LOCUS       sequence-417440-        4557 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The plasmid contains a 312 bp DNA fragment that contained 5 
            canonical E-boxes (GCCACGTGCA) spaced by 50 nucleotides and cloned 
            into pGL4.23[luc2/minP] (XhoI/HindIII)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4557)
  AUTHORS   Sporbeck K, Haas ML, Pastor-Maldonado CJ, Schussele DS, Hunter C, 
            Takacs Z, Diogo de Oliveira AL, Franz-Wachtel M, Charsou C, 
            Pfisterer SG, Gubas A, Haller PK, Knorr RL, Kaulich M, Macek B, 
            Eskelinen EL, Simonsen A, Proikas-Cezanne T
  TITLE     The ABL-MYC axis controls WIPI1-enhanced autophagy in lifespan 
            extension.
  JOURNAL   Commun Biol. 2023 Aug 24;6(1):872. doi: 10.1038/s42003-023-05236-9.
  PUBMED    37620393
REFERENCE   2  (bases 1 to 4557)
  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: "Commun 
            Biol."; date: "2023-08-24"; volume: "6(1):872. doi"; pages: " 
            10.1038/s42003-023-05236-9"
COMMENT     Sequence Label: pGL4.23-5xE-box
FEATURES             Location/Qualifiers
     source          1..4557
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        351..382
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      409..418
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             415..2067
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     polyA_signal    2111..2232
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2122..2141)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2176..2195
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2480..2497)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2651..3239)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2731..2750)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3439..4230,4231..4299))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    4404..4452
                     /note="synthetic polyadenylation signal"
     misc_feature    4466..4557
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4506..4525
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctcgcta gcctcgagag ttcccacccc gcccagaccc
       61 cttgccacgt gcaactctgc ctattaaact tacagagtag ttcccacccc gcccagaccc
      121 cttgccacgt gcaactctgc ctattaaact tacagagtag ttcccacccc gcccagaccc
      181 cttgccacgt gcaactctgc ctattaaact tacagagtag ttcccacccc gcccagaccc
      241 cttgccacgt gcaactctgc ctattaaact tacagagtag ttcccacccc gcccagaccc
      301 cttgccacgt gcaactctgc ctattaaact tacagagtaa gcttagacac tagagggtat
      361 ataatggaag ctcgacttcc agcttggcaa tccggtactg ttggtaaagc caccatggaa
      421 gatgccaaaa acattaagaa gggcccagcg ccattctacc cactcgaaga cgggaccgcc
      481 ggcgagcagc tgcacaaagc catgaagcgc tacgccctgg tgcccggcac catcgccttt
      541 accgacgcac atatcgaggt ggacattacc tacgccgagt acttcgagat gagcgttcgg
      601 ctggcagaag ctatgaagcg ctatgggctg aatacaaacc atcggatcgt ggtgtgcagc
      661 gagaatagct tgcagttctt catgcccgtg ttgggtgccc tgttcatcgg tgtggctgtg
      721 gccccagcta acgacatcta caacgagcgc gagctgctga acagcatggg catcagccag
      781 cccaccgtcg tattcgtgag caagaaaggg ctgcaaaaga tcctcaacgt gcaaaagaag
      841 ctaccgatca tacaaaagat catcatcatg gatagcaaga ccgactacca gggcttccaa
      901 agcatgtaca ccttcgtgac ttcccatttg ccacccggct tcaacgagta cgacttcgtg
      961 cccgagagct tcgaccggga caaaaccatc gccctgatca tgaacagtag tggcagtacc
     1021 ggattgccca agggcgtagc cctaccgcac cgcaccgctt gtgtccgatt cagtcatgcc
     1081 cgcgacccca tcttcggcaa ccagatcatc cccgacaccg ctatcctcag cgtggtgcca
     1141 tttcaccacg gcttcggcat gttcaccacg ctgggctact tgatctgcgg ctttcgggtc
     1201 gtgctcatgt accgcttcga ggaggagcta ttcttgcgca gcttgcaaga ctataagatt
     1261 caatctgccc tgctggtgcc cacactattt agcttcttcg ctaagagcac tctcatcgac
     1321 aagtacgacc taagcaactt gcacgagatc gccagcggcg gggcgccgct cagcaaggag
