Team:Edinburgh/BioBricks

From 2010.igem.org

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<p>The light-sensing and light-production BioBricks used in the <a href="http://2010.igem.org/Team:Edinburgh/Bacterial">bacterial BRIDGEs component</a> of the project are listed below. A few are <b>resubmissions</b> of existing parts (noted in their description), whilst others are composite devices with promoters and reporter systems.</p>
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<p>The light-sensing and light-production BioBricks used in the <a href="http://2010.igem.org/Team:Edinburgh/Bacterial">bacterial BRIDGEs component</a> of the project are listed below. A few are <b>resubmissions</b> of existing parts (noted as such in their descriptions), whilst others are composite devices with promoters and reporter systems.</p>
<p>The BioBricks for <a href="http://2010.igem.org/Team:Edinburgh/Bacterial/Red_light_producer">red light production</a> are two variants of mutated firefly luciferase.</p>
<p>The BioBricks for <a href="http://2010.igem.org/Team:Edinburgh/Bacterial/Red_light_producer">red light production</a> are two variants of mutated firefly luciferase.</p>

Revision as of 14:50, 27 October 2010







BioBricks: Genomic BRIDGEs


For this part of the project we will have one main BioBrick, which will be the construct required for the two-step markerless insertion, encompassing cat (chloramphenicol resistance) and sacB (prevents growth on sucrose).

Chloramphenicol resistance has already been well characterised in the registry, so our characterisation has mainly focused on sacB.

In addition we will probably be submitting the construct with the up- and down- stream sequences of useful genes which can be removed, e.g. tnaA, which produces indole - when 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 used in the bacterial BRIDGEs component of the project 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.


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 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 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 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 the LovTAP reporter system from Lausanne 2009 in a new plasmid.


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



Throughout this wiki there are words in bold that indicate a relevance to human aspects. It will become obvious that human aspects are a part of almost everything in iGEM.