Team:Cambridge/Oligos

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>BBa_G00101 Part-only sequence (20 bp)
>BBa_G00101 Part-only sequence (20 bp)
attaccgcctttgagtgagc
attaccgcctttgagtgagc
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===The Vibrio Lux operon===
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[http://www.ncbi.nlm.nih.gov/nuccore/5726577 AF170104-1]
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===The Phosphoreum Lux operon===
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[http://www.ncbi.nlm.nih.gov/nuccore/166797857 DQ988873]
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[http://partsregistry.org/cgi/partsdb/dna.cgi?part_name=BBa%20I0500 Pbad Promoter Part]
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[http://www.fasebj.org/cgi/reprint/7/11/1016 Phosphoreum LuxL gene upstream of LuxC]
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[[Team:Cambridge/PhosphoreumOligos | The phosphoreum oligos we ordered]]
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===Notes on alterations to the SLOCK pHK724 clone===
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*The sequence which SLOCK sent us is just the sequence which is found on [http://www.ncbi.nlm.nih.gov/nuccore/AF170104.1 NCBI] (found by sequence alignment of the two sequences)
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*We do not know exactly what is in the plasmid, maybe some information can be found in the references [[Team:Cambridge/References/ProjectBioluminescence/Luciferase#Plasmid_experiment | here]]
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*From the annotated start of the lux ICDABEG operon from NCBI there are no restriction sites in conflict with the BioBrick standards.
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*Editing ideas:
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**Remove I and G regions from the operon (are these useful?)
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**Attach suffices and prefixes
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**Is this sufficient to then put into a BioBrick? what Chassis should we use?
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*A good [http://www.bio.cmu.edu/courses/03441/TermPapers/97TermPapers/lux/operon.html summary] of the lux operon with quality references
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*How to design a [http://partsregistry.org/Ribosome_Binding_Sites/Design RBS]
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[[Team:Cambridge/OligoOrderVibrio13.08.10 | Oligo Order for Vibrio Fischeri on 13.08.10]]
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[[Team:Cambridge/OligoOrderVibrio17.08.10 | Oligo Order for Vibrio Fischeri on 17.08.12]]
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===Firefly Oligos to be ordered===
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*[[Team:Cambridge/mutagenesisprimers17.08.10 | Mutagenesis Oligos to be ordered before DNA 2.0 comes through]]
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*[[Team:Cambridge/biobrickingoligosfirefly | Oligos to biobrick our DNA 2.0 order]]
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===Our first attempts at oligo design (a little excessive)===
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[[Team:Cambridge/Biobrickingluxoperon | Oligos to BioBrick the whole lux operon]] (never ordered as too expensive)
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[[Team:Cambridge/luxICDABEoperon | luxICDABE operon sequence]] from NCBI
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[[Team:Cambridge/LuxR | LuxR sequence]] from NCBI
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==Thioesterase==
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ygcB gene from E. Coli K12 to over-express thioesterase:
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GTGAATACAACGCTGTTTCGATGGCCGGTTCGCGTCTACTATGAAGATACCGATGCCGGTGGTGTGGTGTACCACGCCAGTTACGTCGCTTTTTATGAAA
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GAGCACGCACAGAGATGCTGCGTCATCATCACTTCAGTCAGCAGGCGCTGATGGCTGAACGCGTTGCCTTTGTGGTACGTAAAATGACGGTGGAATATTA
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CGCACCTGCGCGGCTCGACGATATGCTCGAAATACAGACTGAAATAACATCAATGCGTGGCACCTCTTTGGTTTTCACGCAACGTATTGTCAACGCCGAG
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AATACTTTGCTGAATGAAGCAGAGGTTCTGGTTGTTTGCGTTGACCCACTCAAAATGAAGCCTCGTGCGCTTCCCAAGTCTATTGTCGCGGAGTTTAAGC
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AGTGA
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Forward Primer:
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gaattcgcggccgcttctagaGTGAATACAACGCTGTTTC
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Complement (for Gibson):
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GAAACAGCGTTGTATTCACtctagaagcggccgcgaattc
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Bacward primer:
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ctgcagcggccgctactagtaTCACTGCTTAAACTCCG
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Complement:
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CGGAGTTTAAGCAGTGAtactagtagcggccgctgcag
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{{:Team:Cambridge/Templates/footer}}
{{:Team:Cambridge/Templates/footer}}

Latest revision as of 10:27, 18 August 2010

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