Basic Vector Information
LIC-pIVEX-LC2 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-pIVEX-LC2 vector Sequence
LOCUS 40924_1764 5236 bp DNA circular SYN 17-DEC-2018
DEFINITION Expression vector LIC-pIVEX-LC2, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5236)
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 5236)
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 5236)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 5236)
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
(12-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..5236
/mol_type="other DNA"
/organism="synthetic DNA construct"
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"
RBS 670..692
/label=RBS
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
misc_feature 699..701
/label=start codon
/note="start codon"
misc_feature 711..718
/label=PmeI recognition site
/note="PmeI recognition site"
misc_feature 719..1385
/label=LIC stuffer fragment
/note="LIC stuffer fragment"
misc_feature 1386..1393
/label=PmeI recognition site
/note="PmeI recognition site"
CDS 1399..1419
/codon_start=1
/label=TEV site
/note="tobacco etch virus (TEV) protease recognition and
cleavage site"
/translation="ENLYFQG"
CDS 1429..2412
/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 2428..2445
/codon_start=1
/label=6xHis
/note="6xHis affinity tag"
/translation="HHHHHH"
misc_feature 2446..2451
/label=double stop codon (TAA TAA)
/note="double stop codon (TAA TAA)"
terminator 2553..2600
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
promoter complement(2966..2994)
/label=tet promoter
/note="E. coli promoter for tetracycline efflux protein
gene"
primer_bind complement(3015..3031)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(3039..3055)
/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(3063..3093)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(3108..3129)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(3417..4005)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4179..5036)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(5037..5141)
/label=AmpR promoter
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