Team:Edinburgh/Sequences

It may or may not be useful to have annotated nucleotide sequences of various stuff that we use. Some of this information can be retrieved from the Registry, but it may be useful to have it in a more nicely usable form. Copying DNA sequences from the Registry using the web-browser's normal copy function is strangely broken...

Note that any biobricks we submit to the Registry will still have to be properly documented there.

We have BBa_K523000 to BBa_K523999 available this year.

Biobrick prefix, suffix, scar

• gaattcgcggccgcttctag - prefix, short
• gaattcgcggccgcttctagag - prefix, long
• tactagtagcggccgctgcag - suffix
• tactag - scar, short
• tactagag - scar, long

Restriction sites

• gaattc - EcoRI (prefix 1)
• tctaga - XbaI (prefix 2)
• actagt - SpeI (suffix 1)
• ctgcag - PstI (suffix 2)

• gcggccgc - NotI (in both prefix and suffix)

• agatct - BglII (not part of RFC10, we use it internally after the prefix)

• tacgta - SnaBI (might become relevant)

pSB1C3 vector

• This is the official sequence from the Registry ([http://partsregistry.org/wiki/index.php?title=Part:pSB1C3 here])
• Red is CmlR coding sequence ([http://en.wikipedia.org/wiki/Chloramphenicol_acetyltransferase chloramphenicol acetyltransferase])
• Blue is restriction sites for the 4 main enzymes (SpeI, PstI, EcoRI, XbaI)

tactagtagcggccgctgcagtccggcaaaaaagggcaaggtgtcaccaccctgccctttttctttaaaaccgaaaagattacttcgcgttatgcaggct tcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggata acgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccacaggctccgcccccctgacga gcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcct gttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgt aggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaag acacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactac ggctacactagaagaacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccg ctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctca gtggaacgaaaactcacgttaagggattttggtcatgagattatcaaaaaggatcttcacctagatccttttaaattaaaaatgaagttttaaatcaatc taaagtatatatgagtaaacttggtctgacagctcgaggcttggattctcaccaataaaaaacgcccggcggcaaccgagcgttctgaacaaatccagat ggagttctgaggtcattactggatctatcaacaggagtccaagcgagctcgatatcaaattacgccccgccctgccactcatcgcagtactgttgtaatt cattaagcattctgccgacatggaagccatcacaaacggcatgatgaacctgaatcgccagcggcatcagcaccttgtcgccttgcgtataatatttgcc catggtgaaaacgggggcgaagaagttgtccatattggccacgtttaaatcaaaactggtgaaactcacccagggattggctgagacgaaaaacatattc tcaataaaccctttagggaaataggccaggttttcaccgtaacacgccacatcttgcgaatatatgtgtagaaactgccggaaatcgtcgtggtattcac tccagagcgatgaaaacgtttcagtttgctcatggaaaacggtgtaacaagggtgaacactatcccatatcaccagctcaccgtctttcattgccatacg aaattccggatgagcattcatcaggcgggcaagaatgtgaataaaggccggataaaacttgtgcttatttttctttacggtctttaaaaaggccgtaata tccagctgaacggtctggttataggtacattgagcaactgactgaaatgcctcaaaatgttctttacgatgccattgggatatatcaacggtggtatatc cagtgatttttttctccattttagcttccttagctcctgaaaatctcgataactcaaaaaatacgcccggtagtgatcttatttcattatggtgaaagtt ggaacctcttacgtgcccgatcaactcgagtgccacctgacgtctaagaaaccattattatcatgacattaacctataaaaataggcgtatcacgaggca gaatttcagataaaaaaaatccttagctttcgctaaggatgatttctggaattcgcggccgcttctagag

Plac-LacZα insert (clone 2)

