LOCUS       Exported                3650 bp ds-DNA     circular SYN 13-SEP-2021
DEFINITION  Pichia pastoris vector for methanol-inducible expression of a 
            secreted protein. For other reading frames, use pPIC6-alpha A or 
            pPIC6-alpha C.
ACCESSION   .
VERSION     .
KEYWORDS    pPIC6-B
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3650)
  AUTHORS   Invitrogen (Life Technologies)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 3650)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     Linearize with SacI, PmeI, or BstXI for integration at the AOX1 
            locus.
FEATURES             Location/Qualifiers
     source          1..3650
                     /organism="synthetic DNA construct"
                     /lab_host="Pichia pastoris"
                     /mol_type="other DNA"
     promoter        2..940
                     /gene="Pichia pastoris AOX1"
                     /note="Pichia pastoris AOX1 AOX1 promoter"
                     /note="AOX1 promoter;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"
                     /note="MF-alpha-1"
                     /note="alpha-factor secretion signal;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    1206..1280
                     /note="MCS"
                     /note="MCS;multiple cloning site"
     CDS             1279..1308
                     /codon_start=1
                     /product="Myc (human c-Myc oncogene) epitope tag"
                     /note="Myc"
                     /translation="EQKLISEEDL"
     CDS             1324..1341
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     terminator      1421..1667
                     /gene="Pichia pastoris AOX1"
                     /note="Pichia pastoris AOX1 AOX1 terminator"
                     /note="AOX1 terminator;transcription terminator for AOX1"
     promoter        1682..2095
                     /gene="S. cerevisiae TEF1"
                     /note="S. cerevisiae TEF1 TEF1 promoter"
                     /note="TEF1 promoter;promoter for EF-1-alpha"
     promoter        2102..2149
                     /note="EM7 promoter"
                     /note="EM7 promoter;synthetic bacterial promoter"
     CDS             2168..2566
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /note="Aspergillus terreus BSD"
                     /note="BSD;confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     terminator      2660..2907
                     /gene="S. cerevisiae CYC1"
                     /note="S. cerevisiae CYC1 CYC1 terminator"
                     /note="CYC1 terminator;transcription terminator for CYC1"
     rep_origin      complement(2982..3570)
                     /direction=LEFT
                     /note="ori"
                     /note="ori;high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"