Team:Edinburgh/BioBricks

From 2010.igem.org







BioBricks: Genomic BRIDGEs


For the BRIDGE protocol we have one main BioBrick, which is the construct required for the two-step markerless insertion, a composite construct of cat (chloramphenicol resistance) and sacB (prevention of growth on sucrose).

In addition, we submitted the up- and downstream sequences of an example of a useful gene to be removed from the E. coli genome using the BRIDGE protocol: tnaA, which produces indole, such that when it is removed E. coli no longer smells!


Name Type Link Description
cat Translational Unit BBa_K322210 Chloramphenicol acetyltransferase (chloramphenicol resistance gene).
sacB Translational Unit BBa_K322921 Bacillus subtilis levansucrase, lethal to E. coli in the presence of sucrose.
cat-sacB construct Composite BBa_K322922 Construct necessary for BRIDGE protocol.
tnaA upstream region Other BBa_K322705 Upstream region of E. coli tryptophanase locus, used for targetting genes to this locus using BRIDGE.
tnaA downstream region Other BBa_K322706 Downstream region of E. coli tryptophanase locus, used for targetting genes to this locus using BRIDGE.


BioBricks: Bacterial BRIDGEs


The light-sensing and light-production BioBricks of various emission / absorbance wavelengths developed in the FORTH light communication framework are listed below. A few are resubmissions of existing parts (noted as such in their descriptions), whilst others are composite devices with promoters and reporter systems.

The BioBricks for red light production are two variants of mutated firefly luciferase (S284T and 356K) and their composite reporter systems.


Name Type Link Description
Firefly luciferase S284T mutant Coding BBa_K322246 Firefly luciferase from Photinus pyralis mutated towards the red spectrum.
S284T mutant luciferase under lac promoter Composite BBa_K322247 Lac promoter followed by firefly luciferase S284T mutant.
Firefly luciferase 356K mutant Coding BBa_K322211 Firefly luciferase S284T mutant from Photinus pyralis re-mutated towards the red spectrum.
356K mutant luciferase under lac promoter Composite BBa_K322212 Lac promoter followed by firefly luciferase 356K mutant.


The BioBricks for blue light production are corrected variants of the LuxAB, LuxCDE, and LumP proteins from Edinburgh 2009 in various combinations, contained in new plasmids and with various reporter systems.


Name Type Link Description
luxAB Translational Unit BBa_K322139 Bacterial luciferase producing blue light (from X. luminescens).
luxCDE Composite BBa_K322312 Codes for a fatty acid reductase complex that makes the fatty acids necessary for LuxAB.
lumP Translational Unit BBa_K322007 Lumazine protein to shift the wavelength of LuxAB to blue.


Name Type Link Description
luxAB under lac promoter Composite BBa_K322140 Lac promoter followed by luxAB.
luxAB and luxCDE under lac promoter Composite BBa_K322141 Lac promoter followed by luxAB and luxCDE.
luxAB and lumP Composite BBa_K322149 luxAB and lumP composite part.
luxAB and lumP under lac promoter Composite BBa_K322150 Lac promoter followed by luxAB and lumP


The BioBricks for green light production are a modified version of the wildtype firefly luciferase from Ljubljana 2007 and a codon-optimised variant with brighter output.


Name Type Link Description
Firefly luciferase Coding BBa_K322237 Ljubljana 2007's BBa_I712019 in pSB1C3, from Photinus pyralis.
Wildtype luciferase under lac promoter Composite BBa_K322238 Lac promoter followed by wildtype luciferase.
Codon optimised bright firefly luciferase Translational Unit BBa_K322451 Firefly luciferase from Photinus pyralis mutated for increased bioluminescence.


The BioBricks for red light sensing are the updated Cph8 systems from UT Austin 2004 and Harvard 2008 in a variety of combinations, contained in a new plasmid and with two different reporter systems.


Name Type Link Description
Phycocyanobilin synthesis operon Translational Unit BBa_K322122 Harvard 2008's BBa_K098010 in pSB1C3.
Phycocyanobilin synthesis operon without terminator Translational Unit BBa_K322123 BBa_K098010 without terminator, allowing addition of cph8.
cph8 with RBS Translational Unit BBa_K322124 Light sensing protein for red light (BBa_I15010 in pSB1C3).
cph8 and reporter system (lacZ) Composite BBa_K322125 Red light sensor with lacZ reporter system.
cph8 and reporter system (EYFP) Composite BBa_K322126 Red light sensor with EYFP reporter system.
Phycocyanobilin synthesis genes with cph8 Composite BBa_K322127 The entire phycocyanobilin - cph8 system, without reporter system.
Phycocyanobilin synthesis genes with cph8 and EYFP reporter system Composite BBa_K322128 The entire phycocyanobilin - cph8 system, with EYFP reporter system.


The BioBrick for blue light sensing is a LovTAP reporter system based on work by Lausanne 2009.


Name Type Link Description
LovTAP and reporter system (RFP) Composite BBa_K322999 Blue light sensor with RFP reporter system.


The BioBricks for the green light sensing are the CcaS-PhoR light sensor and the associated parts necessary to create the desired reporter system.


Name Type Link Description
phoA promoter Regulatory BBa_K322115 Promoter activated by the CcaS-PhoR pathway.
phoA promoter with lacZ Composite BBa_K322117 phoA promoter coupled to lacZ reporter.
CcaS-PhoR Composite BBa_K322119 Green light sensor.
CcaS-PhoR and reporter system (lacZ) Composite BBa_K322120 Green light sensor with lacZ reporter system.



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