• Assigned code <partinfo>BBa_K523000</partinfo>
• A modified version (seems to lack about 45 bases at the start) of [http://partsregistry.org/wiki/index.php?title=Part:BBa_J33207 BBa_J33207]
• Sanger sequencing seemed to confirm that all was well with clone 2, though in a few regions the chromatograms were poor.
• Sequence below includes BioBrick prefix and suffix (blue) as well as the BglII site (bold), which is partly in the prefix
• lacZα is in red.
• Both forward and reverse sequencing reactions failed to give good info about the suffix; so I have simply added the standard suffix. However we've not conclusively proved that the SpeI site exists; this should be verified by digest.

gaattcgcggccgcttctagagatctgctggggcaaaccagcgtggaccgcttgctgcaactctctcagggccaggcggtgaagggcaatcagctgttgc ccgtctcactggtgaaaagaaaaaccaccctggcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggt ttcccgactggaaagcgggcagtgagcgcaacgcaattaatgtgagttagctcactcattaggcaccccaggctttacactttatgcttccggctcgtat gttgtgtgaaattgtgagcggataacaatttcacacaggaaacagctatgaccatgattacggattcactggccgtcgttttacaacgtcgtgactggga aaaccctggcgttacccaacttaatcgccttgcagcacatccccctttcgccagctggcgtaatagcgaagaggcccgcaccgatcgcccttcccaacag ttgcgcagcctgaatggcgaatggcgctttgcctggtttccggcaccagaagcggtgccggaaagctggctggagtgatactagtagcggccgctgcag

OmpR-EYFP (clone 2)

• This is [http://partsregistry.org/wiki/index.php?title=Part:BBa_K216011 BBa_K216011].
• Sequence below includes BioBrick prefix and suffix (blue)
• Note the missing NotI site in the suffix (it's still [http://dspace.mit.edu/bitstream/handle/1721.1/59800/BBFRFC54.pdf?sequence=1 RFC54] compliant)
• We have a plate of this in the walk in cold room. They glow under blue light.
• The part consists of a promoter plus EYFP (Enhanced Yellow Fluorescent Protein) gene.

gaattcgcggccgcttctagagtcccttgcatttacattttgaaacatctatagcgataaatgaaacatcttaaaagttttagtatcatattcgtgttgg attattctgcatttttggggagaatggacttactagagaaagaggagaaatactagatggtgagcaagggcgaggagctgttcaccggggtggtgcccat cctggtcgagctggacggcgacgtaaacggccacaagttcagcgtgtccggcgagggcgagggcgatgccacctacggcaagctgaccctgaagttcatc tgcaccaccggcaagctgcccgtgccctggcccaccctcgtgaccaccttcggctacggcctgcaatgcttcgcccgctaccccgaccacatgaagctgc acgacttcttcaagtccgccatgcccgaaggctacgtccaggagcgcaccatcttcttcaaggacgacggcaactacaagacccgcgccgaggtgaagtt cgagggcgacaccctggtgaaccgcatcgagctgaagggcatcgacttcaaggaggacggcaacatcctggggcacaagctggagtacaactacaacagc cacaacgtctatatcatggccgacaagcagaagaacggcatcaaggtgaacttcaagatccgccacaacatcgaggacggcagcgtgcagctcgccgacc actaccagcagaacacccccatcggcgacggccccgtgctgctgcccgacaaccactacctgagctaccagtccgccctgagcaaagaccccaacgagaa gcgcgatcacatggtcctgctggagttcgtgaccgccgccgggatcactctcggcatggacgagctgtacaagtaataatactagtagctgcag

malS using primers f1/r1 (colony 1)

• Prefix and suffix in blue.
• Pre-coding sequence in CAPITALS and contains native RBS.
• BglII site in bold.
• Double stop codon is present.
• This has been sequenced and looks good.

