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FHRE-Luc vector (V009791#)

Basic Vector Information

Basic Vector Information
Vector Name FHRE-Luc Antibiotic Resistance Ampicillin
Length 5094 bp Type Mammalian Expression, Luciferase
Copy Number High Copy Cloning Method Restriction Enzyme
5' Primer RVprimer3

FHRE-Luc vector Vector Map

FHRE-Luc5094 bp6001200180024003000360042004800synthetic polyadenylation signalpause siteSV40 promoterKozak sequenceSV40 poly(A) signalL4440oriAmpRAmpR promoterf1 ori

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

Copy Sequence

Download GeneBank File(.gb)

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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