LOCUS PWJ716.GB 6141 BP DS-DNA CIRCULAR SYN 17-FEB-1999 DEFINITION K. lactis URA3 gene cloned into a S. cerevisiae CEN/ARS vector ACCESSION Not submitted as of 4/2002 KEYWORDS cloning-free allele replacement SOURCE - REFERENCE 1 AUTHORS Naz Erdeniz, Uffe H. Mortensen, and Rodney Rothstein TITLE Cloning-Free PCR-Based Allele Replacement Methods JOURNAL Genome Research 7(12):1174-83 FEATURES Location/Qualifiers CDS 388..1059 /note="TRP1" primer_bind complement(3356..3379) /note="d-Kl" primer_bind 1731..1754 /note="c-Kl" primer_bind complement(3211..3251) /note="b adaptamer" primer_bind 1931..1970 /note="a adaptamer" CDS complement(2145..2948) /note="K.lactis-URA3" misc_structure 5559..6076 /note="ARSH1" misc_structure 5559..5659 /note="CEN" primer_bind 2396..2420 /note="KLi-3'" primer_bind complement(2819..2843) /note="KLi-5'" CDS complement(>4567..5427) /note="AMP-R (bla)" conflict 5131..5131 /note="This position was C in Naz's strider file, but G in the pRS414 sequence. Bob 2/99" BASE COUNT 1703 A 1367 C 1409 G 1662 T 0 OTHER ORIGIN - 1 GCCGGGAGCA GACAAGCCCG TCAGGGCGCG TCAGCGGGTG TTGGCGGGTG TCGGGGCTGG 61 CTTAACTATG CGGCATCAGA GCAGATTGTA CTGAGAGTGC ACCATAAACG ACATTACTAT 121 ATATATAATA TAGGAAGCAT TTAATAGACA GCATCGTAAT ATATGTGTAC TTTGCAGTTA 181 TGACGCCAGA TGGCAGTAGT GGAAGATATT CTTTATTGAA AAATAGCTTG TCACCTTACG 241 TACAATCTTG ATCCGGAGCT TTTCTTTTTT TGCCGATTAA GAATTAATTC GGTCGAAAAA 301 AGAAAAGGAG AGGGCCAAGA GGGAGGGCAT TGGTGACTAT TGAGCACGTG AGTATACGTG 361 ATTAAGCACA CAAAGGCAGC TTGGAGTATG TCTGTTATTA ATTTCACAGG TAGTTCTGGT 421 CCATTGGTGA AAGTTTGCGG CTTGCAGAGC ACAGAGGCCG CAGAATGTGC TCTAGATTCC 481 GATGCTGACT TGCTGGGTAT TATATGTGTG CCCAATAGAA AGAGAACAAT TGACCCGGTT 541 ATTGCAAGGA AAATTTCAAG TCTTGTAAAA GCATATAAAA ATAGTTCAGG CACTCCGAAA 601 TACTTGGTTG GCGTGTTTCG TAATCAACCT AAGGAGGATG TTTTGGCTCT GGTCAATGAT 661 TACGGCATTG ATATCGTCCA ACTGCATGGA GATGAGTCGT GGCAAGAATA CCAAGAGTTC 721 CTCGGTTTGC CAGTTATTAA AAGACTCGTA TTTCCAAAAG ACTGCAACAT ACTACTCAGT 781 GCAGCTTCAC AGAAACCTCA TTCGTTTATT CCCTTGTTTG ATTCAGAAGC AGGTGGGACA 841 GGTGAACTTT TGGATTGGAA CTCGATTTCT GACTGGGTTG