LOCUS       sequence-467808-        5497 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Measure the activity of the reduced differentially methylated region
            (rDMR) of Cyr61 lacking Tead-binding motif with luciferase assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5497)
  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 5497)
  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-Cyr61-rDMRdeltaTEAD
FEATURES             Location/Qualifiers
     source          1..5497
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        535..731
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      582..717
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      761..770
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             767..2419
                     /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(833..851)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2460..2581
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2471..2490)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2525..2544
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2829..2846)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3000..3588)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3080..3099)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3759..4550,4551..4619))
                     /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     4382..4401
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4620..4724)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4751..5206
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4838..4857)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5048..5069
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5337..5385
                     /note="synthetic polyadenylation signal"
     misc_feature    5399..5490
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5439..5458
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt agaactactc taggtgccct acttgggtga cagtgaactg
       61 ttaggaaagt ggctgccctt gcaaatcttt agggaacaat ggtggcatat aagctgtcag
      121 cttggagggc gaagtactga acacgggcat ttgtgaagca gaaaaaggaa gcagaaattg
      181 aatttgttaa cctaaaacaa tgatttattt ttacaaggat aaagcctaag agttacccca
      241 ttaccccgaa caagtgcggc acttgtttct tattttagct tcggggtgat tctgtgtgtg
      301 gtgcaagtag aggtggggaa ccttttagta acctctgacc acagctgcat ggatgaagtg
      361 tatctaaaaa ttagagaagc tgcttcaata ttttaccatt ggctactagc agaagcgagg
      421 ggaaattcct gggacctatt gggtcgctgg tttgttaata gatgagcttg aagcaaagac
      481 ttgtgttaat cctaataagc cagtgaggaa tttcaaggct cgagatctgc gatctgcatc
      541 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
      601 ccagttccgc ccattctccg ccccatcgct gactaatttt ttttatttat gcagaggccg
      661 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
      721 gcttttgcaa aaagcttggc attccggtac tgttggtaaa gccaccatgg aagacgccaa
      781 aaacataaag aaaggcccgg cgccattcta tccgctggaa gatggaaccg ctggagagca
      841 actgcataag gctatgaaga gatacgccct ggttcctgga acaattgctt ttacagatgc
      901 acatatcgag gtggacatca cttacgctga gtacttcgaa atgtccgttc ggttggcaga
      961 agctatgaaa cgatatgggc tgaatacaaa tcacagaatc gtcgtatgca gtgaaaactc
     1021 tcttcaattc tttatgccgg tgttgggcgc gttatttatc ggagttgcag ttgcgcccgc
     1081 gaacgacatt tataatgaac gtgaattgct caacagtatg ggcatttcgc agcctaccgt
     1141 ggtgttcgtt tccaaaaagg ggttgcaaaa aattttgaac gtgcaaaaaa agctcccaat
     1201 catccaaaaa attattatca tggattctaa aacggattac cagggatttc agtcgatgta
     1261 cacgttcgtc acatctcatc tacctcccgg ttttaatgaa tacgattttg tgccagagtc
     1321 cttcgatagg gacaagacaa ttgcactgat catgaactcc tctggatcta ctggtctgcc
     1381 taaaggtgtc gctctgcctc atagaactgc ctgcgtgaga ttctcgcatg ccagagatcc
     1441 tatttttggc aatcaaatca ttccggatac tgcgatttta agtgttgttc cattccatca
     1501 cggttttgga atgtttacta cactcggata tttgatatgt ggatttcgag tcgtcttaat
     1561 gtatagattt gaagaagagc tgtttctgag gagccttcag gattacaaga ttcaaagtgc
     1621 gctgctggtg ccaaccctat tctccttctt cgccaaaagc actctgattg acaaatacga
     1681 tttatctaat ttacacgaaa ttgcttctgg tggcgctccc ctctctaagg aagtcgggga
     1741 agcggttgcc aagaggttcc atctgccagg tatcaggcaa ggatatgggc tcactgagac
     1801 tacatcagct attctgatta cacccgaggg ggatgataaa ccgggcgcgg tcggtaaagt
     1861 tgttccattt tttgaagcga aggttgtgga tctggatacc gggaaaacgc tgggcgttaa
     1921 tcaaagaggc gaactgtgtg tgagaggtcc tatgattatg tccggttatg taaacaatcc
     1981 ggaagcgacc aacgccttga ttgacaagga tggatggcta cattctggag acatagctta
     2041 ctgggacgaa gacgaacact tcttcatcgt tgaccgcctg aagtctctga ttaagtacaa
     2101 aggctatcag gtggctcccg ctgaattgga atccatcttg ctccaacacc ccaacatctt
     2161 cgacgcaggt gtcgcaggtc ttcccgacga tgacgccggt gaacttcccg ccgccgttgt
     2221 tgttttggag cacggaaaga cgatgacgga aaaagagatc gtggattacg tcgccagtca
     2281 agtaacaacc gcgaaaaagt tgcgcggagg agttgtgttt gtggacgaag taccgaaagg
     2341 tcttaccgga aaactcgacg caagaaaaat cagagagatc ctcataaagg ccaagaaggg
     2401 cggaaagatc gccgtgtaat tctagagtcg gggcggccgg ccgcttcgag cagacatgat
     2461 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     2521 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     2581 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     2641 ttaaagcaag taaaacctct acaaatgtgg taaaatcgat aaggatccgt cgaccgatgc
     2701 ccttgagagc cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg
     2761 ccgcacttat gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct
     2821 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     2881 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     2941 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     3001 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     3061 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     3121 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     3181 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     3241 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     3301 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     3361 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     3421 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3481 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     3541 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3601 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     3661 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     3721 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     3781 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     3841 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     3901 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     3961 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     4021 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     4081 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     4141 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     4201 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     4261 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     4321 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     4381 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     4441 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     4501 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     4561 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     4621 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     4681 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     4741 gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     4801 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     4861 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     4921 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     4981 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     5041 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     5101 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     5161 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttgcca ttcgccattc
     5221 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagccc
     5281 aagctaccat gataagtaag taatattaag gtacgggagg tacttggagc ggccgcaata
     5341 aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaatcg atagtactaa
     5401 catacgctct ccatcaaaac aaaacgaaac aaaacaaact agcaaaatag gctgtcccca
     5461 gtgcaagtgc aggtgccaga acatttctct atcgata
//
