Double terminator consisting of BBa_B0010 and BBa_B0012
It's a simple coding device of HpNikR. The protein is synthesized when IPTG is present.
pUreA was synthesized starting from the genome sequence of H. pylori, instead the other parts coming from iGEM kit.
This part was built as a ligation product between the promoter of our design, pUreA, RBS (B0034), RFP (LVA tag) and the terminator B0015.
This is a nickel sensitive system driven by the promoter pnikR (BBa_K1151003) with a GFP reporter. Normally the production of GFP is switched on, it is possible to repress the GFP expression using nickel ions in medium and in presence of HpNikR (BBa_K1151000) as trascription factor.
This is a system, driven by the strong promoter pureA (BBa_K1151005), for the production of LuxI (BBa_C0061) by the addiction of nickel ions in medium and in presence of HpNikR (BBa_K1151000) as trascription factor.
Derived from the TE terminator of T7 bacteriophage between Genes 1.3 and 1.4 <genbank>V01146</genbank>.
Transcription terminator for the E.coli RNA polymerase.
Suggested by Sri Kosuri and Drew Endy as a high efficiency terminator. The 5' end cutoff was placed immediately after the TAA stop codon and the 3' end cutoff was placed just prior to the RBS of Gene 1.4 (before AAGGAG). Use anywhere transcription should be stopped when the gene of interest is upstream of this terminator.
Transcriptional terminator consisting of a 64 bp stem-loop.
Nickel-responsive pleiotropic regulator (HpNikR)
It was amplified by PCR from the genomic sequence of Helicobacter pylori G27.
The HpNikR protein is a pleiotropic regulator from Helicobacter pylori. In presence of nickel it can acts as an activator or a repressor depending of the specific promoter that contains its operator sites. Each monomer consists in 133aa for a molecular weight of approximately 17.2 kDa. Apo-HpNikR is a tetramer able to bind up to six Ni2+ ions and then a conformational change allows it to activate or repress trascription.
It was inserted in a pET-15b plasmid vector, in order to have a better storage.
This promoter derived from ureAB operon of Helicobacter pylori. It can be linked by HpNikR (BBa_K1151000) in nickel presence for promoting trascription of the downstream gene
This part was modified from a plasmid provided by the Murray lab at Caltech.
This part contains the fluorescent protein mCherry attached to a degradation tag. It will decrease the amount of fluorescent protein over time.
We modified the AAV sequence to LVA with PCR.
This part derives from H. pylori genomic sequence, and it was amplified by PCR.
This promoter derived from Helicobacter pylori, it is costitutively ON and repressed by HpNikR (BBa_K1151000) in nickel presence.
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.
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.
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
green fluorescent protein derived from jellyfish Aequeora victoria wild-type GFP (SwissProt: P42212
GFP (mut3b) [note that this part does not have a barcode]
V. fischeri <genbank>AF170104</genbank>
Synthesizes 3OC6HSL, which binds to LuxR. The lux cassette of V. fischeri contains a left and a right promoter. The right promoter gives weak constitutive expression of downstream genes.This expression is up-regulated by the action of the Lux repressor, LuxR. Two molecules of LuxR protein form a complex with two molecules the signalling compound HSL. This complex binds to a palindromic site on the promoter, increasing the rate of transcription.
An LVA tail (sequence: AANDENYALVA) was added to increase protein degradation. .
Vinay S Mahajan, Voichita D. Marinescu, Brian Chow, Alexander D Wissner-Gross and Peter Carr
Conversion of the iGEM parts registry to SBOL2.1