LOCUS       Exported                4558 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Extracellular expression vector pKanB alpha, complete sequence.
ACCESSION   EU285586
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4558)
  AUTHORS   Lin-Cereghino J, Hashimoto MD, Moy A, Castelo J, Orazem CC, Kuo P, 
            Xiong S, Gandhi V, Hatae CT, Chan A, Lin-Cereghino GP.
  TITLE     Direct selection of Pichia pastoris expression strains using new 
            G418 resistance vectors
  JOURNAL   Yeast 25 (4), 293-299 (2008)
  PUBMED    18327886
REFERENCE   2  (bases 1 to 4558)
  AUTHORS   Lin-Cereghino G, Lin-Cereghino J.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-NOV-2007) Biological Sciences, University of the 
            Pacific, 3601 Pacific Ave, Stockton, CA 95211, USA
REFERENCE   3  (bases 1 to 4558)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4558
                     /organism="Extracellular expression vector pKanB alpha"
                     /mol_type="other DNA"
                     /label=used for expressing proteins in Pichia pastoris
                     /note="used for expressing proteins in Pichia pastoris"
                     /db_xref="taxon:495221"
     misc_feature    1..940
                     /label=AOXI promoter and 5UTR
                     /note="AOXI promoter and 5UTR"
     promoter        2..940
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     CDS             941..1207
                     /codon_start=1
                     /gene="MF-alpha-1"
                     /product="N-terminal secretion signal from S. cerevisiae 
                     alpha-factor"
                     /label=-alpha-factor secretion signal
                     /note="Cleavage by the Kex2 protease occurs after the 
                     dibasic KR sequence. The EA dipeptides are then removed by 
                     dipeptidyl aminopeptidase A."
                     /translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLE
                     GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKREAEA"
     misc_feature    941..1193
                     /label=MAT alpha prepro signal peptide for secretion
                     /note="MAT alpha prepro signal peptide for secretion"
     misc_feature    1193..1264
                     /label=multiple cloning site
                     /note="multiple cloning site"
     CDS             1279..1308
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1324..1341
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1421..1667
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     CDS             2226..3041
                     /codon_start=1
                     /product="kanamycin/G418 resistance protein"
                     /label=kanamycin/G418 resistance protein
                     /note="selectable marker"
                     /protein_id="ABY67234.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             3060..3089
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3105..3122
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      3201..3447
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     terminator      3568..3815
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     rep_origin      complement(3890..4478)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"