Team:BCCS-Bristol/Wetlab/BioBricks

From 2010.igem.org

Biobricks

Biobricks Used in Preliminary Experiments

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.


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

BBa_K216005 - PyeaR promoter

Nitrate- and nitrite-sensitive promoter. This will be combined with BBa_E0840 or BBa_E0240 to create a nitrate-sensitive system which signals through expression of GFP.

Vector used: pSB1A2. Ampicillin resistance used as selective marker.


BBa_E0840 - GFP generator

GFP plus strong RBS and termination sequences.

Vector used: pSB1A2. Ampicillin resistance used as selective marker.


BBa_E0240 - GFP generator

GFP plus weaker RBS and termination sequences.

Vector used: pSB1A2. Ampicillin resistance used as selective marker.


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.


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.