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| attaccgcctttgagtgagc | | attaccgcctttgagtgagc |
| | | |
| + | ===The Vibrio Lux operon=== |
| + | [http://www.ncbi.nlm.nih.gov/nuccore/5726577 AF170104-1] |
| + | |
| + | ===The Phosphoreum Lux operon=== |
| + | [http://www.ncbi.nlm.nih.gov/nuccore/166797857 DQ988873] |
| + | |
| + | [http://partsregistry.org/cgi/partsdb/dna.cgi?part_name=BBa%20I0500 Pbad Promoter Part] |
| + | |
| + | [http://www.fasebj.org/cgi/reprint/7/11/1016 Phosphoreum LuxL gene upstream of LuxC] |
| + | |
| + | [[Team:Cambridge/PhosphoreumOligos | The phosphoreum oligos we ordered]] |
| | | |
| ===Notes on alterations to the SLOCK pHK724 clone=== | | ===Notes on alterations to the SLOCK pHK724 clone=== |
| + | *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) |
| + | *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]] |
| + | *From the annotated start of the lux ICDABEG operon from NCBI there are no restriction sites in conflict with the BioBrick standards. |
| + | *Editing ideas: |
| + | **Remove I and G regions from the operon (are these useful?) |
| + | **Attach suffices and prefixes |
| + | **Is this sufficient to then put into a BioBrick? what Chassis should we use? |
| + | *A good [http://www.bio.cmu.edu/courses/03441/TermPapers/97TermPapers/lux/operon.html summary] of the lux operon with quality references |
| + | *How to design a [http://partsregistry.org/Ribosome_Binding_Sites/Design RBS] |
| + | |
| + | [[Team:Cambridge/OligoOrderVibrio13.08.10 | Oligo Order for Vibrio Fischeri on 13.08.10]] |
| + | |
| + | [[Team:Cambridge/OligoOrderVibrio17.08.10 | Oligo Order for Vibrio Fischeri on 17.08.12]] |
| + | |
| + | ===Firefly Oligos to be ordered=== |
| + | *[[Team:Cambridge/mutagenesisprimers17.08.10 | Mutagenesis Oligos to be ordered before DNA 2.0 comes through]] |
| + | *[[Team:Cambridge/biobrickingoligosfirefly | Oligos to biobrick our DNA 2.0 order]] |
| + | |
| + | ===Our first attempts at oligo design (a little excessive)=== |
| + | |
| + | [[Team:Cambridge/Biobrickingluxoperon | Oligos to BioBrick the whole lux operon]] (never ordered as too expensive) |
| + | |
| + | [[Team:Cambridge/luxICDABEoperon | luxICDABE operon sequence]] from NCBI |
| + | |
| + | [[Team:Cambridge/LuxR | LuxR sequence]] from NCBI |
| + | |
| + | ==Thioesterase== |
| + | |
| + | ygcB gene from E. Coli K12 to over-express thioesterase: |
| + | GTGAATACAACGCTGTTTCGATGGCCGGTTCGCGTCTACTATGAAGATACCGATGCCGGTGGTGTGGTGTACCACGCCAGTTACGTCGCTTTTTATGAAA |
| + | GAGCACGCACAGAGATGCTGCGTCATCATCACTTCAGTCAGCAGGCGCTGATGGCTGAACGCGTTGCCTTTGTGGTACGTAAAATGACGGTGGAATATTA |
| + | CGCACCTGCGCGGCTCGACGATATGCTCGAAATACAGACTGAAATAACATCAATGCGTGGCACCTCTTTGGTTTTCACGCAACGTATTGTCAACGCCGAG |
| + | AATACTTTGCTGAATGAAGCAGAGGTTCTGGTTGTTTGCGTTGACCCACTCAAAATGAAGCCTCGTGCGCTTCCCAAGTCTATTGTCGCGGAGTTTAAGC |
| + | AGTGA |
| + | |
| + | Forward Primer: |
| + | |
