SAM:salicylic acid carboxyl methyltransferase; converts salicylic acid to methyl salicylate (winter
ketoisocaproate decarboxylase (THI3); converts alpha-ketoisocaproate to 3-methylbutanal
This device will make isoamyl alcohol. It codes for an enzymatic pathway that takes in leucine and converts it to isoamyl alcohol.
Out of time constraints, we did not get rid of the EcoRI cut sites in THI3. So...don't use EcoRI when making assemblies with this part!
The coding regions (BAT2 and THI3) in this part came from S. cerevisae.
This is the wintergreen generating device hooked up to a constitutive promoter with a different RBS (B0032, different from 45100).
This part is an intermediate to the protein generator BBa_45300. This part is almost identical to BBa_J45299 except that is uses as its RBS B0032 instead of B0030.
SAM:jasmonic acid carboxyl methyltransferase; converts jasmonic acid to jasmine odor
This ORF encodes the enzyme responsible for catalyzing the following reaction: Jasmonic Acid--->Methyl Jasmonate It is initially found in ''Arabidopsis thaliana'' but has been transferred successfully to the E. coli genome by Seo, et al. The product, methyl jasmonate has a jasmine smell to it.
Km: 38.5 ??M; Kcat:25 S−1 for Jasmonic Acid--->Methyl Jasmonate with the aid of JAMT: Jasmonic acid carboxyl methyltransferase
If the part becomes physically available, it would probably be donated by Seo, et al.
Stationary phase dependent GFP generator
This part is a composite part consisting of <biobrick>J45992</biobrick/> (a stationary phase promoter) and <biobrick>E0840</biobrick/> (a GFP generator). Thus, this device creates green fluorescent protein in stationary phase.
This part was mainly designed to test whether J45250 would smell like banana only in stationary phase.
J45992 was PCRed out of the E. coli genome, while E0840 was constructed at the Registry of Standard Biological Parts.
Exponential phase transcriptional control device
This composite regulator device produces a transcriptional signal in early growth phase but not in stationary phase.
It is a composite part of <partinfo>BBa_J45992</partinfo> and <partinfo>BBa_Q04401</partinfo>.
Exponential phase wintergreen odor biosynthetic system
SAM:benzoic acid carboxyl methyltransferase; converts benzoic acid to methyl benzoate (floral odor)
BAMT catalyzes the transfer of a methyl group from SAM (S-adenosyl methyltransferase) to benzoic acid, creating methyl benzoate. Methyl benzoate produces a "pleasant" smell.
Genbank accession number(s): AF198492 This part is from A. majus.
Stationary-phase-dependent banana odor generator
This part is an ATF1-1148 protein generator under control by the osmY stationary phase promoter. It is almost identical to J45270, except that: 1) The promoter is the truncated version of osmY and 2) it uses the B0030 RBS instead of B0032 (which is in J45270).
SAM:benzoic acid/salicylic acid carboxyl methyltransferase I; converts salicylic acid to methyl sali
Codes for BSMT enzyme to convert Benzoic acid/salicylic acid to methyl benzoate and methyl salicylate. This part is phBSMT1 from Petunia x hybrida.
Species: Petunia x hybrida from Genbank; (also available: A. thaliana and N. suaveolens)
This system designed by the MIT 2006 iGEM team generates mint smell by biosynthesizing salicylic acid from chassis-produced metabolites (chorismate) and then converts salicylic acid into methyl salicylate (wintergreen scent). It uses a stationary phase promoter followed by an inverter (see <bbpart>BBa_J45996<bbpart> for details) to only express the necessary enzymes during exponential phase.
We recommend use of strain <bbpart>BBa_J45999</bbpart> as it does not produce indole. Indole is responsible for the foul smell of wt e.coli, and masks the wintergreen scent produced by J45800.
This part is composed of <bbpart>BBa_J45320</bbpart> and <bbpart>BBa_J45180</bbpart>
Wintergreen odor enzyme (BMST1) generator
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.
Derived from the early part J45098 and sister to J45099.
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 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 Conserved Forward Primer: -99.97 kcal/mol
This part was PCRed out of E. coli's genome.
E. coli strain YYC912; Odor-free chassis
Escherichia coli strain YYC912. Biosafety level 1 organism.
This strain was obtained from the E.coli Genetic Resources: CGSC, The Coli Genetic Stock Center http://cgsc.biology.yale.edu/. Strain YYC912.
Exponential phase dependent GFP generator
This composite part consists of <bbrick>J45992</bbrick> (a stationary phase promoter), <bbrick>Q04401</bbrick> (an inverter), and E0840 (a green fluorescent protein generator). This device expresses GFP in the exponential phase of cell growth.
This device was constructed to test whether the J45180 device would work.
J45992 was PCRed out of the E. coli genome, while Q04401 and E0840 were developed by others in the Registry.
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.
Protein generator converting PoPS to the protein ATF1. ATFI encodes the enzyme alcohol acetyltransferase I (AATase I), which produces acetate esters from acetyl CoA and alcohols.
This part is the ATF sequence BBa_45J006 with a single point mutation at base pair 1148 (T -> C). Despite the mutation, the enzyme is still functional (kinetic studies have not been done to determine realitive functionality).
This part was cloned from S. cerevisiae.
alcohol acetyltransferase I; converts isoamyl alcohol to isoamyl acetate (banana odor)
This part is the ATF sequence (BBa_45J006) with a single point mutation at base pair 1176 (T -> C).
It has a single point mutation at base pair 1176 (T -> C).
SAM:salicylic acid carboxyl methyltransferase; converts salicylic acid to methyl salicylate (winterg
we need an MTA or to sequence the coding region for this enzyme
Protein generator converting PoPS to the prot BSMT. The BSMT enzyme converts benzoic acid/salicylic acid to methyl benzoate and methyl salicylate.
This part is phBSMT1 from Petunia x hybrida (GenBank). See part design and discussion for more information.
This part is an intermediate to the protein generator BBa_45400.
This part has EcoRI and SpeI sites in it. Use caution when conducting standard assembly with the part.
This part was made via standard assembly of J45200 and J45400.
branched-chain amino acid transaminase (BAT2); converts leucine to alpha-ketoisocaproate
This part is an intermediate to the protein generator BBa_45200.
Stationary phase banana odor biosynthetic system
This system designed by the MIT 2006 iGEM team generates mint smell by biosynthesizing salicylic acid from chassis-produced metabolites (chorismate) and then converts salicylic acid into methyl salicylate (wintergreen scent).
We recommend use of strain <bbpart>BBa_J45999</bbpart> as it does not produce indole. Indole is responsible for the foul smell of wt e.coli, and masks the wintergreen scent produced by J45600.
This system is composed of devices <bbpart>J45120</bbpart> and <bbpart>J45320</bbpart>.
Full-length stationary phase osmY promoter
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
isochorismate pyruvate-lyase and isochorismate synthase (pchBA); converts chorismate to salicylate
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.
pchBA is found in pseudomonas. The plasmid was kindly sent to the MIT iGEM team by Dr. Cornelia Riemmann from the Universite de Lausanne.
This system designed by the MIT 2006 iGEM team and generates mint smell by biosynthesizing salicylic acid from chassis-produced metabolites (chorismate) and then converts salicylic acid into methyl salicylate (wintergreen scent).