LOCUS       40924_35439        3820 bp DNA     circular SYN 18-DEC-2018
DEFINITION  E.coli-K.pastoris shuttle vector pPpT4_Alpha_S, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3820)
  AUTHORS   Naatsaari L, Mistlberger B, Ruth C, Hajek T, Hartner FS, Glieder A.
  TITLE     Deletion of the Pichia pastoris KU70 Homologue Facilitates Platform 
            Strain Generation for Gene Expression and Synthetic Biology
  JOURNAL   PLoS ONE 7 (6), E39720 (2012)
  PUBMED    22768112
REFERENCE   2  (bases 1 to 3820)
  AUTHORS   Naeaetsaari L, Mistlberger B, Ruth C, Hajek T, Hartner F, Glieder A.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-JAN-2012) Institute of Molecular Biotechnology, Graz 
            University of Technology, Petersgasse 14/2, Graz, Styria 8010, 
            Austria
REFERENCE   3  (bases 1 to 3820)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3820)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE"; 
            date: "2012"; volume: "7"; issue: "6"; pages: "E39720"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (30-JAN-2012) Institute of Molecular Biotechnology, Graz University 
            of Technology, Petersgasse 14/2, Graz, Styria 8010, Austria"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: SeqMan v. 7.0.0
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..3820
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        139..1071
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     CDS             1085..1351
                     /label=alpha-factor secretion signal
                     /note="N-terminal secretion signal from S. cerevisiae 
                     alpha-factor"
     terminator      1372..1618
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     regulatory      1633..2190
                     /label=ILV5 promoter
                     /note="ILV5 promoter"
                     /regulatory_class="promoter"
     regulatory      2191..2254
                     /label=EM72 synthetic promoter
                     /note="EM72 synthetic promoter"
                     /regulatory_class="promoter"
     CDS             2256..2627
                     /label=BleoR
                     /note="antibiotic-binding protein"
     regulatory      2632..3107
                     /label=AOD terminator
                     /note="AOD terminator"
                     /regulatory_class="terminator"
     rep_origin      3154..3742
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agatccaatt cccgctttga ctgcctgaaa tctccatcgc ctacaatgat gacatttgga
       61 tttggttgac tcatgttggt attgtgaaat agacgcagat cgggaacact gaaaaataca
      121 cagttattat tcatttaaat aacatccaaa gacgaaaggt tgaatgaaac ctttttgcca
      181 tccgacatcc acaggtccat tctcacacat aagtgccaaa cgcaacagga ggggatacac
      241 tagcagcaga ccgttgcaaa cgcaggacct ccactcctct tctcctcaac acccactttt
      301 gccatcgaaa aaccagccca gttattgggc ttgattggag ctcgctcatt ccaattcctt
      361 ctattaggct actaacacca tgactttatt agcctgtcta tcctggcccc cctggcgagg
      421 ttcatgtttg tttatttccg aatgcaacaa gctccgcatt acacccgaac atcactccag
      481 atgagggctt tctgagtgtg gggtcaaata gtttcatgtt ccccaaatgg cccaaaactg
      541 acagtttaaa cgctgtcttg gaacctaata tgacaaaagc gtgatctcat ccaagatgaa
      601 ctaagtttgg ttcgttgaaa tgctaacggc cagttggtca aaaagaaact tccaaaagtc
      661 ggcataccgt ttgtcttgtt tggtattgat tgacgaatgc tcaaaaataa tctcattaat
      721 gcttagcgca gtctctctat cgcttctgaa ccccggtgca cctgtgccga aacgcaaatg
      781 gggaaacacc cgctttttgg atgattatgc attgtctcca cattgtatgc ttccaagatt