gaattcgcggccgcttctagagatctGAAATCGCAGCAATAAGGACTCATCCGCCatgaaactcgccgcctgttttctgacactccttcctggcttcgcc gttgccgccagctggacttctccggggtttcccgcctttagcgaacaggggacaggaacatttgtcagccacgcgcagttgcccaaaggtacgcgtccac taacgctaaattttgaccaacagtgctggcagcctgcggatgcgataaaactcaatcagatgctttccctgcaaccttgtagcaacacgccgcctcaatg gcgattgttcagggacggcgaatatacgctgcaaatagacacccgctccggtacgccaacattgatgatttccatccagaacgccgccgaaccggtagca agcctggtccgtgaatgcccgaaatgggatggattaccgctcacagtggatgtcagcgccactttcccggaaggagccgccgtacgggattattacagcc agcaaattgcgatagtgaagaacggtcaaataatgttacaacccgctgccaccagcaacggtttactcctgctggaacgggcagaaactgacacatccgc ccctttcgactggcataacgccacggtttactttgtgctgacagatcgtttcgaaaacggcgatcccagtaatgaccagagttacggacgtcataaagac ggtatggcggaaattggcacttttcacggcggcgatttacgcggcctgaccaacaaactggattacctccagcagttgggcgttaatgctttatggataa gcgccccatttgagcaaattcacggctgggtcggcggcggtacaaaaggcgatttcccgcattatgcctaccacggttattacacacaggactggacgaa tcttgatgccaatatgggcaacgaagccgatctacggacgctggttgatagcgcacatcagcgcggtattcgtattctctttgatgtcgtgatgaaccac accggctatgccacgctggcggatatgcaggagtatcagtttggcgcgttatatctttctggtgacgaagtgaaaaaatcgctgggtgaacgctggagcg actggaaacctgccgccgggcaaacctggcatagctttaacgattacattaatttcagcgacaaaacaggctgggataaatggtggggaaaaaactggat cagaacggatatcggcgattacgacaatcctggattcgacgatctcactatgtcgctagcctttttgccggatatcaaaaccgaatcaactaccgcttct ggtctgccggtgttctataaaaacaaaatggatacccacgccaaagccattgacggctatacgccgcgcgattacttaacccactggttaagtcagtggg tccgcgactatgggattgatggttttcgggtcgataccgccaaacatgttgagttgcccgcctggcagcaactgaaaaccgaagccagcgccgcgcttcg cgaatggaaaaaagctaaccccgacaaagcattagatgacaaacctttctggatgaccggtgaagcctggggccacggcgtgatgcaaagtgactactat cgccacggcttcgatgcgatgatcaatttcgattatcaggagcaggcggcgaaagcagtcgactgtctggcgcagatggatacgacctggcagcaaatgg cggagaaattgcagggtttcaacgtgttgagctacctctcgtcgcatgatacccgcctgttccgtgaagggggcgacaaagcagcagagttattactatt agcgccaggcgcggtacaaatcttttatggtgatgaatcctcgcgtccgttcggtcctacaggttctgatccgctgcaaggtacacgttcggatatgaac tggcaggatgttagcggtaaatctgccgccagcgtcgcgcactggcagaaaatcagccagttccgcgcccgccatcccgcaattggcgcgggcaaacaaa cgacacttttgctgaagcagggctacggctttgttcgtgagcatggcgacgataaagtgctggtcgtctgggcagggcaacagtaataatactagtagcg
gccgctgcag

malS using primers f2/r2 (colony 1)

• This is essentially as above but starting at the start codon, and with no stop codon, and adding two bases at the end of the BioBrick, which (together with the T at the start of the suffix) code for a glycine.
• This has been sequenced and looks good.

