Exponential phase wintergreen odor biosynthetic system
Exponential phase dependent wintergreen odor generator
This part converts salicylic acid into methyl salicylate (a mint-smelling compound) in the exponential phase of cell growth. The part consists of a stationary phase promoter (J45992), connected to an inverter (Q04401) as well as the methyl salicylate protein generator (J45119). This part replaces the defective part J45180, which used the truncated version of the stationary phase promoter (J45993). This part did not function as it was designed due to an imbalance between the strength of the stationary phase promoter and the strength of the promoter in the inverter (R0040).
We needed the full osmY promoter strength to be relatively comparable to the R0040 promoter strength.
This part is phBSMT1 from Petunia x hybrida (GenBank). See part design and discussion for more information.
Protein generator converting PoPS to the prot BSMT. The BSMT enzyme converts benzoic acid/salicylic acid to methyl benzoate and methyl salicylate.
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.
RBS.3 (medium) -- derivative of BBa_0030
Weak1 RBS based on Ron Weiss thesis. Strength is considered relative to <bb_part>BBa_B0030</bb_part>, <bb_part>BBa_B0031</bb_part>, <bb_part>BBa_B0033</bb_part>.
Varies from -6 to +1 region from original sequence to accomodate BioBricks suffix ("RBS-2" in figure 4-14 of thesis). Contact info for this part: Brian Chow
Vinay S Mahajan, Voichita D. Marinescu, Brian Chow, Alexander D Wissner-Gross and Peter Carr IAP, 2003.
Transcriptional terminator consisting of a 64 bp stem-loop.
Promoter (lacI regulated, lambda pL hybrid)
represillator of Elowitz and Leibler (2000)
Inverting regulatory region controlled by LacI (<bb_part>BBa_C0010</bb_part>, <bb_part>BBa_C0011</bb_part>, etc.) The PLlac 0-1 promoter is a hybrid regulatory region consisting of the promoter P(L) of phage lambda with the cI binding sites replaced with lacO1. The hybrid design allows for strong promotion that can nevertheless be tightly repressed by LacI, the Lac inhibitor (i.e. repressor) (<bb_part>BBa_C0010</bb_part>) ([LUTZ97]). The activity of the promoter can be regulated over a >600-fold range by IPTG in E.Coli DH5-alpha-Z1 (same paper reference).
hybrid promoter design to create strong promoter that is, at the same time, highly repressible. note that the upstream operator installed in this hybrid is slightly different than the one in the original source (Lutz and Bujard, 1997). the most upstream operator region is slightly truncated in the represillator version, so that both operators in the hybrid are the same sequence. see references for details. also, the sequence has been truncated after the transcriptional start site. 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 increase transcription. This part is incompatible with environments containing lactose or lactose analogs.
Neelaksh Varshney, Grace Kenney, Daniel Shen, Samantha Sutton
Exponential phase transcriptional control device
It is a composite part of <partinfo>BBa_J45992</partinfo> and <partinfo>BBa_Q04401</partinfo>.
This composite regulator device produces a transcriptional signal in early growth phase but not in stationary phase.
isochorismate pyruvate-lyase and isochorismate synthase (pchBA); converts chorismate to salicylate
pchBA is found in pseudomonas. The plasmid was kindly sent to the MIT iGEM team by Dr. Cornelia Riemmann from the Universite de Lausanne.
pchBA codes for proteins used to generate salicylic acid. It doesn't make a fusion protein, even though the coding regions overlap; the RBS for pchA is located at the end of the coding region of pchB.
Wintergreen odor enzyme (BMST1) generator
Derived from the early part J45098 and sister to J45099.
This composite part include a ribosome binding site (B0032 different from J45099), the BSMT coding region, and a terminator. This part is attached to different promoters.
Full-length stationary phase osmY promoter
This part was PCRed out of E. coli's genome.
osmY's promoter is active in stationary phase and under high osmotic pressure conditions.
