Copper binding paired with Polystyrene binding
BioBricks found in the Registry of Standard Biological Parts were used to make this composite sequence.
Contains circularly permuted OmpX protein that enables E.coli to bind to a polystyrene surface. This sequence also contains the surface display protein OmpC, which has been modified to include a copper binding protein (CutA) on the extracellular loop of the membrane protein. This system is designed to be used for removing copper from solution by adding E. coli bound to a polystyrene surface and containing the copper binding mechanism to the solution.
The OmpC BioBrick would need to be mutated to remove EcoRI and a SpeI sites. There were no other special design considerations.
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.
Lutz, R., Bujard, H., Nucleic Acids Research (1997) 25, 1203-1210.
Sequence for pTet inverting regulator driven by the TetR protein.
BBa_R0040 TetR-Regulated Promoter is based on a cI promoter. It has been modified to include two TetR binding sites and the BioBrick standard assembly head and tail restriction sites.
June Rhee, Connie Tao, Ty Thomson, Louis Waldman
CPX Terminal Surface Display Protein with Polystyrene-Binding Peptide
Rice JJ, Schohn A, Bessette PH, Boulware KT, and Daugherty PS. Bacterial display using circularly permuted outer membrane protein OmpX yields high affinity peptide ligands. Protein Sci 2006 Apr; 15(4) 825-36. doi:10.1110/ps.051897806 pmid:16600968
Omp C is native to E.coli See Xu Z, Lee SY. (1999). Display of Polyhistidine Peptides on the Escherichia coli Cell Surface by Using Outer Membrane Protein C as an Anchoring Motif. Applied and Environmental Microbiology. 65(11): 5142-5147
Membrane display OmpC protein with inserted copper-binding protein that is expressed on the surface of Escherichia coli in an extracellular loop.
note: copper-binding protein was inserted in a Pst1 restriction enzyme site ater it was filled in using DNA Polymerase. Two nucletides were inserted at the end of the copper-binding sequence to avoid throwing off the reading frame.
Conversion of the iGEM parts registry to SBOL2.1