Yeast Sequencing Report Analysis of 41 kb of the DNA sequence from the right arm of chromosome II of Schizosaccharomyces pombe Manuel Sa ´nchez 1 , Jose ´ L. Revuelta 1 , Francisco del Rey 1 , Rhian Gwilliam 2 , Jason Skelton 2 , Carol Churcher 2 , Marie-Ade `le Rajandream 2 , Valerie Wood 2 , Bart Barrell 2 , Rachel Lyne 2 , Richard Reinhardt 3 , Katja Borzym 3 , Alfred Beck 3 , Sergio Moreno 1 and Angel Domı ´nguez 1 * 1 Departamento de Microbiologı ´a y Gene ´tica, Instituto de Microbiologı ´a Bioquı ´mica/CSIC. Universidad de Salamanca, 37071 Salamanca, Spain 2 Sanger Centre, Wellcome Trust Genome Campus, Hinxton, Cambridge CB10 1SA, UK 3 Max Planck Institut fu ¨r Molekulare Genetik, Ihnestrasse 73, D-14195 Berlin, Germany * Correspondence to: A. Domı ´nguez, Departamento de Microbiologı ´a y Gene ´tica, Instituto de Microbiologı ´a Bioquı ´mica/CSIC, Universidad de Salamanca, 37071 Salamanca, Spain. E-mail: ado@gugu.usal.es Received: 28 February 2001 Accepted: 24 April 2001 Abstract We report the complete sequence of cosmid c18A7 (41 046 bp insert), located on the right arm of chromosome II of the Schizosaccharomyces pombe genome. The sequence, which partially overlaps with cosmids SPBC4F6 and SPBC336, contains 16 open reading frames (ORFs) capable of coding for proteins of at least 100 amino acid residues in length (one partial) and one small nucleolar RNA (snoRNA). Four known genes were found: swi10 (encoding a mating-type switching protein also involved in nucleotide excision repair); dim1 (encoding a dimethyladenosine transferase); arf1 (encoding ADP-ribosylation factor 1); and pol3 (cdc6) the partial fragment, encoding the 125 kDa catalytic subunit of the DNA polymerase type B. Six ORFs similar to known proteins were found. They include a transporter of the major facilitator superfamily class, a vacuolar sorting protein, an asparagine synthase, a nuclear protein, a reticulum oxidoreductin and a heat shock protein. Each protein product of the other six ORFs has conserved domains and can be assigned a molecular, but not a biological, function. The sequence has been submitted to the EMBL database under Accession No. AL080287. Copyright # 2001 John Wiley & Sons, Ltd. Keywords: genome sequencing; Schizosaccharomyces pombe; chromosome II; swi10; arf1; dim1; pol3; enterobactin transporter; vacuolar sorting protein; asparagine synthase; oxidoreductin 1-Lb; LIM domain protein; heat shock protein; DNA helicases Introduction As participants in the European Schizosacchar- omyces pombe Genome Sequencing Project, we have been involved in the sequencing of 120 kb contained in three different cosmids. We have previously presented the sequence and a computer analysis of one cosmid, SP32F12 (Sa ´ nchez et al., 1999). Here we describe the results obtained for SPBC18A7. This fragment (41 046 bp) corresponds to the entire insert of cosmid c18A7 and is located on the right arm of chromosome II (Hoheisel et al., 1993). Cosmid c18A7 overlaps at one end with cosmid c4F6 (Accession No. AL031534) and at the other with c336 (Accession No. AL121815). ORFs identified from both cosmids are annotated with the systematic gene names of SPBC4F6 and SPBC336. Materials and methods Cosmids, plasmids and strains The DNA coordinator R. Gwilliam, The Sanger Centre, Hinxton, Cambridge, provided cosmid c18A7. This contains a 41 kb insert of chromosome II obtained by Sau3A partial digestion of Sz. pombe DNA (strain 972 h x ) and cloned into the BamHI site of the cosmid vector Lawrist4 (Hoheisel et al., 1993). The insert of cosmid c18A7 partially overlaps the insert of cosmids c4F6 (Hinxton) and c336 (Berlin). The pBluescript KS+ (Stratagene) phage- mid was used as vector for all subsequent sub- cloning and sequencing steps. The Escherichia coli strain used as host for transformation and amplifica- tion of plasmids was DH5a supE44 DlacU169(ø80 Yeast Yeast 2001; 18: 1111–1116. DOI: 10.1002 / yea.760 Copyright # 2001 John Wiley & Sons, Ltd.