Truncated Forward Primer: 5'- GTT TCT TCG AAT TCG CGG CCG CTT CTA GGCT TAT GTT TTC GCT GAT ATC - 3' Total Length: 49 bp Annealing Length: 21 bp GC Content: 38.1% Melting Temperature: 49.5 degrees C hairpin deltaG: -2.66 kcal/mol self dimer deltaG: -99.97 kcal/mol Full Forward Primer: 5'- GTT TCT TCG AAT TCG CGG CCG CTT CTA GCTG GCA CAG GAA CGT TAT C - 3' Total Length: 47 bp Annealing Length: 19 bp GC Content: 52.6% Melting Temperature: 53.4 degrees C hairpin deltaG: -3.99 kcal/mol self dimer deltaG: -98.3 kcal/mol Reverse Primer: 5'-GTT TCT TCC TGC AGC GGC CGC TAC TAG TAT TGT TAA ATA TAG ATC ACA ATT TTG- 3' Total Length: 54 bp Annealing Length: 25 bp GC Content: 20.0% Melting Temperature: 46.4 degrees C hairpin deltaG: -2.86 kcal/mol self dimer deltaG: -99.44 kcal/mol heterodimer deltaG with Forward Primer: -99.44 kcal/mol heterodimer deltaG with Conserved Forward Primer: -99.97 kcal/mol
Currently entered as a placeholder. This is a modified version of the tetR inverter Q04400 with a single base switch in the RBS: AAAGAGGAGAAA to AAAGAGGGGAAA
SAM:benzoic acid/salicylic acid carboxyl methyltransferase I; converts salicylic acid to methyl sali
Species: Petunia x hybrida from Genbank; (also available: A. thaliana and N. suaveolens)
Codes for BSMT enzyme to convert Benzoic acid/salicylic acid to methyl benzoate and methyl salicylate. This part is phBSMT1 from Petunia x hybrida.
Lutz, R., Bujard, H., Nucleic Acids Research (1997) 25, 1203-1210.
Sequence for pTet inverting regulator driven by the TetR protein.
BBa_R0040 TetR-Regulated Promoter is based on a cI promoter. It has been modified to include two TetR binding sites and the BioBrick standard assembly head and tail restriction sites.
June Rhee, Connie Tao, Ty Thomson, Louis Waldman
Double terminator consisting of BBa_B0010 and BBa_B0012
tetracycline repressor from transposon Tn10 (+LVA)
Elowitz, M. B. Transport, Assembly, and Dynamics in Systems of Interacting Proteins. Thesis, Princeton Univ., Princeton (1999)
Coding region for the TetR protein without the Ribosome Binding Site. Modified with an LVA tail for rapid degradation of the protein and faster fall time for the emission. TetR binds to the pTet regulator (BBa_R0040). aTc (anhydrotetracycline) binds to TetR and inhibits its operation.
References (unparsed) here: Elowitz, M. B. Transport, Assembly, and Dynamics in Systems of Interacting Proteins. Thesis, Princeton Univ., Princeton (1999). Lutz R, Bujard H., Independent and tight regulation of transcriptional units in Escherichia coli via the LacR/O, the TetR/O and AraC/I1-I2 regulatory elements. Nucleic Acids Res. 1997 Mar 15;25(6):1203-10. PMID: 9092630 References (unparsed) here: Elowitz, M. B. Transport, Assembly, and Dynamics in Systems of Interacting Proteins. Thesis, Princeton Univ., Princeton (1999). Lutz R, Bujard H., Independent and tight regulation of transcriptional units in Escherichia coli via the LacR/O, the TetR/O and AraC/I1-I2 regulatory elements. Nucleic Acids Res. 1997 Mar 15;25(6):1203-10. PMID: 9092630 BBa_C0040 TetR Protein is based on the TetR sequence from Elowitz's repressilator. It has been modified to include a rapid degradation LVA tail, and includes the BioBrick standard assembly head and tail restriction sites. The RBS has been removed. The stop codon has been changed from TAA to a double stop codon TAATAA.
June Rhee, Connie Tao, Ty Thomson, Louis Waldman.
This is a single bp change from B0034. It's strength is approximately 35% that of B0034.
Conversion of the iGEM parts registry to SBOL2.1