rc s .,ors*i4n1~ 06

Nucleic Acids Research, Vol. 18, No. 23 7147

The MYO1 gene from Saccharomyces cerevisiae: its complete nucleotide sequence Frank P.Sweeney, Felicity Z.Watts, Michael J.Pocklington and Elisha Orr Department of Genetics, Leicester University, University Road, Leicester, UK Submitted October 30, 1990

We have previously published the sequence of the 5' end of the yeast MYOJ gene (Watts et al., 1987). This gene was isolated from a X47.1 genomic library screened with a probe containing the active thiol region of the nematode myosin UNC54 (Karn .

.

et al., s 1983). Here we present its complete nucleotide sequence achieved by the dideoxy chain termination method (Sanger,

1977). Templates cloned in pUC 18 and Ml3mpl 8 were primed

with universal M 13 primers or synthetic primers for the A A +1 xv o+venA Tho o1onn+e, The o.,hlv whole gene was independenty strand. complementary

sequenced at least twice. The gene is 5553 bp long and encodes a type II myosin heavy chain of 213 kDa (Sweeney et al.,* 1990,s s in press). Putative translation start and stop simaIs are underlined.

-195

CATCATTTAGCCCAAAAGGTAATTGCGTAAACATTTGTAATTTCTTGGTGTATTTGTTTTG&TTCATTGTTTA?GAC

46

ACAAATAGGAACTGTCCACCCGTTGGTTCCACAGAGGTCSAATCTATCGCAGGTACGAATATCCGTSTATTTCCAG TTAACCCATCGACTTTTGACAAAGTAGAGAATATGTCTSGTGTAACCCATTTCAATGAGCCGTCTGTCCTTTAT^AiCCTA

-:5 TCAGTACSGTAGACAGGACAACAACTAGCTAbACCtGGCGGGCAGTCTTGCAGT TcATTC TCW TAs CCAGATGAGA.AAGAAGTTTTCGTAAA&GGGGAGCTGATGAGTACCGATATCAACAAGAATATATTTACAGGCCAAGAAGE 126 206

366

TTTATATGSAGACCATATAACASCT TACCACAACAAGCATAACAGGTTATCA =GACACAGGAG&ArCTCCC

525

ATTTTAGTCACTGGTGA&TCCGGTGCGGGCAAGAAAATACGAAAGAATGTCTACAATATCTAGCATCTATAACTTC

TGGCTCTCCTTCCAATATAGCTCCTGTTAGTGGTAGTTCTATTGTAWGAACTTCGAzATGAAAxSCTACAAAGTMCC ~~~~~~~~~~606

686 TTTKATGAACATGGTATGATCAATGGTGCGCATATCGAGTGGTACC"TTTAGGAAATCMGAAGTGTCATCAAMASC CTATCTTAGGTCTTTTGGTAATGCACAGCTGTACGAAATAACAACTCTTCAAGATTCGGTAzATTCATAAACGAAGAA ~~~~~~~~~~~~~~~~~~~~766 146 GAAAGAA&GAAATTATCATATATTTTACCAACTATTATCCGGTTTAGACGATTCTGAGTTGAAAAATCTACGCCTTJAATI 926

1. Karn,J., Brenner,S. and Barnett,L. (1983) Proc. Natl. Acad. Sci. USA 80,

TSTTGTTTTAGAACAGAGCGAATATTAGGACATTCAGGGATTATATAAGTTACCGAAGTGCAACTGAC ATTCGAAAATAATTCCTTTGAACAATTATGCATCAACTSASACAAATGAAASTACAGCAGTTCTTTAATAACCATATG ~~~~~~1406 1486 1566

1646

17 26

1806 ~~1886

12046 2126 2206

2366 2446 2526

2606 2 6 86

2766

2846

4253-4257. § *2926 2. Sanger,F., Niclden,S. and Coulson,A.R. (1977) Proc. Nal. Acad. Sci. USA 3086 3166 3246 74, 5,v63 5,v67.

3. Sweeney,F.P., Pocklington,M.J. and Orr,E. (1990) J. Muscle Res. & Cell MofL. in press.

4. Watts,F.Z., Shiels,G. and Orr,E. (1987) EMBO J. 6, 3499-3505.

CTAGGAACGTAAAAGATTACAAATTTTATCCAATTCTA&CCAGGATATTATACCAGGAATCGACGTAGAGAATTAfAAA

GAACTGCTCTCGGCATTAAGTATTATTGGGTTTTCAAAAGATCAAATAATGAGCATATTTCAAGTAGTGGCAATTASTTT ATTAsSCGGTAACATTGAGTTCGTATCAGACAGAGCAGAACAAGCATCTTTCAAAJAATGTTAGCGCCATTTGTAGCA 1016 ATTTAGGCGTGGACGAAAAAGATTTCCAAACTGCCATATTAAGGCCTAGATCAAAAGCCGGAAA&GAGTGGGTTTCACAG 1166 1246 TCCAAAAACTCAACAAAGCTAAGTTCACTCTTGAATGCCTTATCAAGAAATCTCTATGGCGGTTGTTCGGSATATATS 13 26 GGATATGATTAATSAAALACTSTGGAC CATGGGAGTG CAACSTTSGAATTACASSGTGASSGSSGATATGCTGGTTTTGAA