GAAGGCAAGA GAGCCCCGAA 901 AGCTTACATT TTATGTTAGC TGGTGGACTG ACGCCAGAAA ATGTTGGTGA TGCGCTTAGA 961 TTAAATGGCG TTATTGGTGT TGATGTAAGC GGAGGTGTGG AGACAAATGG TGTAAAAGAC 1021 TCTAACAAAA TAGCAAATTT CGTCAAAAAT GCTAAGAAAT AGGTTATTAC TGAGTAGTAT 1081 TTATTTAAGT ATTGTTTGTG CACTTGCCTA TGCGGTGTGA AATACCGCAC AGATGCGTAA 1141 GGAGAAAATA CCGCATCAGG AAATTGTAAA CGTTAATATT TTGTTAAAAT TCGCGTTAAA 1201 TTTTTGTTAA ATCAGCTCAT TTTTTAACCA ATAGGCCGAA ATCGGCAAAA TCCCTTATAA 1261 ATCAAAAGAA TAGACCGAGA TAGGGTTGAG TGTTGTTCCA GTTTGGAACA AGAGTCCACT 1321 ATTAAAGAAC GTGGACTCCA ACGTCAAAGG GCGAAAAACC GTCTATCAGG GCGATGGCCC 1381 ACTACGTGAA CCATCACCCT AATCAAGTTT TTTGGGGTCG AGGTGCCGTA AAGCACTAAA 1441 TCGGAACCCT AAAGGGAGCC CCCGATTTAG AGCTTGACGG GGAAAGCCGG CGAACGTGGC 1501 GAGAAAGGAA GGGAAGAAAG CGAAAGGAGC GGGCGCTAGG GCGCTGGCAA GTGTAGCGGT 1561 CACGCTGCGC GTAACCACCA CACCCGCCGC GCTTAATGCG CCGCTACAGG GCGCGTCGCG 1621 CCATTCGCCA TTCAGGCTGC GCAACTGTTG GGAAGGGCGA TCGGTGCGGG CCTCTTCGCT 1681 ATTACGCCAG CTGGCGAAAG GGGGATGTGC TGCAAGGCGA TTAAGTTGGG TAACGCCAGG 1741 GTTTTCCCAG TCACGACGTT GTAAAACGAC GGCCAGTGAG CGCGCGTAAT ACGACTCACT 1801 ATAGGGCGAA TTGGGTACCG GGCCCCCCCT CGAGGTCGAC GGTATCGATA AGCTTGATAT 1861 CGAATTCCTG CAGCCCCTAG ATGCATGCTC GAGCGGCCGC CAGTGTGATG GATATCTGCA 1921 GAATTCGGCT CATGGTGGTC AGCTGGAATT CGATGATGTA GTTTCTGGTT TTTAAACTCT 1981 CACATAATGA AAGAGAGAGA GAGAAGCAAA CAAAAAATTT TCAGTTCAAT ACAACAGATC 2041 ACGTGATCTT TTGTAAGATG AAGTTGAAGT GAGTGTTGCA CCGTGCCAAT GCAGGTGGCT 2101 ATTAGATTAA ATATGTGATT TGTTCTATTA AGTTTCCTGT ATAATTAATG GGGAGCGCTG 2161 ATTCTCTTTT GGTACGCTTC CCATCCAGCA TTTCTGTATC TTTCACCTTC AACCTTAGGA 2221 TCTCTACCCT TGGCGAAAAG TCCTCTGCCA ACAATGATGA TATCTGATCC ACCACTTACA 2281 ACTTCGTCGA CGGTTCTGTA CTGCTGACCC AATGCATCGC CTTTGTCGTC TAAACCTACA 2341 CCTGGGGTCA TGATTAGCCA ATCAAACCCT TCTTCTCTTC CTCCCATATC GTTCTGAGCA 2401 ATGAACCCAA TAACGAAATC TTTATCACTC TTTGCAATAT CAACGGTACC CTTAGTATAT 2461 TCACCGTGTG CTAGAGAACC CTTGGAAGAC AATTCAGCAA GCATCAATAA TCCCCTTGGT 2521 TCTTTGGTGA CCTCTTGCGC ACCTTGTTTC AAGCCAGCAA CAATACCAGC ACCAGTAACC 2581 CCGTGGGCGT TGGTGATATC AGACCATTCT GCGATACGGT AAACGCCCGA TGTATATTGT 