LIC-pDEST-LC1 vector (V009871)

Basic Vector Information

      • Vector Name:
      • LIC-pDEST-LC1
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 6366 bp
      • Type:
      • Expression vector
      • Source/Author:
      • Dortay H, Akula UM, Westphal C, Sittig M, Mueller-Roeber B.

LIC-pDEST-LC1 vector Vector Map

LIC-pDEST-LC16366 bp3006009001200150018002100240027003000330036003900420045004800510054005700600063006xHis taginfrared fluorescent proteinTEV recognition sitePmeI recognition siteLIC stuffer fragmentPmeI recognition siteT7 terminatortet promoterAmpR promoterampRpBR322 originbomropT7 promoterRBSstart codon

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-pDEST-LC1 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                6366 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector LIC-pDEST-LC1, complete sequence.
ACCESSION   JF327846
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6366)
  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 6366)
  AUTHORS   Dortay H, Mueller-Roeber B.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-FEB-2011) Molecular Biology, University of Potsdam, 
            Germany, Karl-Liebknecht-Str. 24-25, Haus 20, Potsdam, Brandenburg 
            14476, Germany
REFERENCE   3  (bases 1 to 6366)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6366
                     /organism="Expression vector LIC-pDEST-LC1"
                     /mol_type="other DNA"
                     /label=Escherichia coli expression vector
                     /note="Escherichia coli expression vector"
                     /db_xref="taxon:1029635"
     misc_feature    10..27
                     /label=6xHis tag
                     /note="6xHis tag"
     CDS             10..27
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    37..1020
                     /label=infrared fluorescent protein
                     /note="infrared fluorescent protein"
     CDS             37..1020
                     /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    1021..1041
                     /label=TEV recognition site
                     /note="TEV recognition site"
     CDS             1021..1041
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     misc_feature    1060..1067
                     /label=PmeI recognition site
                     /note="PmeI recognition site"
     misc_feature    1068..1734
                     /label=LIC stuffer fragment
                     /note="LIC stuffer fragment"
     misc_feature    1735..1742
                     /label=PmeI recognition site
                     /note="PmeI recognition site"
     regulatory      1764..1892
                     /regulatory_class="terminator"
                     /note="T7 terminator"
     terminator      1820..1867
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(2233..2261)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     promoter        2374..2478
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2479..3339
                     /codon_start=1
                     /gene="ampR"
                     /product="beta-lactamase"
                     /label=ampR
                     /protein_id="AEE81070.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            2479..3339
                     /gene="ampR"
                     /label=ampR
     rep_origin      3483..4157
                     /label=pBR322 origin
                     /note="pBR322 origin"
     rep_origin      3510..4098
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    4284..4426
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(4528..4719)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        6284..6302
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      6284..6300
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     RBS             6334..6356
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     regulatory      6349..6355
                     /regulatory_class="ribosome_binding_site"
     misc_feature    6364..6366
                     /label=start codon
                     /note="start codon"

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