LOCUS       sequence-443482-        5129 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Firefly-luciferase reporter expression driven by proximal 330-bp of 
            mouse Tph2 promoter point mutation at RRE site 1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5129)
  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 5129)
  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)-mut1-Luc
FEATURES             Location/Qualifiers
     source          1..5129
                     /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      492..501
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             498..2150
                     /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(564..582)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2191..2312
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2202..2221)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2256..2275
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2560..2577)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2731..3319)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2811..2830)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3490..4281,4282..4350))
                     /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     4113..4132
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4351..4455)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4482..4937
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4569..4588)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4779..4800
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5068..5116
                     /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 ctgatcgagc
      241 aataaaattg tatcagaaga gtatcaaagg aatgatgggc ctatgggcat ttcatttcca
      301 tatatttctt acttaatgaa gaacattcta gaaggttagc atctgagcta agttcaagcc
      361 tcgaatttca aaagcctgaa tatccttcta catggaaaca ccatgggttt tggaagcaga
      421 cttcttatga aagcccccgg gctcgagatc tgcgatctaa gtaagcttgg cattccggta
      481 ctgttggtaa agccaccatg gaagacgcca aaaacataaa gaaaggcccg gcgccattct
      541 atccgctgga agatggaacc gctggagagc aactgcataa ggctatgaag agatacgccc
      601 tggttcctgg aacaattgct tttacagatg cacatatcga ggtggacatc acttacgctg
      661 agtacttcga aatgtccgtt cggttggcag aagctatgaa acgatatggg ctgaatacaa
      721 atcacagaat cgtcgtatgc agtgaaaact ctcttcaatt ctttatgccg gtgttgggcg
      781 cgttatttat cggagttgca gttgcgcccg cgaacgacat ttataatgaa cgtgaattgc
      841 tcaacagtat gggcatttcg cagcctaccg tggtgttcgt ttccaaaaag gggttgcaaa
      901 aaattttgaa cgtgcaaaaa aagctcccaa tcatccaaaa aattattatc atggattcta
      961 aaacggatta ccagggattt cagtcgatgt acacgttcgt cacatctcat ctacctcccg
     1021 gttttaatga atacgatttt gtgccagagt ccttcgatag ggacaagaca attgcactga
     1081 tcatgaactc ctctggatct actggtctgc ctaaaggtgt cgctctgcct catagaactg
     1141 cctgcgtgag attctcgcat gccagagatc ctatttttgg caatcaaatc attccggata
     1201 ctgcgatttt aagtgttgtt ccattccatc acggttttgg aatgtttact acactcggat
     1261 atttgatatg tggatttcga gtcgtcttaa tgtatagatt tgaagaagag ctgtttctga
     1321 ggagccttca ggattacaag attcaaagtg cgctgctggt gccaacccta ttctccttct
     1381 tcgccaaaag cactctgatt gacaaatacg atttatctaa tttacacgaa attgcttctg
     1441 gtggcgctcc cctctctaag gaagtcgggg aagcggttgc caagaggttc catctgccag
     1501 gtatcaggca aggatatggg ctcactgaga ctacatcagc tattctgatt acacccgagg
     1561 gggatgataa accgggcgcg gtcggtaaag ttgttccatt ttttgaagcg aaggttgtgg
     1621 atctggatac cgggaaaacg ctgggcgtta atcaaagagg cgaactgtgt gtgagaggtc
     1681 ctatgattat gtccggttat gtaaacaatc cggaagcgac caacgccttg attgacaagg
     1741 atggatggct acattctgga gacatagctt actgggacga agacgaacac ttcttcatcg
     1801 ttgaccgcct gaagtctctg attaagtaca aaggctatca ggtggctccc gctgaattgg
     1861 aatccatctt gctccaacac cccaacatct tcgacgcagg tgtcgcaggt cttcccgacg
     1921 atgacgccgg tgaacttccc gccgccgttg ttgttttgga gcacggaaag acgatgacgg
     1981 aaaaagagat cgtggattac gtcgccagtc aagtaacaac cgcgaaaaag ttgcgcggag
     2041 gagttgtgtt tgtggacgaa gtaccgaaag gtcttaccgg aaaactcgac gcaagaaaaa
     2101 tcagagagat cctcataaag gccaagaagg gcggaaagat cgccgtgtaa ttctagagtc
     2161 ggggcggccg gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca
     2221 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     2281 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     2341 ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     2401 gtaaaatcga taaggatccg tcgaccgatg cccttgagag ccttcaaccc agtcagctcc
     2461 ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg
     2521 caactcgtag gacaggtgcc ggcagcgctc ttccgcttcc tcgctcactg actcgctgcg
     2581 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     2641 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     2701 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     2761 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     2821 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     2881 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     2941 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3001 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3061 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3121 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3181 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3241 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3301 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3361 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     3421 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     3481 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     3541 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     3601 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     3661 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     3721 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     3781 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     3841 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     3901 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     3961 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     4021 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     4081 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     4141 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4201 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     4261 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     4321 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     4381 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     4441 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgcgccct gtagcggcgc
     4501 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     4561 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4621 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     4681 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4741 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4801 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     4861 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     4921 attaacgctt acaatttgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     4981 ggtgcgggcc tcttcgctat tacgccagcc caagctacca tgataagtaa gtaatattaa
     5041 ggtacgggag gtacttggag cggccgcaat aaaatatctt tattttcatt acatctgtgt
     5101 gttggttttt tgtgtgaatc gatagtact
//
