LOCUS       sequence-209921-        5028 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The core promoter region of FTO with mutant CEBPA binding sites was 
            inserted into pGL3 basic vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5028)
  AUTHORS   Su R, Dong L, Li C, Nachtergaele S, Wunderlich M, Qing Y, Deng X, 
            Wang Y, Weng X, Hu C, Yu M, Skibbe J, Dai Q, Zou D, Wu T, Yu K, Weng
            H, Huang H, Ferchen K, Qin X, Zhang B, Qi J, Sasaki AT, Plas DR, 
            Bradner JE, Wei M, Marcucci G, Jiang X, Mulloy JC, Jin J, He C, Chen
            J
  TITLE     R-2HG Exhibits Anti-tumor Activity by Targeting FTO/m(6)A/MYC/CEBPA 
            Signaling.
  JOURNAL   Cell. 2018 Jan 11;172(1-2):90-105.e23. doi: 
            10.1016/j.cell.2017.11.031. Epub 2017 Dec 14.
  PUBMED    29249359
REFERENCE   2  (bases 1 to 5028)
  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: "Cell."; 
            date: "2018-01-11"; volume: "172(1-2):90-105.e23. doi"; pages: " 
            10.1016/j.cell.2017.11.031. Epub 2017 Dec 14"
COMMENT     Sequence Label: pGL3-basic-FTO promoter_Mut
FEATURES             Location/Qualifiers
     source          1..5028
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      44..499
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(131..150)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     341..362
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    630..678
                     /note="synthetic polyadenylation signal"
     misc_feature    692..783
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     732..751
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     regulatory      1082..1091
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1088..2740
                     /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(1154..1172)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2781..2902
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2792..2811)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2846..2865
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3150..3167)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3321..3909)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3401..3420)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4080..4871,4872..4940))
                     /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     4703..4722
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(4941..5028,1..17))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgcgcc ctgtagcggc
       61 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
      121 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
      181 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
      241 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
      301 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
      361 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
      421 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
      481 atattaacgc ttacaatttg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
      541 tcggtgcggg cctcttcgct attacgccag cccaagctac catgataagt aagtaatatt
      601 aaggtacggg aggtacttgg agcggccgca ataaaatatc tttattttca ttacatctgt
      661 gtgttggttt tttgtgtgaa tcgatagtac taacatacgc tctccatcaa aacaaaacga
      721 aacaaaacaa actagcaaaa taggctgtcc ccagtgcaag tgcaggtgcc agaacatttc
      781 tctatcgata ggtacctcca cccaccctca tcctccccca tcctcccggg tactcaccgt
      841 gccactggcc ctgcagctag ctaccgttgc tatagcgccg acagcgtggc gggcggctgg
      901 ccgagaggag cacgggagaa acatggcagg ctcccgtagc ctcgatcgat cggtagttct
      961 ccttggactc tagcctgttt gctcgcgggg tagcggacta cgctcttcca gctgtcggac
     1021 ctgggaaatt cctcgagatc tgcgatctaa gtaagcttgg cattccggta ctgttggtaa
     1081 agccaccatg gaagacgcca aaaacataaa gaaaggcccg gcgccattct atccgctgga
     1141 agatggaacc gctggagagc aactgcataa ggctatgaag agatacgccc tggttcctgg
     1201 aacaattgct tttacagatg cacatatcga ggtggacatc acttacgctg agtacttcga
     1261 aatgtccgtt cggttggcag aagctatgaa acgatatggg ctgaatacaa atcacagaat
     1321 cgtcgtatgc agtgaaaact ctcttcaatt ctttatgccg gtgttgggcg cgttatttat
     1381 cggagttgca gttgcgcccg cgaacgacat ttataatgaa cgtgaattgc tcaacagtat
     1441 gggcatttcg cagcctaccg tggtgttcgt ttccaaaaag gggttgcaaa aaattttgaa
     1501 cgtgcaaaaa aagctcccaa tcatccaaaa aattattatc atggattcta aaacggatta
     1561 ccagggattt cagtcgatgt acacgttcgt cacatctcat ctacctcccg gttttaatga
     1621 atacgatttt gtgccagagt ccttcgatag ggacaagaca attgcactga tcatgaactc
     1681 ctctggatct actggtctgc ctaaaggtgt cgctctgcct catagaactg cctgcgtgag
     1741 attctcgcat gccagagatc ctatttttgg caatcaaatc attccggata ctgcgatttt
     1801 aagtgttgtt ccattccatc acggttttgg aatgtttact acactcggat atttgatatg
     1861 tggatttcga gtcgtcttaa tgtatagatt tgaagaagag ctgtttctga ggagccttca
     1921 ggattacaag attcaaagtg cgctgctggt gccaacccta ttctccttct tcgccaaaag
     1981 cactctgatt gacaaatacg atttatctaa tttacacgaa attgcttctg gtggcgctcc
     2041 cctctctaag gaagtcgggg aagcggttgc caagaggttc catctgccag gtatcaggca
     2101 aggatatggg ctcactgaga ctacatcagc tattctgatt acacccgagg gggatgataa
     2161 accgggcgcg gtcggtaaag ttgttccatt ttttgaagcg aaggttgtgg atctggatac
     2221 cgggaaaacg ctgggcgtta atcaaagagg cgaactgtgt gtgagaggtc ctatgattat
     2281 gtccggttat gtaaacaatc cggaagcgac caacgccttg attgacaagg atggatggct
     2341 acattctgga gacatagctt actgggacga agacgaacac ttcttcatcg ttgaccgcct
     2401 gaagtctctg attaagtaca aaggctatca ggtggctccc gctgaattgg aatccatctt
     2461 gctccaacac cccaacatct tcgacgcagg tgtcgcaggt cttcccgacg atgacgccgg
     2521 tgaacttccc gccgccgttg ttgttttgga gcacggaaag acgatgacgg aaaaagagat
     2581 cgtggattac gtcgccagtc aagtaacaac cgcgaaaaag ttgcgcggag gagttgtgtt
     2641 tgtggacgaa gtaccgaaag gtcttaccgg aaaactcgac gcaagaaaaa tcagagagat
     2701 cctcataaag gccaagaagg gcggaaagat cgccgtgtaa ttctagagtc ggggcggccg
     2761 gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     2821 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     2881 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     2941 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga
     3001 taaggatccg tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg
     3061 cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag
     3121 gacaggtgcc ggcagcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3181 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     3241 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3301 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3361 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3421 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3481 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3541 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3601 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3661 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3721 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     3781 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3841 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3901 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3961 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     4021 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     4081 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     4141 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     4201 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     4261 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     4321 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     4381 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     4441 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     4501 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     4561 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     4621 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     4681 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     4741 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     4801 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     4861 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     4921 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     4981 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaa
//
