Basic Vector Information
- Vector Name:
- FHRE-Luc
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5094 bp
- Type:
- Mammalian Expression, Luciferase
- Replication origin:
- ori
- Copy Number:
- High Copy
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- RVprimer3
FHRE-Luc 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 ℃.
FHRE-Luc vector Sequence
LOCUS 40924_820 5094 bp DNA circular SYN 17-AUG-2021
DEFINITION Reporter plasmid for FOXO3a. Forkhead responsive element with
luciferase readout. .
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5094)
AUTHORS Brunet A, Bonni A, Zigmond MJ, Lin MZ, Juo P, Hu LS, Anderson MJ,
Arden KC, Blenis J, Greenberg ME
TITLE Akt promotes cell survival by phosphorylating and inhibiting a
Forkhead transcription factor.
JOURNAL Cell 1999 Mar 19;96(6):857-68.
PUBMED 10102273
REFERENCE 2 (bases 1 to 5094)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 5094)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Cell";
date: "1999-03-19"; volume: "96(6)"; pages: "857-68"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5094
/mol_type="other DNA"
/organism="synthetic DNA construct"
polyA_signal 75..123
/label=poly(A) signal
/note="synthetic polyadenylation signal"
misc_feature 137..228
/label=pause site
/note="RNA polymerase II transcriptional pause signal from
the human alpha-2 globin gene"
promoter 367..563
/label=SV40 promoter
/note="SV40 early promoter"
CDS 599..2248
/codon_start=1
/label=luciferase
/note="firefly luciferase"
/translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
polyA_signal complement(2292..2413)
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2661..2678)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2832..3420)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(3594..4451)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(4452..4556)
/label=AmpR promoter
rep_origin 4583..5038
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
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