Basic Vector Information
- Vector Name:
- pLeo645
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7413 bp
- Type:
- Expression vector
- Replication origin:
- ori
- Source/Author:
- Scheller L, Strittmatter T, Fuchs D, Bojar D, Fussenegger M.
- Promoter:
- SV40
pLeo645 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 ℃.
pLeo645 vector Sequence
LOCUS 40924_28087 7413 bp DNA circular SYN 18-DEC-2018
DEFINITION Expression vector pLeo645, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7413)
AUTHORS Scheller L, Strittmatter T, Fuchs D, Bojar D, Fussenegger M.
TITLE Generalized extracellular molecule sensor platform for programming
cellular behavior
JOURNAL Nat. Chem. Biol. 14 (7), 723-729 (2018)
PUBMED 29686358
REFERENCE 2 (bases 1 to 7413)
AUTHORS Scheller L, Strittmatter T.
TITLE Direct Submission
JOURNAL Submitted (06-NOV-2017) D-BSSE, ETH Zurich, Mattenstrasse 26, Basel
4058, Switzerland
REFERENCE 3 (bases 1 to 7413)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 7413)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat. Chem.
Biol."; date: "2018"; volume: "14"; issue: "7"; pages: "723-729"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(06-NOV-2017) D-BSSE, ETH Zurich, Mattenstrasse 26, Basel 4058,
Switzerland"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7413
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 7..215
/label=Psv40
/note="Psv40"
promoter 18..214
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 65..200
/label=SV40 ori
/note="SV40 origin of replication"
regulatory 240..249
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
misc_feature 246..308
/label=igk secretion signal
/note="igk secretion signal"
misc_feature 246..294
/label=secrete
/note="secrete"
misc_feature 309
/label=scfv
/note="scfv"
misc_feature 315..692
/label=anti RR120
/note="anti RR120"
misc_feature 705..1448
/label=mouse EpoR-D1_D2_TM
/note="mouse EpoR-D1_D2_TM"
misc_feature 978..980
/label=F93A
/note="F93A"
misc_feature 1380..1448
/label=transmembrane
/note="transmembrane"
misc_feature 1461..2732
/label=mFGFR 405-end
/note="mFGFR 405-end"
polyA_signal 2786..3010
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 3056..3484
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 3498..3827
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
CDS 3894..4685
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
polyA_signal 4862..4995
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5032..5048)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(5056..5072)
/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(5080..5110)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(5125..5146)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(5434..6019)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6193..7050)
/label=AmpR
/note="beta-lactamase"
promoter complement(7051..7155)
/label=AmpR promoter
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