DNA damage sensor with Gemini Reporter (weak LexA Generator)
Attaches BBa_M36777 to a Gemini actuator so it can report out.
DNA damage sensor with weak LexA generator
LexA gene and promoter from E. coli genome, other parts from registry
Same as our BBa_M36053 DNA damage sensor, but using a weaker 5' UTR in the LexA generator so not as much LexA is generated.
This sensor with a weaker LexA generator is another candidate for level matching with the Gemini actuator we will attach to the damage sensor.
Gemini, a Bifunctional Enzymatic and Fluorescent Reporter of Gene Expression. PLOS One. Lance Martin et al.
A medium strength RBS expressing Gemini, ending with a strong terminator.
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.
Transcriptional terminator consisting of a 64 bp stem-loop.
5' Bicistronic UTR (weak), does not include ATG start
c/o Vivek Mutalik et al. BIOFAB Emeryville
5' UTR based on bicistronic junction architecture. First RBS is strong and drives expression of short leader cistron. Second RBS internal to first cistron control translation of gene of interest. Architecture is optimized so that downstream CDS for gene of interest does not have a chance to form a secondary structure with the 5' UTR. See BIOFAB.org for more information.
Guillaume Cambray et al. c/o BIOFAB Emeryville
E. coli RNA pol transcription terminator designed and tested by Guillaume Cambray at BIOFAB Emeryville. Based on the natural E. coli rnpB T1 terminator.
LexA is a repressor that represses SOS response genes coding for DNA polymerases required for repairing DNA damage.
This is the promoter sequence for the LexA gene. The lexA protein regulates its own promoter. There is a common binding site for the lexA protein found in this promoter that is also found in ~40 other SOS genes.
We know exactly were the lexA protein binds, but we are not entirely sure of the exact deliniation of the promoter itself. Therefore this sequence may contain superfluous information.
amino acids 3-60 from lacZ and amino acids 2-end of GFP (E0043)
This part is a fusion protein between GFP and the alpha fragment of lacZ. It can be measured via fluorescence from GFP or through an assay for beta-galactosidase in a strain containing the omega fragment of lacZ.
The amino acid linker AGGSEGGGSEHHHHHHGSE is between the lacZa and GFP. The 6-his can also facilitate detection on a Western blot, for purification, etc.
Conversion of the iGEM parts registry to SBOL2.1