HpaC- 4-hydroxyphenylacetate 3-monooxygenase reductase subunit
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.
DBT-sulfone monooxygenase (DszA) from R. erythropolis
Obtained from PCR amplification from a plasmid found in an environmental isolate of R. erythropolis shown to be an active desulfurizer.
A monooxygenase enzyme involved in the 4S desulfurization pathway. This protein encoded by this sequence carries out the third step in the pathway, converting DBT-sulfone to 2-hydroxybiphenyl-2-sulfinic acid through addition of an oxygen to the sulfur heteroatom. FMNH2 is consumed by this reaction.
4 internal PstI cut-sites had to be mutated with site-directed mutagenesis.
http://www.ncbi.nlm.nih.gov/nuccore/L37363.1 (GenBank: L37363.1)
This is a DszC coding gene from the DszABC operon in the 4S pathway of Rhodococcus erythropolis IGTS8. It encodes one of the three biodesulfurizing enzymes in the DszABC operon. DszC enzymes have been shown to catalyze dibenzothiophene (DBT)to dibenzothiophene-5-oxide (DBTO) in the first reaction and then from DBTO to DBT sulfoxide (DBTO2) in the second reaction, both in the presence of NADH, FMN and flavin reductases.
2 PstI sites were removed using site-directed mutagenesis
DszB is from a plasmid from an environmental isolate of Rhodococcus shown to be an active desulfurizer, promotor from registry.
Inducible expression (using LacI promotor J04500) of DszB
HBPS desulfinase (DszB) from R. erythropolis (mutated for improved activity)
Amplified from a plasmid in an environmental isolate of Rhodococcus shown to be an active desulfurizer.
DszB - HBPS desulfinase which carries out the last step of the 4S pathway, converting 2-hydroxybiphenyl-2-sulfinic acid (HBPS) to 2-hydroxybiphenyl. The sequence has been mutated Y63F to increase enzyme activity.
Mutations of internal PstI and NotI cut sites in order to make biobrick compatible, as well as an added mutation of Y63F in the protein to increase its activity.
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.
Conversion of the iGEM parts registry to SBOL2.1