Team:UPMC-Paris/Description

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BBa_K2180001 : Zinc Operator - CzrA Repressor Binding Site

http://parts.igem.org/Part:BBa_K2180001
Biobrick based on a previous biobrick made by the iGEM09_Newcastle Team (2009-10-14) :
http://parts.igem.org/Part:BBa_K174015
We added two restriction sites: SacII before and PacI after the regulatory region in order to use it in our project.
This restriction sites allow us to replace a regulatory region with another regulatory region.

BBa_K2180002 : Fructose Operator - FruR Binding Site

http://parts.igem.org/Part:BBa_K2180002
The sequence of the regulatory region comes from this site
We added two restriction sites: SacII before and PacI after the regulatory region in order to use it in our project.
This restriction sites allow us to replace a regulatory region with another regulatory region.
The regulatory region is composed by two FruR binding sites wich are derepressed in the presence of fructose.

BBa_K2180003 : Sucrose Operator - DegU Binding Site

http://parts.igem.org/Part:BBa_K2180003
The sequence of the regulatory region comes from this site
We added two restriction sites SacII before and PacI after the regulatory region in order to use it in our project.
They allow us to replace a regulatory region with another regulatory region.
The regulatory region is composed by two DegU binding (one forward, one reverse) sites which are derepressed in the presence of sucrose.

BBa_K2180004 : Promoter +FUR regulatory sequence + RBS + LacI-LVA + Terminator

http://parts.igem.org/Part:BBa_K2180004
This part is an adaptation for B. subtilis of the biobrick BBa_K1163999 from iGEM13_Evry (2013-09-02):
http://parts.igem.org/Part:BBa_K2180004
This system allow the synthese of the protein Lac I in the absence of Iron.
This is the assembly of our basic parts BBa_K2180007, BBa_K2180009 and BBa_K2180008

BBa_K2180005 : Blue Pigment for B. subtilis

http://parts.igem.org/Part:BBa_K2180005
Biobrick based on a previous biobrick made by the Team iGEM11_Uppsala-Sweden   (2011-09-18) :
http://parts.igem.org/Part:BBa_K592009
This sequence is coding for a blue pigment we use in our detection system as reporter.
We had to improve the codon usage for B. subtilis.

BBa_K2180006 : Promoter PlepA + Operator LacI for B. subtilis

http://parts.igem.org/Part:BBa_K2180006
The part is made of a previous IGEM part made by the team iGEM12_LMU-Munich   (2012-07-16) :
http://parts.igem.org/Part:BBa_K823002
We add two LacI binding sites (Gilbert and Maxam, PNAS, 1973)
This part is made of PlepA promoter, two LacI binding sites and a RBS. There is a PacI restriction site before LacI operator binding sites and a SacII restriction site after them. They allow us to replace the LacI binding sites by other operating sequences. A SalI restriction sites is presents before the PlepA sequence and a NdeI restriction site after the RBS that may be used to insert the whole sequence. We use this part in our project to promote the pigment expression under the control of LacI.

BBa_K2180007 : Promoter PliaG + Fur binding site B. subtilis

http://parts.igem.org/Part:BBa_K2180007
This part is based on a previous IGEM part made by the Team iGEM12_LMU-Munich  (2012-07-16) :
http://parts.igem.org/Part:BBa_K823000
We add the Fur Box site (Baichoo and Helmann, J. Bacteriol., 2002) in order to make this promoter inactive in the absence of Iron.
This part is made of PliaG promoter, a fur (Ferric Uptake Regulator) binding site and a RBS. There is a ClaI restriction site before the Fur Box (Fur Binding Site) and a PscI restriction site after. They allows us to remplace Fur Box by another operating sequence. A BamHI restriction sites is present before the PliaG sequence and a NcoI restriction site after the RBS that may be used to insert the whole sequence. We use this part in our project to promote the LacI + LVA expression under the control of Fur. We had to check the compatibility of the promoter with the Fur Box.

BBa_K2180008 : Double terminator

http://parts.igem.org/Part:BBa_K2180008
This part is based on a previous iGEM part made by the Team Antiquity (2003-07-17) :
http://parts.igem.org/Part:BBa_B0015
This is a double terminator we use in our project for the pigment production. He is made of a transcriptional terminator in forward and reverse direction. We add a PvuII restriction site before the first terminator (forward) and an AvrII restriction site after the second terminator (reverse).

