LOCUS pAsylum 7776 bp DNA circular SYN 17-Jun-2003 DEFINITION . FEATURES Location/Qualifiers rep_origin complement(55..643) /note="MB1 Ori" misc_feature complement(573..597) /note="Ori hairpin" 3'UTR complement(644..718) /note="annotated as Ori in pAsylum" /note="pUC57 AmpR 3' UTR" misc_recomb complement(727..958) /note="Gateway BP Clonase target for removal of bacterial backbone" /note="attP1" regulatory complement(959..1009) /regulatory_class="terminator" /note="rrnB T2" 3'UTR complement(1010..1107) /note="pUC57 AmpR 3'UTR" CDS complement(1108..1482) /note="Zeocin resistance" /product="ShBle" regulatory complement(1488..1493) /regulatory_class="ribosome_binding_site" /note="Shine-Dalgarno" regulatory complement(1491..1594) /regulatory_class="promoter" /gene="bla" /note="AmpR promoter" regulatory complement(1505..1510) /regulatory_class="TATA_box" regulatory complement(1529..1534) /regulatory_class="CAAT_signal" misc_recomb complement(1601..1628) /note="Gateway BP Clonase target for removal of bacterial backbone" /note="attB1.1" misc_feature 1669..1674 /note="Pst1 for yeast-only recircularization" regulatory 1677..2046 /regulatory_class="enhancer" /note="GAL10-UAS" regulatory 2069..2324 /regulatory_class="promoter" /note="PYK1 promoter" regulatory 2277..2283 /regulatory_class="TATA_box" /note="TATA" regulatory 2311..2336 /regulatory_class="promoter" /note="TSS motifs" 5'UTR 2337..2389 /note="5' UTR of highly expressed gene in Niederer21" /note="FYV8 leader" intron 2390..2855 /note="Hoshida16" /note="RPS24A intron" misc_recomb 2855 /note="Sal1" regulatory 2859..2863 /regulatory_class="ribosome_binding_site" /note="ideal yeast Kozak (Cuperus17)" CDS 2864..3952 /product="BKV-IV VP1" regulatory 3982..4410 /regulatory_class="terminator" /note="Ito20" /note="RPL15A terminator" regulatory 4071..4076 /regulatory_class="terminator" /note="efficiency element" regulatory 4105..4115 /regulatory_class="terminator" /note="stability element" misc_RNA complement(4107..4188) /note="SRR5876624" /note="opposing transcript" polyA_site 4116 /note="SRR5876624" /note="polyA" regulatory 4294..4299 /regulatory_class="terminator" /note="cleavage and polyadenylation" regulatory 4333..4337 /regulatory_class="terminator" /note="positioning