LOCUS       sequence-467803-        5549 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Measure the activity of the reduced differentially methylated region
            (rDMR) of Ctgf lacking Tead-binding motif with luciferase assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5549)
  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 5549)
  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-Ctgf-rDMRdeltaTEAD
FEATURES             Location/Qualifiers
     source          1..5549
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        587..783
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      634..769
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      813..822
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             819..2471
                     /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(885..903)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2512..2633
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2523..2542)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2577..2596
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2881..2898)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3052..3640)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3132..3151)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3811..4602,4603..4671))
                     /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     4434..4453
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4672..4776)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4803..5258
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4890..4909)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5100..5121
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5389..5437
                     /note="synthetic polyadenylation signal"
     misc_feature    5451..5542
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5491..5510
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt agcagcatcc atgaggaacc caggtcagcc aattgtatgt
       61 tggtcacttc cccatggtct ctcaccatgc actcagctct ctcctctcag gtaccacatg
      121 cagacacccc cactgacaga cagtgagtgc atagtccatg catgggagca ctcagcgctc
      181 catgttcagg ctcacgaccc attctacagc attggtaaaa ttcattcaga gctttcagcg
      241 ttctatggct ccccagctca tacaactatg aacgtcattt atttattctc agaacacatt
      301 tctggcttct gcaaacacat cagagtcttc ttgcaagcaa gctccaggtg actgctcata
      361 ttccaccata tgttaacaca acactagaaa taaactgccc aaaacactgt gaattatctt
      421 tgttattcag aggtacacaa gaataaattt ctagctttcc atttggttct ttggaatggg
      481 tttagcaatc tgagagaatt tcacacttct tacacctaaa ccttgaggac cctgacaatt
      541 atttttaaaa attccatcag cccgtcagtc ctcgagatct gcgatctgca tctcaattag
      601 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
      661 gcccattctc cgccccatcg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
      721 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
      781 aaaaagcttg gcattccggt actgttggta aagccaccat ggaagacgcc aaaaacataa
      841 agaaaggccc ggcgccattc tatccgctgg aagatggaac cgctggagag caactgcata
      901 aggctatgaa gagatacgcc ctggttcctg gaacaattgc ttttacagat gcacatatcg
      961 aggtggacat cacttacgct gagtacttcg aaatgtccgt tcggttggca gaagctatga
     1021 aacgatatgg gctgaataca aatcacagaa tcgtcgtatg cagtgaaaac tctcttcaat
     1081 tctttatgcc ggtgttgggc gcgttattta tcggagttgc agttgcgccc gcgaacgaca
     1141 tttataatga acgtgaattg ctcaacagta tgggcatttc gcagcctacc gtggtgttcg
     1201 tttccaaaaa ggggttgcaa aaaattttga acgtgcaaaa aaagctccca atcatccaaa
     1261 aaattattat catggattct aaaacggatt accagggatt tcagtcgatg tacacgttcg
     1321 tcacatctca tctacctccc ggttttaatg aatacgattt tgtgccagag tccttcgata
     1381 gggacaagac aattgcactg atcatgaact cctctggatc tactggtctg cctaaaggtg
     1441 tcgctctgcc tcatagaact gcctgcgtga gattctcgca tgccagagat cctatttttg
     1501 gcaatcaaat cattccggat actgcgattt taagtgttgt tccattccat cacggttttg
     1561 gaatgtttac tacactcgga tatttgatat gtggatttcg agtcgtctta atgtatagat
     1621 ttgaagaaga gctgtttctg aggagccttc aggattacaa gattcaaagt gcgctgctgg
     1681 tgccaaccct attctccttc ttcgccaaaa gcactctgat tgacaaatac gatttatcta
     1741 atttacacga aattgcttct ggtggcgctc ccctctctaa ggaagtcggg gaagcggttg
     1801 ccaagaggtt ccatctgcca ggtatcaggc aaggatatgg gctcactgag actacatcag
     1861 ctattctgat tacacccgag ggggatgata aaccgggcgc ggtcggtaaa gttgttccat
     1921 tttttgaagc gaaggttgtg gatctggata ccgggaaaac gctgggcgtt aatcaaagag
     1981 gcgaactgtg tgtgagaggt cctatgatta tgtccggtta tgtaaacaat ccggaagcga
     2041 ccaacgcctt gattgacaag gatggatggc tacattctgg agacatagct tactgggacg
     2101 aagacgaaca cttcttcatc gttgaccgcc tgaagtctct gattaagtac aaaggctatc
     2161 aggtggctcc cgctgaattg gaatccatct tgctccaaca ccccaacatc ttcgacgcag
     2221 gtgtcgcagg tcttcccgac gatgacgccg gtgaacttcc cgccgccgtt gttgttttgg
     2281 agcacggaaa gacgatgacg gaaaaagaga tcgtggatta cgtcgccagt caagtaacaa
     2341 ccgcgaaaaa gttgcgcgga ggagttgtgt ttgtggacga agtaccgaaa ggtcttaccg
     2401 gaaaactcga cgcaagaaaa atcagagaga tcctcataaa ggccaagaag ggcggaaaga
     2461 tcgccgtgta attctagagt cggggcggcc ggccgcttcg agcagacatg ataagataca
     2521 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     2581 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     2641 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca
     2701 agtaaaacct ctacaaatgt ggtaaaatcg ataaggatcc gtcgaccgat gcccttgaga
     2761 gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt
     2821 atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct cttccgcttc
     2881 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     2941 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     3001 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3061 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3121 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3181 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3241 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3301 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3361 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3421 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3481 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     3541 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     3601 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     3661 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     3721 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     3781 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     3841 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     3901 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     3961 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     4021 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4081 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     4141 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     4201 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     4261 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     4321 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     4381 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     4441 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     4501 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     4561 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     4621 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     4681 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     4741 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     4801 tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4861 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4921 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4981 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     5041 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     5101 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     5161 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     5221 taacgcgaat tttaacaaaa tattaacgct tacaatttgc cattcgccat tcaggctgcg
     5281 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc ccaagctacc
     5341 atgataagta agtaatatta aggtacggga ggtacttgga gcggccgcaa taaaatatct
     5401 ttattttcat tacatctgtg tgttggtttt ttgtgtgaat cgatagtact aacatacgct
     5461 ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt
     5521 gcaggtgcca gaacatttct ctatcgata
//