2286

r n EFErENCEcS

EMBL accession no. X53947

3406

GATTGATTTGATCGAAGCAAGGGGCCACGACCGTGTACTACCGTTGTTGGTAGAGGAAGCCGTTTTGCCCAA&TCACTGA

TGGAGTCATTCTACTCTAAACTGATCTCAACTTGGGACCAAAACTCTTCAAAGTTTAAACGTTCAGCATTAAAAAATGGG STTCAT TTTGAAIiGACTATGCTGGGGATGTGAATACACTGTGGAAGGCTGGOTTATCCAAAACAGATC CTTSTAAAC GATMA

TCTCTTGTCTTTGTTGTCTTCTTCACAAAACGATATCATTCAAAACTGTTCCAGCCAGAGGGCGGAAAAAATCTTCTAG TGTGTGGTGTGGAAGCCAACATCTCCAACCAAGAAGTTAAGAAATCAGCTAGGACAGTACCTTCAAGACTACATCATCA

CAACGTGAMAAAGTGAAAACATTTAACAGAAGTTTAATCTTAAG TCAATTACGTTGTAATGGTTGCTAGAGGGTATTA

GACTTGCCAGAGAAGGTTACCCAWTAGGATAGCATTCCAGAATTTTTCCAGCGGTATAGGATCTTGTATCCTGAAAT

TCAACCACCACGACTTTCAGTTCTAAATTAAAAGCCAGTACCAAACAAAACTGTCGATTTCTTCTAACGTCTTTGCAACT

GGATACAAAAGTTTATAAAATTGGAATACTAACTGTTTTTCAAAAGcTGGAGTATTGGTCAGATTTGGAAAAACAAAAAG

ATGTTAACGTGAATAATATTATGATTAAACTAACAGCAACTATACGAGGTTACACAGTAAGAAAAGAAATAACGTACCAT

CTACAAAAATTAAAGAAA&CAAGGGGATTGGTAATACCTTCAGATTATACAATAGACTGGTGAAGGAAGATCCTTGGTT

TAATTTATTTATCAGGATCAAGCCACTTTTAACATCATCCAATGACATGACCAGAACCAAAAAATTCAACGAGCAAATTA ATAAACTGAAGAACGACCTTCAAGAAATGGAATCTAAGAAGAAGTTTTTGGAAGAAAAGAACCAAAAAACTGTAACTGAG TTGGAAAATAGCCAGATCACCAAAASTCACACGAATATAMCT"GAACACTCAAGTACATATATTG"GAAGAGACCAAA

ACGTGTCATATGTGGAAATACGCAGGACTTGCTAATCCAGCGAAAGAGAA.TTAAGAAAAAGGTAGCTATAAAGATAT TAAAACCAGCGATAAACATTACAAAACAATTCATGACCTTGTTTCTAAAGGATG&AATAGCAGGAAAAACTAGAAGTT CGCAAAATCTTGAAGAAGCTCGATCAAAAAATCCAAGGCCTTCAAGAAACTATTAGAGAACGGGAGCGACCTTAGAA AGATGTCATAAACTCGAAGGGGAGAA CATCAAAGAATCTAAATAAGCTAGAAAAC GA GGAA

ATAAATCGTTCAACGATAAGTTAAGTTCTTCAGAAGAAGATCTTGACATAAAAGACGTCACTTTAGGAAAAATTCTAA CATTGCGATATCAAGACTACAATCCCTTGTAACAGAAAATTCAGATTTGCGTTCGAAAAATGAGAATTTCAAGAAAGAMA

GAGCTCTACTTTCCAAGCGA^GATGACGATGTAGCGAACATGGTATACTGCTAATCAAAAGAGACAAG

3486

AATCCAACTTACCGAGTA?AATCTAATATATCAAAAGATTAAAGAAGAATATTCCAACTTCCAAACGAGAAAGAAAGAAG AACAGGACAAAGAAAGAAATAGCCTGGTTGAGTCTCTGAACGATGAGTTAAAAGTTAAAGAATGGAAAGCTCGTTGTCAC