element" rep_origin complement(4418..5762) /note="2 micron Ori (yeast)" CDS 4419..4598 /product="Flp1p (C-term fragment)" 3'UTR 4643..4648 /note="efficiency element" 3'UTR 4846..4851 /note="positioning element" CDS complement(4966..5016) /product="Rep1p (C-term fragment)" regulatory complement(<5763..5831) /regulatory_class="terminator" /note="rrnB T1 terminator" regulatory complement(5840..6083) /regulatory_class="terminator" /note="Ito16" /note="DIT1-d22 terminator" 3'UTR complement(5900..5905) /note="Savinov21" /note="Efficiency element" 3'UTR complement(6007..6024) /note="mRNA stability element" misc_difference complement(6013..6014) /note="d22 mutation deletes TCA between these two bases" intron complement(6029..6041) /note="alternate splice acceptor" misc_difference complement(6033) /note="Ito16 m22 G->A (kills splice acceptor)" intron complement(6033..6079) /note="possible regulatory intron; stop codon of upstream gene must be within 50 bases to avoid nonsense mediated decay" /note="intron" intron complement(6044..6051) /note="lariat" misc_difference complement(6052..6053) /note="d7 mutation deletes TTG between these two bases" CDS complement(6091..7236) /note="formaldehyde dehydrogenase from Candida maltosa; confers formaldehyde resistance; codon optimized cDNA [Split]" /product="FDH1" regulatory complement(7237..7241) /regulatory_class="ribosome_binding_site" /note="ideal yeast Kozak (Cuperus17)" regulatory complement(7242..7248) /regulatory_class="ribosome_binding_site" /note="Shine-Dalgarno" 5'UTR complement(7250..7265) /note="leader of highly expressed gene (Niederer21)" /note="MEH1 leader" mRNA complement(7271..7296) /note="transcription start site motifs" /note="TSS motifs" regulatory complement(7279..7689) /regulatory_class="promoter" /note="FDH1 promoter" regulatory complement(7347..7350) /regulatory_class="TATA_box" /note="TATA" regulatory complement(7386..7405) /regulatory_class="" /note="CAAT" misc_feature 7690..7695 /note="Pst1 for yeast-only recircularization" regulatory complement(7703..7750) /regulatory_class="promoter" /note="EM7" ORIGIN 1 AGCAAAAGGC CAGCAAAAGG CCAGGAACCG TAAAAAGGCC GCGTTGCTGG CGTTTTTCCA 61 TAGGCTCCGC CCCCCTGACG AGCATCACAA AAATCGACGC TCAAGTCAGA GGTGGCGAAA 121 CCCGACAGGA CTATAAAGAT