Basic Vector Information
LIC-pLEXSY-LC1 vector Vector Map
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
LOCUS 40924_1779 9305 bp DNA circular SYN 17-DEC-2018
DEFINITION Expression vector LIC-pLEXSY-LC1, complete sequence.
ACCESSION .
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)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 9305)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE";
date: "2011"; volume: "6"; issue: "4"; pages: "E18900"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(14-FEB-2011) Molecular Biology, University of Potsdam, Germany,
Karl-Liebknecht-Str. 24-25, Haus 20, Potsdam, Brandenburg 14476,
Germany"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..9305
/mol_type="other DNA"
/organism="synthetic DNA construct"
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"
CDS 1106..1123
/codon_start=1
/label=6xHis
/note="6xHis affinity tag"
/translation="HHHHHH"
CDS 1133..2116
/codon_start=1
/label=IFP1.4
/note="bacteriophytochrome-based monomeric infrared
fluorescent protein (Shu et al., 2009)"
/translation="MARDPLPFFPPLYLGGPEITTENCEREPIHIPGSIQPHGALLTAD
GHSGEVLQVSLNAATFLGQEPTVLRGQTLAALLPEQWPALQAALPPGCPDALQYRATLD
WPAAGHLSLTVHRVAELLILEFEPTEAWDSIGPHALRNAMFALESAPNLRALAEVATQT
VRELTGFDRVMLYKFAPDATGEMIAEARREGMQAFLGHRFPASHTPAQARALYTRHLLR
LTADTRAAAVPLDPVLNPQTNAPTPLGGAVLRATSPMHMQYLRNMGVGSSLSVSVVVGG
QLWGLIVCHHQTPYVLPPDLRTTLEELGRKLSGQVQRKEAGMDELYK"
CDS 2117..2137
/codon_start=1
/label=TEV site
/note="tobacco etch virus (TEV) protease recognition and
cleavage 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 complement(6522..6538)
/label=lac operator
/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 complement(6591..6612)
/label=CAP binding site
/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(7662..8519)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(8520..8624)
/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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