LIC-pIVEX-LC1 vector (V009869)

Basic Vector Information

      • Vector Name:
      • LIC-pIVEX-LC1
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 5250 bp
      • Type:
      • Expression vector
      • Source/Author:
      • Dortay H, Akula UM, Westphal C, Sittig M, Mueller-Roeber B.

LIC-pIVEX-LC1 vector Vector Map

LIC-pIVEX-LC15250 bp6001200180024003000360042004800M13 fwdT7 promoterRBSstart codon6xHis tagFactor Xa siteinfrared fluorescent proteinTEV recognition sitePmeI recognition siteLIC stuffer fragmentPmeI recognition siteT7 terminatortet promoterM13 revlac operatorlac promoterCAP binding sitepBR322 originampRAmpR promoter

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-pIVEX-LC1 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                5250 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector LIC-pIVEX-LC1, complete sequence.
ACCESSION   JF327844
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5250)
  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 5250)
  AUTHORS   Dortay H, Mueller-Roeber B.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-FEB-2011) Molecular Biology, University of Potsdam, 
            Germany, Karl-Liebknecht-Str. 24-25, Haus 20, Potsdam, Brandenburg 
            14476, Germany
REFERENCE   3  (bases 1 to 5250)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5250
                     /organism="Expression vector LIC-pIVEX-LC1"
                     /mol_type="other DNA"
                     /db_xref="taxon:1029637"
     primer_bind     381..397
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        620..638
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      620..636
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     RBS             670..692
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     regulatory      686..691
                     /regulatory_class="ribosome_binding_site"
     misc_feature    699..701
                     /label=start codon
                     /note="start codon"
     misc_feature    711..728
                     /label=6xHis tag
                     /note="6xHis tag"
     CDS             711..728
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             738..749
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_feature    756..1739
                     /label=infrared fluorescent protein
                     /note="infrared fluorescent protein"
     CDS             756..1739
                     /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    1740..1760
                     /label=TEV recognition site
                     /note="TEV recognition site"
     CDS             1740..1760
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     misc_feature    1779..1786
                     /label=PmeI recognition site
                     /note="PmeI recognition site"
     misc_feature    1787..2453
                     /label=LIC stuffer fragment
                     /note="LIC stuffer fragment"
     misc_feature    2454..2461
                     /label=PmeI recognition site
                     /note="PmeI recognition site"
     terminator      2567..2614
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     regulatory      2574..2611
                     /regulatory_class="terminator"
                     /note="T7 terminator"
     promoter        complement(2980..3008)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(3029..3045)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3053..3069
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(3077..3107)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3122..3143
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      3372..4045
                     /label=pBR322 origin
                     /note="pBR322 origin"
     rep_origin      complement(3431..4019)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4190..5050)
                     /codon_start=1
                     /gene="ampR"
                     /product="beta-lactamase"
                     /label=ampR
                     /protein_id="AEE81068.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(4190..5050)
                     /gene="ampR"
                     /label=ampR
     promoter        complement(5051..5155)
                     /gene="bla"
                     /label=AmpR promoter

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