BBa_K2180009 : LacI-LVA for B. subtilis

http://parts.igem.org/Part:BBa_K2180009
The sequence is based on a previous IGEM part made by the group Antiquity (2003-01-31) coding for LacI + LVA for E. coli:
http://parts.igem.org/Part:BBa_C0012
This biobrick is the coding sequence of LacI protein + LVA (rapid degradation tail) wich improve the switch time and make it more sensitive.
We had to improve the codon usage for B. subtilis

BBa_K2180010 : Cadmium Operator - MntR Binding Site

http://parts.igem.org/Part:BBa_K2180010
Biobrick based on a previous biobrick made by the iGEM09_Cornell Team (2009-08-10) :
http://partsregistry.org/Part:BBa_K285103:Design
We added two restriction sites: SacII before and PacI after the regulatory region in order to use it in our project. They allow us to replace a regulatory region with another regulatory region. This Region allow MntR to bind to DNA and repress transcription. In the presence of Cadmium transcription is derepressed because of the fixation of cadmium to MntR.
*Other metallic ions may bind to MntR.

BBa_K2180011 : Cadmium Operator - ArsR Binding Site

http://parts.igem.org/Part:BBa_K2180011
Biobrick based on a previous biobrick made by the iGEM09_Newcastle Team (2009-10-14) :
http://parts.igem.org/Part:BBa_K174015
We added two restriction sites: SacII before and PacI after the regulatory region in order to use it in our project. They allow us to replace a regulatory region with another regulatory region. This Region allows ArsR to bind to DNA and repress transcription. In the presence of Heavy Metals such as Arsenic or Cadmium transcription is derepressed because of the fixation to ArsR.
*Other metalic ions may bind to ArsR.

BBa_K2180012 : Promoter PlepA + Operator LacI + Blue Pigment + Double terminator

http://parts.igem.org/Part:BBa_K21800102
This biobrick allow the production of the blue pigment.
This is the assembly of our parts BBa_K2180005, BBa_K2180006 and BBa_K2180008

BBa_K2180013 : Promoter + Zinc Operator - CzrA Repressor Binding Site + Blue Pigment + Terminator

http://parts.igem.org/Part:BBa_K2180013
This biobrick allow the production of the blue pigment in the presence of zinc.
This is the assembly of our parts BBa_K2180001, BBa_K2180006 and BBa_K2180008

BBa_K2180014 : Promoter + Fructose Operator - FruR Binding Site + Blue Pigment + Terminator

http://parts.igem.org/Part:BBa_K2180014
This biobrick allow the production of the blue pigment in the presence of fructose.
This is the assembly of our parts BBa_K2180002, BBa_K2180006 and BBa_K2180008

BBa_K2180015 : Promoter + Sucrose Operator - DegU Binding Site + Blue Pigment + Terminator

http://parts.igem.org/Part:BBa_K2180015
This biobrick allow the production of the blue pigment in the presence of sucrose.
This is the assembly of our parts BBa_K2180003, BBa_K2180006 and BBa_K2180008

BBa_K2180016 : Promoter + Cadmium Operator - MntR Binding Site + Blue Pigment + Terminator

http://parts.igem.org/Part:BBa_K2180016
This biobrick allow the production of the blue pigment in the presence of cadmium.
This is the assembly of our parts BBa_K2180010, BBa_K2180006 and BBa_K2180008

BBa_K2180017 : Promotor + Cadmium Operator - ArsR Binding Site + Blue Pigment + Terminator

http://parts.igem.org/Part:BBa_K2180017
This biobrick allow the production of the blue pigment in the presence of arsenic.
This is the assembly of our parts BBa_K2180011, BBa_K2180006 and BBa_K2180008

BBa_K2180018 : Promoter PlepA +FUR regulatory sequence + RBS + LacI-LVA + Terminator + Pigment + RBS + Operateur Lac I + Promoter PliaG

http://parts.igem.org/Part:BBa_K2180018
This biobrick allow the production of the blue pigment in the presence of iron.
This is the assembly of our parts BBa_K2180002, BBa_K2180006 and BBa_K2180008

BBa_K2180019 : Blue pigment adaptated with B. subtilis' code codon with LVA tail and enzyme restriction site at both extremities.

http://parts.igem.org/Part:BBa_K2180019

Biobrick based on a previous biobrick made by the Team iGEM11_Uppsala-Sweden   (2011-09-18) :
http://parts.igem.org/Part:BBa_K592009
This sequence is coding for a blue pigment we use in our detection system as reporter.
We had to improve the codon usage for B. subtilis.

BBa_K2180020 : Blue pigment adaptated with B. subtilis' code codon with LVA tail

http://parts.igem.org/Part:BBa_K2180020
Biobrick based on a previous biobrick made by the Team iGEM11_Uppsala-Sweden   (2011-09-18) :
http://parts.igem.org/Part:BBa_K592009
This sequence is coding for a blue pigment we use in our detection system as reporter.
We had to improve the codon usage for B. subtilis and had an LVA tail (rapid degradation tail) wich improve the switch time and make it more sensitive.