ACCAGGCGTT TCCCCCTGGA AGCTCCCTCG TGCGCTCTCC 181 TGTTCCGACC CTGCCGCTTA CCGGATACCT GTCCGCCTTT CTCCCTTCGG GAAGCGTGGC 241 GCTTTCTCAT AGCTCACGCT GTAGGTATCT CAGTTCGGTG TAGGTCGTTC GCTCCAAGCT 301 GGGCTGTGTG CACGAACCCC CCGTTCAGCC CGACCGCTGC GCCTTATCCG GTAACTATCG 361 TCTTGAGTCC AACCCGGTAA GACACGACTT ATCGCCACTG GCAGCAGCCA CTGGTAACAG 421 GATTAGCAGA GCGAGGTATG TAGGCGGTGC TACAGAGTTC TTGAAGTGGT GGCCTAACTA 481 CGGCTACACT AGAAGAACAG TATTTGGTAT CTGCGCTCTG CTGAAGCCAG TTACCTTCGG 541 AAAAAGAGTT GGTAGCTCTT GATCCGGCAA ACAAACCACC GCTGGTAGCG GTGGTTTTTT 601 TGTTTGCAAG CAGCAGATTA CGCGCAGAAA AAAAGGATCT CAAGAAGATC CTTTGATCTT 661 TTCTACGGGG TCTGACGCTC AGTGGAACGA AAACTCACGT TAAGGGATTT TGGTCATGCC 721 TGCAGGTACA GGTCACTAAT ACCATCTAAG TAGTTGATTC ATAGTGACTG GATATGTTGT 781 GTTTTACAGT ATTATGTAGT CTGTTTTTTA TGCAAAATCT AATTTAATAT ATTGATATTT 841 ATATCATTTT ACGTTTCTCG TTCAGCTTTT TTGTACAAAG TTGGCATTAT AAAAAAGCAT 901 TGCTTATCAA TTTGTTGCAA CGAACAGGTC ACTATCAGTC AAAATAAAAT CATTATTTTT 961 GTAGAAACGC AAAAAGGCCA TCCGTCAGGA TGGCCTTCTG CTTAATTTGA GATTATCAAA 1021 AAGGATCTTC ACCTAGATCC TTTTAAATTA AAAATGAAGT TTTAAATCAA TCTAAAGTAT 1081 ATATGAGTAA ACTTGGTCTG ACAGTTATCA GTCCTGCTCC TCTGCCACAA AGTGCACACA 1141 GTTGCCTGCT GGATCTCTCA GGGCAAACTC TCTCCCCCAG GGCTGCTCTC CAATCTCTGT 1201 CATGGCAGGG CCACTGGCAT CCCTGAAGTT GGTGGACACC ACCTCACTCC ACTCAGCATA 1261 CAGCTCATCC AGTCCTCTCA CCCACACCCA AGCCAGGGTG TTGTCAGGCA CCACCTGGTC 1321 CTGGACTGCT GAGATGAACA GGGTGACATC ATCTCTGACC ACACCTGCAA AGTCATCCTC 1381 CACAAAATCT CTGGAGAACC CCAACCTGTC AGTCCAGAAC TCAACAGCTC CAGCCACATC 1441 CCTGGCTGTG AGCACTGGGA CAGCACTGGT CAACTTGGCC ATTATGCACC TCCTAACTTT 1501 TTCAATATTA TTGAAGCATT TATCAGGGTT ATTGTCTCAT GAGCGGATAC ATATTTGAAT 1561 GTATTTAGAA AAATAAACAA ATAGGGGTTC CGCGTACTTT GCCAACTTTT TTGTACAAAG 1621 TTGTCCCCTG AAAAATTTAA TTTTGTTGCG GCCGTAGTAG AAGCTTGCCT GCAGGGGATC 1681 AAAAATCATC GCTTCGCTGA TTAATTACCC CAGAAATAAG GCTAAAAAAC TAATCGCATT 1741 ATCATCCTAT GGTTGTTAAT TTGATTCGTT CATTTGAAGG TTTGTGGGGC CAGGTTACTG 1801 CCAATTTTTC CTCTTCATAA CCATAAAAGC TAGTATTGTA GAATCTTTAT TGTTCGGAGC 1861 AGTGCGGCGC GAGGCACATC TGCGTTTCAG GAACGCGACC GGTGAAGACG AGGACGCACG 1921 GAGGAGAGTC TTCCTTCGGA GGGCTGTCAC CCGCTCGGCG GCTTCTAATC CGTACTTCAA 1981 TATAGCAATG AGCAGTTAAG CGTATTACTG AAAGTTCCAA AGAGAAGGTT TTTTTAGGCT 2041 AATCGGCCTC GAATTTGAGC