LOCUS       sequence-443488-        6655 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Firefly-luciferase reporter expression driven by human Tph2 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6655)
  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 6655)
  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: hTph2p-Luc
FEATURES             Location/Qualifiers
     source          1..6655
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     234..253
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(472..576)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     644..662
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(700..722)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     822..841
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1035..1057
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1049..1066
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1050..1066
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      3130..3139
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3136..4788
                     /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(3202..3220)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    4829..4950
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4840..4859)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4894..4913
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5105..5121)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5105..5121)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5118..5140)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5129..5145
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5153..5159,5160..5177,5178..5183))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5198..5219
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5336..5353)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5507..6095)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5587..5606)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6266..6655,1..402,403..471))
                     /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"
ORIGIN
        1 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
       61 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
      121 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
      181 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
      241 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
      301 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
      361 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
      421 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
      481 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
      541 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
      601 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
      661 gcccttttgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
      721 ggagactgtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
      781 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
      841 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
      901 catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
      961 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     1021 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat tgacgcgtat
     1081 tgggatggta ccatttgagc ctttagggta ttctgtcatg taaagtgcgt tgcttctaag
     1141 tcttcttcat tgatagctta aggttcagct ttcccatgac tgcaaagcct ttgatcagtt
     1201 ttttcttctt tcaagctgct aattttttgt tgccattgtt ttccaactta tgcgactctt
     1261 gtgtaccttg aaaaggaatt ttttatcttg ttttaatagg gtttccaggg tgagcaaaag
     1321 tagttaaatc tttaatattc taaatatata ttataaataa attactttta atatcttttt
     1381 ttccctcatt tccacatcca agcagggaca ctatctagtg gattttatcc cttcacatct
     1441 ctggttcttt cattacatct cagtaacatt agctactatt attgtcatta gttcatttgc
     1501 cttatttaaa cttgcttctc ctattagctg attagagctg acccttacag aaaatgtaag
     1561 ccaaataata ttttcagaat tagatcaatt aataattatg tctcttcaag actccagaaa
     1621 caattaaact tcagtcaata ttagttcatc ccgacttgtt ttgaaatttt ttttctatat
     1681 agacaagcag tgccagcacc agattttgtt tagattttca gtcaaacaag cacatttggt
     1741 caagagtttt tcattcttca ggagattggt aacattttcc tgtggcctgg tgaattttat
     1801 gactttctaa ggtttgaata gtaagaatat tgattaatct atgcatgtca agttgctgga
     1861 tttgatctag ttgacattga tatttgacag tatgtttagt cattaaaagc tcaaatgtca
     1921 tagtactctt aacctctgct ttctcctcac cacataacgc acagatctcg tatatttaag
     1981 tagcactttt tatatgaaag ggtcatgtca aagattaaat ttccatgatt tccagtagag
     2041 agaaaaacca caagagtatt aaaatgaatg gctgatatat gggaatctag tttcctttta
     2101 tttctcattt agtaaggaag attccaggat tagtatctga gctatatgta atctacatgt
     2161 ggctaaattg aacccttacc tttcctttgc aatacatttt cctccatata actctgcata
     2221 gaggcatcac aggattaaga agaagccctt ttatgaaagc cattacacat atatacactc
     2281 acacatttgc atgcacaaaa ttagaatatg tcaagtcaga aaaagcttat taacataaaa
     2341 tggagttggt caatgagtaa aaaaaatatg ctgatgggag ggataagatc tagtgttcgg
