The plasmid sequences can be accessed by clicking on the plasmid name (e.g. piG0040) or the plasmid picture.

Construct types:

Regulator Constructs

Producer Constructs

Sensor Constructs

BUFFER Gate Construct

Signal Propagation Construct

Combined Sensor and Producer Constructs

Regulator Constructs

piG0040

LasR 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]. 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].

piG0041

LuxR 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]. 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].

piG0042

RhlR 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]. 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].

piG0042max

RhlR is expressed under the strong constitutive promoter [http://parts.igem.org/Part:BBa_J23100 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].

piG0046

LuxR is expressed under the weak constitutive promoter [http://parts.igem.org/Part:BBa_J23109 BBa_J23109] while the expression level is influenced by the RBS [http://parts.igem.org/Part:BBa_B0034 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].

piG0047

LuxR is expressed under the medium strong constitutive promoter [http://parts.igem.org/Part:BBa_J23111 BBa_J23111 ] while the expression level is influenced by the RBS [http://parts.igem.org/Part:BBa_B0034 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

piG0049

LasI 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]. 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].

piG0049max

LasI 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 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].

piG0050

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^[16].

piG0050max

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 (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].

piG0051

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^[16].

piG0051max

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 (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

piG0058

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].

piG0059

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].

piG0060

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].

piG0062

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].

piG0065

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. A biologically neutral spacer sequence was designed using the web application R2oDNA^[34]. The p15A is present at a copy number of approximately 15 to 25^[35].

piG0066

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. A biologically neutral spacer sequence was designed using the web application R2oDNA^[34]. The p15A is present at a copy number of approximately 15 to 25^[35].

piG0109

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. A biologically neutral spacer sequence was designed using the web application R2oDNA^[34]. 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.

piG0110

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. A biologically neutral spacer sequence was designed using the web application R2oDNA^[34]. 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

piG0067

The directional terminator [http://parts.igem.org/Part:BBa_B0015 BBa_B0015] blocks transcription of sfGFP under the strong constitutive promoter [http://parts.igem.org/Part:BBa_J23100 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

piG0071

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. A biologically neutral spacer sequence was designed using the web application R2oDNA^[34]. The p15A is present at a copy number of approximately 15 to 25^[35].

Combined Sensor and Producer Constructs

piG0096

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). A biologically neutral spacer sequence was designed using the web application R2oDNA^[34]. The p15A is present at a copy number of approximately 15 to 25^[35].

piG0097

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 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^[34]. The p15A is present at a copy number of approximately 15 to 25^[35].

piG0099

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^[35].