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Team:BCCS-Bristol/Wetlab/BioBricks
From 2010.igem.org
iGEM 2010
Biobricks
Biobricks Used in Preliminary Experiments
[http://partsregistry.org/Part:BBa_I13522 BBa_I13522] - pTet GFP
GFP under the control of a constitutive promoter. This was used to visualise E.coli on soil under a fluorescence microscope during the preliminary testing stage.
Vector used: pSB1A2. Ampicillin resistance used as selective marker.
[http://partsregistry.org/Part:BBa_K216009 BBa_K216009] - PyeaR with lacZ'
Nitrate- and nitrite-sensitive promoter upstream of lacZ' reporter. Used to characterize the promoter through a Miller assay.
Vector used: pSB1A2. Ampicillin resistance used as selective marker.
Biobricks to be used in the Project
[http://partsregistry.org/Part:BBa_K216005 BBa_K216005] - PyeaR promoter
Nitrate- and nitrite-sensitive promoter. This will be combined with [http://partsregistry.org/Part:BBa_E0840 BBa_E0840] or [http://partsregistry.org/Part:BBa_E0240 BBa_E0240] to create a nitrate-sensitive system which signals through expression of GFP.
Vector used: pSB1A2. Ampicillin resistance used as selective marker.
[http://partsregistry.org/Part:BBa_E0840 BBa_E0840] - GFP generator
GFP plus strong RBS and termination sequences.
Vector used: pSB1A2. Ampicillin resistance used as selective marker.
[http://partsregistry.org/Part:BBa_E0240 BBa_E0240] - GFP generator
GFP plus weaker RBS and termination sequences.
Vector used: pSB1A2. Ampicillin resistance used as selective marker.
[http://partsregistry.org/Part:BBa_J04450 BBa_J04450] - RFP coding device
RFP under the control of a constitutive promoter. This will be included in the E.coli chassis as a background fluorescence to aid in sensing and identifying the GFP signal, plus to act as a reference point for analysis of expression level of the GFP signal.
Vector used: pSB1K3. Kanamycin resistance used as selective marker.
[http://partsregistry.org/Part:BBa_I732017 BBa_I732017] - lacZ full-length (RBS+ TERM-)
Full length copy of lacZ which will be combined with PyeaR to further categorise the promoter used in our Nitrate sensor system.