TCGGTACCTT TAGTGTCTTC CTAAAAAAAA AAAAAGGCTC 2101 GCCATCAAAA CGATATTCGT TGGCTTTTTT TTCTGAATTA TAAATACTCT TTGGTAACTT 2161 TTCATTTCCA AGAACCTCTT TTTTCCAGTT ATATCATGGT CCCCTTTCAA AGTTATTCTC 2221 TACTCTTTTT CATATTCATT CTTTTTCATC CTTTGGTTTT TTATTCTTAA CTTGTTTATT 2281 ATTCTCTCTT GTTTCTATTT ACAAGACACC AATCAAAACA AAATAACAAT TAAATAATTA 2341 TAGTTAAAAG GGACTGTTTG ATCATTACCC TTTTTCTGGA AAGCGGATAG TATGTTAAAA 2401 AGTGCTCAGA TGAAAGATGA CGTCCCATAT TCCATGTTAA AGTTGCCATG AGTTAGTATA 2461 AAAGGCAATA AAGGTACCAT AACGAGAAAA GAAAGTAAAA GAAGAAATGT TTTGCAAATT 2521 TAAGGTGGCG TAACAGCAAC TATTGGCTAA TATATTAATG GCAATCATTG TGAAAAGAAA 2581 TGTGAAATCC GTATCGTAAT TGGTTAAATT TATAACTCAA ATTTAAACAC TCGAATAGCT 2641 GACCCGTAAT ATTTTATCCC GCTTTAAGAG TTCAACGCTA AAAAATGATA TCTAGCTAAC 2701 CATTATGCTA CAGGAACCTT TTTTGGACTA TTATCCCTAT CATGAAAACT GGAAGGATCT 2761 TTCAAGGACG CCATTTCCAA AGATTTCCCC CACACTGGAA CCTATTTTAC TAACAATTTT 2821 TAAAATGGTT TTTTACCTAA ATTGTTTTTT CTCAGTCGAC AAGATGGCTC CAACCAAAAG 2881 AAAAGGCGAA TGTCCTGGAG CGGCGCCTAA GAAACCTAAA GAGCCCGTAC AAGTTCCCAA 2941 GCTGTTGATC AAAGGCGGCG TTGAAGTGCT GGAGGTTAAG ACTGGGGTTG ACGCGATTAC 3001 AGAAGTTGAA TGCTTCCTAA ACCCCGAAAT GGGAGATCCG GATAATGATC TAAGAGGATA 3061 CAGTCTGAGA TTAACGGCGG AAACTGCGTT TGATTCCGAC TCACCTGACA GAAAGATGTT 3121 ACCTTGTTAC AGTACCGCAA GAATCCCGCT TCCAAACCTG AACGAGGATT TGACCTGTGG 3181 CAACCTACTT ATGTGGGAGG CCGTAACTGT CAAAACGGAA GTAATCGGCA TTACATCAAT 3241 GCTTAATCTA CATGCGGGAT CTCAGAAAGT TCACGAAAAT GGAGGCGGCA AACCTATACA 3301 GGGGAGTAAC TTTCATTTCT TTGCTGTTGG CGGCGACCCA CTGGAAATGC AAGGAGTACT 3361 TATGAACTAC AGAACGAAGT ACCCAGAGGG AACTGTGACC CCGAAGAATC CGACTGCGCA 3421 GTCACAAGTT ATGAACACGG ATCATAAAGC CTATTTGGAC AAGAACAATG CGTACCCTGT 3481 AGAGTGCTGG ATACCAGATC CATCAAAGAA CGAGAATACC CGTTACTTCG GCACTTATAC 3541 GGGCGGCGAG AATGTTCCAC CAGTGTTGCA TGTTACAAAC ACAGCTACAA CTGTACTATT 3601 AGACGAACAG GGCGTCGGAC CTCTGTGCAA GGCAGACTCG TTATACGTCT CTGCGGCAGA 3661 CATCTGTGGT TTGTTCACTA ATTCAAGTGG CACCCAGCAG TGGCGTGGCT TGCCACGTTA 3721 CTTCAAGATA AGACTTAGGA AAAGATCAGT CAAGAATCCT TACCCAATCA GCTTCTTGTT 3781 ATCAGATTTG ATCAATAGGC GTACTCAAAG GGTTGACGGG CAGCCTATGT ACGGGATGGA 3841 GAGCCAAGTG GAAGAAGTTA GGGTTTTCGA TGGGACAGAA CAGCTACCTG GGGACCCGGA 3901 TATGATTAGA TACATAGATA GGCAGGGTCA ACTACAGACT AAGATGGTCT AATGAGATCT 3961 