LOCUS       40924_35409        7107 bp DNA     circular SYN 18-DEC-2018
DEFINITION  E.coli-K.pastoris shuttle vector pPpGUT1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7107)
  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 7107)
  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 7107)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7107)
  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..7107
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1..900
                     /label=GUT1 promoter
                     /note="GUT1 promoter"
                     /regulatory_class="promoter"
     CDS             901..2766
                     /codon_start=1
                     /product="glycerol kinase"
                     /label=glycerol kinase
                     /note="GUT1"
                     /protein_id="AFJ20664.1"
                     /translation="MGKDYTPLVATIDIGTTSTRAILFDYHGQEVAKHQIEYSTSAQDD
                     IKRKRSQIISSEGISLTVSDDLEVESVDNKAGPTLQFPQPGWVECRPSHILANAVQCLA
                     ACLVTMENKNLDRDEKNKYKLISIGVANMRETTVVWSKKTGKPLYNGIVWNDTRNNDIV
                     DEYTAKYSEKEREEMRTLCGCPISTYFSATKFRWLLKHVPEVKQAYDNADGDLMFGTID
                     SWLIYHLTNEKSHVTDVTNASRTNFMNIETNKYDDRLLKFWDVDTSKVILPEIRSSAEV
                     YGHFKVPHLESIGYVESYLTDDALALLETIEGAPLAGCLGDQSASLVGQLAVRKGDAKC
                     TYGTGAFLLYNTGDQTLISEHGALTTVGYWFPGLDESEDGKHSSKPQYALEGSIAVAGS
                     VVQWLRDNLRLISKAQDVGPLASQVDNSGGVVFVPAFSGLFAPYWDSNSRGTIFGLTQY
                     TSASHIARAALEGVCFQTRAILKAMISDAGASADFLEESSKATGHNPLSVLAVDGGMSK
                     SDEMMQIQADILGPCVTVRRSINPECTALGAAIAAGFGVPKEDRIWGSLKECTEAILEG
                     NKMYLAAGNTSLDFKATLSDEVRRKEWRLWENAIAKAKGWLKDTA"
     regulatory      2767..3266
                     /label=GUT1 terminator
                     /note="GUT1 terminator"
                     /regulatory_class="terminator"
     promoter        3273..4204
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     terminator      4232..4478
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     misc_feature    4492..5184
                     /note="integration sequence for recombination into target
                     locus"
     regulatory      5185..5242
                     /label=EM72 synthetic promoter
                     /note="EM72 synthetic promoter"
                     /regulatory_class="promoter"
     CDS             5243..6100
                     /label=AmpR
                     /note="beta-lactamase"
     terminator      6110..6297
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     rep_origin      6461..7049
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ataccgaaag gtaaacaact tcggggaatt gccctggtta aacatttatt aagcgagata
       61 aataggggat agcgagatag ggggcggaga agaagaaggg tgttaaattg ctgaaatctc
      121 tcaatctgga agaaacggaa taaattaact ccttcctgag ataataagat ccgactctgc
      181 tatgacccca cacggtactg acctcggcat accccattgg atctggtgcg aagcaacagg
      241 tcctgaaacc tttatcacgt gtagtagatt gaccttccag caaaaaaagg cattatatat
      301 tttgttgttg aaggggtgag gggaggtgca ggtggttctt ttattcgtct tgtagttaat
      361 tttcccgggg ttgcggagcg tcaaaagttt gcccgatctg atagcttgca agatgccacc
      421 gcttatccaa cgcacttcag agagcttgcc gtagaaagaa cgttttcctc gtagtattcc
      481 agcacttcat ggtgaagtcg ctatttcacc gaaggggggg tattaaggtt gcgcaccccc
      541 tccccacacc ccagaatcgt ttattggctg ggttcaatgg cgtttgagtt agcacatttt
      601 ttccttaaac accctccaaa cacggataaa aatgcatgtg catcctgaaa ctggtagaga
      661 tgcgtactcc gtgctccgat aataacagtg gtgttggggt tgctgttagc tcacgcactc
      721 cgtttttttt caaccagcaa aattcgatgg ggagaaactt ggggtacttt gccgactcct
      781 ccaccatact ggtatataaa taatactcgc ccacttttcg tttgctgctt ttatatttca
      841 aggactgaaa aagactcttc ttctactttt tcacactata ccacagatat atctactata
