LOCUS       Exported                7238 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  E.coli-K.pastoris shuttle vector pPpHIS4, complete sequence.
ACCESSION   JQ519697
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7238)
  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 7238)
  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 7238)
  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..7238
                     /organism="Escherichia coli-Pichia pastoris shuttle vector
                     pPpHIS4"
                     /mol_type="other DNA"
                     /db_xref="taxon:1182038"
     regulatory      139..1068
                     /regulatory_class="promoter"
                     /note="AOX1 promoter"
     regulatory      1097..1356
                     /regulatory_class="terminator"
                     /note="AOX1 terminator"
     terminator      1097..1343
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     regulatory      1363..2222
                     /regulatory_class="promoter"
                     /note="ADH1 promoter"
     promoter        1506..2210
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             2223..4757
                     /codon_start=1
                     /product="HIS4"
                     /label=HIS4
                     /note="HIS4; codon optimized"
                     /protein_id="AFJ20662.1"
                     /translation="MTFPLLPAYASVAEFDNSLSLVGKAVFPYAADQLHNLIKFTQSTE
                     LQVNVQVESSVTEDQFEELIDNLLKLYNNGINEVILDLDLAERVVQRMIPGARVIYRTL
                     VDKVASLPANASIAVPFSSPLGDLKSFTNGGSRTVYAFSETAKLVDVTSTVASGIIPII
                     DARQLTTEYELSEDVKKFPVSEILLASLTTDRPDGLFTTLVADSSNYSLGLVYSSKKSI
                     PEAIRTQTGVYQSRRHGLWYKGATSGATQKLLGIELDCDGDCLKFVVEQTGVGFCHLER
                     TSCFGQSKGLRAMEATLWDRKSNAPEGSYTKRLFDDEVLLNAKIREEADELAEAKSKED
                     IAWECADLFYFALVRCAKYGVTLDEVERNLDMKSLKVTRRKGDAKPGYTKEQPKEESKP
                     KEVPSEGRIELCKIDVSKASSQEIEDALRRPIQKTEQIMELVKPIVDNVRQNGDKALLE
                     LTAKFDGVALKTPVLEAPFPEELMQLPDNVKRAIDLSIDNVRKFHEAQLTETLQVETCP
                     GVVCSRFARPIEKVGLYIPGGTAILPSTSLMLGVPAKVAGCKEIVFASPPKKDGTLTPE
                     VIYVAHKVGAKCIVLAGGAQAVAAMAYGTETVPKCDKIFGPGNQFVTAAKMMVQNDTSA
                     LCSIDMPAGPSEVLVIADKYADPDFVASDLLSQAEHGIDSQVILLAVDMTDKELARIED
                     AVHNQAVQLPRVEIVRKCIAHSTTLSVATYEQALEMSNQYAPEHLILQIENASSYVDQV
                     QHAGSVFVGAYSPESCGDYSSGTNHTLPTYGYARQYSGVNTATFQKFITSQDVTPEGLK
                     HIGQAVMDLAAVEGLDAHRNAVKVRMEKLGLI"
     regulatory      4758..5289
                     /regulatory_class="terminator"
                     /note="TIF51A terminator"
     regulatory      5290..5347
                     /regulatory_class="promoter"
                     /note="EM72 synthetic promoter"
     CDS             5354..6214
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /note="confers resistance to ampicillin"
                     /protein_id="AFJ20663.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     regulatory      6221..6520
                     /regulatory_class="terminator"
                     /note="ADH1 terminator"
     terminator      6221..6408
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     rep_origin      6546..7218
                     /label=pUC origin of replication
                     /note="pUC origin of replication"
     rep_origin      6572..7160
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"