Basic Vector Information
- Vector Name:
- LIC-pDEST-LC2
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6349 bp
- Type:
- Expression vector
- Replication origin:
- ori
- Source/Author:
- Dortay H, Akula UM, Westphal C, Sittig M, Mueller-Roeber B.
- Promoter:
- tet
LIC-pDEST-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-pDEST-LC2 vector Sequence
LOCUS 40924_1754 6349 bp DNA circular SYN 17-DEC-2018
DEFINITION Expression vector LIC-pDEST-LC2, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6349)
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 6349)
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 6349)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6349)
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..6349
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 10..17
/label=PmeI recognition site
/note="PmeI recognition site"
misc_feature 18..684
/label=LIC stuffer fragment
/note="LIC stuffer fragment"
misc_feature 685..692
/label=PmeI recognition site
/note="PmeI recognition site"
CDS 698..718
/codon_start=1
/label=TEV site
/note="tobacco etch virus (TEV) protease recognition and
cleavage site"
/translation="ENLYFQG"
CDS 728..1711
/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 1718..1735
/codon_start=1
/label=6xHis
/note="6xHis affinity tag"
/translation="HHHHHH"
misc_feature 1736..1738
/label=stop codon (TAG)
/note="stop codon (TAG)"
terminator 1803..1850
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
promoter complement(2216..2244)
/label=tet promoter
/note="E. coli promoter for tetracycline efflux protein
gene"
promoter 2357..2461
/label=AmpR promoter
CDS 2462..3319
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 3493..4081
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature complement(4267..4409)
/label=bom
/note="basis of mobility region from pBR322"
CDS complement(4514..4702)
/codon_start=1
/label=rop
/note="Rop protein, which maintains plasmids at low copy
number"
/translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
promoter 6267..6285
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
RBS 6317..6339
/label=RBS
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
misc_feature 6347..6349
/label=start codon
/note="start codon"
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