      901 atgggaaaag actatacacc actagttgct accatcgata ttggtactac ctccaccaga
      961 gctattcttt ttgactacca cggtcaggaa gtggccaagc accagatcga gtactctacc
     1021 tctgctcagg atgatatcaa aagaaagcgt tctcagatca tctcttccga aggtatttcc
     1081 ctgacagttt ctgacgactt ggaagttgag tccgttgaca ataaggctgg tccaactttg
     1141 caatttcctc agccaggctg ggttgaatgt cgtccaagtc acatcttggc caacgccgtt
     1201 cagtgtcttg ctgcttgttt agtcaccatg gagaacaaga acttggaccg agatgaaaaa
     1261 aataaataca agctcatatc tatcggtgtt gccaacatga gagagaccac ggttgtttgg
     1321 tccaagaaga caggaaagcc tctttacaac ggtattgtgt ggaacgatac cagaaacaac
     1381 gatattgttg acgagtacac cgccaagtac tctgagaagg agagagagga aatgcgaacc
     1441 ttgtgtggtt gtccaatctc cacctacttt tctgcaacca agttcagatg gttactgaag
     1501 catgttcctg aggtcaaaca ggcctatgac aatgctgatg gggatctaat gtttggtact
     1561 attgactctt ggttgattta ccacttgact aacgaaaaat cccacgtcac tgatgttacc
     1621 aatgcctcca gaaccaactt catgaacatt gaaaccaaca aatatgacga cagacttttg
     1681 aaattctggg acgtcgatac ttccaaagtc atccttccag aaatcagatc ttccgcagaa
     1741 gtctacggac acttcaaggt cccacacttg gagtctattg gatatgttga gtcttacctt
     1801 actgatgacg cccttgctct tctggaaact attgaaggtg ctcctttggc tggatgtctg
     1861 ggtgaccaat ctgcctcttt ggttggacag ttggctgtca gaaagggtga tgccaaatgt
     1921 acatatggta ccggtgcttt cttgctgtac aacactggtg atcagacttt gatttctgag
     1981 cacggtgctt tgaccactgt gggatattgg ttcccaggtt tggatgagtc cgaagatggc
     2041 aaacactctt ctaagccaca gtatgctttg gagggatcga ttgctgtcgc tggatctgtc
     2101 gtgcaatggt taagagataa ccttcgtttg atttccaagg ctcaggacgt cggaccattg
     2161 gcttctcaag ttgacaactc tggaggtgtg gtatttgttc cagcattttc aggattgttt
     2221 gccccttact gggattccaa ctccagagga accattttcg gtctgaccca atacacctca
     2281 gcttctcata ttgctagagc tgctttggaa ggtgtctgtt tccaaactag agccattttg
     2341 aaggccatga tcagcgatgc aggagcttct gctgactttt tggaggaatc atccaaggcc
     2401 actggccaca accctctgtc agttcttgcc gtggacggag gtatgtccaa atcagacgag
     2461 atgatgcaga tccaagctga tattttgggt ccatgtgtca ctgttagacg ttccatcaac
     2521 cctgaatgta ctgcactggg agctgccatt gctgccggtt ttggtgtccc taaggaagat
     2581 agaatttggg gttccttgaa ggaatgtacc gaggccattc ttgagggtaa caagatgtac
     2641 ttggctgcag ggaacacttc tttggacttc aaggccacat tgagcgacga ggtcagaaga
     2701 aaggaatgga gattgtggga aaatgccatt gcaaaggcaa agggctggct taaggacact
     2761 gcttaagaag agcagctgta attatattat catgttaggt cattagacaa tgtattgtag
     2821 ttatatatgt aatgataatg aagaatggaa atcgatggat atcgcggatt taatcacttc
     2881 ctcatctctt tctttcagat tgcactcaac gtaggcagca gcagtccatg accagtgtga
     2941 aagactccat cactattgac agagggctgg cacagaatgt tgaaaagatc catggagtta
     3001 accctgtgtt tttgatagac aaaattctca gggaaaaaat cttagatagt ctgttttgga
     3061 aaaaacactg ctttctacta gatattctgg aactgcaaga tcttgcaagt gaagtggaga
     3121 ttattggcac atacgacacc aatcaaaact ctagagcaac cgactatatt tgtctgttac
     3181 tgaaaatgct acagttacag ccgaaagacg agatagtgat atacatgttg acccaaccct
     3241 atttcaagta tgtgacagct ctggcaattt aaataacatc caaagacgaa aggttgaatg
     3301 aaaccttttt gccatccgac atccacaggt ccattctcac acataagtgc caaacgcaac
     3361 aggaggggat acactagcag cagaccgttg caaacgcagg acctccactc ctcttctcct
     3421 caacacccac ttttgccatc gaaaaaccag cccagttatt gggcttgatt ggagctcgct
     3481 cattccaatt ccttctatta ggctactaac accatgactt tattagcctg tctatcctgg
     3541 cccccctggc gaggttcatg tttgtttatt tccgaatgca acaagctccg cattacaccc
     3601 gaacatcact ccagatgagg gctttctgag tgtggggtca aatagtttca tgttccccaa
     3661 atggcccaaa actgacagtt taaacgctgt cttggaacct aatatgacaa aagcgtgatc
     3721 tcatccaaga tgaactaagt ttggttcgtt gaaatgctaa cggccagttg gtcaaaaaga
     3781 aacttccaaa agtcggcata ccgtttgtct tgtttggtat tgattgacga atgctcaaaa
     3841 ataatctcat taatgcttag cgcagtctct ctatcgcttc tgaaccccgg tgcacctgtg