     1381 gtaggtgagg ccgtggccaa acgcttccac ctaccaggca tccgccaggg ctacggcctg
     1441 acagaaacaa ccagcgccat tctgatcacc cccgaagggg acgacaagcc tggcgcagta
     1501 ggcaaggtgg tgcccttctt cgaggctaag gtggtggact tggacaccgg taagacactg
     1561 ggtgtgaacc agcgcggcga gctgtgcgtc cgtggcccca tgatcatgag cggctacgtt
     1621 aacaaccccg aggctacaaa cgctctcatc gacaaggacg gctggctgca cagcggcgac
     1681 atcgcctact gggacgagga cgagcacttc ttcatcgtgg accggctgaa gagcctgatc
     1741 aaatacaagg gctaccaggt agccccagcc gaactggaga gcatcctgct gcaacacccc
     1801 aacatcttcg acgccggggt cgccggcctg cccgacgacg atgccggcga gctgcccgcc
     1861 gcagtcgtcg tgctggaaca cggtaaaacc atgaccgaga aggagatcgt ggactatgtg
     1921 gccagccagg ttacaaccgc caagaagctg cgcggtggtg ttgtgttcgt ggacgaggtg
     1981 cctaaaggac tgaccggcaa gttggacgcc cgcaagatcc gcgagattct cattaaggcc
     2041 aagaagggcg gcaagatcgc cgtgtaataa ttctagagtc ggggcggccg gccgcttcga
     2101 gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     2161 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     2221 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
     2281 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga taaggatccg
     2341 tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat
     2401 gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc
     2461 ggcagcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     2521 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     2581 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     2641 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     2701 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     2761 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     2821 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     2881 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     2941 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     3001 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     3061 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     3121 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     3181 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     3241 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     3301 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     3361 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagc ggccgcaaat
     3421 gctaaaccac tgcagtggtt accagtgctt gatcagtgag gcaccgatct cagcgatctg
     3481 cctatttcgt tcgtccatag tggcctgact ccccgtcgtg tagatcacta cgattcgtga
     3541 gggcttacca tcaggcccca gcgcagcaat gatgccgcga gagccgcgtt caccggcccc
     3601 cgatttgtca gcaatgaacc agccagcagg gagggccgag cgaagaagtg gtcctgctac
     3661 tttgtccgcc tccatccagt ctatgagctg ctgtcgtgat gctagagtaa gaagttcgcc
     3721 agtgagtagt ttccgaagag ttgtggccat tgctactggc atcgtggtat cacgctcgtc
     3781 gttcggtatg gcttcgttca actctggttc ccagcggtca agccgggtca catgatcacc
     3841 catattatga agaaatgcag tcagctcctt agggcctccg atcgttgtca gaagtaagtt
     3901 ggccgcggtg ttgtcgctca tggtaatggc agcactacac aattctctta ccgtcatgcc
     3961 atccgtaaga tgcttttccg tgaccggcga gtactcaacc aagtcgtttt gtgagtagtg
     4021 tatacggcga ccaagctgct cttgcccggc gtctatacgg gacaacaccg cgccacatag
     4081 cagtactttg aaagtgctca tcatcgggaa tcgttcttcg gggcggaaag actcaaggat
     4141 cttgccgcta ttgagatcca gttcgatata gcccactctt gcacccagtt gatcttcagc
     4201 atcttttact ttcaccagcg tttcggggtg tgcaaaaaca ggcaagcaaa atgccgcaaa
     4261 gaagggaatg agtgcgacac gaaaatgttg gatgctcata ctcgtccttt ttcaatatta
     4321 ttgaagcatt tatcagggtt actagtacgt ctctcaagga taagtaagta atattaaggt
     4381 acgggaggta ttggacaggc cgcaataaaa tatctttatt ttcattacat ctgtgtgttg
     4441 gttttttgtg tgaatcgata gtactaacat acgctctcca tcaaaacaaa acgaaacaaa
     4501 acaaactagc aaaataggct gtccccagtg caagtgcagg tgccagaaca tttctct
//
