LOCUS 40924_48278 7596 bp DNA circular SYN 18-DEC-2018 DEFINITION Shuttle vector pYT3, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7596) AUTHORS Tan X, Waterham HR, Veenhuis M, Cregg JM. TITLE The Hansenula polymorpha PER8 gene encodes a novel peroxisomal integral membrane protein involved in proliferation JOURNAL J. Cell Biol. 128 (3), 307-319 (1995) PUBMED 7844145 REFERENCE 2 (bases 1 to 7596) AUTHORS Baerends RJS., van Dijk R, Cregg JM, Veenhuis M. TITLE Direct Submission JOURNAL Submitted (08-FEB-2001) Eukaryotic Microbiology, Groningen Biomolecular Sciences and Biotechnology Institute, Kerklaan 30, Haren, Gn 9751 NN, the Netherlands REFERENCE 3 (bases 1 to 7596) TITLE Direct Submission REFERENCE 4 (bases 1 to 7596) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J. Cell Biol."; date: "1995"; volume: "128"; issue: "3"; pages: "307-319" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (08-FEB-2001) Eukaryotic Microbiology, Groningen Biomolecular Sciences and Biotechnology Institute, Kerklaan 30, Haren, Gn 9751 NN, the Netherlands" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..7596 /mol_type="other DNA" /organism="synthetic DNA construct" rep_origin 28..2724 /note="derived from Hansenula polymorpha; confers autonomous replication activity (HARS) in Hansenula polymorpha" RBS 1139..1147 /label=Shine-Dalgarno sequence /note="full consensus sequence for ribosome-binding sites upstream of start codons in E. coli; complementary to a region in the 3' end of the 16S rRNA (Chen et al., 1994)" CDS complement(3218..4309) /label=LEU2 /note="3-isopropylmalate dehydrogenase, required for leucine biosynthesis" promoter complement(4310..4717) /label=LEU2 promoter primer_bind complement(4989..5005) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(5013..5029) /label=lac operator /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(5037..5067) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(5082..5103) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(5391..5979) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(6153..7010) /label=AmpR /note="beta-lactamase" promoter complement(7011..7115) /label=AmpR promoter ORIGIN 1 acggccagtg aattcgagct cggtacccga cagaagaaat tgctcaactc taatgtgcgc 61 catgtttaat ctcaatcagc ttattagtat ctcgtatatc ttatttcaat acctatcaca 121 tattaacgtt ggattgacat attgcataga ctatgtagaa atgctatctg attacgatca 181 cgcctggact ttgtttttcg atttttcaac ttctgatcag cttctatgtt ttaaaaaaac 241 gaggctctag tttacgtaat ctacacggtt gcagaaaatt agaacaatct ttcaaacttc 301 tgatcaccat aaaccttgtt gtcgagatgc aaatggttct ttattcatca ttccgcctat 361 acgatacctg tcgtgacaga atattgtcac cagtacacaa gcttgtctca aattcgtttc 421 acaaaagagg agggattgct tgtttgtgtg atcatctttg actaaccgaa taagtgaccc 481 atgggtctgc ttgatggggt agatctgatg ttggggagtt tcaatgtata aaattcgata 541 acggggctat ctctattttg tagcacttat aatggtatct gctcaaaaga gattcatggt 601 agcattatct caatacccat cctaagcttt agcagcatgt ttttagatgg gggtagaatg 661 cttgcctaat taaacatgaa aacttatata cttccatgaa actcttcgag gctctagtta 721 tcctcaaatc tcggattacc taccactccc tcggccaaaa ggcgccaaaa cttactgctt 781 ggtataccgt aattgtgagg tctacctttt caaggagcag tcgttagatg ttccggtact 841 tagggacttc actaaacaaa tgcacattat cagtgttaat ttagctatca aaaaaaatat 901 tgcactaata gtcctctcaa agtctcaata tttataaccg tagttgataa tagatagtga 961 gtgttttttg agcttcatta aaatacgaaa actagaaggt atgcgaacta gtcttttcta 1021 ttatgactat ttgcgcaacg ttacatcaaa ataatatatg cttcaaatat tcagctatta 1081 agcatctgac ggaaaatatt cacactatat ctaggctggt cattggcgga ctgggtatac 1141 ctccttagta aaggtttagg gcgaaagaga gggcagaagt tatttagtct