LIC-pLEXSY-LC1 vector (V009865)

Basic Vector Information

      • Vector Name:
      • LIC-pLEXSY-LC1
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 9305 bp
      • Type:
      • Expression vector
      • Source/Author:
      • Dortay H, Akula UM, Westphal C, Sittig M, Mueller-Roeber B.

LIC-pLEXSY-LC1 vector Vector Map

LIC-pLEXSY-LC19305 bp4008001200160020002400280032003600400044004800520056006000640068007200760080008400880092005' SSU 18S rRNAstart codon6xHis taginfrared fluorescent proteinTEV recognition sitePmeI recognition siteLIC stuffer fragmentPmeI recognition sitenourseothricin resistance marker3' SSU 18S rRNAT3 promoterM13 revlac operatorlac promoterCAP binding siteoriampRAmpR promoterf1 oriM13 fwdT7 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-pLEXSY-LC1 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                9305 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector LIC-pLEXSY-LC1, complete sequence.
ACCESSION   JF327852
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9305)
  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 9305)
  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 9305)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9305
                     /organism="Expression vector LIC-pLEXSY-LC1"
                     /mol_type="other DNA"
                     /label=Leishmania tarentolae expression vector
                     /note="Leishmania tarentolae expression vector"
                     /db_xref="taxon:1029641"
     rRNA            1..686
                     /product="18S ribosomal RNA"
                     /label=5' SSU 18S rRNA
                     /note="5' SSU 18S rRNA"
     misc_feature    1094..1096
                     /label=start codon
                     /note="start codon"
     misc_feature    1106..1123
                     /label=6xHis tag
                     /note="6xHis tag"
     CDS             1106..1123
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    1133..2116
                     /label=infrared fluorescent protein
                     /note="infrared fluorescent protein"
     CDS             1133..2116
                     /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    2117..2137
                     /label=TEV recognition site
                     /note="TEV recognition site"
     CDS             2117..2137
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     misc_feature    2156..2163
                     /label=PmeI recognition site
                     /note="PmeI recognition site"
     misc_feature    2164..2830
                     /label=LIC stuffer fragment
                     /note="LIC stuffer fragment"
     misc_feature    2831..2838
                     /label=PmeI recognition site
                     /note="PmeI recognition site"
     CDS             4209..4733
                     /codon_start=1
                     /product="nourseothricin resistance marker"
                     /label=nourseothricin resistance marker
                     /protein_id="AEE81079.1"
                     /translation="MKISVIPEQVAETLDAENHFIVREVFDVHLSDQGFELSTRSVSPY
                     RKDYISDDDSDEDSACYGAFIDQELVGKIELNSTWNDLASIEHIVVSHTHRGKGVAHSL
                     IEFAKKWALSRQLLGIRLETQTNNVPACNLYAKCGFTLGGIDLFTYKTRPQVSNETAMY
                     WYWFSGAQDDA"
     rRNA            5357..6441
                     /product="18S ribosomal RNA"
                     /label=3' SSU 18S rRNA
                     /note="3' SSU 18S rRNA"
     promoter        complement(6459..6477)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6498..6514)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6522..6538
                     /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(6546..6576)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6591..6612
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6900..7488)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7659..8519)
                     /codon_start=1
                     /gene="ampR"
                     /product="beta-lactamase"
                     /label=ampR
                     /protein_id="AEE81078.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(7659..8519)
                     /gene="ampR"
                     /label=ampR
     promoter        complement(8520..8624)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      8651..9107
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     9248..9264
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        9274..9292
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"

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