IGEM:UCSF/2007/Plasmids
Expected sequences
Below you have the sequences of the different parts in the right order, as well as the adaptor sequences between parts. You will need to arrange them accordingly to your particular constructs, so that you will have 1-2-3-4-5-6-7-8.
1-Promoter sequence (variable, depends on particular acceptor)
2-ATG-XhoI-“A” and 2 adaptor bases: ATGCTCGAGGGAGCT
3-mCherry:
GTGAGCAAGGGCGAGGAGGATAACATGGCCATCATCAAGGAGTTCATGCGCTTCAAGGTGCACATGGAGGGCTCCGTGAACGGCCACG AGTTCGAGATCGAGGGCGAGGGCGAGGGCCGCCCCTACGAGGGCACCCAGACCGCCAAGCTGAAGGTGACCAAGGGTGGCCCCCTGCC CTTCGCCTGGGACATCCTGTCCCCTCAGTTCATGTACGGCTCCAAGGCCTACGTGAAGCACCCCGCCGACATCCCCGACTACTTGAAG CTGTCCTTCCCCGAGGGCTTCAAGTGGGAGCGCGTGATGAACTTCGAGGACGGCGGCGTGGTGACCGTGACCCAGGACTCCTCCCTGC AGGACGGCGAGTTCATCTACAAGGTGAAGCTGCGCGGCACCAACTTCCCCTCCGACGGCCCCGTAATGCAGAAGAAGACCATGGGCTG GGAGGCCTCCTCCGAGCGGATGTACCCCGAGGACGGCGCCCTGAAGGGCGAGATCAAGCAGAGGCTGAAGCTGAAGGACGGCGGCCAC TACGACGCTGAGGTCAAGACCACCTACAAGGCCAAGAAGCCCGTGCAGCTGCCCGGCGCCTACAACGTCAACATCAAGTTGGACATCA CCTCCCACAACGAGGACTACACCATCGTGGAACAGTACGAACGCGCCGAGGGCCGCCACTCCACCGGCGGCATGGACGAGCTGTACAAG
4-Adaptor bases, followed by “B” and another adaptor base: GGTAGTTCCCTA
5-Effectors
For YopH:
aacttatcattaagcgatcttcatcgtcaggtatctcgattggtgcagcaagagagcggtgattgtaccgggaaattaagaggtaacgt tgctgccaataaagaaactacctttcaaggtttgaccatagccagtggagccagagagtcagaaaaagtatttgctcaaactgtactaa gccacgtagcaaatgttgttctaactcaagaagataccgctaagctattgcaaagtacggtaaagcataatttgaataattatgactta agaagtgtcggcaatggtaatagtgtacttgtcagtttacgtagtgaccaaatgacactacaagacgccaaagtgctgttggaggccgc attgcgacaagagtcgggagcgagggggcatgtatcatctcattcacattcagcccttcacgcaccgggaaccccggtgcgtgaaggac tgcgttcacatctagaccccagaactccaccgttgccaccgcgtgaacgaccacacacttctggccatcacggggctggcgaagccaga gccaccgcaccaagcactgtttctccttatggcccagaagcgcgcgcagaactcagcagccgcctcaccacattgcgcaatacgctggc gccagcaacgaatgatccgcgttacttacaagcctgcggcggtgaaaagctaaaccgatttagagatattcaatgctgtcggcaaaccg cagtacgcgccgatcttaatgccaattacatccaggtcggtaacactcgtaccatagcgtgccagtatccgctacaatctcaacttgaa agccatttccgtatgctggcagaaaaccgaacgccagtgttggctgttttagcgtccagttctgagatagccaatcaaagattcggtat gccagattatttccgccagagtggtacctatggcagtatcactgtagagtctaaaatgactcagcaagttggtctcggtgacgggatta