LOCUS       sequence-470263-        5649 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Measure the activity of the reduced differentially methylated region
            (rDMR) of Myc lacking Tead-binding motif with luciferase assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5649)
  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 5649)
  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-rDMRdeltaTEAD
FEATURES             Location/Qualifiers
     source          1..5649
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    535..656
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(546..565)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     600..619
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(904..921)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1075..1663)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1155..1174)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1834..2625,2626..2694))
                     /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     2457..2476
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2695..2799)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2826..3281
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2913..2932)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3123..3144
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    3412..3460
                     /note="synthetic polyadenylation signal"
     misc_feature    3474..3565
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     3514..3533
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        4259..4455
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4306..4441
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      4485..4494
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4491..5649,1..494)
                     /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(4557..4575)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
ORIGIN
        1 gaggcgaact gtgtgtgaga ggtcctatga ttatgtccgg ttatgtaaac aatccggaag
       61 cgaccaacgc cttgattgac aaggatggat ggctacattc tggagacata gcttactggg
      121 acgaagacga acacttcttc atcgttgacc gcctgaagtc tctgattaag tacaaaggct
      181 atcaggtggc tcccgctgaa ttggaatcca tcttgctcca acaccccaac atcttcgacg
      241 caggtgtcgc aggtcttccc gacgatgacg ccggtgaact tcccgccgcc gttgttgttt
      301 tggagcacgg aaagacgatg acggaaaaag agatcgtgga ttacgtcgcc agtcaagtaa
      361 caaccgcgaa aaagttgcgc ggaggagttg tgtttgtgga cgaagtaccg aaaggtctta
      421 ccggaaaact cgacgcaaga aaaatcagag agatcctcat aaaggccaag aagggcggaa
      481 agatcgccgt gtaattctag agtcggggcg gccggccgct tcgagcagac atgataagat
      541 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
      601 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
      661 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa
      721 gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga tccgtcgacc gatgcccttg
      781 agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat cgtcgccgca
      841 cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc gctcttccgc
      901 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
      961 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     1021 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     1081 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     1141 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     1201 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     1261 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     1321 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     1381 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     1441 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     1501 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     1561 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     1621 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     1681 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     1741 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     1801 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     1861 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     1921 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     1981 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     2041 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     2101 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     2161 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     2221 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     2281 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     2341 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     2401 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     2461 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     2521 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     2581 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     2641 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     2701 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     2761 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     2821 acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     2881 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     2941 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     3001 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     3061 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     3121 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     3181 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     3241 atttaacgcg aattttaaca aaatattaac gcttacaatt tgccattcgc cattcaggct
     3301 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agcccaagct
     3361 accatgataa gtaagtaata ttaaggtacg ggaggtactt ggagcggccg caataaaata
     3421 tctttatttt cattacatct gtgtgttggt tttttgtgtg aatcgatagt actaacatac
     3481 gctctccatc aaaacaaaac gaaacaaaac aaactagcaa aataggctgt ccccagtgca
     3541 agtgcaggtg ccagaacatt tctctatcga taggtaccga gctcttacgc gtggtttccg
     3601 caccaaccag agctggataa ctctagactt gcttcccttg ctgtgccccc tccagcagac
     3661 agccacgacg atgcccctca acgtgaactt caccaacagg aactatgacc tcgactacga
     3721 ctccgtacag ccctctgcga cgaggaagag aatttctatc accagcaaca gcagagcgag
     3781 ctgcagccgc ccgcgcccag tgaggatatc tggaagaaat tcgagctgct tcccaccccg
     3841 cccctgtccc cgagccgccg ctccgggctc tgctctccat cctatgttgc ggtcgctacg
     3901 tccttctccc caagggaaga cgatgacggc ggcggtggca acttctccac cgccgatcag
     3961 ctggagatga tgaccgagtt acttggagga gacatggtga accagagctt catctgcgat
     4021 cctgacgacg agaccttcat caagaacatc atcatccagg actgtatgtg gagcggtttc
     4081 tcagccgctg ccaagctggt ctcggagaag ctggcctcct accaggctgc gcgcaaagac
     4141 agcaccagcc tgagccccgc ccgcgggcac agcgtctgct ccacctccag cctgtacctg
     4201 caggacctca ccgccgccgc gtccgagtgc attgacccct cactcgagat ctgcgatctg
     4261 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     4321 ccgcccagtt ccgcccattc tccgccccat cgctgactaa ttttttttat ttatgcagag
     4381 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4441 ctaggctttt gcaaaaagct tggcattccg gtactgttgg taaagccacc atggaagacg
     4501 ccaaaaacat aaagaaaggc ccggcgccat tctatccgct ggaagatgga accgctggag
     4561 agcaactgca taaggctatg aagagatacg ccctggttcc tggaacaatt gcttttacag
     4621 atgcacatat cgaggtggac atcacttacg ctgagtactt cgaaatgtcc gttcggttgg
     4681 cagaagctat gaaacgatat gggctgaata caaatcacag aatcgtcgta tgcagtgaaa
     4741 actctcttca attctttatg ccggtgttgg gcgcgttatt tatcggagtt gcagttgcgc
     4801 ccgcgaacga catttataat gaacgtgaat tgctcaacag tatgggcatt tcgcagccta
     4861 ccgtggtgtt cgtttccaaa aaggggttgc aaaaaatttt gaacgtgcaa aaaaagctcc
     4921 caatcatcca aaaaattatt atcatggatt ctaaaacgga ttaccaggga tttcagtcga
     4981 tgtacacgtt cgtcacatct catctacctc ccggttttaa tgaatacgat tttgtgccag
     5041 agtccttcga tagggacaag acaattgcac tgatcatgaa ctcctctgga tctactggtc
     5101 tgcctaaagg tgtcgctctg cctcatagaa ctgcctgcgt gagattctcg catgccagag
     5161 atcctatttt tggcaatcaa atcattccgg atactgcgat tttaagtgtt gttccattcc
     5221 atcacggttt tggaatgttt actacactcg gatatttgat atgtggattt cgagtcgtct
     5281 taatgtatag atttgaagaa gagctgtttc tgaggagcct tcaggattac aagattcaaa
     5341 gtgcgctgct ggtgccaacc ctattctcct tcttcgccaa aagcactctg attgacaaat
     5401 acgatttatc taatttacac gaaattgctt ctggtggcgc tcccctctct aaggaagtcg
     5461 gggaagcggt tgccaagagg ttccatctgc caggtatcag gcaaggatat gggctcactg
     5521 agactacatc agctattctg attacacccg agggggatga taaaccgggc gcggtcggta
     5581 aagttgttcc attttttgaa gcgaaggttg tggatctgga taccgggaaa acgctgggcg
     5641 ttaatcaaa
//
