Basic Vector Information
- Vector Name:
- pLeo1246
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5848 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Scheller L, Schmollack M, Bertschi A, Mansouri M
- Promoter:
- SV40
pLeo1246 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 ℃.
pLeo1246 vector Sequence
LOCUS 62056_13990 5848 bp DNA circular SYN 30-JUN-2020
DEFINITION Cloning vector pLeo1246, complete sequence.
ACCESSION MT267307
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5848)
AUTHORS Scheller L, Schmollack M, Bertschi A, Mansouri M, Saxena P,
Fussenegger M.
TITLE Phosphoregulated orthogonal signal transduction in mammalian cells
JOURNAL Nat Commun 11 (1), 3085 (2020)
PUBMED 32555187
REFERENCE 2 (bases 1 to 5848)
AUTHORS Scheller L.
TITLE Direct Submission
JOURNAL Submitted (29-MAR-2020) STI IBI-STI LPDI, EPFL, AI 3123 (Batiment
AI), Station 19, Lausanne 1015, Switzerland
REFERENCE 3 (bases 1 to 5848)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat
Commun"; date: "2020"; volume: "11"; issue: "1"; pages: "3085"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(29-MAR-2020) STI IBI-STI LPDI, EPFL, AI 3123 (Batiment AI), Station
19, Lausanne 1015, Switzerland"
FEATURES Location/Qualifiers
source 1..5848
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(4..861)
/label=AmpR
/note="beta-lactamase"
promoter complement(862..966)
/label=AmpR promoter
misc_feature 1035..1238
/label=SV40 early promoter
/note="SV40 early promoter"
promoter 1288..1306
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature 1335..1340
/label=Kozak+ATG
/note="Kozak+ATG"
CDS 1338..2408
/codon_start=1
/transl_table=11
/product="ORK"
/label=ORK
/protein_id="QKE44346.1"
/translation="MTSGSQVQLVESGGGLVQAGGSLRLSCTASGRTGTIYSMAWFRQA
PGKEREFLATVGWSSGITYYMDSVKGRFTISRDKGKNTVYLQMDSLKPEDTAVYYCTAT
RAYSVGYDYWGQGTQVTVSSASGGGGSASLMQRLDGLVNYADALRERSHEFMNKLHVIL
GLLHLKSYKQLEDYILKTANNYQEEIGSLLGKIKSPVIAGFLISKINRATDLGHTLILN
SESQLPDSGSEDQVATLITTLGNLIENALEALGPEPGGEISVTLHYRHGWLHCEVNDDG
PGIAPDKIDHIFDKGVSTKGSERGVGLALVKQQVENLGGSIAVESEPGIFTQFFVQIPW
DGERSNRASASGSTGV"
misc_feature 1347..1709
/label=aCaffVHH
/note="aCaffVHH"
misc_feature 1428..1451
/label=CDR1
/note="CDR1"
misc_feature 1500..1523
/label=CDR2
/note="CDR2"
misc_feature 1641..1676
/label=CDR3
/note="CDR3"
misc_feature 1716..1730
/label=scfv
/note="scfv"
misc_feature 1737..2378
/label=DcuS truncation
/note="DcuS truncation"
misc_feature 1737..1739
/label=leu330
/note="leu330"
misc_feature 1749..1751
/label=leucin 334
/note="leucin 334"
misc_feature 1761..1763
/label=V338
/note="V338"
misc_feature 1764..1766
/label=asn339
/note="asn339"
misc_feature 1767..1784
/label=alpha helix
/note="alpha helix"
misc_feature 1785..2378
/label=Kinase domain 346-538
/note="Kinase domain 346-538"
polyA_signal 2445..2669
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2715..3143
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 3157..3486
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
CDS 3553..4344
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
polyA_signal 4521..4654
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4691..4707)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(4715..4731)
/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(4739..4769)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(4784..4805)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(5093..5678)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
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