tggcagatatgtatactttaacgattcgtgaagcgggtcaaaaaacaatctctgttcctgtggttcatgttggcaattggcccgatcag accgcagtcagctctgaagttaccaaggcactcgcttcactggtagatcaaacagcagaaacaaaacgcaatatgtatgaaagcaaagg aagttcagcggtaggagatgactccaaattacggccggtaatacattgccgtgcgggtgttggccgtactgcgcaactgattggcgcaa tgtgcatgaatgatagtcgtaatagtcagttaagcgtagaagatatggtcagccaaatgcgagtacaaagaaatggtattatggtacaa aaagatgagcaacttgatgttctgattaagttggctgaaggacaagggcgaccattattaaatagc
For YopJ:
atcggaccaatatcacaaataaatatctccggtggcttatcagaaaaagagaccagttctttaatcagtaatgaagagcttaaaaatat cataacacagttggaaactgatatatcggatggatcctggttccataaaaattattcacgtatggatgtagaagtcatgcccgcattgg taatccaggcgaacaataaatatccggaaatgaatcttaatcttgttacatctccattggacctttcaatagaaataaaaaacgtcata gaaaatggagttagatcttcccgcttcataattaacatgggggaaggtggaatacatttcagtgtaattgattacaaacatataaatggg aaaacatctctgatattgtttgaaccagcaaactttaacagtatggggccagcgatgctggcaataaggacaaaaacggctattgaacgt tatcaattacctgattgccatttctccatggtggaaatggatattcagcgaagctcatctgaatgtggtatttttagtttggcactggca aaaaaactttacatcgagagagatagcctgttgaaaatacatgaagataatataaaaggtatattaagtgatggtgaaaatcctttaccc cacgataagttggacccgtatctcccggtaactttttacaaacatactcaaggtaaaaaacgtcttaatgaatatttaaatactaacccg cagggagttggtactgttgttaacaaaaaaaatgaaaccatcgttaatagatttgataacaataaatccattgtagatggaaaggaatta tcagtttcggtacataaaaagagaatagctgaatataaaacacttctcaaagta
For OspF:
CCAATTAAAAAGCCTTGTTTGAAATTGAATTTAGATTCTTTGAACGTTGTTAGATCAGAAATTCCACAAATGTTATCTGCTAATGAAAGAT TGAAAAATAATTTTAACATTTTATATAATCAAATAAGACAGTACCCAGCTTATTATTTTAAAGTTGCTTCTAATGTTCCTACTTACTCTGA TATATGTCAATTTTTCTCAGTTATGTATCAAGGTTTTCAGATAGTTAATCATTCTGGTGATGTTTTTATTCATGCTTGTAGAGAAAATCCA CAATCTAAAGGTGATTTTGTTGGTGACAAGTTCCATATTTCTATAGCTAGAGAACAAGTTCCATTGGCATTTCAAATTTTATCTGGTTTGTT ATTTTCTGAGGACTCTCCTATTGACAAATGGAAGATAACTGATATGAATAGAGTTTCTCAACAGTCAAGAGTTGGTATTGGTGCTCAATTTAC TTTGTATGTTAAATCTGATCAAGAATGTTCTCAGTATTCAGCTTTATTGTTGCATAAAATTAGACAATTCATAATGTGCTTGGAATCTAATTT GTTAAGATCTAAGATTGCTCCAGGTGAATATCCTGCATCTGATGTTAGACCAGAGGATTGGAAGTACGTTTCATATAGAAATGAATTGAGATC TGACAGAGATGGTTCTGAAAGACAAGAGCAGATGTTAAGAGAAGAACCTTTTTATAGATTGATGATTGAG
6-Adaptor bases, followed by “C” followed by another adaptor: GGTAGCGAT
7-Zippers: For RR:
