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Team:Cambridge/Oligos
From 2010.igem.org
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===Our first attempts at oligo design (a little excessive)=== | ===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/Biobrickingluxoperon | Oligos to BioBrick the whole lux operon]] (never ordered as too expensive) |
| - | + | ||
===luxICDABE operon=== | ===luxICDABE operon=== | ||
Revision as of 14:22, 17 August 2010
Biobrick forward primer
>BBa_G00100 Part-only sequence (20 bp) tgccacctgacgtctaagaa
Biobrick backwards primer
>BBa_G00101 Part-only sequence (20 bp) attaccgcctttgagtgagc
The Vibrio Lux operon
The Phosphoreum Lux operon
Pbad Promoter Part Phosphoreum LuxL gene upstream of LuxC
The phosphoreum oligos we ordered
Notes on alterations to the SLOCK pHK724 clone
- The sequence which SLOCK sent us is just the sequence which is found on 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 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 summary of the lux operon with quality references
- How to design a RBS
Oligo Order for Vibrio Fischeri on 13.08.10
Firefly Oligos to be ordered
Mutagenesis Oligos to be ordered before DNA 2.0 comes through Oligos to biobrick our DNA 2.0 order
Our first attempts at oligo design (a little excessive)
Oligos to BioBrick the whole lux operon (never ordered as too expensive)
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
Thioesterase
ygcB gene from E. Coli K12 to over-express thioesterase: GTGAATACAACGCTGTTTCGATGGCCGGTTCGCGTCTACTATGAAGATACCGATGCCGGTGGTGTGGTGTACCACGCCAGTTACGTCGCTTTTTATGAAA GAGCACGCACAGAGATGCTGCGTCATCATCACTTCAGTCAGCAGGCGCTGATGGCTGAACGCGTTGCCTTTGTGGTACGTAAAATGACGGTGGAATATTA CGCACCTGCGCGGCTCGACGATATGCTCGAAATACAGACTGAAATAACATCAATGCGTGGCACCTCTTTGGTTTTCACGCAACGTATTGTCAACGCCGAG AATACTTTGCTGAATGAAGCAGAGGTTCTGGTTGTTTGCGTTGACCCACTCAAAATGAAGCCTCGTGCGCTTCCCAAGTCTATTGTCGCGGAGTTTAAGC AGTGA
