LOCUS       sequence-209927-        7125 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MYC mRNA CDS region with wild type m6A motifs was inserted into 
            pMIR-REPORT vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7125)
  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 7125)
  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: pMIR-REPORT-MYC CDS_WT
FEATURES             Location/Qualifiers
     source          1..7125
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             533..2182
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIVPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(599..617)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     primer_bind     complement(2993..3010)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2993..3009)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3002..3024)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3218..3237)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3399..3715
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3566..3701
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3812..4411
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3812..3831)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4308..4328
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4641..4722
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4678..4697)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4810..4914
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4915..4983,4984..5775)
                     /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     complement(5133..5152)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5946..6534
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6435..6454
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6688..6705
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6822..6843
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6858..6863,6864..6881,6882..6888)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6896..6912
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     6901..6923
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6920..6936
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6920..6936
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        6956..6974
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     6956..6973
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     6986..7005
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6986..7004
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtggatcca ccatggaaga
      541 cgccaaaaac ataaagaaag gcccggcgcc attctatcct ctagaggatg gaaccgctgg
      601 agagcaactg cataaggcta tgaagagata cgccctggtt cctggaacaa ttgcttttac
      661 agatgcacat atcgaggtga acatcacgta cgcggaatac ttcgaaatgt ccgttcggtt
      721 ggcagaagct atgaaacgat atgggctgaa tacaaatcac agaatcgtcg tatgcagtga
      781 aaactctctt caattcttta tgccggtgtt gggcgcgtta tttatcggag ttgcagttgc
      841 gcccgcgaac gacatttata atgaacgtga attgctcaac agtatgaaca tttcgcagcc
      901 taccgtagtg tttgtttcca aaaaggggtt gcaaaaaatt ttgaacgtgc aaaaaaaatt
      961 accaataatc cagaaaatta ttatcatgga ttctaaaacg gattaccagg gatttcagtc
     1021 gatgtacacg ttcgtcacat ctcatctacc tcccggtttt aatgaatacg attttgtacc
     1081 agagtccttt gatcgtgaca aaacaattgc actgataatg aattcctctg gatctactgg
     1141 gttacctaag ggtgtggccc ttccgcatag aactgcctgc gtcagattct cgcatgccag
     1201 agatcctatt tttggcaatc aaatcgttcc ggatactgcg attttaagtg ttgtcccatt
     1261 ccatcacggt tttggaatgt ttactacact cggatatttg atatgtggat ttcgagtcgt
     1321 cttaatgtat agatttgaag aagagctgtt tttacgatcc cttcaggatt acaaaattca
     1381 aagtgcgttg ctagtaccaa ccctattttc attcttcgcc aaaagcactc tgattgacaa
     1441 atacgattta tctaatttac acgaaattgc ttctgggggc gcacctcttt cgaaagaagt
     1501 cggggaagcg gttgcaaaac gcttccatct tccagggata cgacaaggat atgggctcac
     1561 tgagactaca tcagctattc tgattacacc cgagggggat gataaaccgg gcgcggtcgg
     1621 taaagttgtt ccattttttg aagcgaaggt tgtggatctg gataccggga aaacgctggg
     1681 cgttaatcag agaggcgaat tatgtgtcag aggacctatg attatgtccg gttatgtaaa
     1741 caatccggaa gcgaccaacg ccttgattga caaggatgga tggctacatt ctggagacat
     1801 agcttactgg gacgaagacg aacacttctt catagttgac cgcttgaagt ctttaattaa
     1861 atacaaagga tatcaggtgg cccccgctga attggaatcg atattgttac aacaccccaa
     1921 catcttcgac gcgggcgtgg caggtcttcc cgacgatgac gccggtgaac ttcccgccgc
     1981 cgttgttgtt ttggagcacg gaaagacgat gacggaaaaa gagatcgtgg attacgtcgc
     2041 cagtcaagta acaaccgcga aaaagttgcg cggaggagtt gtgtttgtgg acgaagtacc
     2101 gaaaggtctt accggaaaac tcgacgcaag aaaaatcaga gagatcctca taaaggccaa
     2161 gaagggcgga aagtccaaat tgctcgagtg atgaaagctg cgcactagtg acccctttaa
     2221 ctcaagactg cctcccgctt tgtgtgcccc gctccagcag cctcccgcga cgatgcccct
     2281 caacgttagc ttcaccaaca ggaactatga cctcgactac gactcggtgc agccgtattt
     2341 ctactgcgac gaggaggaga acttctacca gcagcagcag cagagcgagc tgcagccccc
     2401 ggcgcccagc gaggatatct ggaagaaatt cgagctgctg cccaccccgc ccctgtcccc
     2461 tagccgccgc tccgggctct gctcgccctc ctacgttgcg gtcacaccct tctcccttcg