Ggttctggttctggttctggttctctggaaatccgtgcggcgttcctggaaaaagaaaacaccgcgctgcgtacccgtgcggcggaactgcg taaacgtgttggtcgttgccgtaacatcgtttctaaatacgaaacccgttacggtccgctg
For EE:
Ggttccggtagcggttccggttctatcactattagggctgcttttttggaaaaagaaaatactgctttgagaactgaaattgctgaattgga aaaagaagttggtagatgtgaaaatattgtttctaaatatgaaactagatatggtccattg
For RR(T):
Ggatctggatcgggatctgatccagatttggaaattagagctgcttttttgagacaaagaaatactgctttgagaactgaagttgctgaatt ggaacaagaagttcaaagattggaaaatgaagtttctcaatatgaaactagatatggtccattgggtggtggtaaa
For EE(T):
ggatctggatcgggatctgatccagatttggaaattgaagctgcttttttggaaagagaaaatactgctttggaaactagagttgctgaatt gagacaaagagttcaaagattgagaaatagagtttctcaatatagaactagatatggtccattgggtggtggtaaa
8-Final adaptors, followed by “D”, followed by BamHI, 3 STOPS in three different frames, then NotI, the Adh terminator and SacI:
TAATGCGGGATCCTGAGTAAATAAGCGGCCGCGCGAATTTC
TTATGATTTATGATTTTTATTATTAAATAAGTTATAAAAAAAAATAAGTGTAT
ACAAATTTTAAAGTGACTCTTAGGTTTTAAAACGAAAATTCTTATTCTTGAGT
AACTCTTTCCTGTGGTCAGGTTGCTTTCTCAGGTATAGCATGAGGTCGCTCTT
ATTGACCACACCGAGCTC
pFig1
TTGAATGATCAACCAAACGCCGATATAGTCACTATTGGTGATAAAATTAAAATGCAGCTATTGAGAACCCATCCTAACAACATTAAATTGTCTATTAATGGAATTGGTAAAGATGCCCCAACTTTCCAAGAATCTGCTTTGATTGCCGATAAGTTATTGAGTGTCATGAAGGCCGGCACTTACCCAAAGATTTCCATTTTCTACAATGACCCAGTGTCTTCCCTATCTTTTGAACCATCTGAAAAACCGATCTTTAACGCCAAGACCATTGAACAATCCCCATCATTCGGCAAATTTGAGATCGACACGGACGCAAACGTTCCAAGAGATTTGTTTGAATATACTTTGGCTAACCAAATGTTGACAGCAATGGCTCAAGGTTATGCTGCTGAAATTTCCGCCAGAAGAAACGCTATGGATAACGCTTCCAAGAATGCCGGTGATATGATCAATCGTTACTCTATCTTGTACAACAGAACAAGACAAGCTGTCATTACTAATGAACTGGTTGATATTATTACTGGTGCTTCCTCTTTGGGATGATAAAAAAATCACCCTGCATTGCCTCTTTATTTGACGTTGTTTTTGTAGAACATGAAACGAATTTTGACTTGATGAGACGAAGTATATATCCAAAGAATACCTTAAATAGAAAAGGAAAGATAATAAATACTAAACACTACTATATATTCAGGTAAAATACAAAAATTATAACATTTTTAAAACTTTTTTTTTTTGAAAGTCCTTCTCGCTTTAGGATTTTTTCCCATTAAGATTATGATGGTTTCATGTATGTGTCAGTTAAAAAAAATATGGCTAAGTAGCAATGAAACGAACCAAGAAAATGAAAACAATATATAGTGCTGTTGAAATAACAAAGACATTGGTATATATTTGTAAAATGTCTGTTAAATGTTTTTTATCTCAGGTTCTTGCTTGTCTTTGGTAGAAGAAATTATAGTAAACAAACAAACAAACAAACAAAAAAAAAAAAAAAAAA
pAdh
