Basic Vector Information
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 Exported 5094 bp ds-DNA circular SYN 17-AUG-2021
DEFINITION Reporter plasmid for FOXO3a. Forkhead responsive element with
luciferase readout. .
ACCESSION .
VERSION .
KEYWORDS FHRE-Luc
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)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Cell";
date: "1999-03-19"; volume: "96(6)"; pages: "857-68"
FEATURES Location/Qualifiers
source 1..5094
/organism="synthetic DNA construct"
/mol_type="other DNA"
polyA_signal 75..123
/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"
primer_bind 177..196
/label=RVprimer3
/note="pGL3 vector, forward primer"
promoter 367..563
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 414..549
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 476..495
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
regulatory 593..602
/regulatory_class="other"
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 599..2251
/codon_start=1
/gene="luc+"
/product="firefly luciferase"
/label=luciferase
/note="enhanced luc+ version of the luciferase gene"
/translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
primer_bind complement(665..683)
/label=LucNrev
/note="Firefly luciferase, reverse primer"
polyA_signal 2292..2413
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2303..2322)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 2357..2376
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
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"
primer_bind complement(2912..2931)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3591..4451)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 4214..4233
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4452..4556)
/gene="bla"
/label=AmpR promoter
rep_origin 4583..5038
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4670..4689)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4880..4901
/label=F1ori-F
/note="F1 origin, forward primer"
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