LOCUS       sequence-467802-        5656 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Measure the activity of the reduced differentially methylated region
            (rDMR) of Myc with luciferase assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5656)
  AUTHORS   Wu BK, Mei SC, Chen EH, Zheng Y, Pan D
  TITLE     YAP induces an oncogenic transcriptional program through 
            TET1-mediated epigenetic remodeling in liver growth and 
            tumorigenesis.
  JOURNAL   Nat Genet. 2022 Aug;54(8):1202-1213. doi: 
            10.1038/s41588-022-01119-7. Epub 2022 Jul 14.
  PUBMED    35835915
REFERENCE   2  (bases 1 to 5656)
  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"; doi: 
            "10.1038/s41588-022-01119-7"; journalName: "Nat Genet"; date: 
            "2022-08"; volume: "54"; issue: "8"; pages: "1202-1213"
COMMENT     Sequence Label: pGL3-Myc-rDMR
FEATURES             Location/Qualifiers
     source          1..5656
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        694..890
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      741..876
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      920..929
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             926..2578
                     /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(992..1010)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2619..2740
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2630..2649)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2684..2703
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2988..3005)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3159..3747)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3239..3258)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3918..4709,4710..4778))
                     /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     4541..4560
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4779..4883)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4910..5365
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4997..5016)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5207..5228
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5496..5544
                     /note="synthetic polyadenylation signal"
     misc_feature    5558..5649
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5598..5617
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt ggtttccgca ccaaccagag ctggataact ctagacttgc
       61 ttcccttgct gtgccccctc cagcagacag ccacgacgat gcccctcaac gtgaacttca
      121 ccaacaggaa ctatgacctc gactacgact ccgtacagcc ctatttcatc tgcgacgagg
      181 aagagaattt ctatcaccag caacagcaga gcgagctgca gccgcccgcg cccagtgagg
      241 atatctggaa gaaattcgag ctgcttccca ccccgcccct gtccccgagc cgccgctccg
      301 ggctctgctc tccatcctat gttgcggtcg ctacgtcctt ctccccaagg gaagacgatg
      361 acggcggcgg tggcaacttc tccaccgccg atcagctgga gatgatgacc gagttacttg
      421 gaggagacat ggtgaaccag agcttcatct gcgatcctga cgacgagacc ttcatcaaga
      481 acatcatcat ccaggactgt atgtggagcg gtttctcagc cgctgccaag ctggtctcgg
      541 agaagctggc ctcctaccag gctgcgcgca aagacagcac cagcctgagc cccgcccgcg
      601 ggcacagcgt ctgctccacc tccagcctgt acctgcagga cctcaccgcc gccgcgtccg
      661 agtgcattga cccctcactc gagatctgcg atctgcatct caattagtca gcaaccatag
      721 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
      781 cccatcgctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
      841 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagcttggca
      901 ttccggtact gttggtaaag ccaccatgga agacgccaaa aacataaaga aaggcccggc
      961 gccattctat ccgctggaag atggaaccgc tggagagcaa ctgcataagg ctatgaagag
     1021 atacgccctg gttcctggaa caattgcttt tacagatgca catatcgagg tggacatcac
     1081 ttacgctgag tacttcgaaa tgtccgttcg gttggcagaa gctatgaaac gatatgggct
     1141 gaatacaaat cacagaatcg tcgtatgcag tgaaaactct cttcaattct ttatgccggt
     1201 gttgggcgcg ttatttatcg gagttgcagt tgcgcccgcg aacgacattt ataatgaacg
     1261 tgaattgctc aacagtatgg gcatttcgca gcctaccgtg gtgttcgttt ccaaaaaggg
     1321 gttgcaaaaa attttgaacg tgcaaaaaaa gctcccaatc atccaaaaaa ttattatcat
     1381 ggattctaaa acggattacc agggatttca gtcgatgtac acgttcgtca catctcatct
     1441 acctcccggt tttaatgaat acgattttgt gccagagtcc ttcgataggg acaagacaat
     1501 tgcactgatc atgaactcct ctggatctac tggtctgcct aaaggtgtcg ctctgcctca
     1561 tagaactgcc tgcgtgagat tctcgcatgc cagagatcct atttttggca atcaaatcat
     1621 tccggatact gcgattttaa gtgttgttcc attccatcac ggttttggaa tgtttactac
     1681 actcggatat ttgatatgtg gatttcgagt cgtcttaatg tatagatttg aagaagagct
     1741 gtttctgagg agccttcagg attacaagat tcaaagtgcg ctgctggtgc caaccctatt
     1801 ctccttcttc gccaaaagca ctctgattga caaatacgat ttatctaatt tacacgaaat
     1861 tgcttctggt ggcgctcccc tctctaagga agtcggggaa gcggttgcca agaggttcca
     1921 tctgccaggt atcaggcaag gatatgggct cactgagact acatcagcta ttctgattac
     1981 acccgagggg gatgataaac cgggcgcggt cggtaaagtt gttccatttt ttgaagcgaa
     2041 ggttgtggat ctggataccg ggaaaacgct gggcgttaat caaagaggcg aactgtgtgt
     2101 gagaggtcct atgattatgt ccggttatgt aaacaatccg gaagcgacca acgccttgat
     2161 tgacaaggat ggatggctac attctggaga catagcttac tgggacgaag acgaacactt
     2221 cttcatcgtt gaccgcctga agtctctgat taagtacaaa ggctatcagg tggctcccgc
     2281 tgaattggaa tccatcttgc tccaacaccc caacatcttc gacgcaggtg tcgcaggtct
     2341 tcccgacgat gacgccggtg aacttcccgc cgccgttgtt gttttggagc acggaaagac
     2401 gatgacggaa aaagagatcg tggattacgt cgccagtcaa gtaacaaccg cgaaaaagtt
     2461 gcgcggagga gttgtgtttg tggacgaagt accgaaaggt cttaccggaa aactcgacgc
     2521 aagaaaaatc agagagatcc tcataaaggc caagaagggc ggaaagatcg ccgtgtaatt
     2581 ctagagtcgg ggcggccggc cgcttcgagc agacatgata agatacattg atgagtttgg
     2641 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     2701 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     2761 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     2821 caaatgtggt aaaatcgata aggatccgtc gaccgatgcc cttgagagcc ttcaacccag
     2881 tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct
     2941 ttatcatgca actcgtagga caggtgccgg cagcgctctt ccgcttcctc gctcactgac
     3001 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     3061 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     3121 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3181 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3241 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3301 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3361 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3421 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3481 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3541 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     3601 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3661 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3721 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3781 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     3841 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     3901 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     3961 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     4021 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     4081 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     4141 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     4201 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     4261 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     4321 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     4381 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     4441 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     4501 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     4561 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     4621 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     4681 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     4741 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     4801 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     4861 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt
     4921 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     4981 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     5041 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     5101 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     5161 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     5221 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     5281 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     5341 aacaaaatat taacgcttac aatttgccat tcgccattca ggctgcgcaa ctgttgggaa
     5401 gggcgatcgg tgcgggcctc ttcgctatta cgccagccca agctaccatg ataagtaagt
     5461 aatattaagg tacgggaggt acttggagcg gccgcaataa aatatcttta ttttcattac
     5521 atctgtgtgt tggttttttg tgtgaatcga tagtactaac atacgctctc catcaaaaca
     5581 aaacgaaaca aaacaaacta gcaaaatagg ctgtccccag tgcaagtgca ggtgccagaa
     5641 catttctcta tcgata
//