| + | gaattcgcggccgcttctagaGTGAATACAACGCTGTTTC |
| + | |
| + | Complement (for Gibson): |
| + | |
| + | GAAACAGCGTTGTATTCACtctagaagcggccgcgaattc |
| + | |
| + | Bacward primer: |
| + | |
| + | ctgcagcggccgctactagtaTCACTGCTTAAACTCCG |
| + | |
| + | Complement: |
| + | |
| + | CGGAGTTTAAGCAGTGAtactagtagcggccgctgcag |
| + | |
| | | |
| | | |
- | ===luxICDABE operon===
| |
- | gtcgaccttcctggttcagagcctcatatccatattacgaccacttcaat
| |
- | gggcgatcaaaaagtacattacatgattttttgcccaacagaaaaagcct
| |
- | ctcatttagagcagcttattcgtcaagatttcatggaaatgtatgagatc
| |
- | aattttccacaaaaaacagagtaacaaagagaaaaacctataggtataaa
| |
- | gctttacttacgtacttaatttttaaagtatgggcaatcaattgctcctg
| |
- | ttaaaattgctttagaaatactttggcagcggtttgttgtattgagtttc
| |
- | atttgcgcattggttaaatggaaagtgacagtacgctcactgcagcctaa
| |
- | tatttttgaaatatcccaagagctttttccttcgcatgcccacgctaaac
| |
- | attctttttctcttttggttaaatcgttgtttgatttattatttgctata
| |
- | tttatttttcgataattatcaactagagaaggaacaattaatggtatgtt
| |
- | catacacgcatgtaaaaataaactatctatatagttgtctttttctgaat
| |
- | gtgcaaaactaagcattccgaagccattgttagccgtatgaatagggaaa
| |
- | ctaaacccagtgataagacctgatgttttcgcttctttaattacatttgg
| |
- | agattttttatttacagcattgttttcaaatatattccaattaattggtg
| |
- | aatgattggagttagaataatctactataggatcatattttattaaatta
| |
- | gcgtcatcataatattgcctccattttttagggtaattatctagaattga
| |
- | aatatcagatttaaccatagaatgaggataaatgatcgcgagtaaataat
| |
- | attcacaatgtaccattttagtcatatcagataagcattgattaatatca
| |
- | ttattgcttctacaagctttaattttattaattattctgtatgtgtcgtc
| |
- | ggcatttatgtttttcatacccatctctttatccttacctattgtttgtc
| |
- | gcaagttttgcgtgttatatatcattaaaacggtaatggattgacatttg
| |
- | attctaataaattggatttttgtcacactattgtatcgctgggaatacaa
| |
- | ttacttaacataagcacctgtaggatcgtacaggtttacgcaagaaaatg
| |
- | gtttgttatagtcgaataaacgcaagggaggttggtatgactataatgat
| |
- | aaaaaaatcggattttttggcaattccatcggaggagtataaaggtattc
| |
- | taagtcttcgttatcaagtgtttaagcaaagacttgagtgggacttagtt
| |
- | gtagaaaataaccttgaatcagatgagtatgataactcaaatgcagaata
| |
- | tatttatgcttgtgatgatactgaaaatgtaagtggatgctggcgtttat
| |
- | tacctacaacaggtgattatatgctgaaaagtgtttttcctgaattgctt
| |
- | ggtcaacagagtgctcccaaagatcctaatatagtcgaattaagtcgttt
| |
- | tgctgtaggtaaaaatagctcaaagataaataactctgctagtgaaatta
| |
- | caatgaaactatttgaagctatatataaacacgctgttagtcaaggtatt
| |
- | acagaatatgtaacagtaacatcaacagcaatagagcgatttttaaagcg
| |
- | tattaaagttccttgtcatcgtattggagacaaagaaattcatgtattag
| |
- | gtgatactaaatcggttgtattgtctatgcctattaatgaacagtttaaa
| |
- | aaagcagtcttaaattaatgttgttaaatcattaatttattttaaatact
| |
- | aagtatattatagggaaaataatgaataaatgtattccaatgataattaa
| |
- | tggaatgattcaagattttgataattatgcatataaagaagttaaactaa
| |
- | ataatgataatagagtaaaattatctgtcattactgaaagttcagtttca
| |
- | aaaacattaaatatcaaagatagaattaatctaaatttaaatcagattgt
| |
- | gaattttttatataccgttggtcaacgatggaaaagtgaagaatataatc
| |
- | ggcgacgaacctatatccgtgagttaaaaacatatcttggttattctgat
| |
- | gaaatggcaagattagaagcgaattggattgcaatgttattgtgctctaa
