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Team:Edinburgh/BioBricks
From 2010.igem.org
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 | Coding | BBa_K322921 | Bacillus subtilis levansucrase, lethal to E. coli in presence of sucrose. |
| tnaA upstream region | Protein Domain | BBa_K322705 | Upstream region of E. coli tryptophanase locus, used for targetting genes to this locus using BRIDGE. |
| tnaA downstream region | Protein Domain | BBa_K322706 | Downstream region of E. coli tryptophanase locus, used for targetting genes to this locus using BRIDGE. |
| cat-sacB construct | Composite | BBa_K322922 | Construct necessary for BRIDGE protocol. |
BioBricks: Bacterial BRIDGEs
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| 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. |
| 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 | Coding | BBa_K322451 | Firefly luciferase from Photinus pyralis mutated for increased bioluminescence. |
| Name | Type | Link | Description |
|---|---|---|---|
| luxAB | Translational Unit | BBa_K322139 | Bacterial luciferase producing blue light (from X. luminescens). |
| luxCDE | Protein Domain | 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. |
| 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 | Signalling | BBa_K322149 | luxAB and lumP composite part. |
| luxAB and lumP under lac promoter | Composite | BBa_K322150 | Lac promoter followed by luxAB and lumP |
| 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 (chimeric Cph1 light receptor / EnvZ protein) for red light. |
| 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. |
| LovTAP and reporter system (RFP) | Composite | BBa_K322999 | Blue light sensor with RFP reporter system. |
