This is half of a planned part which would contain all three PHA synthase genes required to produce [http://en.wikipedia.org/wiki/Polyhydroxybutyrate polyhydroxybutyrate] (PHB) in cells: ''phaA'', ''phaB1'', ''phaC1''. This half contains only ''phaA'': <partinfo>BBa_K156012</partinfo>. It was designed to be ligated upstream of part BBa_K338004.
When ligated upstream of BBa_K338004, the completed construct was designed to express all three PHA synthase genes required to make PHB oligomers from soybean oil. The three genes would be transcribed polycistronically on a single mRNA transcript under the IPTG-inducible control of the <partinfo>BBa_K215000</partinfo> promoter. Naturally, each gene is preceded by a standard RBS (<partinfo>B0034</partinfo>) and the transcript finishes with a strong terminator (<partinfo>B0015</partinfo>), for a total size of about 3500bp. Note that these three genes should only cause the production of PHB ''oligomers'' in cells, not hardened plastic. A crosslinking agent is required to link the oligomers and form the final plastic product. Over-expression of the ''phaC1'' gene could cause some crosslinking, but this has not been experimentally verified.
phaB BioPlastic polyhydroxybutyrate synthesis pathway (origin PCC6803 slr1993)
This composite part is made of a standard ribosomal binding site and a part that codes for an enzyme that catalyzes the formation of 3-hydroxybutyryl-CoA from acetoacetyl-CoA in polyhydroxyalkanoate synthesis, and a terminator.
A composite of RBS(b0034) and phaC(BBa_K342001).
phaC is a Bio brick registered by 10_ISNA Lyon.
Plastic Intermediate: phaC1 + Terminator
A construction intermediate used to produce <partinfo>BBa_K338004</partinfo>. Contains the third of three genes required to synthesize polyhydroxybutyrate (PHB).
Designed to be ligated downstream of <partinfo>K338022</partinfo> and <partinfo>K338023</partinfo>.
The Registry of Standard Biological Parts
Promoter + RBS + phaA + double terminator
This device is a synthetic gene that generates PhaA, an enzyme that catalyzes the synthesis of acetoacetyl-coenzyme A from two molecules of acetyl-coenzyme A. The enzyme can also act as a thiolase, catalyzing the reverse reaction and generating two-carbon units from the four-carbon product of fatty acid oxidation.
This part is a composite part made of a standard ribosomal binding site, a part that codes for an acetyl-CoA acetyltransferase (PhaA), and a terminator (B0015).
This part codes for an enzyme that catalyzes the formation of 3-hydroxybutyryl-CoA from acetoacetyl-CoA in polyhydroxyalkanoate synthesis.
This part was codon-optimized for expression in E. coli. The following restriction enzyme sites were also removed: EcoRI, NotI, XbaI, SpeI, PstI, NheI, AvrII, ApoI, MfeI, NsiI, SbfI.
This coding region is native to Ralstonia eutropha H16. The original sequence was obtained from GenBank (VERSION NC_008313.1, GI:113866031).
This composite part is made of a standard ribosomal binding site and a part that codes for an enzyme that catalyzes the formation of 3-hydroxybutyryl-CoA from acetoacetyl-CoA in polyhydroxyalkanoate synthesis.
This is a construct of an P3HB (bio plastic) producing operon. RBS(BBa_b0034) is assembled on each phaC, phaA, phaB gene.
phaA and phaB gene is cloned from pGEM phbCAB operon which carries Ralstronia Eutropha gene.
phaC1 (Poly(3-hydroxybutyrate) polymerase)
This part codes for a polyhydroxybutyrate (PHB) synthase, which catalyzes the polymerization of hydroxybutyryl-CoA.
phaE BioPlastic polyhydroxybutyrate synthesis pathway (origin PCC6803 slr1829)
E. coli codon optimized, Ile ATT -> ATC, avoided rare Ile ATA
A construction intermediate used to produce <partinfo>BBa_K338003</partinfo>. Contains the first of three genes required to synthesize polyhydroxybutyrate (PHB).
