LOCUS       sequence-443481-        5137 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Firefly-luciferase reporter expression driven by proximal 330-bp of 
            mouse Tph2 promoter point mutation at PRE site 4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5137)
  AUTHORS   Park I, Choi M, Kim J, Jang S, Kim D, Kim J, Choe Y, Geum D, Yu SW, 
            Choi JW, Moon C, Choe HK, Son GH, Kim K
  TITLE     Role of the circadian nuclear receptor REV-ERBalpha in dorsal raphe 
            serotonin synthesis in mood regulation.
  JOURNAL   Commun Biol. 2024 Aug 15;7(1):998. doi: 10.1038/s42003-024-06647-y.
  PUBMED    39147805
REFERENCE   2  (bases 1 to 5137)
  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: "2024-08-15"; volume: "7(1):998. doi"; pages: " 
            10.1038/s42003-024-06647-y"
COMMENT     Sequence Label: mTph2p(330bp)-mut4-Luc
FEATURES             Location/Qualifiers
     source          1..5137
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..92
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     41..60
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     regulatory      500..509
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             506..2158
                     /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(572..590)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2199..2320
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2210..2229)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2264..2283
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2568..2585)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2739..3327)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2819..2838)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3498..4289,4290..4358))
                     /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     4121..4140
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4359..4463)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4490..4945
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4577..4596)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4787..4808
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5076..5124
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc
       61 cagtgcaagt gcaggtgcca gaacatttct ctatcgatag gtaccgagct cttacgcgtc
      121 tcttgcttaa gaattttcca ttcttcaaag gattgtcaag ctttcctgtg gctttctaaa
      181 gttggaaaag tacaaatata atcttgtcta tgcctgtcaa attgctgggt ctgatcaggt
      241 catagatgga gagcaataaa attgtatcag aagagtatca aaggaatgat gggcctatgg
      301 gcatttcata tatttcttac ttaatgaaga acattctaga aggttagcat ctgagctaag
      361 ttcaagcctc gaatttcaaa agcctgaata tccttctaca tggaaacacc atgggttttg
      421 gaagcagact tcttatgaaa gcccccgggc tcgagatctg cgatctaagt aagcttggca
      481 ttccggtact gttggtaaag ccaccatgga agacgccaaa aacataaaga aaggcccggc
      541 gccattctat ccgctggaag atggaaccgc tggagagcaa ctgcataagg ctatgaagag
      601 atacgccctg gttcctggaa caattgcttt tacagatgca catatcgagg tggacatcac
      661 ttacgctgag tacttcgaaa tgtccgttcg gttggcagaa gctatgaaac gatatgggct
      721 gaatacaaat cacagaatcg tcgtatgcag tgaaaactct cttcaattct ttatgccggt
      781 gttgggcgcg ttatttatcg gagttgcagt tgcgcccgcg aacgacattt ataatgaacg
      841 tgaattgctc aacagtatgg gcatttcgca gcctaccgtg gtgttcgttt ccaaaaaggg
      901 gttgcaaaaa attttgaacg tgcaaaaaaa gctcccaatc atccaaaaaa ttattatcat
      961 ggattctaaa acggattacc agggatttca gtcgatgtac acgttcgtca catctcatct
     1021 acctcccggt tttaatgaat acgattttgt gccagagtcc ttcgataggg acaagacaat
     1081 tgcactgatc atgaactcct ctggatctac tggtctgcct aaaggtgtcg ctctgcctca
     1141 tagaactgcc tgcgtgagat tctcgcatgc cagagatcct atttttggca atcaaatcat
     1201 tccggatact gcgattttaa gtgttgttcc attccatcac ggttttggaa tgtttactac
     1261 actcggatat ttgatatgtg gatttcgagt cgtcttaatg tatagatttg aagaagagct
     1321 gtttctgagg agccttcagg attacaagat tcaaagtgcg ctgctggtgc caaccctatt
     1381 ctccttcttc gccaaaagca ctctgattga caaatacgat ttatctaatt tacacgaaat
     1441 tgcttctggt ggcgctcccc tctctaagga agtcggggaa gcggttgcca agaggttcca
     1501 tctgccaggt atcaggcaag gatatgggct cactgagact acatcagcta ttctgattac
     1561 acccgagggg gatgataaac cgggcgcggt cggtaaagtt gttccatttt ttgaagcgaa
     1621 ggttgtggat ctggataccg ggaaaacgct gggcgttaat caaagaggcg aactgtgtgt
     1681 gagaggtcct atgattatgt ccggttatgt aaacaatccg gaagcgacca acgccttgat
     1741 tgacaaggat ggatggctac attctggaga catagcttac tgggacgaag acgaacactt
     1801 cttcatcgtt gaccgcctga agtctctgat taagtacaaa ggctatcagg tggctcccgc
     1861 tgaattggaa tccatcttgc tccaacaccc caacatcttc gacgcaggtg tcgcaggtct
     1921 tcccgacgat gacgccggtg aacttcccgc cgccgttgtt gttttggagc acggaaagac
     1981 gatgacggaa aaagagatcg tggattacgt cgccagtcaa gtaacaaccg cgaaaaagtt
     2041 gcgcggagga gttgtgtttg tggacgaagt accgaaaggt cttaccggaa aactcgacgc
     2101 aagaaaaatc agagagatcc tcataaaggc caagaagggc ggaaagatcg ccgtgtaatt
     2161 ctagagtcgg ggcggccggc cgcttcgagc agacatgata agatacattg atgagtttgg
     2221 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     2281 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     2341 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     2401 caaatgtggt aaaatcgata aggatccgtc gaccgatgcc cttgagagcc ttcaacccag
     2461 tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct
     2521 ttatcatgca actcgtagga caggtgccgg cagcgctctt ccgcttcctc gctcactgac
     2581 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     2641 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     2701 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     2761 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     2821 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     2881 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     2941 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3001 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3061 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3121 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     3181 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3241 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3301 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3361 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     3421 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     3481 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     3541 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     3601 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     3661 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     3721 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     3781 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     3841 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     3901 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     3961 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     4021 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     4081 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     4141 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     4201 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     4261 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     4321 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     4381 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     4441 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt
     4501 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     4561 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     4621 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     4681 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     4741 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     4801 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     4861 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     4921 aacaaaatat taacgcttac aatttgccat tcgccattca ggctgcgcaa ctgttgggaa
     4981 gggcgatcgg tgcgggcctc ttcgctatta cgccagccca agctaccatg ataagtaagt
     5041 aatattaagg tacgggaggt acttggagcg gccgcaataa aatatcttta ttttcattac
     5101 atctgtgtgt tggttttttg tgtgaatcga tagtact
//
