This section of DNA contains the sequence from 300 base pairs upstream of the Nitrilase/cyanide hydratase and apolipoprotein N-acyltransferase gene to 30 base pairs downstream into the gene from Pseudomonas fluorescens PfO-1. Our hope is that this region of DNA will contain the promoter for the nitrilase gene. At the time that this part was submitted, it was not tested or sequenced yet.
We isolated this segment of DNA from the Pseudomonas fluorescens PfO-1 genome using a PCR reaction.
This section of DNA comes from the Pseudomonas fluorescens PfO-1 genome (NC_007492). The GeneID for the nitrilase gene that we working with is 3715134, and the protein ID for the nitrilase gene is YP_349417.1. We took the region 300 base pairs upstream of the nitrilase gene and 10 base pairs downstream into the gene.
This is the promoter activated by the transcriptional regulator DntR [BB# here] in response to detection of environmental [salicylate].
'''''This does not physically exist as a basic part due to time constraints; it was cloned as part of composite part which incorporates all of the components necessary for expression of a reporter gene under the control of DntR'''''. For this reason, the ends of the supplied sequence may not accurately represent the boundaries of the functional unit.
Cloned from a historical plasmid based on pQF52 carrying genomic sequences from ''Burkholderia cepacia''.
Cell free Chassis: Mass production of aqueous vesicles in oil phase
Method to mass produce vesicles of uniform shape and volume Vesicle characterization Content of the vesicles can be chosen freely. Environment inside vesicles is aquious. Vesicle shape: spherical Vesicle diameter: 15 ??m Vesicle volume: ~12 pl Vesicle population has a very narrow distribution in size. Production efficiency: 5-15 vesicles per second Vesicle stability: not fully characterized, depends on surfactant used, at least 2 hours; fusion of vesicles observed after >24 hours Optical properties: transparent in brightfield microscopy; for observation of fluorescence the surfactant used should be non-fluorescent Vesicle isolation through output hole. Vesicle content is released when vesicles transfered to aqueous Protocol You will need a silicone waver with the engraved microfluidic system, and a micropump system Setup of the microfluidic system The microfluidic system consists of a flow chamber made of Polydimethylsiloxane (PDMS) and a pump system that controls the flow rates of the various liquids into the chamber. Droplets are created within a defined space in the chamber and are propagated along a grid that allows containment and imaging. Two types of chambers have been used, differing in the geometry of the space where droplets were produced. One featured T-junction, and the other a V-junction. Production of flow chambers: * mix PDMS and curing agent in 10:1 ratio * degas and pour on wafer with etched microstructures * polymerize on heat plate at 150 ??C for 30 min * add unpolymerized PDMS mixture to points on microstructure where microtube inlets are to be pierced * polymerize on heat plate at 150 ??C for 30 minutes * remove polymerized PDMS from wafer, cut to fit onto glass cover slide (24 x 60 mm), and use clean needles (0.8 mm) or laser cutter (Trotec Speedy 100TM) to pierce tube inlets * ionize PDMS and glass slide in plasma chamber for 30 sec to make it reactive * align PDMS on glass slide and seal * seal irreversibly by heating on plate at 60??C for 6 hours The pumping system (ceDOSYS SP-4) allows control of syringes filled with aqueous material and mineral oil treated with surfactant(1% Span 80), respectively. The syringes access the chamber via the tubing inlets. Two inlets are used to pump in material in the aqueous phase; the remaining one is used for the oil phase. The flow rates of the syringes are controlled via a ceDOSYS user interface software. Control via pump system: * two syringes are loaded with 1ml each of material in the aqueous phase; during the first trial, distilled water * another is filled with a 1ml solution of 0.5% span 80 in oil * use flow rate on ceDOSYS interface to flood the chamber first with oil phase * gradually introduce aqueous phase and modify rates of both phases until the shear stress breaks the aqueous phase into droplets at the T- or V- junctions in the respective chambers.
does not apply. part contains no sequence
magnetite tight-binding transmembrane protein interfacing magnetic field and mechanobiology.
PCR product from Magnetospirillum magneticum AMB-1
Stimulated by CafA found in cafein addicted students
Also activated by sucrose Inhibited by nicotine
Encodes for oxidoreductase and promotes the reduction/oxidation of iron (FeII and FeII).
Digested using Pst I, Xba I inserted into BBa_E0422.
Ore is a consensus sequence localized in some promoters that respond to oleic acid concentration. Its sequence is CGGN3TNA/(R)N8-12CCG
We amplificated this part from p2UG2XORE plasmid
This part is in BBb Format. It is flanked by BamHI and BglII sites instead of XbaI and SpeI. More information about the BBb Format is available at: [http://openwetware.org/wiki/Template:AndersonLab:BBb_Standard BBb Standard Description Page] <!-- Add more about the biology of this part here
This is the naturally-occurring (possibly stationary-phase only) promoter for the XylR gene (which encodes the transcriptional regulator XylR).
Ore is a consensus sequence localized in some promoters that respond to oleic acid concentration. Its sequence is CGGN3TNA/(R)N8-12CCG. In this case we have 2 ore consensus sequences.
Downstream to ORE sequence there is CYC TATA
We amplified this part from p2UG2XORE plasmid
Iron sensitive promoter (test delete later)
This is a promoter that is sensitive to iron (Fe++) concentration (test only)
made a base substitution to remove a EcoRI site
This is the naturally-occurring (possibly stationary-phase only) promoter for the DntR gene (which encodes the transcriptional regulator DntR) back-to-back with the DntR-regulated promoter.
This does not physically exist as a basic part due to time constraints; it was cloned as part of composite part which incorporates all of the components necessary for expression of a reporter gene under the control of DntR. For this reason, the ends of the supplied sequence may not accurately represent the boundaries of the functional unit.
nitrogenase iron protein, vnfH Rhodopseudomonas palustris CGA009
only the plasmid contain rep gene can be used in magnetic bacteria AMB-1
This promoter encoding for a thiolase involved in beta-oxidation of fatty acids.
This promoter contains one only ora consensus sequence that respond to oleic acid. Its sequence is CGGN3TNA/(R)N8-12CCG
We amplified this part from genomic DNA yeast.