3726

AG TATTCAATTCTTGACAACAAGAATAAAAAAAAATACTATGACCTTCAATTAGCGTTTACTGAAA TAACTAGGAATCTAGAGAATGAAATTCAAGAGAAGAAGAACTTAATTTCTAGATTGAGATTCACTGAAACAAGACTAGCA TCTTCGTCTTTTGAGGACCAAAAGATTAAGGCACAAATGAAGAAATTAAAAAATTGATCCAGGATATGGACCCTAGTAT TCCTTTGGACAGTATTCTAAATGAGCCGCTAGATA&CTGCCTGACAAAGAGTCTGATATTAACAAATGGTAATGCTTCGG TCGATTATTTAAAAAGACAATTGGATASCGAAACAAGAGCTCACTACGATGCAGAAAATGCCATATCSGCTTTACACAGT AATTTAGAAAGATCCAAGGGGAAAGCTCCCTGTCATCTTCT0ATATTTACAAACT0AAGTTCGAAGCCAGTGAAGAAAG AGTCAAATCCTTGGAAGACAAGCTAAAAACCATGCCTTTACGTGATCGAACAAATTTACCTGTCGGACATATTATAAGA ACCGTGATAGCATTTCAAATATGAAGAAGAAATTCGGTATTATAAACTTGAAAACTACAAGCTCCAGAAATATTAAmT GAATCAAATGGAAA'TTGAGCCAACTCACTCTTGACTTGAGGCAATCAAAATCCAAAGAAGCCCTACTTCCATTGAAGAG AGTCCAAGATGTAAAGAAAATCCTGAGTGATGTTTTGGCCCATTTAAAGGAGCGATTGAGCGCTGTAGAGATAGATCCC

3566 3806

3886 3966 4046

4126

4206

4286 4366

4446 4S26 4606 4686 4766

4846 4926 5006 5086 5166

5246

AATATACGGATGAGATTAATAGATTAAAGGAAGAATTGAACTGCTCTTTAAAAGCTGAA.ACAAACTTAAAAAAGAATTT GCAACCCTTAAGTACAAACTGGAAACTTCGACTAATGATTCTGAAGCAAAAATCTCCTCTOMGCAGCTCGATCATTATA CAAAAGTGGTGGAAATGTTGACAACGAAAAAGATGCTATTTCTCTTGCAGAAAAAGAACTTTATCAGAAATACGAGGCAC TCAATACTGAATGCGAGTCTCTTAAAGGAAAGATAGTGTCTTTACTAAAATCAAGCAGGAACTGGAATCTGACTTAmAT CAAAAGACTGATGCGCTATCAAGTTCCACACAAAGAATAAAGAGATTACTGAAAAAATCATATCTGGAGGAAATAC AACTGCAAATGGAACAAAATTCAAGGAATGGGAGTTGGTTAAGACATTACAGGCCAGCTGCAATGGATATAAAGATAAGT TGATGATGAAGCAGAAGAATATTGATTTATATGAAGAAAATCAAACTTTACAAAAGCTCAACACCGAGTTACAGCTTCM CTAAAAAATTTGCATGAAACATTATCAGACACTACTGAAAAAAACGCATGGTTATCAAAGATTCATGAATGGAAAATA GGmAGCCTAGAAACGGACTTGAAATATGAAGAAATGAAAMGAACAAAGATTTGGAGAGAGCAGTGGAAGAGTTACAAA

CTAAGAACTCCCAACAAACAGATGTAATAGAACTAGCGAATAAAAATAGAAGCGAATTTGAGCAAGCTACTTTGAAATAT

5326

GAGGCTCAAATCTCTGACTTAGAAAAGTACATTTCTCAACAAGAGCTTGAGATGAAAAAATGGATTAGAGATAATTCTTC

5486

ATGAATCTACCATGATAGGCTCGAAAAATATTGATAGTAACAATGCACAGAGTAAMAATTTCAGTZMCGAACTTTTTTT

5406 5166

5646 S726

5806 5886

TTACCGCGACAAAGTGCAGGAAATGGCCCAAGAAATTG&GAATTCTGGAAAGCGGTTAGATGCAGATGATCTGAGCCGTT

TAAAAATTTGGAAGACTTATATCCTTTATTTAAAAITAASTJGATGAGATGTTTTGTATATACAGAATGAGAATATGCA

TTATTAACAGAAAAAGCGACAGAGCGTTlTATCfAAGTGGCATGAACTCTTTCGACCCGTATCCTTCGATAGTAGTCTT AUAGTGGTCATAAATAAGAAAATAACCGATTCAATAGATAAGCAGCGGAAATAAAGTCTTTTTCCTATCTTGAAATCTC GAG CAAAATAAAAGTAATAAAGTACTGCTAAAGTTGTTCAAAACCAACCCTTCTTCTTGGGACTATCGTTSTSGGAG CTTGTACCCAATCCATATGCC?TCAGTA