LOCUS       sequence-354598-        6072 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Luciferase reporter for ARE/NRF2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6072)
  AUTHORS   Grigolon G, Araldi E, Erni R, Wu JY, Thomas C, La Fortezza M, Laube 
            B, Pohlmann D, Stoffel M, Zarse K, Carreira EM, Ristow M, Fischer F
  TITLE     Grainyhead 1 acts as a drug-inducible conserved transcriptional 
            regulator linked to insulin signaling and lifespan.
  JOURNAL   Nat Commun. 2022 Jan 10;13(1):107. doi: 10.1038/s41467-021-27732-4.
  PUBMED    35013237
REFERENCE   2  (bases 1 to 6072)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2022-01-10"; volume: "13(1):107. doi"; pages: " 
            10.1038/s41467-021-27732-4"
COMMENT     Sequence Label: pGL4.27_ARE/NRF2-SPE
FEATURES             Location/Qualifiers
     source          1..6072
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(1..792,793..861))
                     /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    966..1014
                     /note="synthetic polyadenylation signal"
     misc_feature    1028..1119
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     1068..1087
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        1258..1289
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      1316..1325
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1322..2971
                     /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"
     CDS             2975..3094
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    3146..3267
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3157..3176)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3211..3230
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3461..3818
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3461..3652
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3484..3504)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3669..3804
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3731..3750
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3849..4886
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4865..4873
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    4910..4958
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5114..5131)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5285..5873)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5365..5384)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttaccagtgc ttgatcagtg aggcaccgat ctcagcgatc tgcctatttc gttcgtccat
       61 agtggcctga ctccccgtcg tgtagatcac tacgattcgt gagggcttac catcaggccc
      121 cagcgcagca atgatgccgc gagagccgcg ttcaccggcc cccgatttgt cagcaatgaa
      181 ccagccagca gggagggccg agcgaagaag tggtcctgct actttgtccg cctccatcca
      241 gtctatgagc tgctgtcgtg atgctagagt aagaagttcg ccagtgagta gtttccgaag
      301 agttgtggcc attgctactg gcatcgtggt atcacgctcg tcgttcggta tggcttcgtt
      361 caactctggt tcccagcggt caagccgggt cacatgatca cccatattat gaagaaatgc
      421 agtcagctcc ttagggcctc cgatcgttgt cagaagtaag ttggccgcgg tgttgtcgct
      481 catggtaatg gcagcactac acaattctct taccgtcatg ccatccgtaa gatgcttttc
      541 cgtgaccggc gagtactcaa ccaagtcgtt ttgtgagtag tgtatacggc gaccaagctg
      601 ctcttgcccg gcgtctatac gggacaacac cgcgccacat agcagtactt tgaaagtgct
      661 catcatcggg aatcgttctt cggggcggaa agactcaagg atcttgccgc tattgagatc
      721 cagttcgata tagcccactc ttgcacccag ttgatcttca gcatctttta ctttcaccag
      781 cgtttcgggg tgtgcaaaaa caggcaagca aaatgccgca aagaagggaa tgagtgcgac
      841 acgaaaatgt tggatgctca tactcgtcct ttttcaatat tattgaagca tttatcaggg
      901 ttactagtac gtctctcaag gataagtaag taatattaag gtacgggagg tattggacag
      961 gccgcaataa aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgaatcga
     1021 tagtactaac atacgctctc catcaaaaca aaacgaaaca aaacaaacta gcaaaatagg
     1081 ctgtccccag tgcaagtgca ggtgccagaa catttctctg gcctaactgg ccggtacctc
     1141 acagtgactc agcatttaaa cacagtgact cagcatttaa acacagtgac tcagcattaa
     1201 agagctcgct agcctcgagg atatcaagat ctggcctcgg cggccaagct tagacactag
     1261 agggtatata atggaagctc gacttccagc ttggcaatcc ggtactgttg gtaaagccac
     1321 catggaagat gccaaaaaca ttaagaaggg cccagcgcca ttctacccac tcgaagacgg
     1381 gaccgccggc gagcagctgc acaaagccat gaagcgctac gccctggtgc ccggcaccat
     1441 cgcctttacc gacgcacata tcgaggtgga cattacctac gccgagtact tcgagatgag
     1501 cgttcggctg gcagaagcta tgaagcgcta tgggctgaat acaaaccatc ggatcgtggt
     1561 gtgcagcgag aatagcttgc agttcttcat gcccgtgttg ggtgccctgt tcatcggtgt
     1621 ggctgtggcc ccagctaacg acatctacaa cgagcgcgag ctgctgaaca gcatgggcat
     1681 cagccagccc accgtcgtat tcgtgagcaa gaaagggctg caaaagatcc tcaacgtgca
     1741 aaagaagcta ccgatcatac aaaagatcat catcatggat agcaagaccg actaccaggg
     1801 cttccaaagc atgtacacct tcgtgacttc ccatttgcca cccggcttca acgagtacga
     1861 cttcgtgccc gagagcttcg accgggacaa aaccatcgcc ctgatcatga acagtagtgg