     2401 gagcacaata atttattttc ttttgtattt taaaataact ggaagagtgg aattggaatg
     2461 tttctaacac aaaaagaaat gataaatgct tgaggcaatg gatatcttga ttaccttatt
     2521 tgatcattac acattgtacg cttgtgtcaa aatatcacat gtgccttata aatgtgtaca
     2581 actattagtt atccataaaa attaaaaatt aaaaaatccg taaaatggtt taagcattca
     2641 gcagtgctga tctttcttaa attatttttc taattttgga aagaaagcac aaaatctttg
     2701 aattcacaat tgcttaaaga ctgaggttaa cttgccagtg gcaggcttga gagatgagag
     2761 aactaacgtc agaggataga tggtttcttg tacaaataac acccccttat gtattgttct
     2821 ccaccacccc cgcccaaaaa gctactcgac ctatgaaaca aatcacacta tgagcacaga
     2881 taaccccagg cttcaggtct gtaatctgac tgtggccatc ggcaaccaga aatgagtttc
     2941 tttctaatca gtcttgcatc agtctccagt cattcatata aaggagcccg gggatgggag
     3001 gattcgcatt gctcttcagc accagggttc tggacagcgc cccaagcagg cagctgatcg
     3061 cacgcccctt cctctcaatc tcgagatctg cgatctaagt aagcttggca ttccggtact
     3121 gttggtaaag ccaccatgga agacgccaaa aacataaaga aaggcccggc gccattctat
     3181 ccgctggaag atggaaccgc tggagagcaa ctgcataagg ctatgaagag atacgccctg
     3241 gttcctggaa caattgcttt tacagatgca catatcgagg tggacatcac ttacgctgag
     3301 tacttcgaaa tgtccgttcg gttggcagaa gctatgaaac gatatgggct gaatacaaat
     3361 cacagaatcg tcgtatgcag tgaaaactct cttcaattct ttatgccggt gttgggcgcg
     3421 ttatttatcg gagttgcagt tgcgcccgcg aacgacattt ataatgaacg tgaattgctc
     3481 aacagtatgg gcatttcgca gcctaccgtg gtgttcgttt ccaaaaaggg gttgcaaaaa
     3541 attttgaacg tgcaaaaaaa gctcccaatc atccaaaaaa ttattatcat ggattctaaa
     3601 acggattacc agggatttca gtcgatgtac acgttcgtca catctcatct acctcccggt
     3661 tttaatgaat acgattttgt gccagagtcc ttcgataggg acaagacaat tgcactgatc
     3721 atgaactcct ctggatctac tggtctgcct aaaggtgtcg ctctgcctca tagaactgcc
     3781 tgcgtgagat tctcgcatgc cagagatcct atttttggca atcaaatcat tccggatact
     3841 gcgattttaa gtgttgttcc attccatcac ggttttggaa tgtttactac actcggatat
     3901 ttgatatgtg gatttcgagt cgtcttaatg tatagatttg aagaagagct gtttctgagg
     3961 agccttcagg attacaagat tcaaagtgcg ctgctggtgc caaccctatt ctccttcttc
     4021 gccaaaagca ctctgattga caaatacgat ttatctaatt tacacgaaat tgcttctggt
     4081 ggcgctcccc tctctaagga agtcggggaa gcggttgcca agaggttcca tctgccaggt
     4141 atcaggcaag gatatgggct cactgagact acatcagcta ttctgattac acccgagggg
     4201 gatgataaac cgggcgcggt cggtaaagtt gttccatttt ttgaagcgaa ggttgtggat
     4261 ctggataccg ggaaaacgct gggcgttaat caaagaggcg aactgtgtgt gagaggtcct
     4321 atgattatgt ccggttatgt aaacaatccg gaagcgacca acgccttgat tgacaaggat
     4381 ggatggctac attctggaga catagcttac tgggacgaag acgaacactt cttcatcgtt
     4441 gaccgcctga agtctctgat taagtacaaa ggctatcagg tggctcccgc tgaattggaa
     4501 tccatcttgc tccaacaccc caacatcttc gacgcaggtg tcgcaggtct tcccgacgat
     4561 gacgccggtg aacttcccgc cgccgttgtt gttttggagc acggaaagac gatgacggaa
     4621 aaagagatcg tggattacgt cgccagtcaa gtaacaaccg cgaaaaagtt gcgcggagga
     4681 gttgtgtttg tggacgaagt accgaaaggt cttaccggaa aactcgacgc aagaaaaatc
     4741 agagagatcc tcataaaggc caagaagggc ggaaagatcg ccgtgtaatt ctagagtcgg
     4801 ggcggccggc cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca
     4861 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     4921 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     4981 caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
     5041 aaaatcgata aggatccgtc gaccgtcgag atcccaatgg cgcgccgagc ttggcgtaat
     5101 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     5161 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     5221 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     5281 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctgttcc gcttcctcgc
     5341 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     5401 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     5461 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5521 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     5581 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     5641 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5701 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5761 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5821 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5881 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5941 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     6001 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     6061 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6121 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6181 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     6241 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     6301 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6361 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagaa ccacgctcac
     6421 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6481 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     6541 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     6601 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagt
//
