LIC-pPICZ-LC2 vector (V009862)

Basic Vector Information

      • Vector Name:
      • LIC-pPICZ-LC2
      • Length:
      • 4938 bp
      • Type:
      • Expression vector
      • Source/Author:
      • Dortay H, Akula UM, Westphal C, Sittig M, Mueller-Roeber B.

LIC-pPICZ-LC2 vector Vector Map

LIC-pPICZ-LC24938 bp60012001800240030003600420048005' promoter regionstart codonPmeI recognition siteLIC stuffer fragmentPmeI recognition siteTEV recognition siteinfrared fluorescent protein6xHis tagterminatorEM7 promoterzeocin resistance markerCYC1 terminatorpBR322 origin

Plasmid Resuspension Protocol:

1. Centrifuge at 5,000×g for 5 min.

2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.

3. Close the tube and incubate for 10 minutes at room temperature.

4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.

5.Store the plasmid at -20 ℃.

LIC-pPICZ-LC2 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

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"

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