LOCUS       sequence-443556-        5138 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 3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5138)
  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 5138)
  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)-mut3-Luc
FEATURES             Location/Qualifiers
     source          1..5138
                     /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      501..510
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             507..2159
                     /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(573..591)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2200..2321
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2211..2230)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2265..2284
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2569..2586)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2740..3328)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2820..2839)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3499..4290,4291..4359))
                     /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     4122..4141
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4360..4464)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4491..4946
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4578..4597)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4788..4809
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5077..5125
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc
       61 cagtgcaagt gcaggtgcca gaacatttct ctatcgatag gtaccgagct cttacgcgtc
      121 tcttgcttaa gaattttcca ttcttcaaag gattgtcaag ctgtggcttt ctaaagttgg
      181 aaaagtacaa atataatctt gtctatgcct gtcaaattgc tgggtctgat caggtcatag
      241 atggagagca ataaaattgt atcagaagag tatcaaagga atgatgggcc tatgggcatt
      301 tcatttccat atatttctta cttaatgaag aacattctag aaggttagca tctgagctaa
      361 gttcaagcct cgaatttcaa aagcctgaat atccttctac atggaaacac catgggtttt
      421 ggaagcagac ttcttatgaa agcccccggg ctcgagatct gcgatctaag taagcttggc
      481 attccggtac tgttggtaaa gccaccatgg aagacgccaa aaacataaag aaaggcccgg
      541 cgccattcta tccgctggaa gatggaaccg ctggagagca actgcataag gctatgaaga
      601 gatacgccct ggttcctgga acaattgctt ttacagatgc acatatcgag gtggacatca
      661 cttacgctga gtacttcgaa atgtccgttc ggttggcaga agctatgaaa cgatatgggc
      721 tgaatacaaa tcacagaatc gtcgtatgca gtgaaaactc tcttcaattc tttatgccgg
      781 tgttgggcgc gttatttatc ggagttgcag ttgcgcccgc gaacgacatt tataatgaac
      841 gtgaattgct caacagtatg ggcatttcgc agcctaccgt ggtgttcgtt tccaaaaagg
      901 ggttgcaaaa aattttgaac gtgcaaaaaa agctcccaat catccaaaaa attattatca
      961 tggattctaa aacggattac cagggatttc agtcgatgta cacgttcgtc acatctcatc
     1021 tacctcccgg ttttaatgaa tacgattttg tgccagagtc cttcgatagg gacaagacaa
     1081 ttgcactgat catgaactcc tctggatcta ctggtctgcc taaaggtgtc gctctgcctc
     1141 atagaactgc ctgcgtgaga ttctcgcatg ccagagatcc tatttttggc aatcaaatca
     1201 ttccggatac tgcgatttta agtgttgttc cattccatca cggttttgga atgtttacta
     1261 cactcggata tttgatatgt ggatttcgag tcgtcttaat gtatagattt gaagaagagc
     1321 tgtttctgag gagccttcag gattacaaga ttcaaagtgc gctgctggtg ccaaccctat
     1381 tctccttctt cgccaaaagc actctgattg acaaatacga tttatctaat ttacacgaaa
     1441 ttgcttctgg tggcgctccc ctctctaagg aagtcgggga agcggttgcc aagaggttcc
     1501 atctgccagg tatcaggcaa ggatatgggc tcactgagac tacatcagct attctgatta
     1561 cacccgaggg ggatgataaa ccgggcgcgg tcggtaaagt tgttccattt tttgaagcga
     1621 aggttgtgga tctggatacc gggaaaacgc tgggcgttaa tcaaagaggc gaactgtgtg
     1681 tgagaggtcc tatgattatg tccggttatg taaacaatcc ggaagcgacc aacgccttga
     1741 ttgacaagga tggatggcta cattctggag acatagctta ctgggacgaa gacgaacact
     1801 tcttcatcgt tgaccgcctg aagtctctga ttaagtacaa aggctatcag gtggctcccg
     1861 ctgaattgga atccatcttg ctccaacacc ccaacatctt cgacgcaggt gtcgcaggtc
     1921 ttcccgacga tgacgccggt gaacttcccg ccgccgttgt tgttttggag cacggaaaga
     1981 cgatgacgga aaaagagatc gtggattacg tcgccagtca agtaacaacc gcgaaaaagt
     2041 tgcgcggagg agttgtgttt gtggacgaag taccgaaagg tcttaccgga aaactcgacg
     2101 caagaaaaat cagagagatc ctcataaagg ccaagaaggg cggaaagatc gccgtgtaat
     2161 tctagagtcg gggcggccgg ccgcttcgag cagacatgat aagatacatt gatgagtttg
     2221 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     2281 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     2341 attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct
     2401 acaaatgtgg taaaatcgat aaggatccgt cgaccgatgc ccttgagagc cttcaaccca
     2461 gtcagctcct tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgtcttc
     2521 tttatcatgc aactcgtagg acaggtgccg gcagcgctct tccgcttcct cgctcactga
     2581 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     2641 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     2701 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     2761 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2821 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2881 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     2941 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3001 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3061 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3121 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3181 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3241 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3301 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     3361 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     3421 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     3481 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     3541 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     3601 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     3661 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     3721 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     3781 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     3841 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     3901 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     3961 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     4021 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     4081 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     4141 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     4201 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     4261 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     4321 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     4381 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     4441 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg
     4501 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     4561 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     4621 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     4681 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     4741 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     4801 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     4861 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     4921 taacaaaata ttaacgctta caatttgcca ttcgccattc aggctgcgca actgttggga
     4981 agggcgatcg gtgcgggcct cttcgctatt acgccagccc aagctaccat gataagtaag
     5041 taatattaag gtacgggagg tacttggagc ggccgcaata aaatatcttt attttcatta
     5101 catctgtgtg ttggtttttt gtgtgaatcg atagtact
//
