PoPS in -> GFP out Strong RBS (efficiency=0.6).
OmpR Promotor will be activated upon illumination with red light
Positively regulated, OmpR-controlled promoter. This promoter is taken from the upstream region of ompC. Phosphorylated OmpR binds to the three operator sites and activates transcription.
The promoter includes the three OmpR binding sites: C1, C2, and C3. The promoter starts at -107 and goes up to the transcriptional start site at +1. An alternate version, BBa_R0083, cuts out the C2 and C3 sites. The efficacy of this promoter versus the truncated ompC upstream region (BBa_R0083) or the ompF upstream region (BBa_R0084) is currently unknown.
Stephen Lee, Roshan Kumar, Joe Levine, Ziyan Chu (Polkadorks, IAP 2004)
RBS.1 (strong) -- modified from R. Weiss
Strong RBS based on Ron Weiss thesis. Strength is considered relative to <bb_part>BBa_B0031</bb_part>, <bb_part>BBa_B0032</bb_part>, <bb_part>BBa_B0033</bb_part>.
Varies from -6 to +1 region from original sequence to accomodate BioBricks suffix ("orig" in figure 4-14 of Ron Weiss thesis). No secondary structures are formed in the given RBS region. Users should check for secondary structures induced in the RBS by upstream and downstream elements in the +50 to -50 region, as such structures will greatly affect the strength of the RBS. Contact info Brian Chow
Vinay S Mahajan, Voichita D. Marinescu, Brian Chow, Alexander D Wissner-Gross and Peter Carr IAP, 2003.
green fluorescent protein derived from jellyfish Aequeora victoria wild-type GFP (SwissProt: P42212
GFP (mut3b) [note that this part does not have a barcode]
modified E. coli thr terminator, replaced all A-T pairs in stem with C-G pairs
Artificial terminator, estimated %T~>90% *8bp stem, 6nt loop *Bidirectional, estimated reverse %T~>90%
Bidirectional, with the reverse estimated to be less effective than the forward. Has a polyA tail of 9 residues.
Phycobilin synthesis operon without terminator
This is derived from K098010 but PCR was used to remove the terminator, so that cph8 could be added at the end of the operon.
Phycobilin is chromophores (light-capturing molecules) found in cyanobacteria and in the chloroplasts of red algae, glaucophytes and some cryptomonads (though not in green algae and higher plants)， used in light sensor.
The actual forward sequence has the same changes at the start as seen in clone H11 described above. The reverse sequence is as expected. Features same as for the one above apart from the terminator.
Chris French and Richard Edward Partridge-Hicks
Red light sensor Cph8 (from Coliroid) with RBS
BBa_I15010 (Photosensing Chimaera (Cph8) (not working, but amplified out of the whole construct))
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: 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.
Conversion of the iGEM parts registry to SBOL2.1