2641 AATTTGACTG TGTTACCGAT ATCGGCGAAT TTTCTGTCCT CAAATATCAA GAACTTGTAT 2701 TTCTCTGCCA ATGCTTTCAA TGGAACGACA GTACCCTCAT AACTGAAATC ATCCAAGATA 2761 TCAACGTGTG TTTTCAAAAG GCAAATGTAT GGACCCAACG TTTCAACAAG TTTCAATAGC 2821 TCATCAGTCG AACGAACGTC AAGAGAAGCA CACAAATTGG TCTTCTTTTC ATCCATTAAA 2881 CGTAAAAGTT TCGATGCAAC CGGACTTGCA TGAGTCTCAG CTCTACTGGT ATATGATTTT 2941 GTGGACATGG TGCAACTAAT TGACGGGAGT GTATTGACGC TGGCGTACTG GCTTTCACAA 3001 AATGGCCCAA TCACAACCAC ATCTTAGATA GTTGAAATGA CTTTAGATAA CATCAATTGA 3061 GATGAGCTTA ATCATGTCAA AGCTAAAAGT GTCACCATGA ACGACAATTC TTAAGCAAAT 3121 CACGTGATAT AGATCCACGA ATAACCACCA TTTGATGCTC GAGGCAAGTA ATGTGTGTAA 3181 AAAAATGCGT TACCACCATC CAATGCAGAC CGATCTTCTA CCCAGAATCA CGATCCCCGG 3241 GAATTGCCAT GAAGCCGAAT TCCAGCACAC TGGCGGCCGT TACTAGTGGA TCCGAGCTCG 3301 GTACCAAGCT GGGGGATCCA CTAGTTCTAG AGCGGCCGCC ACCGCGGTGG AGCTCCAGCT 3361 TTTGTTCCCT TTAGTGAGGG TTAATTGCGC GCTTGGCGTA ATCATGGTCA TAGCTGTTTC 3421 CTGTGTGAAA TTGTTATCCG CTCACAATTC CACACAACAT AGGAGCCGGA AGCATAAAGT 3481 GTAAAGCCTG GGGTGCCTAA TGAGTGAGGT AACTCACATT AATTGCGTTG CGCTCACTGC 3541 CCGCTTTCCA GTCGGGAAAC CTGTCGTGCC AGCTGCATTA ATGAATCGGC CAACGCGCGG 3601 GGAGAGGCGG TTTGCGTATT GGGCGCTCTT CCGCTTCCTC GCTCACGACT CGCTGCGCTC 3661 GGTCGTTCGC CTGCGGCGAG CGGTATCAGC TCACTCAAAG GCGGTAATAC GGTTATCCAC 3721 AGAATCAGGG GATAACGCAG GAAAGAACAT GTGAGCAAAA GGCCAGCAAA AGGCCAGGAA 3781 CCGTAAAAAG GCCGCGTTGC TGGCGTTTTT CCATAGGCTC CGCCCCCCTG ACGAGCATCA 3841 CAAAAATCGA CGCTCAAGTC AGAGGTGGCG AAACCCGACA GGACTATAAA GATACCAGGC 3901 GTTTCCCCCT GGAAGCTCCC TCGTGCGCTC TCCTGTTCCG ACCCTGCCGC TTACCGGATA 3961 CCTGTCCGCC TTTCTCCCTT CGGGAAGCGT GGCGCTTTCT CATAGCTCAC GCTGTAGGTA 4021 TCTCAGTTCG GTGTAGGTCG TTCGCTCCAA GCTGGGCTGT GTGCACGAAC CCCCCGTTCA 4081 GCCCGACCGC TGCGCCTTAT CCGGTAACTA TCGTCTTGAG TCCAACCCGG TAAGACACGA 4141 CTTATCGCCA CTGGCAGCAG CCACTGGTAA CAGGATTAGC AGAGCGAGGT ATGTAGGCGG 4201 TGCTACAGAG TTCTTGAAGT GGTGGCCTAA CTACGGCTAC ACTAGAAGGA CAGTATTTGG 4261 TATCTGCGCT CTGCTGAAGC CAGTTACCTT CGGAAAAAGA GTTGGTAGCT CTTGATCCGG 4321 CAAACAAACC ACCGCTGGTA GCGGTGGTTT TTTTGTTTGC AAGCAGCAGA TTACGCGCAG 4381 AAAAAAAGGA