LOCUS       Exported                3820 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  E.coli-K.pastoris shuttle vector pPpT4_Alpha_S, complete sequence.
ACCESSION   JQ519691
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3820)
  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 3820)
  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 3820)
  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..3820
                     /organism="Escherichia coli-Pichia pastoris shuttle vector 
                     pPpT4_Alpha_S"
                     /mol_type="other DNA"
                     /db_xref="taxon:1182044"
     regulatory      141..1071
                     /regulatory_class="promoter"
                     /note="AOX1 promoter"
     misc_feature    1085..1351
                     /note="N-terminal prepro signal; derived from Saccharomyces
                     cerevisiae alpha factor"
     regulatory      1372..1631
                     /regulatory_class="terminator"
                     /note="AOX1 terminator"
     terminator      1372..1618
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     regulatory      1633..2190
                     /regulatory_class="promoter"
                     /note="ILV5 promoter"
     regulatory      2191..2254
                     /regulatory_class="promoter"
                     /note="EM72 synthetic promoter"
     CDS             2256..2630
                     /codon_start=1
                     /product="zeocin resistance protein"
                     /label=zeocin resistance protein
                     /protein_id="AFJ20655.1"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     regulatory      2632..3107
                     /regulatory_class="terminator"
                     /note="AOD terminator"
     rep_origin      3128..3800
                     /label=pUC origin of replication
                     /note="pUC origin of replication"
     rep_origin      3154..3742
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"