| |
- | aagtgctttgtatgacattgttaattatgatttgggctctatacacgtat
| |
- | tagatgaatggcttccacgtggtgattgctatgttaaagcacaaccgaaa
| |
- | ggtgtttctgttcacttgttagctggtaatgttccattatcaggagtgac
| |
- | atctattttgcgtgctattttaacaaaaaatgagtgcattattaaaactt
| |
- | cgtcttcagatccttttactgcaaacgctttagtttccagttttattgat
| |
- | gttaatgcagaccatccaataaccaaatcaatgtctgttatgtattggcc
| |
- | gcatgatgaagatatgactctatctcaaagaataatgaatcatgccgacg
| |
- | tggttattgcttggggtggagacgaggcgattaaatgggcggtaaaatat
| |
- | tcaccaccgcatgtcgatattctgaaatttggaccaaagaaaagcttaag
| |
- | tattattgaagctcctaaagatatagaagccgcagcaatgggggttgctc
| |
- | atgatatttgtttctatgaccagcaagcctgcttctctactcaagacgtt
| |
- | tattatataggagataatttacctttatttttaaatgaacttgaaaaaca
| |
- | gctagatcgatacgcgaaaattttaccaaaaggttcaaatagttttgatg
| |
- | aaaaagcggcgtttactcttactgaaaaagaaagtctatttgctggatat
| |
- | gaagtgagaaagggagataagcaagcttggttaatagtcgtatcacctac
| |
- | aaatagctttggaaatcaaccgctatcacgaagtgtgtatgttcatcaag
| |
- | tatctgatattaaagagataattccttttgttaataaaaatagaacacaa
| |
- | actgtttctatttatccttgggaagcgtcattaaaatatcgagataaatt
| |
- | agcaagaagtggagttgaaagaattgttgaatcaggcatgaataatattt
| |
- | tcagagttggaggggctcatgattcattatctcctctccagtacctagtt
| |
- | aggtttgtatcgcatgagagaccatttaattatacgacaaaagatgttgc
| |
- | ggttgaaatcgaacaaacacgttacttagaggaagataagtttttagttt
| |
- | ttgtcccatagttaaaggaaattatatgaaagatgaaagtgcttttttta
| |
- | cgattgatcacatcatcaagcttgataatggtcagtctatccgagtttgg
| |
- | gaaacactccctaaaaagaacgtaccagagaaaaaacatacaatacttat
| |
- | tgcttcgggttttgctagaagaatggatcattttgcaggtcttgctgagt
| |
- | atttatctactaacggttttcatgtcattcgctacgattctttgcatcat
| |
- | gttggattaagcagtggatgtataaatgaatttacgatgtcgattggaaa
| |
- | aaatagcctgcttacagtcgtagattggcttacagatcatggtgtcgaac
| |
- | gaatagggctgattgctgctagtttgtcagcgagaatcgcctatgaggta
| |
- | gtaaataaaattaaattatcatttttaattacggccgtaggtgtcgttaa
| |
- | tcttagagatacattagaaaaagcattggagtatgactatttgcaattac
| |
- | ctatttcagagttaccagaagatcttgactttgaaggtcataatttagga
| |
- | tcggaggtctttgttacagattgctttaaacataattgggacacgttaga
| |
- | ctcgacacttaatagtgttaaaggattagcgattccatttattgctttta
| |
- | ctgcaaacgatgatagttgggtaaagcaaagtgaagttatagagctcatt
| |
- | gatagcattgaatctagtaattgtaagctctattcgctaattggaagttc
| |
- | acatgatcttggggaaaatttggttgtattaagaaatttttatcaatcag
| |
- | tgacgaaggcagccttagcattagatgatggtttattggatttagagata
| |
- | gacattattgaacctcgatttgaggacgttacaagtattactgttaagga
| |
- | gcgtagattaaaaaatgaaattgaaaatgaattattagaattggcttaaa
| |
- | taaacagaatcaccaaaaaggaatagagtatgaagtttggaaatatttgt
| |
- | ttttcgtatcaaccaccaggtgaaactcataagcaagtaatggatcgctt
| |
- | tgttcggcttggtatcgcctcagaagaggtagggtttgatacatattgga
| |
- | ccttagaacatcattttacagagtttggtcttacgggaaatttatttgtt
| |
- | gctgcggctaacctgttaggaagaactaaaacattaaatgttggcactat
| |
- | gggggttgttattccgacagcacacccagttcgacagttagaagacgttt
| |
- | tattattagatcaaatgtcgaaaggtcgttttaattttggaaccgttcga
| |
- | gggctataccataaagattttcgagtatttggtgttgatatggaagagtc