gaattcgcggccgcttctagagatctatgaaactcgccgcctgttttctgacactccttcctggcttcgccgttgccgccagctggacttctccggggtt tcccgcctttagcgaacaggggacaggaacatttgtcagccacgcgcagttgcccaaaggtacgcgtccactaacgctaaattttgaccaacagtgctgg cagcctgcggatgcgataaaactcaatcagatgctttccctgcaaccttgtagcaacacgccgcctcaatggcgattgttcagggacggcgaatatacgc tgcaaatagacacccgctccggtacgccaacattgatgatttccatccagaacgccgccgaaccggtagcaagcctggtccgtgaatgcccgaaatggga tggattaccgctcacagtggatgtcagcgccactttcccggaaggagccgccgtacgggattattacagccagcaaattgcgatagtgaagaacggtcaa ataatgttacaacccgctgccaccagcaacggtttactcctgctggaacgggcagaaactgacacatccgcccctttcgactggcataacgccacggttt actttgtgctgacagatcgtttcgaaaacggcgatcccagtaatgaccagagttacggacgtcataaagacggtatggcggaaattggcacttttcacgg cggcgatttacgcggcctgaccaacaaactggattacctccagcagttgggcgttaatgctttatggataagcgccccatttgagcaaattcacggctgg gtcggcggcggtacaaaaggcgatttcccgcattatgcctaccacggttattacacacaggactggacgaatcttgatgccaatatgggcaacgaagccg atctacggacgctggttgatagcgcacatcagcgcggtattcgtattctctttgatgtcgtgatgaaccacaccggctatgccacgctggcggatatgca ggagtatcagtttggcgcgttatatctttctggtgacgaagtgaaaaaatcgctgggtgaacgctggagcgactggaaacctgccgccgggcaaacctgg catagctttaacgattacattaatttcagcgacaaaacaggctgggataaatggtggggaaaaaactggatcagaacggatatcggcgattacgacaatc ctggattcgacgatctcactatgtcgctagcctttttgccggatatcaaaaccgaatcaactaccgcttctggtctgccggtgttctataaaaacaaaat ggatacccacgccaaagccattgacggctatacgccgcgcgattacttaacccactggttaagtcagtgggtccgcgactatgggattgatggttttcgg gtcgataccgccaaacatgttgagttgcccgcctggcagcaactgaaaaccgaagccagcgccgcgcttcgcgaatggaaaaaagctaaccccgacaaag cattagatgacaaacctttctggatgaccggtgaagcctggggccacggcgtgatgcaaagtgactactatcgccacggcttcgatgcgatgatcaattt cgattatcaggagcaggcggcgaaagcagtcgactgtctggcgcagatggatacgacctggcagcaaatggcggagaaattgcagggtttcaacgtgttg agctacctctcgtcgcatgatacccgcctgttccgtgaagggggcgacaaagcagcagagttattactattagcgccaggcgcggtacaaatcttttatg gtgatgaatcctcgcgtccgttcggtcctacaggttctgatccgctgcaaggtacacgttcggatatgaactggcaggatgttagcggtaaatctgccgc cagcgtcgcgcactggcagaaaatcagccagttccgcgcccgccatcccgcaattggcgcgggcaaacaaacgacacttttgctgaagcagggctacggc tttgttcgtgagcatggcgacgataaagtgctggtcgtctgggcagggcaacagggtactagtagcggccgctgcag

bglX using primers f1/r1 (colony 1)

• Prefix and suffix in blue.
• Pre-coding sequence in CAPITALS and contains native RBS.
• BglII site in bold.
• Double stop codon is present.
• This has been sequenced and looks good. A silent mutation from the expected sequence is indicated as a capital letter.