aactaatagc 1201 agtactttct tagactaagc ctgagtggat gcagctcaag actttaatgg cagcagggtg 1261 tagatctatg tgagcaataa aaaaaattgc gctatttaaa tttcacaaat tttaaatttg 1321 attgagtcat aatcccttga tatagttcat tcaatcaatt gtgataatca gaacacaact 1381 tgactgagcc gtgtgaaaac actgaagcat atagtgtgca gataaggcaa tgatgtattt 1441 gctaactagc taaagacaat aaatagtaaa gataatagtg ctcaaggaaa aggatactca 1501 aaaaactcat ttgataaaat caagataggc atgcaacgtg tggagcccct gttaagtcac 1561 tatgtagtta tcaattattc agattgctca atctcgacat tgaagagtga tacaatgcta 1621 gaaaggaagc tattaacgct cgtgaaaaaa aagatctaca gccacacata ttagtttaat 1681 tggttcgaat ccgaaaaaac aacacttgtt gcaattcgta actctgagct attgaagagt 1741 cggacgaaat ggcatcagga gttcatccat gtaaacacta tcctgatgtc ttggcagtat 1801 gtaatcgagc tttaatttca ttagtggata atacccgtac tttaaagtta aataagcatt 1861 caacatatca acgtcggatt aaggaccaca ttcaaaaagg tcacttgatt attgagcaaa 1921 ctcctgccaa gcattgtaag acctattgtt gatcgacaat ccttcccaaa gctacactta 1981 ccccatgctt attgaaaatg aaattcaaaa acttatgaga tacaagactg agatggttat 2041 tctaataatg cgttcagtcc ccaattaggg tctggccttc atggttggag aaaaaactac 2101 gtagggatac cattactttg atgttgcctc atatgagatg agaacaagta gttgctgata 2161 tgagatagtc agagattgat catggctgtg cggcttatcc tgatatgaag gtttgtgggg 2221 ttttagtaca tgcatattat gtgcaccaga tcctgacgat actaatctgg atcatatacg 2281 actaacaaaa tgacaaggaa gggtttctca tattcagctt ataagtaatt tagttagatc 2341 aacgattaac tattcgatac ctagtcacaa ttttgaagga taaataatga ctgtagacat 2401 gacttattat agctatgttt tgaacagtca tcaagccgag tatactactc cattagccaa 2461 acatggacgc tccagcatga attgcctttg ttggcagttc aagaaaggaa acgccgttgc 2521 acttgatgca ccgtgcaatc cggtagattc ttgtttgaga attcattttc cttctttgct 2581 atcacataat acactagttt ttttttagga tagtagttaa ttccttgtta cggccggcac 2641 tccagaatga aaacccgtct gcaataaaat tcatgacacc tcacactcat ttgtatgtag 2701 tttttttatc aacaagtatt ccagggggat cctctagagt cgactacgtc gtaaggccgt 2761 ttctgacaga gtaaaattct tgagggaact ttcaccatta tgggaaatgg ttcaagaagg 2821 tattgactta aactccatca aatggtcagg tcattgagtg ttttttattt gttgtatttt 2881 ttttttttta gagaaaatcc tccaatatca aattaggaat cgtagtttca tgattttctg 2941 ttacacctaa ctttttgtgt ggtgccctcc tccttgtcaa tattaatgtt aaagtgcaat 3001 tctttttcct tatcacgttg agccattagt atcaatttgc ttacctgtat tcctttacat 3061 cctccttttt ctccttcttg ataaatgtat gtagattgcg tatatagttt cgtctaccct 3121 atgaacatat tccattttgt aatttcgtgt cgtttctatt atgaatttca tttataaagt 3181 ttatgtacaa atatcataaa aaaagagaat ctttttaagc aaggattttc ttaacttctt 3241 cggcgacagc atcaccgact tccgtggtac tgttggaacc acctaaatca ccagttctga 3301 tacctgcatc caaaaccttt ttaactgcat cttcaatggc cttaccttct tcaggcaagt 3361 tcaatgacaa tttcaacatc attgcagcag acaagatagt ggcgataggg ttgaccttat 3421 tctttggcaa atctggagca gaaccgtggc atggttcgta caaaccaaat gcggtgttct 3481 tgtctggcaa agaggccaag gacgcagatg gcaacaaacc caaggaacct gggataacgg 3541 aggcttcatc ggagatgata tcaccaaaca tgttgctggt gattataata ccatttaggt 3601 gggttgggtt cttaactagg atcatggcgg cagaatcaat caattgatgt tgaaccttca 3661 atgtagggaa ttcgttcttg atggtttcct ccacagtttt tctccataat cttgaagagg 3721 ccaaaacatt agctttatcc aaggaccaaa taggcaatgg tggctcatgt tgtagggcca 3781 tgaaagcggc cattcttgtg attctttgca cttctggaac ggtgtattgt tcactatccc 3841 aagcgacacc atcaccatcg tcttcctttc tcttaccaaa gtaaatacct cccactaatt 3901 ctctgacaac aacgaagtca gtacctttag caaattgtgg cttgattgga gataagtcta 3961 aaagagagtc ggatgcaaag ttacatggtc ttaagttggc gtacaattga agttctttac 4021 ggatttttag taaaccttgt tcaggtctaa cactaccggt accccattta ggaccaccca 4081 cagcacctaa caaaacggca tcaaccttct tggaggcttc cagcgcctca tctggaagtg 4141 ggacacctgt agcatcgata gcagcaccac caattaaatg attttcgaaa tcgaacttga 