MutH was isolated from MG1655 genomic DNA and underwent quikchange mutagenesis to become mutH(E56A)
This part contains GFP and the mutH(E56A) protein under the control of an IPTG-inducible promoter. MutH(E56A) has been shown to have mutagenic activity E. coli when expressed.
The IPTG+GFP portion of the plasmid was first constructed followed by the mutH(E56A)
RBS.3 (medium) -- derivative of BBa_0030
Weak1 RBS based on Ron Weiss thesis. Strength is considered relative to <bb_part>BBa_B0030</bb_part>, <bb_part>BBa_B0031</bb_part>, <bb_part>BBa_B0033</bb_part>.
Varies from -6 to +1 region from original sequence to accomodate BioBricks suffix ("RBS-2" in figure 4-14 of thesis). Contact info for this part: Brian Chow
Vinay S Mahajan, Voichita D. Marinescu, Brian Chow, Alexander D Wissner-Gross and Peter Carr IAP, 2003.
IPTG promoter was cloned from registry plasmid BBa_J04500, RBS.3 was designed into the primer for cloning IPTG, and these were cloned into reporter GFP, cutting the front two cut sites.
Contains an IPTG-sensitive promoter (BBa_J04500), with RBS.3 (BBa_00032), and reporter GFP (E0040) downstream. This part was primarily used as the initial plasmid into which mutator genes were cloned and site-directed mutagenesis was conducted. Also, it was intended to be the plasmid in which mutator genes were assembled in combinations before cloning the entire combination out into the AroF-mCherry-LVA backbone.
The back two cut sites may be cut and mutator genes PCR'd from wild-type may then be inserted into this plasmid.
E. coli (MG1655) genomic and quickchanged to E56A
mutagenized mutH useful as a mutagen protein
green fluorescent protein derived from jellyfish Aequeora victoria wild-type GFP (SwissProt: P42212
GFP (mut3b) [note that this part does not have a barcode]
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
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