FixK2 Promoter-RBS-mRFP1-Hsp60 Promoter-RBS-YF1-RBS-FixJ
System for blue light-induced expression of mRFP1 in P. acnes using constitutive Hsp60 promoter from M. bovis (expected to work in P. acnes) to drive expression of blue light-regulated transcription factors YF1 and FixJ.
mRFP with 6x His tag (BBa_K1491008) and a strong RBS (BBa_B0034). It can be used as a reporter.
Constitutive promoter from m. Enternamehere expected to function in p. Acnes.
FixK2 promoter omes from the genome of Bradyrhizobium japonicum.
FixK2 promoter is the wild-type promoter to which phospho-FixJ binds. FixJ in turn can be regulated by YF1, the blue light-sensing protein. This promoter shows very little leaky activity in the absence of FixJ.
The sequence information was provided by Moffat (Moffat et al. 2009). The DNA was synthesized by GenScript.
B0034-YF1-B0034-FixJ, blue light sensor and RR with RBS
The components that made up of YF1 fusion protein comes from Bradyrhizobium japonicum and Bacillus subtilis. FixJ comes from the genome of Bradyrhizobium japonicum.
YF1 works in conjunction with its response regulator, FixJ. This part consists of RBS-coupled YF1 and FixJ without promoter.
This constructs Strong RBS to ensure good expression.
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.
mRFP (BBa_E1010) with 6x His tag for protein purification
Ytva comes from the genome of Bacillus subtilis. FixL derives from the genome of Bradyrhizobium japonicum.
YF1 is a fusion protein of YtvA (B.subtilis) and FixL(Br.japonicum). It contains the N-terminal region of YtvA (1-127) and C-terminal region of FixL (258-505), interconnected by a helical linker. This fusion protein is designed by Andreas Moglich and Keith Moffat. As shown in previous research, Illumination of the fusion kinase YF1 reduced net kinase activity by ∼1000-fold in vitro. YF1 also controls gene expression in a light-dependent manner in vivo.
The sequence information was provided in Moglich and Moffat's article. The physical DNA was synthesized by GenScript. Codon-optimization may enhance its expression.
FixJ comes from the genome of Bradyrhizobium japonicum.
FixJ comes from the genome of Bradyrhizobium japonicum. It's the wild-type response regulator to YF1, the blue light-sensing fusion protein.
The sequence was information was provided by Moffat (Moffat et al. 2009). The DNA was synthesized by GenScript.
Conversion of the iGEM parts registry to SBOL2.1