Methyl Salicylate Production under TetR regulation
KU_Leuven 2013 requested the BBa_J45700 brick from MIT 2006 and recloned into the pSB1C3 standard plasmid backbone (BBa_K1060003). In this part we removed the lac promoter in front of the pchBA gene to get a constitutive expression of all genes needed for methyl salicylate production. We did this by amplifying the For the assembly of this part we refer to the original MIT brick BBa_J45700 => "We assembled a complete wintergreen odor biosynthetic system (BBa_J45700) by combining the salicylic acid (BBa_J45320) and wintergreen odor generators (BBa_J45120). BBa_J45700 catalyzes the conversion of the cellular metabolite chorismate to methyl salicylate or wintergreen odor."
no EcoRI, XbaI, NotI, SpeI or PstI sites present
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.
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
RBS.1 (strong) -- modified from R. Weiss
Strong RBS based on Ron Weiss thesis. Strength is considered relative to <bb_part>BBa_B0031</bb_part>, <bb_part>BBa_B0032</bb_part>, <bb_part>BBa_B0033</bb_part>.
Varies from -6 to +1 region from original sequence to accomodate BioBricks suffix ("orig" in figure 4-14 of Ron Weiss thesis). 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 Brian Chow
Double terminator consisting of BBa_B0010 and BBa_B0012
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.
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.
This is a copy of the Part:B0014. Only here the part is cloned in the standard vector pSB1C3 instead of the pSB1AK3.
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.
Transcriptional terminator consisting of a 64 bp stem-loop.
Double terminator consisting of BBa_B0012 and BBa_B0011
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.
Conversion of the iGEM parts registry to SBOL2.1