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
//