GAAGAACCTA GGGAACCATG GTAAGCTGGT TGATGGAAAA TATAATTTTA TTGGGCAAAC 4021 TTTTGTTTAT CTGATGTGTT TTATACTATT ATCTTTTTAA TTAATGATTC TATATACAAA 4081 CCTGTATATT TTTTCTTTAA CCAATTTTTT TTTTTATAGA CCTAGAGCTG TACTTTTATT 4141 CTGCTATCAA GCAAACCCCT ACCCCCTCTT CTCAATCCTC CCCTCAGGCA GAACTTATCT 4201 ACCTGTATCA AGGAGCGGAC GAGGGAGTCC TAATTGTTCT ACGTATACCA ATGCTAGCAG 4261 CTTACATAGG TGGTGGCACT ACCATAATAC TTATATATAA TATCTTTACG CTCAAGTAAA 4321 TTTCTCCGCC GCATAAATTA ACACCATGTT CTTTTTAGTC GCACCTTATT TTCTTGTATT 4381 TTTTTTCTTT CCTTTTCTTC CCGTTTTTCC ACGCGTATGA TCCAATATCA AAGGAAATGA 4441 TAGCATTGAA GGATGAGACT AATCCAATTG AGGAGTGGCA GCATATAGAA CAGCTAAAGG 4501 GTAGTGCTGA AGGAAGCATA CGATACCCCG CATGGAATGG GATAATATCA CAGGAGGTAC 4561 TAGACTACCT TTCATCCTAC ATAAATAGAC GCATATAAGT ACGCATTTAA GCATAAACAC 4621 GCACTATGCC GTTCTTCTCA TGTATATATA TATACAGGCA ACACGCAGAT ATAGGTGCGA 4681 CGTGAACAGT GAGCTGTATG TGCGCAGCTC GCGTTGCATT TTCGGAAGCG CTCGTTTTCG 4741 GAAACGCTTT GAAGTTCCTA TTCCGAAGTT CCTATTCTCT AGCTAGAAAG TATAGGAACT 4801 TCAGAGCGCT TTTGAAAACC AAAAGCGCTC TGAAGACGCA CTTTCAAAAA ACCAAAAACG 4861 CACCGGACTG TAACGAGCTA CTAAAATATT GCGAATACCG CTTCCACAAA CATTGCTCAA 4921 AAGTATCTCT TTGCTATATA TCTCTGTGCT ATATCCCTAT ATAACCTACC CATCCACCTT 4981 TCGCTCCTTG AACTTGCATC TAAACTCGAC CTCTACATTT TTTATGTTTA TCTCTAGTAT 5041 TACTCTTTAG ACAAAAAAAT TGTAGTAAGA ACTATTCATA GAGTGAATCG AAAACAATAC 5101 GAAAATGTAA ACATTTCCTA TACGTAGTAT ATAGAGACAA AATAGAAGAA ACCGTTCATA 5161 ATTTTCTGAC CAATGAAGAA TCATCAACGC TATCACTTTC TGTTCACAAA GTATGCGCAA 5221 TCCACATCGG TATAGAATAT AATCGGGGAT GCCTTTATCT TGAAAAAATG CACCCGCAGC 5281 TTCGCTAGTA ATCAGTAAAC GCGGGAAGTG GAGTCAGGCT TTTTTTATGG AAGAGAAAAT 5341 AGACACCAAA GTAGCCTTCT TCTAACCTTA ACGGACCTAC AGTGCAAAAA GTTATCAAGA 5401 GACTGCATTA TAGAGCGCAC AAAGGAGAAA AAAAGTAATC TAAGATGCTT TGTTAGAAAA 5461 ATAGCGCTCT CGGGATGCAT TTTTGTAGAA CAAAAAAGAA GTATAGATTC TTTGTTGGTA 5521 AAATAGCGCT CTCGCGTTGC ATTTCTGTTC TGTAAAAATG CAGCTCAGAT TCTTTGTTTG 5581 AAAAATTAGC GCTCTCGCGT TGCATTTTTG TTTTACAAAA ATGAAGCACA GATTCTTCGT 5641 TGGTAAAATA GCGCTTTCGC GTTGCATTTC TGTTCTGTAA AAATGCAGCT CAGATTCTTT 5701 GTTTGAAAAA TTAGCGCTCT CGCGTTGCAT TTTTGTTCTA CAAAATGAAG CACAGATGCT 5761 TCAGCGTTCA CCGACAAACA ACAGATAAAA CGAAAGGCCC AGTCTTTCGA CTGAGCCTTT 5821 CGTTTTATTT GGGATCCTCA GGGCCATTGC AGTGTCTCGA AACCATTAAT