     3901 ccgaaacgca aatggggaaa cacccgcttt ttggatgatt atgcattgtc tccacattgt
     3961 atgcttccaa gattctggtg ggaatactgc tgatagccta acgttcatga tcaaaattta
     4021 actgttctaa cccctacttg acagcaatat ataaacagaa ggaagctgcc ctgtcttaac
     4081 cttttttttt atcatcatta ttagcttact ttcataattg cgactggttc caattgacaa
     4141 gcttttgatt ttaacgactt ttaacgacaa cttgagaaga tcaaaaaaca actaattatt
     4201 gaaagaattc actagtggcg cgccgcggcc gccaagagga tgtcagaatg ccatttgcct
     4261 gagagatgca ggcttcattt ttgatacttt tttatttgta acctatatag tataggattt
     4321 tttttgtcat tttgtttctt ctcgtacgag cttgctcctg atcagcctat ctcgcagcag
     4381 atgaatatct tgtggtaggg gtttgggaaa atcattcgag tttgatgttt ttcttggtat
     4441 ttcccactcc tcttcagagt acagaagatt aagtgagacc ttcgtttgtg cgcgttgtat
     4501 attcggttgg ttttcccttc tgtgagagtt tatcagttgt tagagccgtt attgaatgac
     4561 tatagaaaac tgcgaataag aaagcacggc aacgctgact tgacctacgt ggatcagctg
     4621 gtagatgagt tattaactga ggagaagttt tgtgggttaa cactacccag attggtcaac
     4681 cgattgcgac tggaggatga cggtctacta gaccctagag aaagtgaact acagagcgag
     4741 ttagaaaatg aggaagatga tattgggggt tgaaattggg ctaatttatt atgaccatta
     4801 caattatatt agatacatgt acataggcag gacttgatcg ttacatcctt aggatccttt
     4861 tctcttctac ttatatgctt agaaagccct agcagatggt ctgggtttat ctctgccaac
     4921 tcttggtcca cctctgattg gatctttctt ggccacttta ttcttaggct tgaccaatga
     4981 gtctgtcaaa aggctatcta aattcaaatt atccggcaag atcacgatac gaacgttgtt
     5041 tcctcttata tttataaact ccaggctgac cgggtttctg tttctaactg tcattttaac
     5101 gtttttcagg ttgatattca tagacggcgt caccgagatg atggtaccgc tgacaattgt
     5161 tccgttcttt agttcaactt gcacgtgttg acactttata cttccggctc gtataatacg
     5221 acaaggtgta aggaggacta aaatgagtat tcaacatttc cgtgtcgccc ttattccctt
     5281 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     5341 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     5401 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     5461 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     5521 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     5581 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     5641 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     5701 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     5761 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     5821 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     5881 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     5941 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     6001 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     6061 acagatcgct gagataggtg cctcactgat taagcattgg taacctaggg cgaatttctt
     6121 atgatttatg atttttatta ttaaataagt tataaaaaaa ataagtgtat acaaatttta
     6181 aagtgactct taggttttaa aacgaaaatt cttattcttg agtaactctt tcctgtaggt
     6241 caggttgctt tctcaggtat agcatgaggt cgctcttatt gaccacacct ctaccggcat
     6301 gccgagcaaa tgcctgcaaa tcgctcccca tttcacccaa ttgtagatat gctaactcca
     6361 gcaatgagtt gatgaatctc ggtgtgtatt ttatgtcctc agaggacaac tgcaggtacc
     6421 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     6481 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     6541 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     6601 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     6661 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     6721 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     6781 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     6841 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     6901 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     6961 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     7021 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     7081 tggccttttg ctggcctttt gctcaca
//
