Basic Vector Information
- Vector Name:
- LIC-pPICZ-LC2
- Antibiotic Resistance:
- Bleomycin
- Length:
- 4938 bp
- Type:
- Expression vector
- Replication origin:
- ori
- Source/Author:
- Dortay H, Akula UM, Westphal C, Sittig M, Mueller-Roeber B.
- Promoter:
- AOX1
LIC-pPICZ-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-pPICZ-LC2 vector Sequence
LOCUS 40924_1794 4938 bp DNA circular SYN 17-DEC-2018
DEFINITION Expression vector LIC-pPICZ-LC2, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4938)
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 4938)
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 4938)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 4938)
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..4938
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 2..935
/label=AOX1 promoter
/note="inducible promoter, regulated by methanol"
misc_feature 945..947
/label=start codon
/note="start codon"
misc_feature 957..964
/label=PmeI recognition site
/note="PmeI recognition site"
misc_feature 965..1631
/label=LIC stuffer fragment
/note="LIC stuffer fragment"
misc_feature 1632..1639
/label=PmeI recognition site
/note="PmeI recognition site"
CDS 1645..1665
/codon_start=1
/label=TEV site
/note="tobacco etch virus (TEV) protease recognition and
cleavage site"
/translation="ENLYFQG"
CDS 1675..2658
/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 2665..2682
/codon_start=1
/label=6xHis
/note="6xHis affinity tag"
/translation="HHHHHH"
misc_feature 2683..2685
/label=stop codon (TGA)
/note="stop codon (TGA)"
terminator 2762..3008
/label=AOX1 terminator
/note="transcription terminator for AOX1"
promoter 3023..3434
/label=TEF1 promoter
/note="promoter for EF-1-alpha"
promoter 3442..3489
/label=EM7 promoter
/note="synthetic bacterial promoter"
CDS 3508..3879
/codon_start=1
/label=BleoR
/note="antibiotic-binding protein"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
terminator 3948..4195
/label=CYC1 terminator
/note="transcription terminator for CYC1"
rep_origin complement(4270..4858)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
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