Simple inducible (LacI) ethylene generator
See part documentation for enzyme coding sequences BBa_K417001 and BBa_K417002 for details on the origin of the sequences.
A simpler version of BBa_K417006 which expresses ACC synthase and ACC oxidase (EFE) to produce ethylene. This version does not express the SAM synthetase enzyme (BBa_K417000) and relies on S-adenosyl methionine generated by the chassis.
Native LacI promoter (BBa_R0010) was chosen since it facilitates easier assembly compared with the shorter alternative BBa_R0011, although it would be worth testing both these promoters. It is unknown if the order of the coding sequences is important in this device.
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.
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.
Transcriptional terminator consisting of a 64 bp stem-loop.
1-aminocyclopropane-1-carboxylate (ACC) synthase
Malus domestica (Apple), codon optimized for E. coli, with the C-terminal amino acids -YYNVPEVNGGSQSSHLSHSRRQSLTKWVSRLSFDDRGPIPGR* removed
This is the protein coding sequence of 1-aminocyclopropane-1-carboxylate synthase, which catalyses the formation of 1-aminocyclopropane-1-carboxylate and methylthioadenosine from S-adenosyl methionine (AdoMet). (S-adenosyl-L-methionine <=> 1-aminocyclopropane-1-carboxylate + methylthioadenosine). This enzyme is commonly referred to as ACC synthase.
Removal of the C-terminal amino acids. According to White, M. F., Vasquez, J., Yang, S. F., & Kirsch, J. F. (1994). PNAS, 91(26), 12428-32. (http://www.ncbi.nlm.nih.gov/pubmed/7809054) - this truncation has been reported to have no effect on specific activity.
1-aminocyclopropane-1-carboxylate (ACC) oxidase
Solanum lycopersicum (Tomato) (Lycopersicon esculentum) codon optimized for E. coli.
This is the protein coding sequence of 1-aminocyclopropane-1-carboxylate (ACC) oxidase, which catalyses the formation of ethylene from 1-aminocyclopropane-1-carboxylate under the presence of ascorbate and O2. (1-aminocyclopropane-1-carboxylate + ascorbate + O2 = ethylene + cyanide + dehydroascorbate + CO2 + 2 H2O) This enzyme is commonly referred to as ACC oxidase or ethylene forming enzyme (EFE).
Codon optimization for E. coli and removal of biobrick restriction sites.
Conversion of the iGEM parts registry to SBOL2.1