LOCUS       sequence-354578-        6217 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase reporter for GRHL.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6217)
  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 6217)
  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: "Nat 
            Commun."; date: "2022-01-10"; volume: "13(1):107. doi"; pages: " 
            10.1038/s41467-021-27732-4"
COMMENT     Sequence Label: pGL4.27_GRHL-SPE
FEATURES             Location/Qualifiers
     source          1..6217
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(137..928,929..997))
                     /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    1102..1150
                     /note="synthetic polyadenylation signal"
     misc_feature    1164..1255
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     1204..1223
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        1539..1570
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      1597..1606
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1603..3252
                     /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             3256..3375
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    3427..3548
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3438..3457)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3492..3511
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3742..4099
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3742..3933
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3765..3785)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3950..4085
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4012..4031
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4130..5167
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             5146..5154
                     /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    5191..5239
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5395..5412)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5566..6154)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5646..5665)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
       61 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagcgg ccgcaaatgc
      121 taaaccactg cagtggttac cagtgcttga tcagtgaggc accgatctca gcgatctgcc
      181 tatttcgttc gtccatagtg gcctgactcc ccgtcgtgta gatcactacg attcgtgagg
      241 gcttaccatc aggccccagc gcagcaatga tgccgcgaga gccgcgttca ccggcccccg
      301 atttgtcagc aatgaaccag ccagcaggga gggccgagcg aagaagtggt cctgctactt
      361 tgtccgcctc catccagtct atgagctgct gtcgtgatgc tagagtaaga agttcgccag
      421 tgagtagttt ccgaagagtt gtggccattg ctactggcat cgtggtatca cgctcgtcgt
      481 tcggtatggc ttcgttcaac tctggttccc agcggtcaag ccgggtcaca tgatcaccca
      541 tattatgaag aaatgcagtc agctccttag ggcctccgat cgttgtcaga agtaagttgg
      601 ccgcggtgtt gtcgctcatg gtaatggcag cactacacaa ttctcttacc gtcatgccat
      661 ccgtaagatg cttttccgtg accggcgagt actcaaccaa gtcgttttgt gagtagtgta
      721 tacggcgacc aagctgctct tgcccggcgt ctatacggga caacaccgcg ccacatagca
      781 gtactttgaa agtgctcatc atcgggaatc gttcttcggg gcggaaagac tcaaggatct
      841 tgccgctatt gagatccagt tcgatatagc ccactcttgc acccagttga tcttcagcat
      901 cttttacttt caccagcgtt tcggggtgtg caaaaacagg caagcaaaat gccgcaaaga
      961 agggaatgag tgcgacacga aaatgttgga tgctcatact cgtccttttt caatattatt
     1021 gaagcattta tcagggttac tagtacgtct ctcaaggata agtaagtaat attaaggtac
     1081 gggaggtatt ggacaggccg caataaaata tctttatttt cattacatct gtgtgttggt
     1141 tttttgtgtg aatcgatagt actaacatac gctctccatc aaaacaaaac gaaacaaaac
     1201 aaactagcaa aataggctgt ccccagtgca agtgcaggtg ccagaacatt tctctggcct
     1261 aactggccgg taccaaaccg gtttcgtgat caaaccggtt tcttgaaaac cggtttacga
     1321 aaccggtttt aaaccggttt aaaccggttt taaaaccggt tttagcaaac cggttttcag
     1381 agaaaccggt ttctcgtcgc ctagaataaa ccggtttctc gtcgcctaga ataaaccggt
     1441 ttctcgtcgc ctagaatctc gtcgcctaga ataaaccggt ttgagctcgc tagcctcgag
     1501 gatatcaaga tctggcctcg gcggccaagc ttagacacta gagggtatat aatggaagct
     1561 cgacttccag cttggcaatc cggtactgtt ggtaaagcca ccatggaaga tgccaaaaac
     1621 attaagaagg gcccagcgcc attctaccca ctcgaagacg ggaccgccgg cgagcagctg
     1681 cacaaagcca tgaagcgcta cgccctggtg cccggcacca tcgcctttac cgacgcacat
     1741 atcgaggtgg acattaccta cgccgagtac ttcgagatga gcgttcggct ggcagaagct
     1801 atgaagcgct atgggctgaa tacaaaccat cggatcgtgg tgtgcagcga gaatagcttg
     1861 cagttcttca tgcccgtgtt gggtgccctg ttcatcggtg tggctgtggc cccagctaac
     1921 gacatctaca acgagcgcga gctgctgaac agcatgggca tcagccagcc caccgtcgta
     1981 ttcgtgagca agaaagggct gcaaaagatc ctcaacgtgc aaaagaagct accgatcata
     2041 caaaagatca tcatcatgga tagcaagacc gactaccagg gcttccaaag catgtacacc
     2101 ttcgtgactt cccatttgcc acccggcttc aacgagtacg acttcgtgcc cgagagcttc
