This biobrick is composed of BBa_K887040, BBa_B0034, BBa_K887020,BBa_B003 and BBa_K887023.
This part contains three zinc fingers and following enzymes. These three enzymes will change pyruvate to 2-ketoisoalerate(the Intermediate of isobutanol). With additional zinc finger, these fusion protein can bind to the DNA program according to the order.These strategy can enhance productivity theoretically.
Nothing special, the only problem is this part is a little big.
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.
plac is a promoter regulated by lacI repressor.B0034 is and strong RBS which can be bond by ribosome XD to initiate translation.
It's a combination of the zinc finger (PBSII) and the enzyme (ilvC) that can make the isobutanol pathway more efficient.
This is a part with HivC, which is a zinc finger binding domain, and combined with ilvD, which is the third enzyme in the isobutanol synthesis pathway. By this way, we can rearrange this fusion protein in order on the DNA program, which include 5'GAT GCT GCA'3 sequence, and we treat others enzyme in isobutanol synthesis pathway by the same method, we can make this reaction be more efficient. As a result, we can get higher yield easily.
The Plac insert was PCR'd from the MG1655 strain of E.coli K12.
Inverting regulatory region controlled by LacI (<bb_part>BBa_C0010</bb_part>, <bb_part>BBa_C0011</bb_part>, etc.) The pLac regulatory region is a 243 base-pair sequence with standard BioBrick prefix and suffix sections on its ends. It contains two protein binding sites: CAP, which is generally present in E.coli and is assocciated with cell health and availability of glucose., and LacI, the Lac inhibitor <bb_part>BBa_C0010</bb_part> which binds in an dimerized cooperative manner to inhibit the transcription of the protein that follows. In the presence of lactose or IPTG, an analog of lactose, LacI is unable to correctly bind and inhibit transcription. This allows <bb_part>BBa_R0010</bb_part> to be used as a inverter or as a detector of lactose or IPTG.
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 this regulator. This part is incompatible with environments containing lactose or lactose analogs.
ilvC (ketol-acid reductoisomerase NAD(P)-binding)
It comes from Escherichia coli str. K-12 substr. MG1655
This is the coding region of ilvC from Escherichia coli str. K-12 substr(MG1655). It encodes ketol-acid reductoisomerase NAD(P)-binding which is part of the isobutanol biosynthesis pathway.
Because of the four RE site, EcoR1, Xba1, Spe1 and Pst1, before and after the coding gene, we have to delete those RE site in kivd. The RE site in the part has been modified by point mutation.
This is the coding sequence of ilvD form Escherichia coli str. K-12 substr. MG1655. It encodes dihydroxyacid dehydratase which is part of isobutanol biosynthesis pathway.
Because of the four RE site, EcoR1, Xba1, Spe1 and Pst1, before and after the coding sequence, those RE site should not exist in coding sequence. There is one Pst1 in original sequence of ilvD. However, the RE site in ilvD has been modified by point mutation.
AlsS (alpha-acetolactate synthase protein)
This is the coding region of AlsS from Bacillus subtilis. It encodes alpha-acetolactate synthase protein which is part of isobutanol biosynthesis pathway.
Because of the four RE site, EcoR1, Xba1, Spe1 and Pst1, before and after the coding sequence, those RE site should not exist in coding sequence. There are two Pst1 site, one Xba1 site and one EcoR1 site in original sequence of AlsS. However, those RE site in AlsS has been modified by point mutation.
Conversion of the iGEM parts registry to SBOL2.1