Types | DnaRegion
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Roles | engineered_region
Translational_Unit
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Sequences | BBa_K322124_sequence (Version 1)
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Description
Cph8 is a combination of cph1 and envZ which, together with PCB, form a transmembrane, histidine kinase-based light sensing system. It contains the photoreceptor from Cph1 and the histidine kinase and response-regulator from EnvZ???OmpR, producing a particularly strongresponse to light.
Features: cph8 coding sequence 16 to 2250, RBS 6 to 10.
Notes
Notes: we were unable to revive BioBrick I15010 with the coding sequence of the cph8 red light sensor fusion protein, so we sought to amplify it from BioBrick BBa_M30109. This led to numerous problems. It is clear from the Registry sequence data that M30109 is not what it is supposed to be. It does include cph8 with a ribosome binding site, but the RBS is not joined to cph8 by a standard scar; promoter R0040 may be present, but there appears to be an EcoRI site a little upstream of the ribosome binding site so that, when we cloned our PCR product, we failed to recover the promoter region; also the PstI site in cph8, which was mutated out in I15010, is still present in the version in M30109. We therefore performed corrective PCR to obtain a clone with the correct prefix, a ribosome binding site, and the cph8 coding sequence with the PstI site properly removed, which we present here, cloned in pSB1C3.
Source
BBa_I15010 (Photosensing Chimaera (Cph8) (not working, but amplified out of the whole construct))