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Team:ETH Zurich/lab/sequences
From 2014.igem.org
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[https://static.igem.org/mediawiki/2014/4/49/ETH2014_piG0050.txt piG0050] | [https://static.igem.org/mediawiki/2014/4/49/ETH2014_piG0050.txt piG0050] | ||
| - | LuxI is expressed under the strong constitutive promoter [http://parts.igem.org/Part:BBa_J23100 BBa_J23100 ] | + | LuxI is expressed under the strong constitutive promoter [http://parts.igem.org/Part:BBa_J23100 BBa_J23100 ] while the expression level is influenced by the RBS [http://parts.igem.org/Part:BBa_B0034 BBa_B0034]. LuxI produces the quorum sensing molecule 3OC6-HSL. The pBBR1 origin is present at a copy number of approximately 5, however, the origin is poorly characterized<sup>[[Team:ETH_Zurich/project/references#refLennen|[16]]]</sup>. |
[[File:ETH2014_piG0050_Map.png|link=https://static.igem.org/mediawiki/2014/4/49/ETH2014_piG0050.txt]] | [[File:ETH2014_piG0050_Map.png|link=https://static.igem.org/mediawiki/2014/4/49/ETH2014_piG0050.txt]] | ||
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[https://static.igem.org/mediawiki/2014/5/50/ETH2014_piG0050max.txt piG0050max] | [https://static.igem.org/mediawiki/2014/5/50/ETH2014_piG0050max.txt piG0050max] | ||
| - | LuxI is expressed under the strong constitutive promoter [http://parts.igem.org/Part:BBa_J23100 BBa_J23100 ] | + | LuxI is expressed under the strong constitutive promoter [http://parts.igem.org/Part:BBa_J23100 BBa_J23100 ] while the expression level is increased by an RBS optimised for LuxI ([https://salis.psu.edu/software/forward RBS calculator]). LuxI produces the quorum sensing molecule 3OC6-HSL. The pBBR1 origin is present at a copy number of approximately 5, however, the origin is poorly characterized<sup>[[Team:ETH_Zurich/project/references#refLennen|[16]]]</sup>. |
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[https://static.igem.org/mediawiki/2014/0/00/ETH2014_piG0051.txt piG0051] | [https://static.igem.org/mediawiki/2014/0/00/ETH2014_piG0051.txt piG0051] | ||
| - | RhlI is expressed under the strong constitutive promoter [http://parts.igem.org/Part:BBa_J23100 BBa_J23100 ] | + | RhlI is expressed under the strong constitutive promoter [http://parts.igem.org/Part:BBa_J23100 BBa_J23100 ] while the expression level is influenced by the RBS [http://parts.igem.org/Part:BBa_B0034 BBa_B0034]. RhlI produces the quorum sensing molecule C4-HSL. The pBBR1 origin is present at a copy number of approximately 5, however, the origin is poorly characterized<sup>[[Team:ETH_Zurich/project/references#refLennen|[16]]]</sup>. |
[[File:ETH2014_piG0051_Map.png|link=https://static.igem.org/mediawiki/2014/0/00/ETH2014_piG0051.txt]] | [[File:ETH2014_piG0051_Map.png|link=https://static.igem.org/mediawiki/2014/0/00/ETH2014_piG0051.txt]] | ||
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[https://static.igem.org/mediawiki/2014/9/94/ETH2014_piG0051max.txt piG0051max] | [https://static.igem.org/mediawiki/2014/9/94/ETH2014_piG0051max.txt piG0051max] | ||
| - | RhlI is expressed under the strong constitutive promoter [http://parts.igem.org/Part:BBa_J23100 BBa_J23100 ] | + | RhlI is expressed under the strong constitutive promoter [http://parts.igem.org/Part:BBa_J23100 BBa_J23100 ] while the expression level is increased by an RBS optimised for RhlI ([https://salis.psu.edu/software/forward RBS calculator]). RhlI produces the quorum sensing molecule C4-HSL. The pBBR1 origin is present at a copy number of approximately 5, however, the origin is poorly characterized<sup>[[Team:ETH_Zurich/project/references#refLennen|[16]]]</sup>. |
[[File:ETH2014_piG0051max_Map.png|link=https://static.igem.org/mediawiki/2014/9/94/ETH2014_piG0051max.txt]] | [[File:ETH2014_piG0051max_Map.png|link=https://static.igem.org/mediawiki/2014/9/94/ETH2014_piG0051max.txt]] | ||
Revision as of 20:58, 17 October 2014
Sequences
The plasmid sequences can be accessed by clicking on the plasmid name (e.g. piG0040) or the plasmid picture.
Construct types:
Combined Sensor and Producer Constructs
Regulator Constructs
LasR is expressed under the strong constitutive promoter BBa_J23100 while the expression level is influenced by the RBS BBa_B0034. LasR bound to 3OC12-HSL induces the expression of genes under the control of pLasR. The plasmid pBR322 and its derivatives have a copy number of 15 to 20[15].
