GFP:registry CP/RBS: registry clc-ec1: PCR from genome Pgad: PCR from vector from CUHK 2011 team
This construct was developed by Northwestern's 2012 iGEM team as part of a phytase producing vector. This particular part was constructed in order to test the to make sure that the Pgad/clc system works correctly. We hoped that it would be easier to see GFP for assay purposes. [http://partsregistry.org/wiki/index.php?title=Part:BBa_K912033 The original device] is designed to lyse in the presence of high chloride concentrations. The Pgad promoter in front of the lysis cassette is activated by the combination of high extracellular hydrogen ion concentration and the presence of high intracellular chloride ion concentration.
green fluorescent protein derived from jellyfish Aequeora victoria wild-type GFP (SwissProt: P42212
GFP (mut3b) [note that this part does not have a barcode]
Double terminator consisting of BBa_B0010 and BBa_B0012
Pgad - Intracellular Chloride-Sensing Cassette
The gene sequence was obtained from the cassette from bp 821 to 2071 of GenBank sequence AF005098, which includes PgadR, gadR, Pgad and the starting codon ATG.
Pgad is chloride-sensitive promoter which was first discovered in Lactococcus lactis1, which is a gram-positive bacterium which can live in acidic environment. In this operon, gadC is glutamate-gamma-aminobutyrate antiporter and gadB is glutamate decarboxylase. They are both involved in intracellular pH regulation and co-expressed in the same operon under the control of Pgad2. The gene before Pgad, named gadR, which is constitutively expressed under the control of PgadR, is a positive regulator of Pgad coupled genes while intracellular chloride is level elevated.
Synthesized part by GenScript,all the illegal sites for RFC 10 (EcoRI, XbaI, SpeI, PstI) have been substituted by other codon producing the same amino acid in E. coli.
http://ecocyc.org/ECOLI/sequence-delimited?chromosome=COLI-K12&object=EG12331&up=0&down=0&left=175107&right=176528 genomic sequence.
The ClC-ec1 chloride antiporter pumps protons out of the cell and chloride ions into the cell in acidic conditions.
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.
Conversion of the iGEM parts registry to SBOL2.1