      841 ctggtgggaa tactgctgat agcctaacgt tcatgatcaa aatttaactg ttctaacccc
      901 tacttgacag caatatataa acagaaggaa gctgccctgt cttaaacctt tttttttatc
      961 atcattatta gcttactttc ataattgcga ctggttccaa ttgacaagct tttgatttta
     1021 acgactttta acgacaactt gagaagatca aaaaacaact aattattgaa agaattccga
     1081 aacgatgaga ttcccatcta ttttcaccgc tgtcttgttc gctgcctcct ctgcattggc
     1141 tgcccctgtt aacactacca ctgaagacga gactgctcaa attccagctg aagcagttat
     1201 cggttactct gaccttgagg gtgatttcga cgtcgctgtt ttgcctttct ctaactccac
     1261 taacaacggt ttgttgttca ttaacaccac tatcgcttcc attgctgcta aggaagaggg
     1321 tgtctctctc gagaagagag aggccgaagc ttacgtacct agggcggccg ctcaagagga
     1381 tgtcagaatg ccatttgcct gagagatgca ggcttcattt ttgatacttt tttatttgta
     1441 acctatatag tataggattt tttttgtcat tttgtttctt ctcgtacgag cttgctcctg
     1501 atcagcctat ctcgcagcag atgaatatct tgtggtaggg gtttgggaaa atcattcgag
     1561 tttgatgttt ttcttggtat ttcccactcc tcttcagagt acagaagatt aagtgagacc
     1621 ttcgtttgtg cggatccttc agtaatgtct tgtttctttt gttgcagtgg tgagccattt
     1681 tgacttcgtg aaagtttctt tagaatagtt gtttccagag gccaaacatt ccacccgtag
     1741 taaagtgcaa gcgtaggaag accaagactg gcataaatca ggtataagtg tcgagcactg
     1801 gcaggtgatc ttctgaaagt ttctactagc agataagatc cagtagtcat gcatatggca
     1861 acaatgtacc gtgtggatct aagaacgcgt cctactaacc ttcgcattcg ttggtccagt
     1921 ttgttgttat cgatcaacgt gacaaggttg tcgattccgc gtaagcatgc atacccaagg
     1981 acgcctgttg caattccaag tgagccagtt ccaacaatct ttgtaatatt agagcacttc
     2041 attgtgttgc gcttgaaagt aaaatgcgaa caaattaaga gataatctcg aaaccgcgac
     2101 ttcaaacgcc aatatgatgt gcggcacaca ataagcgttc atatccgctg ggtgactttc
     2161 tcgctttaaa aaattatccg aaaaaatttt ctagagtgtt gttactttat acttccggct
     2221 cgtataatac gacaaggtgt aaggaggact aaaccatggc taaactcacc tctgctgttc
     2281 cagtcctgac tgctcgtgat gttgctggtg ctgttgagtt ctggactgat agactcggtt
     2341 tctcccgtga cttcgtagag gacgactttg ccggtgttgt acgtgacgac gttaccctgt
     2401 tcatctccgc agttcaggac caggttgtgc cagacaacac tctggcatgg gtatgggttc
     2461 gtggtctgga cgaactgtac gctgagtggt ctgaggtcgt gtctaccaac ttccgtgatg
     2521 catctggtcc agctatgacc gagatcggtg aacagccctg gggtcgtgag tttgcactgc
     2581 gtgatccagc tggtaactgc gtgcatttcg tcgcagaaga gcaggactaa caattgacac
     2641 cttacgatta tttagagagt atttattagt tttattgtat gtatacggat gttttattat
     2701 ctatttatgc ccttatattc tgtaactatc caaaagtcct atcttatcaa gccagcaatc
     2761 tatgtccgcg aacgtcaact aaaaataagc tttttatgct cttctctctt tttttccctt
     2821 cggtataatt ataccttgca tccacagatt ctcctgccaa attttgcata atcctttaca
     2881 acatggctat atgggagcac ttagcgccct ccaaaaccca tattgcctac gcatgtatag
     2941 gtgttttttc cacaatattt tctctgtgct ctctttttat taaagagaag ctctatatcg
     3001 gagaagcttc tgtggccgtt atattcggcc ttatcgtggg accacattgc ctgaattggt
     3061 ttgccccgga agattgggga aacttggatc tgattacctt agctgcaggt accactgagc
     3121 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3181 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3241 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3301 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3361 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3421 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3481 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3541 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3601 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     3661 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     3721 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     3781 ttgctggcct tttgctcaca tgttctttcc tgcggtaccc
//