TTAAAACAAGAAGAGGGTTGACTACATCACGATGAGGGGGATCGAAGAAATGATGGTAAATGAAATAGGAAATCAAGGAGCATGAAGGCAAAAGACAAATATAAGGGTCGAACGAAAAATAAAGTGAAAAGTGTTGATATGATGTATTTGGCTTTGCGGCGCCGAAAAAACGAGTTTACGCAATTGCACAATCATGCTGACTCTGTGGCGGACCCGCGCTCTTGCCGGCCCGGCGATAACGCTGGGCGTGAGGCTGTGCCCGGCGGAGTTTTTTGCGCCTGCATTTTCCAAGGTTTACCCTGCGCTAAGGGGCGAGATTGGAGAAGCAATAAGAATGCCGGTTGGGGTTGCGATGATGACGACCACGACAACTGGTGTCATTATTTAAGTTGCCGAAAGAACCTGAGTGCATTTGCAACATGAGTATACTAGAAGAATGAGCCAAGACTTGCGAGACGCGAGTTTGCCGGTGGTGCGAACAATAGAGCGACCATGACCTTGAAGGTGAGACGCGCATAACCGCTAGAGTACTTTGAAGAGGAAACAGCAATAGGGTTGCTACCAGTATAAATAGACAGGTACATACAACACTGGAAATGGTTGTCTGTTTGAGTACGCTTTCAATTCATTTGGGTGTGCACTTTATTATGTTACAATATGGAAGGGAACTTTACACTTCTCCTATGCACATATATTAATTAAAGTCCAATGCTAGTAGAGAAGGGGGGTAACACCCCTCCGCGCTCTTTTCCGATTTTTTTCTAAACCGTGGAATATTTCGGATATCCTTTTGTTGTTTCCGGGTGTACAATATGGACTTCCTCTTTTCTGGCAACCAAACCCATACATCGGGATTCCTATAATACCTTCGTTGGTCTCCCTAACATGTAGGTGGCGGAGGGGAGATATACAATAGAACAGATACCAGACAAGACATAATGGGCTAAACAAGACTACACCAATTACACTGCCTCATTGATGGTGGTACATAACGAACTAATACTGTAGCCCTAGACTTGATAGCCATCATCATATCGAAGTTTCACTACCCTTTTTCCATTTGCCATCTATTGAAGTAATAATAGGCGCATGCAACTTCTTTTCTTTTTTTTTCTTTTCTCTCTCCCCCGTTGTTGTCTCACCATATCCGCAATGACAAAAAAATGATGGAAGACACTAAAGGAAAAAATTAACGACAAAGACAGCACCAACAGATGTCGTTGTTCCAGAGCTGATGAGGGGTATCTCGAAGCACACGAAACTTTTTCCTTCCTTCATTCACGCACACTACTCTCTAATGAGCAACGGTATACGGCCTTCCTTCCAGTTACTTGAATTTGAAATAAAAAAAAGTTTGCTGTCTTGCTATCAAGTATAAATAGACCTGCAATTATTAATCTTTTGTTTCCTCGTCATTGTTCTCGTTCCCTTTCTTCCTTGTTTCTTTTTCTGCACAATATTTCAAGCTATACCAAGCATACAATCAACTATCTCATATACA
pCyc
cagatccgccaggcgtgtatatatagcgtggatggccaggcaactttagtgctgacacatacaggcatatatatatgtgtgcgacgacacatgatcatatggcatgcatgtgctctgtatgtatataaaactcttgttttcttcttttctctaaatattctttccttatacattaggacctttgcagcataaattactatacttctatagacacacaaacacaaatacacacactaaattaata
pSte5
CACGAAGTGACTGACAGAATACTGACATCAGCTGATTTCTCATAGAGCTGTTTCTCTGATAACACGTTGTTTGAACATCGACAAGATGAAAATCTAGAAGTATCAAGTTTCCTTTAAAGGGATATATAACAGATTCTAAAACTGACAGAAATATTTCGAGTGAAGAAGAAGCGTTAAATATTGGATCTTTCCGCAGTTCTACTCTGATACATTTTTGAAGTAGGAGAGTCATTTAGAAGGCGTATTGCTCAATAGTAGAAAGCAGGCCTGTGCACATGAATTAATTAAAAAATATAAAGGTAGTGATTAGACGACACATGTCCATAGGTAACCTGTCATAATTTTGAACAATTTCCCTTCTTTTCTTTTTTTTTTTTGGGTGCGGCGATATGTAGCTTGTTAATTTACACATCATGTACTTTTCTGCATCAAAATATGAAAGGCGATAGTAGCTAAAGAAAATACCGAGAATTTCCTCGAAAAGTTGACGACAAAAGAAAGGCATAAAAAAGTAATTTGAAAATATTTTAAAACTGTTTTAACCCATCTAGCATCCGCGCTAAAAAAGGAAGATACAGGATACAGCGGAAACAACTTTTAA