J23101 + RBS + UirR + B0015 + PlsiR + GFP
UirR and Plsir come from the genomic sequence of the unicellular cyanobacterium Synechocystis sp. PCC6803.
The UV intensity response regulator coding sequence was positioned downstream of J23101, a constitutive promoter family member, and upstream of the double terminator B0015. In this part the promoter associated with UirR was also added, alongside with GFP, so that activity may be reported.
J23101, UirR, and B0015 were ligated to Plsir and GFP after restriction that was followed by an Azure-A-based gel extraction.
The promoter comes from the genome of the unicellular cyanobacterium Synechocystis sp. PCC6803.
The promoter activated by the release of UirR (K1725420) from the transmembrane protein UirS (K1725410) in response to unidirectional UV-A light is upstream of GFP with the B0034 RBS (BBa_I13500). This allows activity in response to UV-A to be measured. For the promoter to be activated UirR and UirS must be present and the cell has to be exposed to UV-A.
The promoter, including the BioBrick prefix and suffix, was ordered as a G-block. It was ligated to I13500 after restriction that was followed by an Azure-A-based gel extraction.
The regulator comes from the genomic sequence of the unicellular cyanobacterium Synechocystis sp. PCC6803.
The UV intensity response regulator coding sequence was positioned downstream of J23101, a constitutive promoter family member, and upstream of the double terminator B0015. This allows the part to be placed in front of other parts with weaker or non-constitutive promoters.
The regulator and J23101 were ligated to B0015 after restriction that was followed by an Azure-A-based gel extraction.
Double terminator consisting of BBa_B0010 and BBa_B0012
Built by Josh as an intermediate in screening plasmid construction
PlsiR (Promoter of light and stress integrating response Regulator)
This promoter originally comes from the unicellular cyanobacterium Synechocystis sp. PCC6803. The promoter is normally off and is activated by the release of UirR (K1725420) from the transmembrane protein UirS (K1725410) in response to unidirectional UV-A light. For the promoter to be activated all three parts must be present and the cell has to be exposed to UV-A.
PlsiR was ordered as a G-block gene fragment.
The UV intensity response regulator coding sequence was placed downstream of J23101, a constitutive promoter family member.
The regulator was ligated to J23101 (which came in the form of an annealed oligo) after restriction that was followed by an Azure-A-based gel extraction.
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.
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.
green fluorescent protein derived from jellyfish Aequeora victoria wild-type GFP (SwissProt: P42212
GFP (mut3b) [note that this part does not have a barcode]
UirR comes from the genomic sequence of the unicellular cyanobacterium Synechocystis sp. PCC6803.
The regulator (K1725420) that participates together with PlsiR (K1725400) and UirS (K1725410) to activate a response to UV-A light has been put downstream of the RBS B0032.
Due to the length of the DNA sequence, the gene was amplified via PCR and the BioBrick suffix and prefix, alongside with the RBS, were added as part of the primers. The last base of the gene was changed from G to A because different stop codons can be more efficient in different species and TAA is especially efficient in E. coli.
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
The regulator comes from the genomic sequence of the unicellular cyanobacterium Synechocystis sp. PCC6803
UirR is a protein that attaches itself to the transmembrane protein UirS (K1725410). After exposure to unidirectional UV-A, UirS releases UirR, which physically interacts with the lsiR promoter (K1725400), activating transcription. All three parts are required for a cell to respond to the UV-A stimulus.
Due to the length of the DNA sequence, the gene was amplified via PCR and the BioBrick suffix and prefix were added as part of the primers. The last base of the gene was changed from G to A because different stop codons can be more efficient in different species and TAA is especially efficient in E. coli.
Conversion of the iGEM parts registry to SBOL2.1