Please refer to http://partsregistry.org/Part:BBa_K559000:Design, both of the genes are gathered from the Registry distribution.
Halorhodopsin is an inward-directed light-driven chloride ion pump originating from Halobacterium. It utilizes light to pump chloride ions against chloride concentration difference into cells from the environment. We added a constitutive working and regulatable pLac promoter in the upstream region of the part in order to use it in our design easily.
Please refer to http://partsregistry.org/Part:BBa_K559000:Design for more detail about halorhodopsin. Please refer to http://partsregistry.org/Part:BBa_J04500:Design for more detail about pLac promoter.
Halorhodopsin comes from part BBa_K559000, which is come from genomic sequence of N.pharaonis (genebank ID:J05199.1)
Halorhodopsin gene allow chloride ion to be pumped in by the level of light intensity. Halorhodopsin gene is under regulation by constitute T7 promoter to allow protein coding to be maximized, and the bidirectional terminator is added for high efficiency termination of both forward and backward transcription
The double terminator wass tested to stop effectively on the transcription of both forward and backward transcription to make our protein generator be a controlled system
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.
The Plac insert was PCR'd from the MG1655 strain of E.coli K12.
Inverting regulatory region controlled by LacI (<bb_part>BBa_C0010</bb_part>, <bb_part>BBa_C0011</bb_part>, etc.) The pLac regulatory region is a 243 base-pair sequence with standard BioBrick prefix and suffix sections on its ends. It contains two protein binding sites: CAP, which is generally present in E.coli and is assocciated with cell health and availability of glucose., and LacI, the Lac inhibitor <bb_part>BBa_C0010</bb_part> which binds in an dimerized cooperative manner to inhibit the transcription of the protein that follows. In the presence of lactose or IPTG, an analog of lactose, LacI is unable to correctly bind and inhibit transcription. This allows <bb_part>BBa_R0010</bb_part> to be used as a inverter or as a detector of lactose or IPTG.
LacI binds to this regulator. This part is incompatible with species containing active LacI coding regions. Lactose and IPTG disable the operation of LacI and this regulator. This part is incompatible with environments containing lactose or lactose analogs.
Double terminator consisting of BBa_B0012 and BBa_B0011
Halorhodopsin come from N.pharaonis, gene bank J05199.1
Halorhodopsin under constitute T7 promoter to regulate the gene transcription to be maximized and no restrict to other regulatory factor
The coding sequence of N.pharaonis J05199.1 is selected from 61nt to 700nt and the T7 promotor placed in front by 3A assembly
Derived from luxICDABEG operon terminator of Vibrio fischeri <genbank>AF170104</genbank>.
Bidirectional transcriptional terminator consisting of a 22 bp stem-loop.
In the naturally-occuring sequence there is a mismatch in the stem of the stem loop. This can be corrected via an A->G mutation (at position 40 -- sequence coordinate/not MFOLD coordinate). The above sequence does not reflect this mutation (but the MFOLD image does). This terminator's location cannot be found using some inverted repeat detectors like PALINDROME because it is too short and contains a mismatch. This one was found with the help of Tom Knight. It lies between two coding regions that point towards eachother.
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.
Conversion of the iGEM parts registry to SBOL2.1