| |
- | tcgagcaattactcaaaatttctaccagatgataatggaaagcttacaga
| |
- | caggaaccattagctctgatagtgattacattcaatttcctaaggttgat
| |
- | gtatatcccaaagtgtactcaaaaaatgtaccaacctgtatgactgctga
| |
- | gtccgcaagtacgacagaatggctagcaatacaagggctaccaatggttc
| |
- | ttagttggattattggtactaatgaaaaaaaagcacagatggaactctat
| |
- | aatgaaattgcgacagaatatggtcatgatatatctaaaatagatcattg
| |
- | tatgacttatatttgttctgttgatgatgatgcacaaaaggcgcaagatg
| |
- | tttgtcgggagtttctgaaaaattggtatgactcatatgtaaatgcgacc
| |
- | aatatctttaatgatagcaatcaaactcgtggttatgattatcataaagg
| |
- | tcaatggcgtgattttgttttacaaggacatacaaacaccaatcgacgtg
| |
- | ttgattatagcaatggtattaaccctgtaggcactcctgagcagtgtatt
| |
- | gaaatcattcaacgtgatattgatgcaacgggtattacaaacattacatg
| |
- | cggatttgaagctaatggaactgaagatgaaataattgcttccatgcgac
| |
- | gctttatgacacaagtcgctcctttcttaaaagaacctaaataaattact
| |
- | tatttgatactagagataataaggaacaagttatgaaatttggattattt
| |
- | tttctaaactttcagaaagatggaataacatctgaagaaacgttggataa
| |
- | tatggtaaagactgtcacgttaattgattcaactaaatatcattttaata
| |
- | ctgcctttgttaatgaacatcacttttcaaaaaatggtattgttggagca
| |
- | cctattaccgcagctggttttttattagggttaacaaataaattacatat
| |
- | tggttcattaaatcaagtaattaccacccatcaccctgtacgtgtagcag
| |
- | aagaagccagtttattagatcaaatgtcagagggacgcttcattcttggt
| |
- | tttagtgactgcgaaagtgatttcgaaatggaattttttagacgtcatat
| |
- | ctcatcaaggcaacaacaatttgaagcatgctatgaaataattaatgacg
| |
- | cattaactacaggttattgtcatccccaaaacgacttttatgattttcca
| |
- | aaggtttcaattaatccacactgttacagtgagaatggacctaagcaata
| |
- | tgtatccgctacatcaaaagaagtcgtcatgtgggcagcgaaaaaggcac
| |
- | tgcctttaacatttaagtgggaggataatttagaaaccaaagaacgctat
| |
- | gcaattctatataataaaacagcacaacaatatggtattgatatttcgga
| |
- | tgttgatcatcaattaactgtaattgcgaacttaaatgctgatagaagta
| |
- | cggctcaagaagaagtgagagaatacttaaaagactatatcactgaaact
| |
- | taccctcaaatggacagagatgaaaaaattaactgcattattgaagagaa
| |
- | tgcagttgggtctcatgatgactattatgaatcgacaaaattagcagtgg
| |
- | aaaaaacagggtctaaaaatattttattatcctttgaatcaatgtccgat
| |
- | attaaagatgtaaaagatattattgatatgttgaaccaaaaaatcgaaat
| |
- | gaatttaccataataaaattaaaggcaatttctatattagattgcctttt
| |
- | taaatttctgttgatattaggtattactggagagggtatgactgtccata
| |
- | ctgaatataaaagaaatcaaatcattgctagttcagaaattgatgatctt
| |
- | atctttatgacgaaaccacaagagtggtcatttgaagagcaaaaagaaat
| |
- | acgggataaattagttcgtgaggctttttattttcactacaatagaaatg
| |
- | aagaatatagaaattattgtatcaatcagcatgtgagtgataatttacac
| |
- | actattgatgaaatacccgtgtttccaacatctgtttttaaatataagaa
| |
- | attacatactgtcacagccgaggacattgaaaattggtatacaagtagtg
| |
- | gaactcgtggagtaaaaagtcatattgcacgtgatcgtcttagcattgaa
| |
- | cgcttgcttggttctgtcaacttcggaatgaaatacgttggagattggtt
| |
- | tgagcatcaaatggaattgataaatttaggaccagatagattcaatacaa
| |
- | ataatatttggtttaaatatgtcatgagtttggtcgagttactttatccg
| |
- | actgaatttacagttgataatgacaaaatagattttgaaaaaacagtaaa
| |
- | acatctatttagaattaagaatagtaaaaaagacatttgcttaattgggc
| |
- | caccattttttgtgtatcttttgtgccaatatatgaaagaaaacaatatt