4201 cattggaacg aacatcagaa atagctttaa gaaccttaat ggcttcggct gtgatttctt 4261 gaccaacgtg gtcacctggc aaaacgacga tcttcttagg ggcagacatt agaatggtat 4321 atccttgaaa tatatatata tatattgctg aaatgtaaaa ggtaagaaaa gttagaaagt 4381 aagacgattg ctaaccacct attggaaaaa acaataggtc cttaaataat attgtcaact 4441 tcaagtattg tgatgcaagc atttagtcat gaacgcttct ctattctata tgaaaagccg 4501 gttccgcggc tctcaccttt cctttttctc ccaatttttc agttgaaaaa ggtatatgcg 4561 tcaggcgacc tctgaaatta acaaaaaatt tccagtcatc gaatttgatt ctgtgcgata 4621 gcgcccctgt gtgttctcgt tatgttgagg aaaaaaataa tggttgctaa gagattcgaa 4681 ctcttgcatc ttacgatacc tgagtattcc cacagttaac tgcggtcaag atatttcttg 4741 aatcaggcgc cttagaccgc tcggccaaac aaccaattac ttgttgagaa atagagtata 4801 attatcctat aaatataacg tttttgaaca cacatgaaca aggaagtaca ggacaattga 4861 ttttgaagag aatgtggatt ttgatgtaat tgttgggatt ccatttttaa taaggcaata 4921 atattaggta tgtagatata ctagaagttc tcctcgacct gcaggcatgc aagcttggcg 4981 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac 5041 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca 5101 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat 5161 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc 5221 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca 5281 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca 5341 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg 5401 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg 5461 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt 5521 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt 5581 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc 5641 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt 5701 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt 5761 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc 5821 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa 5881 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt 5941 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct 6001 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta 6061 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa 6121 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc 6181 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact 6241 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc 6301 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt 6361 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta 6421 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg 6481 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt 6541 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc 6601 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt 6661 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc 6721 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc 6781 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa 6841 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac 6901 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa 6961 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt 7021 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa 7081 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct 7141 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg 7201 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg 7261 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg 7321 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta 7381 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc 7441 atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc 7501 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta 7561 acgccagggt tttcccagtc acgacgttgt aaaacg //