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
//
