LOCUS       sequence-370031-        5408 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for rev-erb-alpha expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5408)
  AUTHORS   Li H, Ou J, Li Y, Xu N, Li Q, Wu P, Peng C, Tang YC, Chang HC
  TITLE     ISX-9 potentiates CaMKIIdelta-mediated BMAL1 activation to enhance 
            circadian amplitude.
  JOURNAL   Commun Biol. 2022 Jul 28;5(1):750. doi: 10.1038/s42003-022-03725-x.
  PUBMED    35902736
REFERENCE   2  (bases 1 to 5408)
  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: "2022-07-28"; volume: "5(1):750. doi"; pages: " 
            10.1038/s42003-022-03725-x"
COMMENT     Sequence Label: pGL3-rev-erb-alpha
FEATURES             Location/Qualifiers
     source          1..5408
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      672..681
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             678..2330
                     /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(744..762)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2371..2492
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2382..2401)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2436..2455
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2740..2757)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2911..3499)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2991..3010)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3670..4461,4462..4530))
                     /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     4293..4312
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4531..4635)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4662..5117
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4749..4768)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4959..4980
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5248..5296
                     /note="synthetic polyadenylation signal"
     misc_feature    5310..5401
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5350..5369
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt ctagtcacca ctaacctcag ggtgaggtca gttctccaat
       61 cacgtgaagc tctcacgttt gctaggtttg cagaaagggc cttttagctt tgatcttcca
      121 cggagcagta aagcttgcca ggccctcccc aggaattcac atgcccttgc catacaaggc
      181 tttccaaaca cgccaccctg actcttcaga aaaccccacc ccagtccact ttcagctcct
      241 ccccgctgcc agcaagcact tttgcaaagc agaaaaggaa taggcacact ccacctacat
      301 tgtcagtaga ctacaaatcc caacaatcct ggcggtgcgc aggcgcacag atctcaacgt
      361 gccggctgct ggaaaagtgt gtcactgggg cacgaggcgc tccctggaat cacatggtac
      421 ctgctccagt gccgcgtgcg gcccgggaac cctgggctgc tggcgcctgc gcagagccct
      481 ctgtcccagg gaaaggctcg ggcaaaaggc ggttgagatt ggcagagtga aatattactg
      541 ctgaggaaac gtagcagggc acacgtctct ctttgcgtcc tggtgcctca tttctccatc
      601 acctacttac ttcctggttg cctcgagatc tgcgatctaa gtaagcttgg cattccggta
      661 ctgttggtaa agccaccatg gaagacgcca aaaacataaa gaaaggcccg gcgccattct
      721 atccgctgga agatggaacc gctggagagc aactgcataa ggctatgaag agatacgccc
      781 tggttcctgg aacaattgct tttacagatg cacatatcga ggtggacatc acttacgctg
      841 agtacttcga aatgtccgtt cggttggcag aagctatgaa acgatatggg ctgaatacaa
      901 atcacagaat cgtcgtatgc agtgaaaact ctcttcaatt ctttatgccg gtgttgggcg
      961 cgttatttat cggagttgca gttgcgcccg cgaacgacat ttataatgaa cgtgaattgc
     1021 tcaacagtat gggcatttcg cagcctaccg tggtgttcgt ttccaaaaag gggttgcaaa
     1081 aaattttgaa cgtgcaaaaa aagctcccaa tcatccaaaa aattattatc atggattcta
     1141 aaacggatta ccagggattt cagtcgatgt acacgttcgt cacatctcat ctacctcccg
     1201 gttttaatga atacgatttt gtgccagagt ccttcgatag ggacaagaca attgcactga
     1261 tcatgaactc ctctggatct actggtctgc ctaaaggtgt cgctctgcct catagaactg
     1321 cctgcgtgag attctcgcat gccagagatc ctatttttgg caatcaaatc attccggata
     1381 ctgcgatttt aagtgttgtt ccattccatc acggttttgg aatgtttact acactcggat
     1441 atttgatatg tggatttcga gtcgtcttaa tgtatagatt tgaagaagag ctgtttctga
     1501 ggagccttca ggattacaag attcaaagtg cgctgctggt gccaacccta ttctccttct
     1561 tcgccaaaag cactctgatt gacaaatacg atttatctaa tttacacgaa attgcttctg
     1621 gtggcgctcc cctctctaag gaagtcgggg aagcggttgc caagaggttc catctgccag
     1681 gtatcaggca aggatatggg ctcactgaga ctacatcagc tattctgatt acacccgagg
     1741 gggatgataa accgggcgcg gtcggtaaag ttgttccatt ttttgaagcg aaggttgtgg
     1801 atctggatac cgggaaaacg ctgggcgtta atcaaagagg cgaactgtgt gtgagaggtc
     1861 ctatgattat gtccggttat gtaaacaatc cggaagcgac caacgccttg attgacaagg
     1921 atggatggct acattctgga gacatagctt actgggacga agacgaacac ttcttcatcg
     1981 ttgaccgcct gaagtctctg attaagtaca aaggctatca ggtggctccc gctgaattgg
     2041 aatccatctt gctccaacac cccaacatct tcgacgcagg tgtcgcaggt cttcccgacg
     2101 atgacgccgg tgaacttccc gccgccgttg ttgttttgga gcacggaaag acgatgacgg
     2161 aaaaagagat cgtggattac gtcgccagtc aagtaacaac cgcgaaaaag ttgcgcggag
     2221 gagttgtgtt tgtggacgaa gtaccgaaag gtcttaccgg aaaactcgac gcaagaaaaa
     2281 tcagagagat cctcataaag gccaagaagg gcggaaagat cgccgtgtaa ttctagagtc
     2341 ggggcggccg gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca
     2401 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     2461 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     2521 ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     2581 gtaaaatcga taaggatccg tcgaccgatg cccttgagag ccttcaaccc agtcagctcc
     2641 ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg
     2701 caactcgtag gacaggtgcc ggcagcgctc ttccgcttcc tcgctcactg actcgctgcg
     2761 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     2821 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     2881 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     2941 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3001 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3061 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3121 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3181 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3241 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3301 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3361 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3421 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3481 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3541 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     3601 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     3661 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     3721 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     3781 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     3841 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     3901 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     3961 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4021 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     4081 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     4141 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     4201 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     4261 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     4321 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4381 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     4441 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     4501 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     4561 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     4621 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgcgccct gtagcggcgc
     4681 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     4741 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4801 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     4861 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4921 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4981 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     5041 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     5101 attaacgctt acaatttgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     5161 ggtgcgggcc tcttcgctat tacgccagcc caagctacca tgataagtaa gtaatattaa
     5221 ggtacgggag gtacttggag cggccgcaat aaaatatctt tattttcatt acatctgtgt
     5281 gttggttttt tgtgtgaatc gatagtacta acatacgctc tccatcaaaa caaaacgaaa
     5341 caaaacaaac tagcaaaata ggctgtcccc agtgcaagtg caggtgccag aacatttctc
     5401 tatcgata
//
