LOCUS       40924_35419        4257 bp DNA     circular SYN 18-DEC-2018
DEFINITION  E.coli-K.pastoris shuttle vector pPpKan_Alpha_S, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4257)
  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 4257)
  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 4257)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4257)
  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..4257
                     /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..3062
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     regulatory      3074..3544
                     /label=AOD terminator
                     /note="AOD terminator"
                     /regulatory_class="terminator"
     rep_origin      3591..4179
                     /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 gacactttat acttccggct
     2221 cgtataatac gacaaggtgt aaggaggact aaaccatggg taaggaaaag actcacgttt
     2281 cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg
     2341 ataatgtcgg gcaatcaggt gcgacaatct atcgattgta tgggaagccc gatgcgccag
     2401 agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca
     2461 gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc
     2521 ctgatgatgc atggttactc accactgcga tccccggcaa aacagcattc caggtattag
     2581 aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt
     2641 tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc
     2701 aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta
     2761 atggctggcc tgttgaacaa gtctggaaag aaatgcataa gcttttgcca ttctcaccgg
     2821 attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat
     2881 taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca
     2941 tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat
     3001 atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt
     3061 tctaacctct aggacacctt acgattattt agagagtatt tattagtttt attgtatgta
     3121 tacggatgtt ttattatcta tttatgccct tatattctgt aactatccaa aagtcctatc
     3181 ttatcaagcc agcaatctat gtccgcgaac gtcaactaaa aataagcttt ttatgctctt
     3241 ctctcttttt ttcccttcgg tataattata ccttgcatcc acagattctc ctgccaaatt
     3301 ttgcataatc ctttacaaca tggctatatg ggagcactta gcgccctcca aaacccatat
     3361 tgcctacgca tgtataggtg ttttttccac aatattttct ctgtgctctc tttttattaa
     3421 agagaagctc tatatcggag aagcttctgt ggccgttata ttcggcctta tcgtgggacc
     3481 acattgcctg aattggtttg ccccggaaga ttggggaaac ttggatctga ttaccttagc
     3541 tgcaggtacc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     3601 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     3661 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     3721 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     3781 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     3841 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     3901 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3961 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     4021 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     4081 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     4141 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     4201 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc ggtaccc
//
