This part is mainly composed of Cph8(BBa_K592018), ho1(BBa_I15008) and PcyA(BBa_I15009), which form the red light sensing system in E.coli. Cph8 is a synthetic red light sensor, consisting of Cph1, a natural red light sensor, and EnvZ, a histidine kinase. The most important part of the light sensor is PCB(phycocyanobilin), without which the red light sensor cannot work. However, PCB is not generated by E.coli naturally. So we need to introduce ho1 and PcyA into E.coli. These two enzymes can change heme into PCB. Here we combine the three main parts together to facilitate parts users.
We need to add a promoter and three RBSs.
RBS (Elowitz 1999) -- defines RBS efficiency
Varies from -6 to +1 region from original sequence to accomodate BioBricks suffix. 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 for this part: Brian Chow
Vinay S Mahajan, Voichita D. Marinescu, Brian Chow, Alexander D Wissner-Gross and Peter Carr IAP, 2003.
Derived from the TE terminator of T7 bacteriophage between Genes 1.3 and 1.4 <genbank>V01146</genbank>.
Transcription terminator for the E.coli RNA polymerase.
Suggested by Sri Kosuri and Drew Endy as a high efficiency terminator. The 5' end cutoff was placed immediately after the TAA stop codon and the 3' end cutoff was placed just prior to the RBS of Gene 1.4 (before AAGGAG). Use anywhere transcription should be stopped when the gene of interest is upstream of this terminator.
Transcriptional terminator consisting of a 64 bp stem-loop.
Overlap extension of synthetic oligonucleotides
One of two requisite genes required for the biosynthesis of phycocyanobilin from heme.
phycocyanobilin:ferredoxin oxidoreductase (PcyA) from synechocystis
Second of two required phycocyanobilin biosynthetic genes.
The cph8 was taken from Tabor's pJT122 plasmid.
This is the red light sensor Cph8 with RBS.
Since Cph8 was considerably longer than the RBS added, sequencing was necessary to confirm the construct.
This part is identical to BBa_I15010, the cph8 red light sensor. We were unable to transform BBa_I15010 from the the 2011 iGEM distribution kit. PCR attempts using VF2 and VR failed to amplify any DNA from the well sample. In the end, we chose to amplify cph8 from Jeffrey Tabor's pJT122 plasmid provided by Chris Voigt. The cph8 in pJT122 contained an internal PstI site, which was removed by point-mutagenesis. The resultant DNA sequence is identical to that of BBa_I15010.
The source DNA (pJT122) contained illegal restriction site (PstI) that had to be removed.
Conversion of the iGEM parts registry to SBOL2.1