Basic Vector Information
- Vector Name:
- pEL-platform
- Antibiotic Resistance:
- Ampicillin
- Length:
- 3707 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Fujii W, Kano K, Sugiura K, Naito K.
pEL-platform 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 ℃.
pEL-platform vector Sequence
LOCUS 40924_17314 3707 bp DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector pEL-platform DNA, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3707)
AUTHORS Fujii W, Kano K, Sugiura K, Naito K.
TITLE Repeatable Construction Method for Engineered Zinc Finger Nuclease
Based on Overlap Extension PCR and TA-cloning
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 3707)
AUTHORS Fujii W, Kano K, Sugiura K, Naito K.
TITLE Direct Submission
JOURNAL Submitted (06-FEB-2013) Contact:Wataru Fujii The University of
Tokyo, Graduate School of Agricultural and Life Sciences; Yayoi
1-1-1-7A-303, Bunkyo-ku, Tokyo 113-8657, Japan URL
:http://www.vm.a.u-tokyo.ac.jp/iden/
REFERENCE 3 (bases 1 to 3707)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 3707)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(06-FEB-2013) Contact:Wataru Fujii The University of Tokyo, Graduate
School of Agricultural and Life Sciences"; volume: " Yayoi
1-1-1-7A-303, Bunkyo-ku, Tokyo 113-8657, Japan URL :http"; pages:
"//www.vm.a.u-tokyo.ac.jp/iden"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..3707
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 67..370
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 371..574
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 620..638
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
CDS 699..722
/codon_start=1
/label=FLAG
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/translation="DYKDDDDK"
CDS 729..749
/codon_start=1
/label=SV40 NLS
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/translation="PKKKRKV"
CDS 831..1418
/codon_start=1
/label=FokI cleavage domain
/note="nonspecific DNA cleavage domain of the FokI
endonuclease (Li et al., 1992)"
/translation="QLVKSELEEKKSELRHKLKYVPHEYIELIEIARNPTQDRILEMKV
MEFFMKVYGYRGEHLGGSRKPDGAIYTVGSPIDYGVIVDTKAYSGGYNLPIGQADEMER
YVEENQTRNKHLNPNEWWKVYPSSVTEFKFLFVSGHFKGNYKAQLTRLNHITNCNGAVL
SVEELLIGGEMIKAGTLTLEEVRRKFNNGEINF"
promoter complement(1622..1640)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 1872..1976
/label=AmpR promoter
CDS 1977..2834
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 3008..3596
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
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