| FIFO
|
| Construction
| Description
| Validation Date
|
   
[http://partsregistry.org/Part:BBa_K136039 pFliL(RBS WT)-ECFP tripart (LVA+)]
| Natural Promoter of E. coli regulated by FlhDC and FliA. The natural rbs of the downstream gene is conserved for a more natural expression of the following gene. The fluorescent marker associated with pFliL is ECFP fused with a LVA tag.
| October 24
|
pFliL(+ RBS WT)-GFP
| Natural Promoter of E. coli regulated by FlhDC and FliA. The natural rbs of the downstream gene is conserved for a more natural expression of the following gene. The fluorescent marker associated with pFliL is GFP.
| October 24
|
pFlgA(+ RBS WT) - YFP Tripart (LVA+)
| Natural Promoter of E. coli regulated by FlhDC and FliA. The natural rbs of the downstream gene is conserved for a more natural expression of the following gene. The fluorescent marker associated with pFlgA is EYFP fused with a LVA tag.
| August 22
|
pFlgA(+ RBS WT) - YFP Tripart (LVA-)
| Natural Promoter of E. coli regulated by FlhDC and FliA. The natural rbs of the downstream gene is conserved for a more natural expression of the following gene. The fluorescent marker associated with pFlgA is EYFP fused without a LVA tag.
| August 22
|
pFlgA(+ RBS WT)-YFP tripart (AAV+)
| Natural Promoter of E. coli regulated by FlhDC and FliA. The natural rbs of the downstream gene is conserved for a more natural expression of the following gene. The fluorescent marker associated with pFlgA is EYFP fused with a AAV tag.
| October 24
|
pFlgA(+ RBS WT)-GFP
| Natural Promoter of E. coli regulated by FlhDC and FliA. The natural rbs of the downstream gene is conserved for a more natural expression of the following gene. The fluorescent marker associated with pFlgA is GFP.
| October 24
|
pFlhB(+ RBS WT)-mRFP tripart (+LVA)
| Natural Promoter of E. coli regulated by FlhDC and FliA. The natural rbs of the downstream gene is conserved for a more natural expression of the following gene. The fluorescent marker associated with pFlhB is mRFP with a LVA tag.
| October 24
|
pFlhB(+ RBS WT)-GFP
| Natural Promoter of E. coli regulated by FlhDC and FliA. The natural rbs of the downstream gene is conserved for a more natural expression of the following gene. The fluorescent marker associated with pFlhB is GFP.
| October 26
|
| OSCILLATION
|
pFliL(+ RBS WT)-EnvZ*
| pFliL is regulated by FlhDC and EnvZ regulates pFlhDC thanks to a negative feedback loop, origin of an oscillation behavior.
| October 26
|
 
pFliL(+ RBS WT)-EnvZ*-GFP generator
| The same construction as before but with a GFP in order to follow the dynamic of expression of pFliL.
| October 29
|
pFlhB(+ RBS WT)-EnvZ*
| pFlhB is regulated by FlhDC and EnvZ regulates pFlhDC thanks to a negative feedback loop, origin of an oscillation behavior.
| October 24
|
pFlhB(+ RBS WT)-EnvZ*-GFP generator
| The same construction as before but with a GFP in order to follow the dynamic of expression of pFlhB.
| October 29
|
araC/pBad-EnvZ*
| A construction made for characterizing the repression power of the mutated EnvZ on our system
| October 27
|
| SYNCHRONISATION
|
pLas - gfp Tripart
| Construction for characterization of the pLas promoter
| August 1
|
pLas - ECFP
| The same as before but with another fluorophore
| August 1
|
strongest promoter-rbs-lasR-dble ter
| A construction for a strong constitutive expression of the Las Regulator in order to get maximal expression in presence of AHL
| August 20
|
Strong promoter-rbs-lasR-dble ter
| A derived construction of the previous one for smaller constitutive expression of LasR in the Bacteria.
| August 20
|
Strongest RBS (1)- LasR activator (+LVA)
| Intermediary construction. We uses different rbs for Las R in order to get various response dynamic to the presence of AHL
| August 13
|
rbs-lasR + dbl ter
| Intermediary construction. We uses different rbs for Las R in order to get various response dynamic to the presence of AHL
| August 19
|
lasR activator + LVA - Double Terminator
| Intermediary construction
| August 1
|
RBS lasI - GFP tripart
| Construction that has to be associated with the FIFO. We will know when LasI is expressed and then when AHL is produced.
| August 13
|
RBS-lasI - ECFP
| The same as the previous one but with another fluorophore.
| August 13
|
| NEW GENE AND PROMOTER BIOBRICKS
|
pFliA
| Natural promoter of E. coli, extracted by PCR and then inserted in a biobrick vector. pFiA is regulated by FlhDC.
| October 04
|
pFliL (+RBS WT)
| Natural promoter with the wild type rbs of its associated gene in E. coli, extracted by PCR and then inserted in a biobrick vector. pFiA is regulated by FlhDC and FliA.
| October 10
|
pFlgA (+RBS WT)
| Natural promoter with the wild type rbs of its associated gene in E. coli, extracted by PCR and then inserted in a biobrick vector. pFiA is regulated by FlhDC and FliA..
| October 10
|
pFlgB (+RBS WT)
| Natural promoter with the wild type rbs of its associated gene in E. coli, extracted by PCR and then inserted in a biobrick vector. pFiA is regulated by FlhDC and FliA.
| October 10
|
pFlhB (+RBS WT)
| Natural promoter with the wild type rbs of its associated gene in E. coli, extracted by PCR and then inserted in a biobrick vector. pFiA is regulated by FlhDC and FliA.
| October 10
|
EnvZ*
| EnvZ is a membrane protein involved into the osmotic response and is interacting with OmpR that will bind the promoter of FlhDC ad will repress it. EnvZ* is a mutated form of the coding sequence, giving a phenotype where pFlhDC is completely repressed. EnvZ has been extracted by PVR for the correct mutant strain.
| September 04
|
FliA
| FliA is a sigma factor involved into the regulation of very various genes, the promoter used in the FIFO were selected to be higly responding to FliA. In order to have the FliA gene possibly used as biobrick, we mutated 2 sites (EcoRI and PstI) inside the FliA sequence by assemby PCR.
| September 04
|
FlhDC
| FlhDC is the main regulator of the flagella genetic system in E. coli. FlhDC regulates all the promoter of the FIFO.
| August 13
|
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