LOCUS       40924_35389        3964 bp DNA     circular SYN 18-DEC-2018
DEFINITION  E.coli-K.pastoris shuttle vector pPpB1_Alpha_S, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3964)
  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 3964)
  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 3964)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3964)
  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..3964
                     /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      1374..1620
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        1783..2487
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     regulatory      2506..2563
                     /label=EM72 synthetic promoter
                     /note="EM72 synthetic promoter"
                     /regulatory_class="promoter"
     CDS             2566..2937
                     /label=BleoR
                     /note="antibiotic-binding protein"
     terminator      2947..3134
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     rep_origin      3298..3886
                     /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 tactagtggc gcgccgcggc cgctcaagag
     1381 gatgtcagaa tgccatttgc ctgagagatg caggcttcat ttttgatact tttttatttg
     1441 taacctatat agtataggat tttttttgtc attttgtttc ttctcgtacg agcttgctcc
     1501 tgatcagcct atctcgcagc agatgaatat cttgtggtag gggtttggga aaatcattcg
     1561 agtttgatgt ttttcttggt atttcccact cctcttcaga gtacagaaga ttaagtgaga
     1621 ccttcgtttg tgcggatcct ttattatgtt acaatatgga agggaacttt acacttctcc
     1681 tatgcacata tattaattaa agtccaatgc tagtagagaa ggggggtaac acccctccgc
     1741 gctcttttcc gatttttttc taaaccgtgg aatatttcgg atatcctttt gttgtttccg
     1801 ggtgtacaat atggacttcc tcttttctgg caaccaaacc catacatcgg gattcctata
     1861 ataccttcgt tggtctccct aacatgtagg tggcggaggg gagatataca atagaacaga
     1921 taccagacaa gacataatgg gctaaacaag actacaccaa ttacactgcc tcattgatgg
     1981 tggtacataa cgaactaata ctgtagccct agacttgata gccatcatca tatcgaagtt
     2041 tcactaccct ttttccattt gccatctatt gaagtaataa taggcgcatg caacttcttt
     2101 tctttttttt tcttttctct ctcccccgtt gttgtctcac catatccgca atgacaaaaa
     2161 aatgatggaa gacactaaag gaaaaaatta acgacaaaga cagcaccaac agatgtcgtt
     2221 gttccagagc tgatgagggg tatctcgaag cacacgaaac tttttccttc cttcattcac
     2281 gcacactact ctctaatgag caacggtata cggccttcct tccagttact tgaatttgaa
     2341 ataaaaaaaa gtttgctgtc ttgctatcaa gtataaatag acctgcaatt attaatcttt
     2401 tgtttcctcg tcattgttct cgttcccttt cttccttgtt tctttttctg cacaatattt
     2461 caagctatac caagcataca atcaactatc tcatatacat ctagagtgtt gacactttat
     2521 acttccggct cgtataatac gacaaggtgt aaggaggact aaaccatggc taaactcacc
     2581 tctgctgttc cagtcctgac tgctcgtgat gttgctggtg ctgttgagtt ctggactgat
     2641 agactcggtt tctcccgtga cttcgtagag gacgactttg ccggtgttgt acgtgacgac
     2701 gttaccctgt tcatctccgc agttcaggac caggttgtgc cagacaacac tctggcatgg
     2761 gtatgggttc gtggtctgga cgaactgtac gctgagtggt ctgaggtcgt gtctaccaac
     2821 ttccgtgatg catctggtcc agctatgacc gagatcggtg aacagccctg gggtcgtgag
     2881 tttgcactgc gtgatccagc tggtaactgc gtgcatttcg tcgcagaaga gcaggactaa
     2941 cctagggcga atttcttatg atttatgatt tttattatta aataagttat aaaaaaaata
     3001 agtgtataca aattttaaag tgactcttag gttttaaaac gaaaattctt attcttgagt
     3061 aactctttcc tgtaggtcag gttgctttct caggtatagc atgaggtcgc tcttattgac
     3121 cacacctcta ccggcatgcc gagcaaatgc ctgcaaatcg ctccccattt cacccaattg
     3181 tagatatgct aactccagca atgagttgat gaatctcggt gtgtatttta tgtcctcaga
     3241 ggacaactgc aggtaccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     3301 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     3361 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     3421 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     3481 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     3541 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     3601 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     3661 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     3721 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     3781 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     3841 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     3901 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcggt
     3961 accc
//