Designed to be ligated upstream of <partinfo>BBa_K338023</partinfo> and <partinfo>BBa_K338024</partinfo>.
phaA BioPlastic polyhydroxybutyrate synthesis pathway (origin PCC6803 slr1994)
This device (a synthetic gene) generates PhaA.
This part is a composite part made of a standard ribosomal binding site and a second part that codes for an acetyl-CoA acetyltransferase (PhaA).
This is a composite part made from existing BioBricks.
A construction intermediate used to produce <partinfo>K338004</partinfo>. Contains the second of three genes required to synthesize polyhydroxybutyrate (PHB).
This part was designed to be ligated between <partinfo>BBa_K3380022</partinfo> and <partinfo>BBa_K338024</partinfo>.
B0034 and phaA (beta-ketothiolase orgin from Ralstonia eutropha)
PhaA was cloned from pGEM phbCAB which carries the synthetic operon genes (phaA,phaB,phaC) of Ralstonia eutropha.
This is half of a planned part which would contain all three PHA synthase genes required to produce [http://en.wikipedia.org/wiki/Polyhydroxybutyrate polyhydroxybutyrate] (PHB) in cells: ''phaA'', ''phaB1'', ''phaC1''. This half contains ''phaB1'' and ''phaC1'': <partinfo>BBa_K156013</partinfo> & <partinfo>BBa_K156014</partinfo>. It was designed to be ligated downstream of part BBa_K338003.
When ligated downstream of BBa_K338003, the completed construct was designed to express all three PHA synthase genes required to make PHB oligomers from soybean oil. The three genes would be transcribed polycistronically on a single mRNA transcript under the IPTG-inducible control of the <partinfo>BBa_K215000</partinfo> promoter. Naturally, each gene is preceded by a standard RBS (<partinfo>B0034</partinfo>) and the transcript finishes with a strong terminator (<partinfo>B0015</partinfo>), for a total size of about 3500bp. Note that these three genes should only cause the production of PHB ''oligomers'' in cells, not hardened plastic. A crosslinking agent is required to link the oligomers and form the final plastic product. Over-expression of the ''phaC1'' gene could cause some crosslinking, but this has not been experimentally verified.
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.
This gene was cloned from pGEM phbCAB. The pGEM phbCAB carries the synthetic operon genes (phaA,phaB,phaC) of Ralstonia eutropha.
P3HB producing construct (without a promoter)
This is an construct for P3HB (bio plastic). It lacks the promoter for the users that wants to try other kinds of promoters.
phaA and phaB was cloned from pGEM phbCAB which carries the gene of Ralstonia Eutropha.
phaC BioPlastic polyhydroxybutyrate synthesis pathway (origin PCC6803 slr1830)
Promoter + RBS + phaB1 + double terminator
This device is a synthetic gene that generates PhaA, an enzyme that catalyzes the formation of 3-hydroxybutyryl-CoA from acetoacetyl-CoA in polyhydroxyalkanoate synthesis.
This part codes for an enzyme that catalyzes the synthesis of acetoacetyl-coenzyme A from two molecules of acetyl-coenzyme A. The enzyme can also act as a thiolase, catalyzing the reverse reaction and generating two-carbon units from the four-carbon product of fatty acid oxidation.
This coding region is native to ''Ralstonia eutropha'' H16. The original sequence was obtained from GenBank (VERSION NC_008313.1, GI:113866031).
RBS(B0034)and phaB (acetoacetyl-CoA reductase orgin from Ralstonia eutropha)
A composite of RBS(BBa_B0034) and phaB (acetoacetyl-CoA reductase orgin from Ralstonia eutropha) BBa_K759005.
phAB(BBa_K750005) was cloned from pGEM phbCAB which carries the synthetic operon genes (phaA,phaB,phaC) 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.
phaA (beta-ketothiolase orgin from 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.