LOCUS       Exported                7107 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  E.coli-K.pastoris shuttle vector pPpGUT1, complete sequence.
ACCESSION   JQ519698
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)
  AUTHORS   .
  TITLE     Direct Submission
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
                     /organism="Escherichia coli-Pichia pastoris shuttle vector
                     pPpGUT1"
                     /mol_type="other DNA"
                     /db_xref="taxon:1182037"
     regulatory      1..900
                     /regulatory_class="promoter"
                     /note="GUT1 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
                     /regulatory_class="terminator"
                     /note="GUT1 terminator"
     regulatory      3275..4204
                     /regulatory_class="promoter"
                     /note="AOX1 promoter"
     regulatory      4233..4491
                     /regulatory_class="terminator"
                     /note="AOX1 terminator"
     terminator      4233..4478
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     misc_feature    4492..5184
                     /note="integration sequence for recombination into target 
                     locus"
     regulatory      5185..5242
                     /regulatory_class="promoter"
                     /note="EM72 synthetic promoter"
     CDS             5243..6103
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /note="confers resistance to ampicillin"
                     /protein_id="AFJ20665.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     regulatory      6110..6409
                     /regulatory_class="terminator"
                     /note="ADH1 terminator"
     terminator      6110..6297
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     rep_origin      6435..7107
                     /label=pUC origin of replication
                     /note="pUC origin of replication"
     rep_origin      6461..7049
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"