phaA and phaB was cloned from pGEM phbCAB which carries the gene of Ralstonia Eutropha.
This is an construct for producing P3HB. A constitutive promoter(BBa_R0040) locates on the upstream of the whole construct and by adding 2 percent gulcose into the media, the bacteria would produce bioplastics.
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.
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
Pha C (poly-β-hydroxybutyrate polymerase)
Pha C is one of the three genes coding for the production of PHB. It code for the poly-β-hydroxybutyrate polymerase, which catalyse the last step of the synthese of poly-3-hydroxybutyrate.
phaA (beta-ketothiolase orgin from Ralstonia eutropha)
This gene was cloned from pGEM phbCAB. The pGEM phbCAB carries the synthetic operon genes (phaA,phaB,phaC) of Ralstonia eutropha.
PhaA codes ???Beta- ketothiolase (acetyl-CoA acetyltransferase)???which catalyzes the conversion of two acetyl-CoA molecules into acetoacetyl-CoA. It is an enzyme that catalyzes the first step of P3(HB) producing pathway.
In the pGEM phbCAB phaA, we found a mutation in the 00th base according to the data base of NCBI. However, it is confirmed that pGEM phbCAB transformed E.Coli produces P3HB.
phaB (acetoacetyl-CoA reductase orgin from Ralstonia eutropha)
PhaB codes ???acetoacetyl-CoA reductase??? which catalyzes the reduction of acetoacetyl-CoA into (R)-3-hydroxybutyryl-CoA (3HB-CoA). This is an enzyme that catalyzes the second step of P3(HB) producing pathway.
We cloned the gene by constructing primers.
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.
Conversion of the iGEM parts registry to SBOL2.1