gaattcgcggccgcttctagagatctGCCACGTCGGGCAACAAAGGAAGAAAAATCCATatgaaatggctatgttcagtaggaatcgcggtgagtctggc cctgcagccagcactggcggatgatttattcggcaaccatccattaacgcccgaagcgcgggatgcgttcgtcaccgaactgcttaagaaaatgacagtt gatgagaaaattggtcagctgcgcttaatcagcgtcggcccggataacccgaaagaggcgatccgcgagatgatcaaagacggtcaggttggggcgattt tcaacaccgtaacccgtcaggatatccgcgccatgcaggatcaggtgatggaattaagccgcctgaaaattcctcttttctttgcttacgacgtgctgca cggtcagcgcacggtgttcccgattagcctcggtctggcctcgtcttttaacctcgatgcagtgaaaacggtcggacgtgtctctgcttatgaagcggca gatgatggcctgaatatgacctgggcaccgatggtcgatgtctcgcgcgatccgcgctggggacgtgcttccgaaggttttggcgaagatacgtatctca cctcaacaatgggtaaaaccatggtggaagcgatgcagggtaaaagcccggcagatcgctactcggtgatgaccagcgtcaaacactttgccgcatacgg cgcggtagaaggcggtaaagagtacaacaccgtcgatatgagtccgcagcgcctgtttaatgattatatgccgccgtacaaagcggggctggacgcaggc agcggcgcggtgatggtggcgctgaactcgctgaacggcacgccagccacctccgattcctggctgctgaaagatgttctgcgcgaccagtggggcttta aaggcatcaccgtttccgatcacggtgcaatcaaagagctgattaaacatggcacggcggcagacccggaagatgcggtgcgcgtggcgctgaaatccgg aatcaacatgagcatgagcgacgagtactactcgaagtatctgcctgggttgattaaatccggcaaagtgacgatggcagagctggacgatgctgcccgc catgtactgaacgttaaatatgatatggggttgtttaacgacccatacagccatttggggccgaaagagtctgacccggtggataccaatgccgaaagcc gcctgcaccgtaaagaagcgcgtgaagtggcgcgcgaaagcttggtgttgctgaaaaaccgtctcgaaacgttaccgctgaaaaaatcggccaccattgc ggtggttgggccactggcggacagtaaacgtgacgtgatgggcagctggtccgcagccggtgttgccgatcaatccgtgaccgtactgaccgggattaaa aatgccgtcggtgaaaacggtaaagtgctgtatgccaaaggggcgaacgttaccagtgacaaaggcattatcgatttcctgaatcagtatgaagaagcgg tcaaagtcgatccgcgttcgccgcaagagatgattgatgaagcggtgcagacggcgaaacaatctgatgtggtggtggctgtagtcggtgaagcacaggg gatggcgcacgaagcctccagccggaccgatatcactattccgcaaagccaacgtgacttgattgcggcgctgaaagccaccggtaaaccgctggtgctg gtgctgatgaacgggcgtccgctggcgctggtgaaagaagatcagcaggctgatgcgattctggaaacctggtttgcggggactgaaggcggtaatgcaa ttgccgatgtattgtttggcgattacaacccgtccggcaagctgccaatgtccttcccgcgttctgtcgggcagatcccggtgtactacagccatctgaa taccggtcgcccgtataatgccgacaagccgaacaaatacacttcgcgttattttgatgaagctaacggggcgttgtatccgttcggctatgggctgagc tacaccactttcaccgtctctgatgtgaaactttctgcgccgaccatgaagcgtgacggcaaagtgactgccagcgtgcaggtgacgaacaccggtaagc gcgagggtgccacggtagtgcagatgtacttgcaggatgtgacTgcttccatgagtcgccctgtgaaacagctgaaaggctttgagaaaatcaccctgaa accgggcgaaactcagactgtcagcttcccgatcgatattgaggcgctgaagttctggaatcaacagatgaaatatgacgccgagcctggcaagttcaat gtctttatcggcactgattccgcacgcgttaagaaaggcgagtttgagttgctgtaataatactagtagcggccgctgcag

bglX using primers f2/r2 (colony 1)

• This is essentially as above but starting at the start codon, and with no stop codon, and adding two bases at the end of the BioBrick, which (together with the T at the start of the suffix) code for a glycine.
• This has been sequenced and looks good. A silent mutation from the expected sequence is indicated as a capital letter.

