Part:BBa_K1065309
light regulated circuit producing amilGFP+EFE at dark
This part includes blue light sensor YF1, its RR fixJ,PfixK2,the reporter amilGFP and EFE+terminators: everything is under the regulation of Plac.
In the presence of blue light (470 nm) the production of the reporter amilGFP and EFE enzyme is inhibited.
In the absence of blue light the device is activated, thus producing amilGFP+EFE. 2-oxoglutarate oxygenase/decarboxylase is an Ethylene Forming Enzyme (EFE) that catalyze Ethylene biosynthesis from 2-oxoglutarate. This enzyme was firstly purified from Pseudomonas siringae pv. phaseolicola PK2, a 2-oxoglutarate-dependent ethylene producing bacterium.
This part was cloned by UNITN-Trento 2013 iGEM team for the creation of an aerobically engineered pathway for the control of fruit ripening: unlike Bba_k1065311, it doesn't have any inverter cassette, so blue light acts like an inhibitor of the circuit rather than the inducer.
Usage and Biology
Safety
This part produces ethylene, a compound that can be inflammable at a concentration between 2.7 to 36%. We characterized this part under the control of an AraC-pBAD promoter. With a air volume/culture volume ratio = 4, we detected about 200 ppm of Ethylene. This concentration is not dangerous and not inflammable. However we suggest to manage this part carefully. (See BBa_K1065001 for more details.)
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 3309
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 770
Illegal NgoMIV site found at 842
Illegal NgoMIV site found at 932
Illegal NgoMIV site found at 950
Illegal NgoMIV site found at 1462
Illegal NgoMIV site found at 1755
Illegal NgoMIV site found at 1849
Illegal AgeI site found at 484
Illegal AgeI site found at 1630
Illegal AgeI site found at 4060 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 1519
Illegal BsaI.rc site found at 383
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