Basic Vector Information
- Vector Name:
- pLeo1227
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6430 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Scheller L, Schmollack M, Bertschi A, Mansouri M
- Promoter:
- SV40
pLeo1227 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 ℃.
pLeo1227 vector Sequence
LOCUS 62056_13965 6430 bp DNA circular SYN 30-JUN-2020
DEFINITION Cloning vector pLeo1227, complete sequence.
ACCESSION MT267302
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6430)
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 6430)
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 6430)
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..6430
/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 1029..1054
/label=DcuR BE2
/note="DcuR BE2"
misc_feature 1073..1098
/label=DcuR BE1
/note="DcuR BE1"
misc_feature 1112..1133
/label=DcuR BE2
/note="DcuR BE2"
misc_feature 1152..1177
/label=DcuR BE1
/note="DcuR BE1"
misc_feature 1191..1216
/label=DcuR BE2
/note="DcuR BE2"
misc_feature 1235..1260
/label=DcuR BE1
/note="DcuR BE1"
promoter 1274..1312
/label=minimal CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 1357..1375
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature 1404..1409
/label=Kozak+ATG
/note="Kozak+ATG"
CDS 1416..2912
/label=SEAP
/note="secreted alkaline phosphatase from human placenta"
polyA_signal 3027..3251
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 3297..3725
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 3739..4068
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
CDS 4135..4926
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
polyA_signal 5103..5236
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5273..5289)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(5297..5313)
/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(5321..5351)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(5366..5387)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(5675..6260)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
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