
[http://2014.igem.org/wiki/images/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 [http://2014.igem.org/Team:ETH_Zurich/expresults#Riboregulators riboregulator] that decreases leakiness of pLuxR. The expression level of LuxI is influenced by the RBS B0034. The p15A is present at a copy number of approximately 15 to 25<sup>[[Team:ETH_Zurich/project/references#refChang|[35]]]</sup>.
+
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 [http://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=http://2014.igem.org/wiki/images/f/fc/ETH2014_piG0097_pluxRRR12y-sfGFP-stRBS-luxI.txt]]
[[File:ETH2014 piG0097 pluxRRR12y-sfGFP-stRBS-luxI Map.png|link=http://2014.igem.org/wiki/images/f/fc/ETH2014_piG0097_pluxRRR12y-sfGFP-stRBS-luxI.txt]]
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[http://2014.igem.org/wiki/images/6/69/ETH2014_piG0099_pluxstRBS-sfGFP-stRBS-luxI.txt piG0099]
[http://2014.igem.org/wiki/images/6/69/ETH2014_piG0099_pluxstRBS-sfGFP-stRBS-luxI.txt piG0099]
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Expression of sfGFP and LuxI is induced when LuxR bound to 3OC6-HSL bind to pLuxR. The expression level of sfGFP and LuxI is influenced by the RBS B0034. The p15A is present at a copy number of approximately 15 to 25<sup>[[Team:ETH_Zurich/project/references#refChang|[35]]]</sup>.
+
Expression of sfGFP and LuxI is induced when LuxR bound to 3OC6-HSL bind to pLuxR. The expression level of sfGFP and 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>.
[[File:ETH2014_piG0099_pluxstRBS-sfGFP-stRBS-luxI_Map.png|link=http://2014.igem.org/wiki/images/6/69/ETH2014_piG0099_pluxstRBS-sfGFP-stRBS-luxI.txt]]
[[File:ETH2014_piG0099_pluxstRBS-sfGFP-stRBS-luxI_Map.png|link=http://2014.igem.org/wiki/images/6/69/ETH2014_piG0099_pluxstRBS-sfGFP-stRBS-luxI.txt]]

Latest revision as of 03:28, 18 October 2014

iGEM ETH Zurich 2014