LOCUS       sequence-370026-        5984 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for Camk2d expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5984)
  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 5984)
  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-Camk2d
FEATURES             Location/Qualifiers
     source          1..5984
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1248..1257
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1254..2906
                     /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(1320..1338)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2947..3068
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2958..2977)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3012..3031
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3316..3333)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3487..4075)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3567..3586)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4246..5037,5038..5106))
                     /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     4869..4888
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5107..5211)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5238..5693
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5325..5344)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5535..5556
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5824..5872
                     /note="synthetic polyadenylation signal"
     misc_feature    5886..5977
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5926..5945
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt ccaccactta caccttaacc catcttacat gctaattttt
       61 tttctccatg ctaattttta agcaacagtg tcagatatgc ttaactgtac acacgttctt
      121 gaactcttaa atggttttcc agatagccac tgagggagta cactcctgtg cccctacttg
      181 gcctgctctt tgagaatgca ggtaaagaac tcccaccctc cctctccctt ctccctcggc
      241 aacgcgaaac aggctaggca cgctcacgtg acgctaatga cgtcagcccc ctctgggtcg
      301 cgtgacccac agccaacctt cctaccgccc caccccctcg ccactggcgt gggacatctc
      361 gcgagaagcc ttcttactat cagaacttgc tgaggttaca ggggagttac tctggagagg
      421 aaggaaggtg tggcagagtg gtgcactttt gggtttcatt atgttagagt ttatgatcaa
      481 gcaaatgctt atggggatcg ccctagtcgg gaaaagggga gagaagggcg aagacgtgac
      541 agtaaaaact gaagcgaaac gtggtgccct gaccatcttt gcccctagct aagggacagc
      601 cgggcgcagc agaggtggcc aaggctggca gccccaggca gtagctgcga gccgggggac
      661 gttggacccg ggtctgggag cagtaaacgc gaaccgcgcc cagatgcaag ggttgcttcc
      721 ctctctcggc ttccccggcc tcgcggccat cccgcccggg tagaggcatc cgggttgtgg
      781 gagcttcgag tggtctctca tctttccgcc cttcgtgggt cactgcgaca tctcagtccc
      841 gtccccttaa gcgcgctccc tgctgctccg cgcatctccc ttcggtctcc ctagacggct
      901 gcgggagagc agcccgaagg gacgcggctg gagtcgctag agggaggtgt ggggccgccg
      961 aggagggggc ttcgctgggc ggggagagga gggaggaggc gccggggagg agccgcggct
     1021 gccagcttcg cccgagccga gcgagcgcac tctgtcgccc cgcgggttcc cagccttgcg
     1081 gccccgtgga gaggagggag cgcgggccac cagcgcctcc tctgcccgct ggaacgaccg
     1141 gctgggtgtt cactcggccg ctgccgcccg ccactccggg cgctgcgttg agctctccgt
     1201 gagatctgcg atctaagtaa gcttggcatt ccggtactgt tggtaaagcc accatggaag
     1261 acgccaaaaa cataaagaaa ggcccggcgc cattctatcc gctggaagat ggaaccgctg
     1321 gagagcaact gcataaggct atgaagagat acgccctggt tcctggaaca attgctttta
     1381 cagatgcaca tatcgaggtg gacatcactt acgctgagta cttcgaaatg tccgttcggt
     1441 tggcagaagc tatgaaacga tatgggctga atacaaatca cagaatcgtc gtatgcagtg
     1501 aaaactctct tcaattcttt atgccggtgt tgggcgcgtt atttatcgga gttgcagttg
     1561 cgcccgcgaa cgacatttat aatgaacgtg aattgctcaa cagtatgggc atttcgcagc
     1621 ctaccgtggt gttcgtttcc aaaaaggggt tgcaaaaaat tttgaacgtg caaaaaaagc
     1681 tcccaatcat ccaaaaaatt attatcatgg attctaaaac ggattaccag ggatttcagt
     1741 cgatgtacac gttcgtcaca tctcatctac ctcccggttt taatgaatac gattttgtgc
     1801 cagagtcctt cgatagggac aagacaattg cactgatcat gaactcctct ggatctactg
     1861 gtctgcctaa aggtgtcgct ctgcctcata gaactgcctg cgtgagattc tcgcatgcca
     1921 gagatcctat ttttggcaat caaatcattc cggatactgc gattttaagt gttgttccat
     1981 tccatcacgg ttttggaatg tttactacac tcggatattt gatatgtgga tttcgagtcg
     2041 tcttaatgta tagatttgaa gaagagctgt ttctgaggag ccttcaggat tacaagattc
     2101 aaagtgcgct gctggtgcca accctattct ccttcttcgc caaaagcact ctgattgaca
     2161 aatacgattt atctaattta cacgaaattg cttctggtgg cgctcccctc tctaaggaag
     2221 tcggggaagc ggttgccaag aggttccatc tgccaggtat caggcaagga tatgggctca
     2281 ctgagactac atcagctatt ctgattacac ccgaggggga tgataaaccg ggcgcggtcg
     2341 gtaaagttgt tccatttttt gaagcgaagg ttgtggatct ggataccggg aaaacgctgg
     2401 gcgttaatca aagaggcgaa ctgtgtgtga gaggtcctat gattatgtcc ggttatgtaa
     2461 acaatccgga agcgaccaac gccttgattg acaaggatgg atggctacat tctggagaca
     2521 tagcttactg ggacgaagac gaacacttct tcatcgttga ccgcctgaag tctctgatta
     2581 agtacaaagg ctatcaggtg gctcccgctg aattggaatc catcttgctc caacacccca
     2641 acatcttcga cgcaggtgtc gcaggtcttc ccgacgatga cgccggtgaa cttcccgccg
     2701 ccgttgttgt tttggagcac ggaaagacga tgacggaaaa agagatcgtg gattacgtcg
     2761 ccagtcaagt aacaaccgcg aaaaagttgc gcggaggagt tgtgtttgtg gacgaagtac
     2821 cgaaaggtct taccggaaaa ctcgacgcaa gaaaaatcag agagatcctc ataaaggcca
     2881 agaagggcgg aaagatcgcc gtgtaattct agagtcgggg cggccggccg cttcgagcag
     2941 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     3001 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     3061 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     3121 aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag gatccgtcga
     3181 ccgatgccct tgagagcctt caacccagtc agctccttcc ggtgggcgcg gggcatgact
     3241 atcgtcgccg cacttatgac tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca
     3301 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     3361 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     3421 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3481 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3541 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3601 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3661 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3721 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3781 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3841 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3901 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     3961 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4021 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4081 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4141 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     4201 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     4261 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     4321 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     4381 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     4441 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     4501 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     4561 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     4621 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     4681 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     4741 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     4801 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     4861 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     4921 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     4981 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     5041 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     5101 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     5161 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     5221 ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     5281 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     5341 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     5401 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     5461 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     5521 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     5581 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     5641 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttgccattc
     5701 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     5761 ccagcccaag ctaccatgat aagtaagtaa tattaaggta cgggaggtac ttggagcggc
     5821 cgcaataaaa tatctttatt ttcattacat ctgtgtgttg gttttttgtg tgaatcgata
     5881 gtactaacat acgctctcca tcaaaacaaa acgaaacaaa acaaactagc aaaataggct
     5941 gtccccagtg caagtgcagg tgccagaaca tttctctatc gata
//