LuxR is expressed under the strong constitutive promoter BBa_J23100 while the expression level is influenced by the RBS BBa_B0034. LuxR bound to 3OC6-HSL induces the expression of genes under the control of pLuxR. The plasmid pBR322 and its derivatives have a copy number of 15 to 20[15].
RhlR is expressed under the strong constitutive promoter BBa_J23100 while the expression level is influenced by the RBS BBa_B0034. RhlR bound to C4-HSL induces the expression of genes under the control of pRhlR. The plasmid pBR322 and its derivatives have a copy number of 15 to 20[15].
RhlR is expressed under the strong constitutive promoter BBa_J23100 while the expression level is influenced by an RBS optimised for RhlR (RBS calculator). RhlR bound to C4-HSL induces the expression of genes under the control of pRhlR. The plasmid pBR322 and its derivatives have a copy number of 15 to 20[15].
LuxR is expressed under the weak constitutive promoter BBa_J23109 while the expression level is influenced by the RBS BBa_B0034. LuxR bound to 3OC6-HSL induces the expression of genes under the control of pLuxR. The plasmid pBR322 and its derivatives have a copy number of 15 to 20[15].
LuxR is expressed under the medium strong constitutive promoter BBa_J23111 while the expression level is influenced by the RBS BBa_B0034. LuxR bound to 3OC6-HSL induces the expression of genes under the control of pLuxR. The plasmid pBR322 and its derivatives have a copy number of 15 to 20[15].
Producer Constructs
LasI is expressed under the strong constitutive promoter BBa_J23100 while the expression level is influenced by the RBS BBa_B0034. LasI produces the quorum sensing molecule 3OC12-HSL. The pBBR1 origin is present at a copy number of approximately 5, however, the origin is poorly characterized [16].
LasI is expressed under the strong constitutive promoter BBa_J23100 while the expression level is increased by an RBS optimised for LasI (RBS calculator). LasI produces the quorum sensing molecule 3OC12-HSL. The pBBR1 origin is present at a copy number of approximately 5, however, the origin is poorly characterized [16].
LuxI is expressed under the strong constitutive promoter BBa_J23100 while the expression level is influenced by the RBS BBa_B0034. LuxI produces the quorum sensing molecule 3OC6-HSL. The pBBR1 origin is present at a copy number of approximately 5, however, the origin is poorly characterized[16].
LuxI is expressed under the strong constitutive promoter BBa_J23100 while the expression level is increased by an RBS optimised for LuxI (RBS calculator). LuxI produces the quorum sensing molecule 3OC6-HSL. The pBBR1 origin is present at a copy number of approximately 5, however, the origin is poorly characterized[16].
RhlI is expressed under the strong constitutive promoter BBa_J23100 while the expression level is influenced by the RBS BBa_B0034. RhlI produces the quorum sensing molecule C4-HSL. The pBBR1 origin is present at a copy number of approximately 5, however, the origin is poorly characterized[16].
RhlI is expressed under the strong constitutive promoter BBa_J23100 while the expression level is increased by an RBS optimised for RhlI (RBS calculator). RhlI produces the quorum sensing molecule C4-HSL. The pBBR1 origin is present at a copy number of approximately 5, however, the origin is poorly characterized[16].
Sensor Constructs
Expression of sfGFP is induced when LasR bound to 3OC12-HSL bind to pLasR. The p15A is present at a copy number of approximately 15 to 25[35].
Expression of sfGFP is induced when LuxR bound to 3OC6-HSL bind to pLuxR. The p15A is present at a copy number of approximately 15 to 25[35].
Expression of sfGFP is induced when RhlR bound to 4C-HSL bind to pRhlR. The p15A is present at a copy number of approximately 15 to 25[35].
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 p15A is present at a copy number of approximately 15 to 25[35].
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 riboregulator that decreases leakiness of pLuxR. The p15A is present at a copy number of approximately 15 to 25[35].
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 riboregulator that decreases leakiness of pLuxR. The p15A is present at a copy number of approximately 15 to 25[35].
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 riboregulator that decreases leakiness of pLuxR. The p15A is present at a copy number of approximately 15 to 25[35]. 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.
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 riboregulator that decreases leakiness of pLuxR. The p15A is present at a copy number of approximately 15 to 25[35]. 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.
BUFFER Gate Construct
The directional terminator BBa_B0015 blocks transcription of sfGFP under the strong constitutive promoter BBa_J23100. Only if the integrase Bxb1 flips B0015 between the attP and the attB sites, transcription of sfGFP is possible. The pBBR1 origin is present at a copy number of approximately 5, however, the origin is poorly characterized[16]
Signal Propagation Construct
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 riboregulator that decreases leakiness of pLuxR. The p15A is present at a copy number of approximately 15 to 25[35].
Combined Sensor and Producer Constructs
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 riboregulator that decreases leakiness of pLuxR. The expression level of LuxI is increased by an RBS optimised for LuxI (RBS calculator).The p15A is present at a copy number of approximately 15 to 25[35].
