LOCUS       sequence-370030-        5555 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for Dbp expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5555)
  AUTHORS   Li H, Ou J, Li Y, Xu N, Li Q, Wu P, Peng C, Tang YC, Chang HC
  TITLE     ISX-9 potentiates CaMKIIdelta-mediated BMAL1 activation to enhance 
            circadian amplitude.
  JOURNAL   Commun Biol. 2022 Jul 28;5(1):750. doi: 10.1038/s42003-022-03725-x.
  PUBMED    35902736
REFERENCE   2  (bases 1 to 5555)
  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: "2022-07-28"; volume: "5(1):750. doi"; pages: " 
            10.1038/s42003-022-03725-x"
COMMENT     Sequence Label: pGL3-Dbp
FEATURES             Location/Qualifiers
     source          1..5555
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      819..828
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             825..2477
                     /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(891..909)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2518..2639
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2529..2548)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2583..2602
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2887..2904)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3058..3646)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3138..3157)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3817..4608,4609..4677))
                     /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     4440..4459
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4678..4782)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4809..5264
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4896..4915)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5106..5127
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5395..5443
                     /note="synthetic polyadenylation signal"
     misc_feature    5457..5548
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5497..5516
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gcccacttct ttctgcctta atctgctcta attgatgttc
       61 tgcttttttt cctcaatgag tctgtgaaca ctcggctcct ttaagaggta cccctcttgc
      121 cccacacccg catccgatag cacgcgcaaa gccatgtgct tcccccttct tcctgcctca
      181 ttggcccgaa gtgggtcacg gtcccgcctc cagcgcctcc tcccattggc caaatatagg
      241 tcagtgtccc gccttcttcc aaaaggcgag gcaactacac aagttcagcc cctcacgcgc
      301 ctccagggtc ttggcatatc ccttcatctc attggctgag cgtctcaagg gctcgagacg
      361 tgattggttc ttgccagaag ccggaggaag gcacgcggct ggggtgcatg gcccaagtgg
      421 agggaggggg cccggaggag gggcgcgcgc tctggcgtcc ggtgttttgg tttagatttt
      481 tttttttccg cgaaagaagt ttgctttggc cacgcctcaa ggctgccacc tcctcctgcc
      541 ccctcctccg cccctgggtg cacacctctg ggtgcagatt gcaaagctcc tttctttgcg
      601 agaagtgcaa aattggctag agccagagac tcgcccactt ggtacagaag gagcgcgcgc
      661 ctgtgtccct tggggaccca cagttgcaaa gagacagctg cttgatttgg tcacccactc
      721 gcccagacta taggagcctc ccgggacagt ctttgagttg cacctttctg caagatctgc
      781 gatctaagta agcttggcat tccggtactg ttggtaaagc caccatggaa gacgccaaaa
      841 acataaagaa aggcccggcg ccattctatc cgctggaaga tggaaccgct ggagagcaac
      901 tgcataaggc tatgaagaga tacgccctgg ttcctggaac aattgctttt acagatgcac
      961 atatcgaggt ggacatcact tacgctgagt acttcgaaat gtccgttcgg ttggcagaag
     1021 ctatgaaacg atatgggctg aatacaaatc acagaatcgt cgtatgcagt gaaaactctc
     1081 ttcaattctt tatgccggtg ttgggcgcgt tatttatcgg agttgcagtt gcgcccgcga
     1141 acgacattta taatgaacgt gaattgctca acagtatggg catttcgcag cctaccgtgg
     1201 tgttcgtttc caaaaagggg ttgcaaaaaa ttttgaacgt gcaaaaaaag ctcccaatca
     1261 tccaaaaaat tattatcatg gattctaaaa cggattacca gggatttcag tcgatgtaca
     1321 cgttcgtcac atctcatcta cctcccggtt ttaatgaata cgattttgtg ccagagtcct
     1381 tcgataggga caagacaatt gcactgatca tgaactcctc tggatctact ggtctgccta
     1441 aaggtgtcgc tctgcctcat agaactgcct gcgtgagatt ctcgcatgcc agagatccta
     1501 tttttggcaa tcaaatcatt ccggatactg cgattttaag tgttgttcca ttccatcacg
     1561 gttttggaat gtttactaca ctcggatatt tgatatgtgg atttcgagtc gtcttaatgt
     1621 atagatttga agaagagctg tttctgagga gccttcagga ttacaagatt caaagtgcgc
     1681 tgctggtgcc aaccctattc tccttcttcg ccaaaagcac tctgattgac aaatacgatt
     1741 tatctaattt acacgaaatt gcttctggtg gcgctcccct ctctaaggaa gtcggggaag
     1801 cggttgccaa gaggttccat ctgccaggta tcaggcaagg atatgggctc actgagacta
     1861 catcagctat tctgattaca cccgaggggg atgataaacc gggcgcggtc ggtaaagttg
     1921 ttccattttt tgaagcgaag gttgtggatc tggataccgg gaaaacgctg ggcgttaatc
     1981 aaagaggcga actgtgtgtg agaggtccta tgattatgtc cggttatgta aacaatccgg
     2041 aagcgaccaa cgccttgatt gacaaggatg gatggctaca ttctggagac atagcttact
     2101 gggacgaaga cgaacacttc ttcatcgttg accgcctgaa gtctctgatt aagtacaaag
     2161 gctatcaggt ggctcccgct gaattggaat ccatcttgct ccaacacccc aacatcttcg
     2221 acgcaggtgt cgcaggtctt cccgacgatg acgccggtga acttcccgcc gccgttgttg
     2281 ttttggagca cggaaagacg atgacggaaa aagagatcgt ggattacgtc gccagtcaag
     2341 taacaaccgc gaaaaagttg cgcggaggag ttgtgtttgt ggacgaagta ccgaaaggtc
     2401 ttaccggaaa actcgacgca agaaaaatca gagagatcct cataaaggcc aagaagggcg
     2461 gaaagatcgc cgtgtaattc tagagtcggg gcggccggcc gcttcgagca gacatgataa
     2521 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     2581 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     2641 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
     2701 aaagcaagta aaacctctac aaatgtggta aaatcgataa ggatccgtcg accgatgccc
     2761 ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc
     2821 gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctcttc
     2881 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     2941 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     3001 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     3061 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     3121 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     3181 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3241 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3301 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3361 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3421 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3481 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     3541 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     3601 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     3661 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     3721 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     3781 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     3841 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     3901 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     3961 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     4021 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     4081 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     4141 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     4201 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     4261 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     4321 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     4381 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     4441 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     4501 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     4561 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     4621 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     4681 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     4741 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     4801 gccacctgac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     4861 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     4921 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     4981 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     5041 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     5101 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     5161 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     5221 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttgccatt cgccattcag
     5281 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagcccaa
     5341 gctaccatga taagtaagta atattaaggt acgggaggta cttggagcgg ccgcaataaa
     5401 atatctttat tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat agtactaaca
     5461 tacgctctcc atcaaaacaa aacgaaacaa aacaaactag caaaataggc tgtccccagt
     5521 gcaagtgcag gtgccagaac atttctctat cgata
//