TCTCAAGAAG ATCCTTTGAT CTTTTCTACG GGGTCTGACG CTCAGTGGAA 4441 CGAAAACTCA CGTTAAGGGA TTTTGGTCAT GAGATTATCA AAAAGGATCT TCACCTAGAT 4501 CCTTTTAAAT TAAAAATGAA GTTTTAAATC AATCTAAAGT ATATATGAGT AAACTTGGTC 4561 TGACAGTTAC CAATGCTTAA TCAGTGAGGC ACCTATCTCA GCGATCTGTC TATTTCGTTC 4621 ATCCATAGTT GCCTGACTCC CCGTCGTGTA GATAACTACG ATACGGGAGG GCTTACCATC 4681 TGGCCCCAGT GCTGCAATGA TACCGCGAGA CCCACGCTCA CCGGCTCCAG ATTTATCAGC 4741 AATAAACCAG CCAGCCGGAA GGGCCGAGCG CAGAAGTGGT CCTGCAACTT TATCCGCCTC 4801 CATCCAGTCT ATTAATTGTT GCCGGGAAGC TAGAGTAAGT AGTTCGCCAG TTAATAGTTT 4861 GCGCAACGTT GTTGCCATTG CTACAGGCAT CGTGGTGTCA CGCTCGTCGT TTGGTATGGC 4921 TTCATTCAGC TCCGGTTCCC AACGATCAAG GCGAGTTACA TGATCCCCCA TGTTGTGCAA 4981 AAAAGCGGTT AGCTCCTTCG GTCCTCCGAT CGTTGTCAGA AGTAAGTTGG CCGCAGTGTT 5041 ATCACTCATG GTTATGGCAG CACTGCATAA TTCTCTTACT GTCATGCCAT CCGTAAGATG 5101 CTTTTCTGTG ACTGGTGAGT ACTCAACCAA GTCATTCTGA GAATAGTGTA TGCGGCGACC 5161 GAGTTGCTCT TGCCCGGCGT CAATACGGGA TAATACCGCG CCACATAGCA GAACTTTAAA 5221 AGTGCTCATC ATTGGAAAAC GTTCTTCGGG GCGAAAACTC TCAAGGATCT TACCGCTGTT 5281 GAGATCCAGT TCGATGTAAC CCACTCGTGC ACCCAACTGA TCTTCAGCAT CTTTTACTTT 5341 CACCAGCGTT TCTGGGTGAG CAAAAACAGG AAGGCAAAAT GCCGCAAAAA AGGGAATAAG 5401 GGCGACACGG AAATGTTGAA TACTCATACT CTTCCTTTTT CAATATTATT GAAGCATTTA 5461 TCAGGGTTAT TGTCTCATGA GCGGATACAT ATTTGAATGT ATTTAGAAAA ATAAACAAAT 5521 AGGGGTTCCG CGCACATTTC CCCGAAAAGT GCCACCTGGG TCCTTTTCAT CACGTGCTAT 5581 AAAAATAATT ATAATTTAAA TTTTTTAATA TAAATATATA AATTAAAAAT AGAAAGTAAA 5641 AAAAGAAATT AAAGAAAAAA TAGTTTTTGT TTTCCGAAGA TGTAAAAGAC TCTAGGGGGA 5701 TCGCCAACAA ATACTACCTT TTATCTTGCT CTTCCTGCTC TCAGGTATTA ATGCCGAATT 5761 GTTTCATCTT GTCTGTGTAG AAGACCACAC ACGAAAATCC TGTGATTTTA CATTTTACTT 5821 ATCGTTAATC GAATGTATAT CTATTTAATC TGCTTTTCTT GTCTAATAAA TATATATGTA 5881 AAGTACGCTT TTTGTTGAAA TTTTTTAAAC CTTTGTTTAT TTTTTTTTCT TCATTCCGTA 5941 ACTCTTCTAC CTTCTTTATT TACTTTCTAA AATCCAAATA CAAAACATAA AAATAAATAA 6001 ACACAGAGTA AATTCCCAAA TTATTCCATC ATTAAAAGAT ACGAGGCGCG TGTAAGTTAC 6061 AGGCAAGCGA TCCGTCCTAA GAAACCATTA TTATCATGAC ATTAACCTAT AAAAATAGGC 6121 GTATCACGAG GCCCTTTCGT C //