     2161 gaccgggaca aaaccatcgc cctgatcatg aacagtagtg gcagtaccgg attgcccaag
     2221 ggcgtagccc taccgcaccg caccgcttgt gtccgattca gtcatgcccg cgaccccatc
     2281 ttcggcaacc agatcatccc cgacaccgct atcctcagcg tggtgccatt tcaccacggc
     2341 ttcggcatgt tcaccacgct gggctacttg atctgcggct ttcgggtcgt gctcatgtac
     2401 cgcttcgagg aggagctatt cttgcgcagc ttgcaagact ataagattca atctgccctg
     2461 ctggtgccca cactatttag cttcttcgct aagagcactc tcatcgacaa gtacgaccta
     2521 agcaacttgc acgagatcgc cagcggcggg gcgccgctca gcaaggaggt aggtgaggcc
     2581 gtggccaaac gcttccacct accaggcatc cgccagggct acggcctgac agaaacaacc
     2641 agcgccattc tgatcacccc cgaaggggac gacaagcctg gcgcagtagg caaggtggtg
     2701 cccttcttcg aggctaaggt ggtggacttg gacaccggta agacactggg tgtgaaccag
     2761 cgcggcgagc tgtgcgtccg tggccccatg atcatgagcg gctacgttaa caaccccgag
     2821 gctacaaacg ctctcatcga caaggacggc tggctgcaca gcggcgacat cgcctactgg
     2881 gacgaggacg agcacttctt catcgtggac cggctgaaga gcctgatcaa atacaagggc
     2941 taccaggtag ccccagccga actggagagc atcctgctgc aacaccccaa catcttcgac
     3001 gccggggtcg ccggcctgcc cgacgacgat gccggcgagc tgcccgccgc agtcgtcgtg
     3061 ctggaacacg gtaaaaccat gaccgagaag gagatcgtgg actatgtggc cagccaggtt
     3121 acaaccgcca agaagctgcg cggtggtgtt gtgttcgtgg acgaggtgcc taaaggactg
     3181 accggcaagt tggacgcccg caagatccgc gagattctca ttaaggccaa gaagggcggc
     3241 aagatcgccg tgaattctca cggcttccct cccgaggtgg aggagcaggc cgccggcacc
     3301 ctgcccatga gctgcgccca ggagagcggc atggatagac accctgctgc ttgcgccagc
     3361 gccaggatca acgtctaagg ccgcgactct agagtcgggg cggccggccg cttcgagcag
     3421 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     3481 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     3541 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     3601 aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag gatccgtttg
     3661 cgtattgggc gctcttccgc tgatctgcgc agcaccatgg cctgaaataa cctctgaaag
     3721 aggaacttgg ttagctacct tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     3781 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3841 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3901 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     3961 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     4021 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     4081 aggcctaggc ttttgcaaaa agctcgattc ttctgacact agcgccacca tgaagaagcc
     4141 cgaactcacc gctaccagcg ttgaaaaatt tctcatcgag aagttcgaca gtgtgagcga
     4201 cctgatgcag ttgtcggagg gcgaagagag ccgagccttc agcttcgatg tcggcggacg
     4261 cggctatgta ctgcgggtga atagctgcgc tgatggcttc tacaaagacc gctacgtgta
     4321 ccgccacttc gccagcgctg cactacccat ccccgaagtg ttggacatcg gcgagttcag
     4381 cgagagcctg acatactgca tcagtagacg cgcccaaggc gttactctcc aagacctccc
     4441 cgaaacagag ctgcctgctg tgttacagcc tgtcgccgaa gctatggatg ctattgccgc
     4501 cgccgacctc agtcaaacca gcggcttcgg cccattcggg ccccaaggca tcggccagta
     4561 cacaacctgg cgggatttca tttgcgccat tgctgatccc catgtctacc actggcagac
     4621 cgtgatggac gacaccgtgt ccgccagcgt agctcaagcc ctggacgaac tgatgctgtg
     4681 ggccgaagac tgtcccgagg tgcgccacct cgtccatgcc gacttcggca gcaacaacgt
     4741 cctgaccgac aacggccgca tcaccgccgt aatcgactgg tccgaagcta tgttcgggga
     4801 cagtcagtac gaggtggcca acatcttctt ctggcggccc tggctggctt gcatggagca
     4861 gcagactcgc tacttcgagc gccggcatcc cgagctggcc ggcagccctc gtctgcgagc
     4921 ctacatgctg cgcatcggcc tggatcagct ctaccagagc ctcgtggacg gcaacttcga
     4981 cgatgctgcc tgggctcaag gccgctgcga tgccatcgtc cgcagcgggg ccggcaccgt
     5041 cggtcgcaca caaatcgctc gccggagcgc agccgtatgg accgacggct gcgtcgaggt
     5101 gctggccgac agcggcaacc gccggcccag tacacgaccg cgcgctaagg aggtaggtcg
     5161 agtttaaact ctagaaccgg tcatggccgc aataaaatat ctttattttc attacatctg
     5221 tgtgttggtt ttttgtgtgt tcgaactaga tgctgtcgac cgatgccctt gagagccttc
     5281 aacccagtca gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact
     5341 gtcttcttta tcatgcaact cgtaggacag gtgccggcag cgctcttccg cttcctcgct
     5401 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     5461 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5521 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5581 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5641 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5701 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5761 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5821 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5881 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5941 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     6001 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     6061 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     6121 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     6181 gtctgacgct cagtggaacg aaaactcacg ttaaggg
//
