Basic Vector Information
LIC-pDEST-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-pDEST-LC1 vector Sequence
LOCUS 40924_1749 6366 bp DNA circular SYN 17-DEC-2018
DEFINITION Expression vector LIC-pDEST-LC1, complete sequence.
ACCESSION .
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)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6366)
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..6366
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 10..27
/codon_start=1
/label=6xHis
/note="6xHis affinity tag"
/translation="HHHHHH"
CDS 37..1020
/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 1021..1041
/codon_start=1
/label=TEV site
/note="tobacco etch virus (TEV) protease recognition and
cleavage 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"
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
/label=AmpR promoter
CDS 2479..3336
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 3510..4098
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature complement(4284..4426)
/label=bom
/note="basis of mobility region from pBR322"
CDS complement(4531..4719)
/codon_start=1
/label=rop
/note="Rop protein, which maintains plasmids at low copy
number"
/translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
promoter 6284..6302
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
RBS 6334..6356
/label=RBS
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
misc_feature 6364..6366
/label=start codon
/note="start codon"
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