     2521 gggagacaac gacggcggtg gcgggagctt ctccacggcc gaccagctgg agatggtgac
     2581 cgagctgctg ggaggagaca tggtgaacca gagtttcatc tgcgacccgg acgacgagac
     2641 cttcatcaaa aacatcatca tccaggactg tatgtggagc ggcttctcgg ccgccgccaa
     2701 gctcgtctca gagaagctgg cctcctacca ggctgcgcgc aaagacagcg gcagcccgaa
     2761 ccccgcccgc ggccacagcg tctgctccac ctccagcttg tacctgcagg atctgagcgc
     2821 cgccgcctca gagtgcatcg acccctcggt ggtcttcccc taccctctca acgacagcag
     2881 ctcgcccaag tcctgcgcct cgcaagactc cagcgccttc aagcttaata aaggatcttt
     2941 tattttcatt ggatctgtgt gttggttttt tgtatgcggc cgctagcttg gcactggccg
     3001 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     3061 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     3121 aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc cttacgcatc
     3181 tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat
     3241 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     3301 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     3361 tttcaccgtc atcaccgaaa cgcgcgagac gaaagattgg tgtggaaagt ccccaggctc
     3421 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3481 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3541 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3601 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     3661 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     3721 agcttgcatg cctgcaggtc ggccgccacg accggtgccg ccaccatccc ctgacccacg
     3781 cccctgaccc ctcacaagga gacgaccttc catgaccgag tacaagccca cggtgcgcct
     3841 cgccacccgc gacgacgtcc cccgggccgt acgcaccctc gccgccgcgt tcgccgacta
     3901 ccccgccacg cgccacaccg tcgacccgga ccgccacatc gagcgggtca ccgagctgca
     3961 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg
     4021 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga
     4081 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga
     4141 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt
     4201 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc
     4261 ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt
     4321 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac
     4381 ctggtgcatg acccgcaagc ccggtgcctg acgcccgccc cacgacccgc agcgcccgac
     4441 cgaaaggagc gcacgacccc atggctccga ccgaagccac ccggggcggc cccgccgacc
     4501 ccgcacccgc ccccgaggcc caccgactct agaggatcat aatcagccat accacatttg
     4561 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     4621 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     4681 atagcatcac aaatttcaca aataaagcat ttttttcact gcaatctcgt gatacgccta
     4741 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     4801 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     4861 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     4921 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     4981 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     5041 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     5101 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     5161 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     5221 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     5281 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     5341 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     5401 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     5461 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     5521 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     5581 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     5641 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     5701 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     5761 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     5821 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     5881 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5941 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     6001 ctaccagcgg tggttttttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     6061 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     6121 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6181 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6241 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     6301 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     6361 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6421 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6481 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     6541 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     6601 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     6661 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     6721 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     6781 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     6841 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     6901 agcggataac aatttcacac aggaaacagc tatgacatga ttacgaattg caacgattta
     6961 ggtgacacta tagaagagaa ggaattaata cgactcacta tagggagaga gagagaatta
     7021 ccctcactaa agggaggaga agcatgaatt caaggtacca gatcttagtt attaatagta
     7081 atcaattacg gggtcattag ttcatagccc atatatggag ttccg
//
