From 2007.igem.org

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Revision as of 15:41, 16 October 2007

Gene Pho80 CODING SEQUENCE

1 ATGGAAAGCA CATCAGGAGA ACGTTCCGAA AATATACATG AGGATCAAGG

     51  GATACCAAAA GTAATTCTGC CCGCTGATTT TAATAAATGC TCTAGAACTG

    101  ACCTAGTGGT GCTCATATCA CGAATGTTAG TATCGCTGAT AGCAATCAAT

    151  GAAAATTCAG CAACAAAGAA ATCTGATGAC CAAATTACTT TAACACGATA

    201  CCATTCTAAG ATTCCTCCAA ACATATCAAT CTTCAACTAT TTCATACGAC

    251  TGACAAAGTT TTCCTCTTTA GAACATTGTG TGCTTATGAC ATCACTCTAT

    301  TATATCGATT TATTGCAAAC TGTGTATCCT GATTTTACGC TTAATTCGTT

    351  GACTGCCCAT AGGTTTTTAT TAACAGCCAC CACAGTCGCA ACAAAAGGCT

    401  TATGTGATTC GTTCTCAACA AACGCCCATT ATGCAAAAGT TGGAGGAGTA

    451  CGATGTCACG AATTGAATAT ACTGGAGAAC GATTTTTTAA AGAGAGTAAA

    501  CTACAGAATC ATTCCGCGGG ATCATAACAT TACGTTATGT AGTATAGAGC
    551  AAAAACAGAA AAAGTTTGTC ATAGATAAAA ACGCATTAGG GTCTCTCGAT

    601  TTGGATTCTT ATTCTTACGT TAATCGTCCA AAAAGTGGAT ATAATGTTCT

    651  AGATAAATAC TATCGAAGAA TAGTTCAGCT GGTGGGTTCC TTTAACGCTT

    701  CACCTGATAA GAGTAGAAAG GTTGATTATG TTCTCCCGCC AAATATTGAT

    751  ATAGTGAGTG AAAGTGGCTC CCAAACTACT CAACTAAAGG GGTCGTCATC

    801  ACCCAATTCT CACTCTTCAC AAAAGCGATA TTCTGAGGCA AAGGACGCAC

    851  ATATCTATAA CAAGCGATCA AAGCCAGATT AA

XbaI cuts in 2 sites

•91 •648

Pho80cds 882 bp e 293aa

XbaI restriction sequence TCTAGA

1)91 bp position 31-32aa SR S=TCT TCC TCA TCG

R=AGA AGG

2)648 bp position 216-217-218aa VLD V=TT GTC GTA GTG

L=TTA TTG CTT CTA CTC CTG

D=GAT GAC

5'3' Frame 1 Met E S T S G E R S E N I H E D Q G I P K V I L P A D F N K C S R T D L V V L I S R Met L V S L I A I N E N S A T K K S D D Q I T L T R Y H S K I P P N I S I F N Y F I R L T K F S S L E H C V L Met T S L Y Y I D L L Q T V Y P D F T L N S L T A H R F L L T A T T V A T K G L C D S F S T N A H Y A K V G G V R C H E L N I L E N D F L K R V N Y R I I P R D H N I T L C S I E Q K Q K K F V I D K N A L G S L D L D S Y S Y V N R P K S G Y N V L D K Y Y R R I V Q L V G S F N A S P D K S R K V D Y V L P P N I D I V S E S G S Q T T Q L K G S S S P N S H S S Q K R Y S E A K D A H I Y N K R S K P D Stop

Virtual Sequence: VIRT14212 ID VIRT14212 Unreviewed; 293 AA. AC VIRT14212; DE Translation of nucleotide sequence generated on ExPASy DE on 10-Jul-2007 by 193.43.101.20. CC -!- This virtual protein sequence will automatically be deleted CC from the server after a few days. CC 9.11 PI. DR SWISS-2DPAGE; VIRT14212; VIRTUAL. SQ SEQUENCE 293 AA; 33227 MW; 72918CAF845AE74C CRC64.

    MESTSGERSE NIHEDQGIPK VILPADFNKC SRTDLVVLIS RMLVSLIAIN ENSATKKSDD
    QITLTRYHSK IPPNISIFNY FIRLTKFSSL EHCVLMTSLY YIDLLQTVYP DFTLNSLTAH
    RFLLTATTVA TKGLCDSFST NAHYAKVGGV RCHELNILEN DFLKRVNYRI IPRDHNITLC
    SIEQKQKKFV IDKNALGSLD LDSYSYVNRP KSGYNVLDKY YRRIVQLVGS FNASPDKSRK
    VDYVLPPNID IVSESGSQTT QLKGSSSPNS HSSQKRYSEA KDAHIYNKRS KPD

PHO80 Primer sequences: Primer Name Primer Sequence (5' to 3') t93c 5'-cgctgattttaataaatgctccagaactgacctagtggtgc-3' t93c_antisense 5'-gcaccactaggtcagttctggagcatttattaaaatcagcg-3' t648c 5'-atcgtccaaaaagtggatataatgtcctagataaatactatcgaagaatag-3' t648c_antisense 5'-ctattcttcgatagtatttatctaggacattatatccactttttggacgat-3' Oligonucleotide information: Primer Name Length (nt.) Tm Duplex Energy at 65°C Energy Cost of Mismatches t93c 41 79.60°C -44.47 kcal/mole 3.0% t93c_antisense 41 79.60°C -46.01 kcal/mole 2.6% t648c 51 78.36°C -46.13 kcal/mole 0.1% t648c_antisense 51 78.36°C -42.85 kcal/mole 1.8%

 Primer-template duplexes:

Primer Name Primer-Template Duplex t93c 5'-cgctgattttaataaatgctccagaactgacctagtggtgc-3'

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cgggcgactaaaattatttacgagatcttgactggatcaccacgagt t93c_antisense gcccgctgattttaataaatgctctagaactgacctagtggtgctca

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3'-gcgactaaaattatttacgaggtcttgactggatcaccacg-5' t648c 5'-atcgtccaaaaagtggatataatgtcctagataaatactatcgaagaatag-3'

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aattagcaggtttttcacctatattacaagatctatttatgatagcttcttatcaag t648c_antisense ttaatcgtccaaaaagtggatataatgttctagataaatactatcgaagaatagttc

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3'-tagcaggtttttcacctatattacaggatctatttatgatagcttcttatc-5'

Primer sequences: Primer Name Primer Sequence (5' to 3') t93g 5'-cgctgattttaataaatgctcgagaactgacctagtggtgc-3' t93g_antisense 5'-gcaccactaggtcagttctcgagcatttattaaaatcagcg-3' Oligonucleotide information: Primer Name Length (nt.) Tm Duplex Energy at 68°C Energy Cost of Mismatches t93g 41 79.60°C -46.43 kcal/mole 1.3% t93g_antisense 41 79.60°C -46.23 kcal/mole 2.2%

 Primer-template duplexes:

Primer Name Primer-Template Duplex t93g 5'-cgctgattttaataaatgctcgagaactgacctagtggtgc-3'

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cgggcgactaaaattatttacgagatcttgactggatcaccacgagt t93g_antisense gcccgctgattttaataaatgctctagaactgacctagtggtgctca

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3'-gcgactaaaattatttacgagctcttgactggatcaccacg-5'