gaattcgcggccgcttctagagatctatgaaatggctatgttcagtaggaatcgcggtgagtctggc cctgcagccagcactggcggatgatttattcggcaaccatccattaacgcccgaagcgcgggatgcgttcgtcaccgaactgcttaagaaaatgacagtt gatgagaaaattggtcagctgcgcttaatcagcgtcggcccggataacccgaaagaggcgatccgcgagatgatcaaagacggtcaggttggggcgattt tcaacaccgtaacccgtcaggatatccgcgccatgcaggatcaggtgatggaattaagccgcctgaaaattcctcttttctttgcttacgacgtgctgca cggtcagcgcacggtgttcccgattagcctcggtctggcctcgtcttttaacctcgatgcagtgaaaacggtcggacgtgtctctgcttatgaagcggca gatgatggcctgaatatgacctgggcaccgatggtcgatgtctcgcgcgatccgcgctggggacgtgcttccgaaggttttggcgaagatacgtatctca cctcaacaatgggtaaaaccatggtggaagcgatgcagggtaaaagcccggcagatcgctactcggtgatgaccagcgtcaaacactttgccgcatacgg cgcggtagaaggcggtaaagagtacaacaccgtcgatatgagtccgcagcgcctgtttaatgattatatgccgccgtacaaagcggggctggacgcaggc agcggcgcggtgatggtggcgctgaactcgctgaacggcacgccagccacctccgattcctggctgctgaaagatgttctgcgcgaccagtggggcttta aaggcatcaccgtttccgatcacggtgcaatcaaagagctgattaaacatggcacggcggcagacccggaagatgcggtgcgcgtggcgctgaaatccgg aatcaacatgagcatgagcgacgagtactactcgaagtatctgcctgggttgattaaatccggcaaagtgacgatggcagagctggacgatgctgcccgc catgtactgaacgttaaatatgatatggggttgtttaacgacccatacagccatttggggccgaaagagtctgacccggtggataccaatgccgaaagcc gcctgcaccgtaaagaagcgcgtgaagtggcgcgcgaaagcttggtgttgctgaaaaaccgtctcgaaacgttaccgctgaaaaaatcggccaccattgc ggtggttgggccactggcggacagtaaacgtgacgtgatgggcagctggtccgcagccggtgttgccgatcaatccgtgaccgtactgaccgggattaaa aatgccgtcggtgaaaacggtaaagtgctgtatgccaaaggggcgaacgttaccagtgacaaaggcattatcgatttcctgaatcagtatgaagaagcgg tcaaagtcgatccgcgttcgccgcaagagatgattgatgaagcggtgcagacggcgaaacaatctgatgtggtggtggctgtagtcggtgaagcacaggg gatggcgcacgaagcctccagccggaccgatatcactattccgcaaagccaacgtgacttgattgcggcgctgaaagccaccggtaaaccgctggtgctg gtgctgatgaacgggcgtccgctggcgctggtgaaagaagatcagcaggctgatgcgattctggaaacctggtttgcggggactgaaggcggtaatgcaa ttgccgatgtattgtttggcgattacaacccgtccggcaagctgccaatgtccttcccgcgttctgtcgggcagatcccggtgtactacagccatctgaa taccggtcgcccgtataatgccgacaagccgaacaaatacacttcgcgttattttgatgaagctaacggggcgttgtatccgttcggctatgggctgagc tacaccactttcaccgtctctgatgtgaaactttctgcgccgaccatgaagcgtgacggcaaagtgactgccagcgtgcaggtgacgaacaccggtaagc gcgagggtgccacggtagtgcagatgtacttgcaggatgtgacTgcttccatgagtcgccctgtgaaacagctgaaaggctttgagaaaatcaccctgaa accgggcgaaactcagactgtcagcttcccgatcgatattgaggcgctgaagttctggaatcaacagatgaaatatgacgccgagcctggcaagttcaat gtctttatcggcactgattccgcacgcgttaagaaaggcgagtttgagttgctgggtactagtagcggccgctgcag

pVIII

• Blue is the leader sequence...

atgaaaaagtctttagtcctcaaagcctctgtagccgttgctaccctcgttccgatgctgtctttcgctgctgagggtgacgatcccgcaaaagcggcct ttaactccctgcaagcctcagcgaccgaatatatcggttatgcgtgggcgatggttgttgtcattgtcggcgcaactatcggtatcaagctgtttaagaa
attcacctcgaaagcaagctga