     1921 cagtaccgga ttgcccaagg gcgtagccct accgcaccgc accgcttgtg tccgattcag
     1981 tcatgcccgc gaccccatct tcggcaacca gatcatcccc gacaccgcta tcctcagcgt
     2041 ggtgccattt caccacggct tcggcatgtt caccacgctg ggctacttga tctgcggctt
     2101 tcgggtcgtg ctcatgtacc gcttcgagga ggagctattc ttgcgcagct tgcaagacta
     2161 taagattcaa tctgccctgc tggtgcccac actatttagc ttcttcgcta agagcactct
     2221 catcgacaag tacgacctaa gcaacttgca cgagatcgcc agcggcgggg cgccgctcag
     2281 caaggaggta ggtgaggccg tggccaaacg cttccaccta ccaggcatcc gccagggcta
     2341 cggcctgaca gaaacaacca gcgccattct gatcaccccc gaaggggacg acaagcctgg
     2401 cgcagtaggc aaggtggtgc ccttcttcga ggctaaggtg gtggacttgg acaccggtaa
     2461 gacactgggt gtgaaccagc gcggcgagct gtgcgtccgt ggccccatga tcatgagcgg
     2521 ctacgttaac aaccccgagg ctacaaacgc tctcatcgac aaggacggct ggctgcacag
     2581 cggcgacatc gcctactggg acgaggacga gcacttcttc atcgtggacc ggctgaagag
     2641 cctgatcaaa tacaagggct accaggtagc cccagccgaa ctggagagca tcctgctgca
     2701 acaccccaac atcttcgacg ccggggtcgc cggcctgccc gacgacgatg ccggcgagct
     2761 gcccgccgca gtcgtcgtgc tggaacacgg taaaaccatg accgagaagg agatcgtgga
     2821 ctatgtggcc agccaggtta caaccgccaa gaagctgcgc ggtggtgttg tgttcgtgga
     2881 cgaggtgcct aaaggactga ccggcaagtt ggacgcccgc aagatccgcg agattctcat
     2941 taaggccaag aagggcggca agatcgccgt gaattctcac ggcttccctc ccgaggtgga
     3001 ggagcaggcc gccggcaccc tgcccatgag ctgcgcccag gagagcggca tggatagaca
     3061 ccctgctgct tgcgccagcg ccaggatcaa cgtctaaggc cgcgactcta gagtcggggc
     3121 ggccggccgc ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact
     3181 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     3241 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     3301 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtaaa
     3361 atcgataagg atccgtttgc gtattgggcg ctcttccgct gatctgcgca gcaccatggc
     3421 ctgaaataac ctctgaaaga ggaacttggt tagctacctt ctgaggcgga aagaaccagc
     3481 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     3541 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     3601 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     3661 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     3721 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     3781 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcgattct tctgacacta
     3841 gcgccaccat gaagaagccc gaactcaccg ctaccagcgt tgaaaaattt ctcatcgaga
     3901 agttcgacag tgtgagcgac ctgatgcagt tgtcggaggg cgaagagagc cgagccttca
     3961 gcttcgatgt cggcggacgc ggctatgtac tgcgggtgaa tagctgcgct gatggcttct
     4021 acaaagaccg ctacgtgtac cgccacttcg ccagcgctgc actacccatc cccgaagtgt
     4081 tggacatcgg cgagttcagc gagagcctga catactgcat cagtagacgc gcccaaggcg
     4141 ttactctcca agacctcccc gaaacagagc tgcctgctgt gttacagcct gtcgccgaag
     4201 ctatggatgc tattgccgcc gccgacctca gtcaaaccag cggcttcggc ccattcgggc
     4261 cccaaggcat cggccagtac acaacctggc gggatttcat ttgcgccatt gctgatcccc
     4321 atgtctacca ctggcagacc gtgatggacg acaccgtgtc cgccagcgta gctcaagccc
     4381 tggacgaact gatgctgtgg gccgaagact gtcccgaggt gcgccacctc gtccatgccg
     4441 acttcggcag caacaacgtc ctgaccgaca acggccgcat caccgccgta atcgactggt
     4501 ccgaagctat gttcggggac agtcagtacg aggtggccaa catcttcttc tggcggccct
     4561 ggctggcttg catggagcag cagactcgct acttcgagcg ccggcatccc gagctggccg
     4621 gcagccctcg tctgcgagcc tacatgctgc gcatcggcct ggatcagctc taccagagcc
     4681 tcgtggacgg caacttcgac gatgctgcct gggctcaagg ccgctgcgat gccatcgtcc
     4741 gcagcggggc cggcaccgtc ggtcgcacac aaatcgctcg ccggagcgca gccgtatgga
     4801 ccgacggctg cgtcgaggtg ctggccgaca gcggcaaccg ccggcccagt acacgaccgc
     4861 gcgctaagga ggtaggtcga gtttaaactc tagaaccggt catggccgca ataaaatatc
     4921 tttattttca ttacatctgt gtgttggttt tttgtgtgtt cgaactagat gctgtcgacc
     4981 gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat
     5041 cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc
     5101 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5161 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5221 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5281 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5341 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5401 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5461 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     5521 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5581 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5641 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5701 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     5761 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5821 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     5881 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     5941 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6001 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagcggccgc aaatgctaaa
     6061 ccactgcagt gg
//
