LOCUS       sequence-339925-        5094 bp    DNA     circular SYN 15-DEC-2025
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)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell"; 
            date: "1999-03-19"; volume: "96(6)"; pages: "857-68"
COMMENT     Sequence Label: FHRE-Luc
FEATURES             Location/Qualifiers
     source          1..5094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     regulatory      593..602
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     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(join(3591..4382,4383..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"
ORIGIN
        1 tcgctattac gccagcccaa gctaccatga taagtaagta atattaaggt acgggaggta
       61 cttggagcgg ccgcaataaa atatctttat tttcattaca tctgtgtgtt ggttttttgt
      121 gtgaatcgat agtactaaca tacgctctcc atcaaaacaa aacgaaacaa aacaaactag
      181 caaaataggc tgtccccagt gcaagtgcag gtgccagaac atttctctat cgataggtac
      241 cgagctctta cgcgtgctag cgtgacagag tgagactctg tctctattta aataaataag
      301 taaataaata aactgggcaa acaatgaaaa tgaaaacatt gtgaaataca aagcagatct
      361 gcgatctgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
      421 ccctaactcc gcccagttcc gcccattctc cgccccatcg ctgactaatt ttttttattt
      481 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
      541 ttggaggcct aggcttttgc aaaaagcttg gcattccggt actgttggta aagccaccat
      601 ggaagacgcc aaaaacataa agaaaggccc ggcgccattc tatccgctgg aagatggaac
      661 cgctggagag caactgcata aggctatgaa gagatacgcc ctggttcctg gaacaattgc
      721 ttttacagat gcacatatcg aggtggacat cacttacgct gagtacttcg aaatgtccgt
      781 tcggttggca gaagctatga aacgatatgg gctgaataca aatcacagaa tcgtcgtatg
      841 cagtgaaaac tctcttcaat tctttatgcc ggtgttgggc gcgttattta tcggagttgc
      901 agttgcgccc gcgaacgaca tttataatga acgtgaattg ctcaacagta tgggcatttc
      961 gcagcctacc gtggtgttcg tttccaaaaa ggggttgcaa aaaattttga acgtgcaaaa
     1021 aaagctccca atcatccaaa aaattattat catggattct aaaacggatt accagggatt
     1081 tcagtcgatg tacacgttcg tcacatctca tctacctccc ggttttaatg aatacgattt
     1141 tgtgccagag tccttcgata gggacaagac aattgcactg atcatgaact cctctggatc
     1201 tactggtctg cctaaaggtg tcgctctgcc tcatagaact gcctgcgtga gattctcgca
     1261 tgccagagat cctatttttg gcaatcaaat cattccggat actgcgattt taagtgttgt
     1321 tccattccat cacggttttg gaatgtttac tacactcgga tatttgatat gtggatttcg
     1381 agtcgtctta atgtatagat ttgaagaaga gctgtttctg aggagccttc aggattacaa
     1441 gattcaaagt gcgctgctgg tgccaaccct attctccttc ttcgccaaaa gcactctgat
     1501 tgacaaatac gatttatcta atttacacga aattgcttct ggtggcgctc ccctctctaa
     1561 ggaagtcggg gaagcggttg ccaagaggtt ccatctgcca ggtatcaggc aaggatatgg
     1621 gctcactgag actacatcag ctattctgat tacacccgag ggggatgata aaccgggcgc
     1681 ggtcggtaaa gttgttccat tttttgaagc gaaggttgtg gatctggata ccgggaaaac
     1741 gctgggcgtt aatcaaagag gcgaactgtg tgtgagaggt cctatgatta tgtccggtta
     1801 tgtaaacaat ccggaagcga ccaacgcctt gattgacaag gatggatggc tacattctgg
     1861 agacatagct tactgggacg aagacgaaca cttcttcatc gttgaccgcc tgaagtctct
     1921 gattaagtac aaaggctatc aggtggctcc cgctgaattg gaatccatct tgctccaaca
     1981 ccccaacatc ttcgacgcag gtgtcgcagg tcttcccgac gatgacgccg gtgaacttcc
     2041 cgccgccgtt gttgttttgg agcacggaaa gacgatgacg gaaaaagaga tcgtggatta
     2101 cgtcgccagt caagtaacaa ccgcgaaaaa gttgcgcgga ggagttgtgt ttgtggacga
     2161 agtaccgaaa ggtcttaccg gaaaactcga cgcaagaaaa atcagagaga tcctcataaa
     2221 ggccaagaag ggcggaaaga tcgccgtgta attctagagt cggggcggcc ggccgcttcg
     2281 agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     2341 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     2401 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     2461 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg ataaggatcc
     2521 gtcgaccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca
     2581 tgactatcgt cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc
     2641 cggcagcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     2701 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     2761 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     2821 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     2881 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     2941 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     3001 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     3061 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     3121 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     3181 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     3241 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     3301 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     3361 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3421 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     3481 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     3541 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     3601 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     3661 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     3721 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     3781 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     3841 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     3901 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     3961 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     4021 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     4081 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     4141 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     4201 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     4261 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     4321 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     4381 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     4441 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     4501 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     4561 tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     4621 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     4681 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     4741 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     4801 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     4861 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     4921 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     4981 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttgc
     5041 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctct
//