ATCACTGAAA 5881 AGATGAAAAG AAAGGCAAAT ATATATTGAT CACTAATTTT CTAAGCTAAA GAATCTATTC 5941 CCCCTCTGTT AAATGGAATT GTGTGAAATA AAATATTATA AAATCAGAAC TTTGGGGGGG 6001 AAACATAAAA AAAGAAAAAG AAAACGAACT AACTAATGTT TAAGTAAAAG AAAAAGGTAG 6061 ACCAATGTAG CGCTCTTACT TTAGCCGGCG TCAGCTCAAG TCTACTACTG CACGTATACA 6121 ATCCCCAGCG TGCATCTCTT CAAATGCCTT ATTGATCGCG GCTAGTGGCT CTCTATGTGT 6181 AATGAACTCT TCGACCTTCA ACTTGCCGTC CAAATAGTTG TTTACAATCC CTGGCAGTTG 6241 GGACCTGCCT TTGACTCCGC CAAATGCAGC TCCCTTCCAC GTCCTCCCCG TAACTAATTG 6301 AAACGGACGT GTGCTGATCT CCTTTCCCGC TGCTGCTACT CCAATGATGA CAGAGGTGCC 6361 CCAGCCTTTA TGACAAGCCT CCAGAGCGTT TCTCATCACG CCAACGTTTC CTGTACAGTC 6421 GAACGTGAAG TCGCAGCCGC CATCCGTCAT CTCAATCAAC TTATCAACTA TCGTCGTCCC 6481 TTCGGGTAGT TTGGTCGGAT TAACGAAAGC GGTCGCTCCC AGCTTTTGGC CCCATTCTTC 6541 TTTCTTGTCA CTGATGTCAA CGAGAATGAT CTGGGCTGCT CCCCTCTCTG CGCATCCCTG 6601 GATTACTGAT AATCCGACGA TTCCTCCGCC GAAGACGGCG ACATTGTCTC CCTTCTGTAC 6661 GTTCGCCGTA ATTGTCGCTG CCCCATAACC AGTTGTTATC CCGCAACCCA GCAGACATGC 6721 TTTGTCGAAC TCCGCTTTGG GGTTAATCGC AACGACGCTT ATGTCTGCCA CAACTGTGTA 6781 TTGACTGAAA GTGGAGCAGC CCATAAAGTG AAGTATCTCT TTCCCTTTAC AGGTAAAACG 6841 ACTTGTCCCG TCAGGCATAA CTCCTTTGCC CTGGGTTGCT CTGATTTTCC CGCATAGATT 6901 CGTTTTTCCG CTTTTACAGA ACTTGCACTC CCCGCATTCG GGAGTGTACA AGGCGATAAC 6961 ATGGTCGCCA ACTTTGACGT TCGTAACTCC TTCTCCGATT GATTCCACAA TTCCGGCCCC 7021 CTCATGCCCC AGTATTACCG GAAATGCGCC CTCTGGATCG ACCCCTGACA AAGTATATGC 7081 ATCTGTGTGA CAGACGCCAG TGTCGTATAA CTTGATACGA ACTTCGTGCG CCTTAGGTGG 7141 CGCAACTGTA ACGTCCTCGA TGCTCAAAGG CTTCGCCGCT TCCCAAGCAA CGGCGGCCTT 7201 GCAAGTGATA GGTTTTCCGA CTGTTGATTC ACTCATCTTG TACCTCCTCT TTTACGACTT 7261 AAATCACTAG TTTTAATGTA AGTATTTTAA TTATTCAATG GTAAGACTAG AAAATGAAAA 7321 GAAGAAGAAG GAGTTGATTT GGTTTTTATA ATCTTTTTTT TTTTTTTCTT CGTTCTATCA 7381 GTTTCATTGA ACTTCTTTGT TTCTCGCGCT GAACGTTGCA GGGAATTGGT TACCCGATTA 7441 GGCTCATCTT TACGGCTTAC TCACGTTTTT TTTTTTAGTT TCGCATAATT TTGACCATGC 7501 TTAATAATTA GGTATCACAA ATAGCGCGGA AACTTATCGC TTAAACTCAT ATAATCACGA 7561 AGTATCTCAC CCGCAATATT GTTTAGAAAA GTATTGGTTC TATCTTGATT ACTAATTCTC 7621 TATTTTGATG TAACGTTGAC ACACTTAAAC TCAGATATTC TCTTTGTGTT GACTATCGCT 7681 TGCTTTTTCC TGCAGTATCG ATGTCGTATT ATACTATGCC GATATACTAT GCCGATGATT 7741 AATTGTCAAC AGAATTCAAG AACATGTTTC TTCGAA //