| |
- | gaatttaaaggaggagatagagtacatattattactggtggaggatggaa
| |
- | atctaatcagaatgactctttagatcgtgctgattttaatcaattattaa
| |
- | tggatactttccaactcgacaagattaatcaaattagagatacctttaat
| |
- | caagttgagcttaatacttgtttttttgaagatgaatttcaaagaaaaca
| |
- | tgttccaccgtgggtatatgctcgggctcttgatcctgaaaccttgaaac
| |
- | ccgtagcagatggtgagatcgggttgttaagttatatggacgcctcatca
| |
- | actgcttaccctgcttttattgttactgatgatatcggtattgtaaaaga
| |
- | aattagagaaccagatccttacccaggggtaactgttgagattgttcggc
| |
- | gcttaaatacacgtgcgcaaaaaggatgcgcgctctctatggctaatgtc
| |
- | atacaaaagaatatcaaggattaagttatgattgttgatggtagagtttc
| |
- | aaaaatagtattagcgtcaataagaaataatatatataaggtatttatta
| |
- | ctgtaaattcaccaataaagttcatcgctggacaatttgtaatggtcacg
| |
- | attaatgggaaaaaatgccctttttcaattgcgaattgcccgacaaaaaa
| |
- | ttacgaaatagaattgcatattggtagttcgaatagagactgctcattgg
| |
- | atattatcgaatattttgtcgatgctcttgttgaggaagtcgcaattgag
| |
- | ttagatgctccccatggaaacgcttggttacggtctgaaagtaataaccc
| |
- | attgctattaattgcgggaggtacaggtttatcatatataaatagcattc
| |
- | taacaaattgcttaaataggaatatacctcaagatatttatctttactgg
| |
- | ggagtaaaagacagttctcttttgtatgaagatgaagagttactaaactt
| |
- | atcactaaacaacaaaaactttcattatattcctgttattgaagataaaa
| |
- | gtgaagaatggatagggagaaaaggcactgttcttgatgctgtcatggaa
| |
- | gattttactgatcttacttattttgatatttatgtttgtggacccttcat
| |
- | gatggctaaaacagcaaaagaaaaattaattgaagagaaaaaagcaaagt
| |
- | cagaacagatgtttgccgatgcttttgcatacgtataaagagaatataaa
| |
- | aagccagattattaatccggcttttttattatttcaatgtgtccaagggt
| |
- | aaccaagttacttcaactcccgcctgagagaacatatcttggctcacttg
| |
- | aattttttcaccccaacgagataagaagtcttcagtttgttcagggcaat
| |
- | atacttttgatattcctgtttggatgatttttgctgcgcagtttggacaa
| |
- | gggaagtgagtaacccaaatatcgcaaccttctaaatcacgcttggcaaa
| |
- | taaaatggcattttcttcagcatgaagtgtttttaaatatttaatttcac
| |
- | gttcatctgtatctgcactgtctgatacaccatgtgggtagccattaaat
| |
- | ccaacagaaacaatacggttatgcttggtgattactgcaccaacttgagt
| |
- | cgac
| |
| | | |
- | ===luxR===
| |
- | ATGAAAAACATAAATGCCGACGACACATACAGAATAATTAATAAAATTAA
| |
- | AGCTTGTAGAAGCAATAATGATATAATCAATGCTTATCTGATATGACTAA
| |
- | AATGGTACATTGTGAATATTATTTATTCGCGATCATTTATCCTCATTCTA
| |
- | TGGTTAAATCTGATATTTCAATCTAGATAATTACCCTAAAAAATGGAGGC
| |
- | AATATTATGATGACGCTAATTTAATAAAATATGATCCTATAGTAGATTAT
| |
- | TCTAACTCCAATCATTCACCAATTAATTGGAATATATTTGAAAACAATGC
| |
- | TGTAAATAAAAAATCTCCAAATGTAATTAAGAAGCGAAAACATCAGGTCT
| |
- | TATCACTGGGTTTAGTTTCCCTATTCATACGGCTAACAATGCCTTCGGAA
| |
- | TGCTTAGTTTTGCACATTCAGAAAAAGACAACTATATAGATAGTTTATTT
| |
- | TTACATGCGTGTATGAACATACCATTAATTGTTCCTTCTCTAGTGTATAA
| |
- | TTATCGAAAAATAAATATAGCAAATAATAAATCAAACAACGATTTAACCA
| |
- | AAAGAGAAAAAGAATGTTTAGCGTGGGCATGCGAAGGAAAAGGCTCTTGG
| |
- | GATATTTCAAAAATATTAGGCTGCAGTGAGCGTACTGTCACTTTCCATTT
| |
- | AACCAATGCGCAAATGAAACTCAATACACCAAACCGCTGCCAAAGTATTT
| |
- | CTAAAGCAATTTTAACAGGAGCAATTGATTGCCCATACTTTAAAAATTAA
| |
| | | |
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