Basic Vector Information
- Vector Name:
- pleo614
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5781 bp
- Type:
- Expression vector
- Replication origin:
- ori
- Source/Author:
- Scheller L, Strittmatter T, Fuchs D, Bojar D, Fussenegger M.
pleo614 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 ℃.
pleo614 vector Sequence
LOCUS 40924_28012 5781 bp DNA circular SYN 18-DEC-2018
DEFINITION Expression vector pleo614, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5781)
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 5781)
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 5781)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 5781)
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..5781
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 8..387
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 388..591
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 636..654
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
sig_peptide 685..747
/label=Ig-kappa leader
/note="leader sequence from mouse immunoglobulin kappa
light
chain"
CDS 754..1077
/codon_start=1
/label=FKBP
/note="human FK506-binding protein FKBP12"
/translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
LKLE"
polyA_signal 1154..1378
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 1424..1852
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 1866..2195
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
CDS 2262..3053
/codon_start=1
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
polyA_signal 3230..3363
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3400..3416)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(3424..3440)
/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(3448..3478)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(3493..3514)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(3802..4387)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4561..5418)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(5419..5523)
/label=AmpR promoter
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