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From 2010.igem.org

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.