LOCUS       Exported                4938 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector LIC-pPICZ-LC2, complete sequence.
ACCESSION   JF327851
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4938)
  AUTHORS   Dortay H, Akula UM, Westphal C, Sittig M, Mueller-Roeber B.
  TITLE     High-Throughput Protein Expression Using a Combination of 
            Ligation-Independent Cloning (LIC) and Infrared Fluorescent Protein 
            (IFP) Detection
  JOURNAL   PLoS ONE 6 (4), E18900 (2011)
  PUBMED    21541323
REFERENCE   2  (bases 1 to 4938)
  AUTHORS   Dortay H, Mueller-Roeber B.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-FEB-2011) Molecular Biology, University of Potsdam, 
            Germany, Karl-Liebknecht-Str. 24-25, Haus 20, Potsdam, Brandenburg 
            14476, Germany
REFERENCE   3  (bases 1 to 4938)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4938
                     /organism="Expression vector LIC-pPICZ-LC2"
                     /mol_type="other DNA"
                     /label=Pichia pastoris expression vector
                     /note="Pichia pastoris expression vector"
                     /db_xref="taxon:1029644"
     regulatory      1..944
                     /regulatory_class="promoter"
                     /note="5 promoter region"
     misc_feature    945..947
                     /label=start codon
                     /note="start codon"
     misc_feature    957..964
                     /label=PmeI recognition site
                     /note="PmeI recognition site"
     misc_feature    965..1631
                     /label=LIC stuffer fragment
                     /note="LIC stuffer fragment"
     misc_feature    1632..1639
                     /label=PmeI recognition site
                     /note="PmeI recognition site"
     misc_feature    1645..1665
                     /label=TEV recognition site
                     /note="TEV recognition site"
     CDS             1645..1665
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     misc_feature    1675..2658
                     /label=infrared fluorescent protein
                     /note="infrared fluorescent protein"
     CDS             1675..2658
                     /codon_start=1
                     /product="bacteriophytochrome-based monomeric infrared 
                     fluorescent protein (Shu et al., 2009)"
                     /label=IFP1.4
                     /translation="MARDPLPFFPPLYLGGPEITTENCEREPIHIPGSIQPHGALLTAD
                     GHSGEVLQVSLNAATFLGQEPTVLRGQTLAALLPEQWPALQAALPPGCPDALQYRATLD
                     WPAAGHLSLTVHRVAELLILEFEPTEAWDSIGPHALRNAMFALESAPNLRALAEVATQT
                     VRELTGFDRVMLYKFAPDATGEMIAEARREGMQAFLGHRFPASHTPAQARALYTRHLLR
                     LTADTRAAAVPLDPVLNPQTNAPTPLGGAVLRATSPMHMQYLRNMGVGSSLSVSVVVGG
                     QLWGLIVCHHQTPYVLPPDLRTTLEELGRKLSGQVQRKEAGMDELYK"
     misc_feature    2665..2685
                     /label=6xHis tag
                     /note="6xHis tag"
     CDS             2665..2682
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    2683..2685
                     /label=stop codon (TGA)
                     /note="stop codon (TGA)"
     regulatory      2686..3027
                     /regulatory_class="terminator"
                     /note="terminator"
     terminator      2762..3008
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        3023..3434
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        3442..3489
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3508..3882
                     /codon_start=1
                     /product="zeocin resistance marker"
                     /label=zeocin resistance marker
                     /protein_id="AEE81077.1"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     terminator      3948..4195
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     rep_origin      4211..4884
                     /label=pBR322 origin
                     /note="pBR322 origin"
     rep_origin      complement(4270..4858)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"