Basic Vector Information
- Vector Name:
- pLeo1286
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6256 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Scheller L, Schmollack M, Bertschi A, Mansouri M
pLeo1286 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 ℃.
pLeo1286 vector Sequence
LOCUS 62056_14030 6256 bp DNA circular SYN 30-JUN-2020
DEFINITION Cloning vector pLeo1286, complete sequence.
ACCESSION MT267315
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6256)
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 6256)
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 6256)
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..6256
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 1..9
/label=SV40 polyA
/note="SV40 polyA"
primer_bind complement(49..65)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(73..89)
/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(97..127)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(142..163)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(451..1036)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(1210..2067)
/label=AmpR
/note="beta-lactamase"
promoter complement(2068..2172)
/label=AmpR promoter
misc_feature 2241..2444
/label=SV40 early promoter
/note="SV40 early promoter"
promoter 2494..2512
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature 2541..2546
/label=Kozak+ATG
/note="Kozak+ATG"
CDS 2544..4022
/codon_start=1
/transl_table=11
/product="Dual-ORK"
/label=Dual-ORK
/protein_id="QKE44354.1"
/translation="MTSGSQVQLVESGGGLVQAGGSLRLSCTASGRTGTIYSMAWFRQA
PGKEREFLATVGWSSGITYYMDSVKGRFTISRDKGKNTVYLQMDSLKPEDTAVYYCTAT
RAYSVGYDYWGQGTQVTVSSASGGGGSASKTEVRKLMQRLDGLVNYADALRERSHEFMN
KLHVILGLLHLKSYKQLEDYILKTANNYQEEIGSLLGKIKSPVIAGFLISKINRATDLG
HTLILNSESQLPDSGSEDQVATLITTLGNLIENALEALGPEPGGEISVTLHYRHGWLHC
EVNDDGPGIAPDKIDHIFDKGVSTKGSERGVGLALVKQQVENLGGSIAVESEPGIFTQF
FVQIPWDGERSNRASASGGGGSASGSQVQLVESGGGLVQAGGSLRLSCTASGRTGTIYS
MAWFRQAPGKEREFLATVGWSSGITYYMDSVKGRFTISRDKGKNTVYLQMDSLKPEDTA
VYYCTATRAYSVGYDYWGQGTQVTVSSASGSTGV"
misc_feature 2553..2915
/label=aCaffVHH
/note="aCaffVHH"
misc_feature 2634..2657
/label=CDR1
/note="CDR1"
misc_feature 2706..2729
/label=CDR2
/note="CDR2"
misc_feature 2847..2882
/label=CDR3
/note="CDR3"
misc_feature 2922..2936
/label=scfv
/note="scfv"
misc_feature 2943..3602
/label=DcuS truncation
/note="DcuS truncation"
misc_feature 3630..3998
/label=aCaffVHH
/note="aCaffVHH"
misc_feature 3717..3740
/label=CDR1
/note="CDR1"
misc_feature 3789..3812
/label=CDR2
/note="CDR2"
misc_feature 3930..3965
/label=CDR3
/note="CDR3"
polyA_signal 4059..4283
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 4329..4757
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 4771..5100
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
CDS 5167..5958
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
polyA_signal 6135..6256
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
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