LOCUS       sequence-443486-        5143 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Firefly-luciferase reporter expression driven by proximal 330-bp of 
            mouse Tph2 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5143)
  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 5143)
  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)-Luc
FEATURES             Location/Qualifiers
     source          1..5143
                     /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      506..515
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             512..2164
                     /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(578..596)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2205..2326
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2216..2235)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2270..2289
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2574..2591)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2745..3333)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2825..2844)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3504..4295,4296..4364))
                     /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     4127..4146
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4365..4469)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4496..4951
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4583..4602)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4793..4814
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5082..5130
                     /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 gcatttcatt tccatatatt tcttacttaa tgaagaacat tctagaaggt tagcatctga
      361 gctaagttca agcctcgaat ttcaaaagcc tgaatatcct tctacatgga aacaccatgg
      421 gttttggaag cagacttctt atgaaagccc ccgggctcga gatctgcgat ctaagtaagc
      481 ttggcattcc ggtactgttg gtaaagccac catggaagac gccaaaaaca taaagaaagg
      541 cccggcgcca ttctatccgc tggaagatgg aaccgctgga gagcaactgc ataaggctat
      601 gaagagatac gccctggttc ctggaacaat tgcttttaca gatgcacata tcgaggtgga
      661 catcacttac gctgagtact tcgaaatgtc cgttcggttg gcagaagcta tgaaacgata
      721 tgggctgaat acaaatcaca gaatcgtcgt atgcagtgaa aactctcttc aattctttat
      781 gccggtgttg ggcgcgttat ttatcggagt tgcagttgcg cccgcgaacg acatttataa
      841 tgaacgtgaa ttgctcaaca gtatgggcat ttcgcagcct accgtggtgt tcgtttccaa
      901 aaaggggttg caaaaaattt tgaacgtgca aaaaaagctc ccaatcatcc aaaaaattat
      961 tatcatggat tctaaaacgg attaccaggg atttcagtcg atgtacacgt tcgtcacatc
     1021 tcatctacct cccggtttta atgaatacga ttttgtgcca gagtccttcg atagggacaa
     1081 gacaattgca ctgatcatga actcctctgg atctactggt ctgcctaaag gtgtcgctct
     1141 gcctcataga actgcctgcg tgagattctc gcatgccaga gatcctattt ttggcaatca
     1201 aatcattccg gatactgcga ttttaagtgt tgttccattc catcacggtt ttggaatgtt
     1261 tactacactc ggatatttga tatgtggatt tcgagtcgtc ttaatgtata gatttgaaga
     1321 agagctgttt ctgaggagcc ttcaggatta caagattcaa agtgcgctgc tggtgccaac
     1381 cctattctcc ttcttcgcca aaagcactct gattgacaaa tacgatttat ctaatttaca
     1441 cgaaattgct tctggtggcg ctcccctctc taaggaagtc ggggaagcgg ttgccaagag
     1501 gttccatctg ccaggtatca ggcaaggata tgggctcact gagactacat cagctattct
     1561 gattacaccc gagggggatg ataaaccggg cgcggtcggt aaagttgttc cattttttga
     1621 agcgaaggtt gtggatctgg ataccgggaa aacgctgggc gttaatcaaa gaggcgaact
     1681 gtgtgtgaga ggtcctatga ttatgtccgg ttatgtaaac aatccggaag cgaccaacgc
     1741 cttgattgac aaggatggat ggctacattc tggagacata gcttactggg acgaagacga
     1801 acacttcttc atcgttgacc gcctgaagtc tctgattaag tacaaaggct atcaggtggc
     1861 tcccgctgaa ttggaatcca tcttgctcca acaccccaac atcttcgacg caggtgtcgc
     1921 aggtcttccc gacgatgacg ccggtgaact tcccgccgcc gttgttgttt tggagcacgg
     1981 aaagacgatg acggaaaaag agatcgtgga ttacgtcgcc agtcaagtaa caaccgcgaa
     2041 aaagttgcgc ggaggagttg tgtttgtgga cgaagtaccg aaaggtctta ccggaaaact
     2101 cgacgcaaga aaaatcagag agatcctcat aaaggccaag aagggcggaa agatcgccgt
     2161 gtaattctag agtcggggcg gccggccgct tcgagcagac atgataagat acattgatga
     2221 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     2281 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     2341 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     2401 cctctacaaa tgtggtaaaa tcgataagga tccgtcgacc gatgcccttg agagccttca
     2461 acccagtcag ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg
     2521 tcttctttat catgcaactc gtaggacagg tgccggcagc gctcttccgc ttcctcgctc
     2581 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     2641 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     2701 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     2761 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     2821 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2881 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     2941 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     3001 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     3061 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     3121 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     3181 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     3241 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     3301 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     3361 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     3421 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     3481 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     3541 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     3601 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     3661 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     3721 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     3781 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     3841 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     3901 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     3961 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     4021 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     4081 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     4141 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     4201 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     4261 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     4321 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     4381 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     4441 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg
     4501 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     4561 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     4621 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     4681 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     4741 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     4801 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     4861 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     4921 aattttaaca aaatattaac gcttacaatt tgccattcgc cattcaggct gcgcaactgt
     4981 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agcccaagct accatgataa
     5041 gtaagtaata ttaaggtacg ggaggtactt ggagcggccg caataaaata tctttatttt
     5101 cattacatct gtgtgttggt tttttgtgtg aatcgatagt act
//
