LOCUS       V010070                 7052 bp    DNA     circular SYN 21-DEC-2021
DEFINITION  Exported.
ACCESSION   V010070
VERSION     V010070
KEYWORDS    pcDNA3/Flag-METTL3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7052)
  AUTHORS   Liu J, Yue Y, Han D, Wang X, Fu Y, Zhang L, Jia G, Yu M, Lu Z, Deng
            X, Dai Q, Chen W, He C
  TITLE     A METTL3-METTL14 complex mediates mammalian nuclear RNA N6-adenosine
            methylation.
  JOURNAL   Nat Chem Biol. 2014 Feb;10(2):93-5. doi: 10.1038/nchembio.1432. Epub
            2013 Dec 6.
   PUBMED   24316715
REFERENCE   2  (bases 1 to 7052)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 7052)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi:
            "10.1038/nchembio.1432"; journalName: "Nat Chem Biol"; date:
            "2014-02"; volume: "10"; issue: "2"; pages: "93-5"
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7052
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     23..44
                     /label="F1ori-F"
                     /note="F1 origin, forward primer"
     primer_bind     complement(163..183)
                     /label="pBABE 3'"
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        207..417
                     /label="SV40 promoter"
                     /note="SV40 early promoter"
     CDS             492..1283
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    1460..1593
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1630..1646)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(1654..1670)
                     /label="lac operator"
                     /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)."
     promoter        complement(1678..1708)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1723..1744)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     primer_bind     complement(1861..1878)
                     /label="L4440"
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2032..2620)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(2794..3651)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(3652..3756)
                     /label="AmpR promoter"
     primer_bind     complement(3831..3850)
                     /label="pRS-marker"
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
     enhancer        4022..4401
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4402..4605
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        4650..4668
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4685..4708
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             4709..6445
                     /gene="METTL3"
                     /label="N6-adenosine-methyltransferase catalytic subunit"
                     /note="N6-adenosine-methyltransferase catalytic subunit
                     from Homo sapiens. Accession#: Q86U44"
     promoter        complement(6464..6482)
                     /label="SP6 promoter"
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    6508..6732
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6778..7052
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
       61 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
      121 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
      181 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
      241 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
      301 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
      361 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
      421 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
      481 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
      541 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
      601 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
      661 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
      721 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
      781 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
      841 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
      901 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
      961 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     1021 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     1081 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     1141 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     1201 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     1261 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     1321 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     1381 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     1441 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     1501 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     1561 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     1621 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     1681 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     1741 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     1801 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     1861 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     1921 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     1981 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     2041 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     2101 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     2161 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     2221 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     2281 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     2341 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     2401 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     2461 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     2521 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     2581 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     2641 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     2701 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     2761 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     2821 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     2881 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     2941 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     3001 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     3061 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     3121 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     3181 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     3241 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     3301 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     3361 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     3421 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     3481 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     3541 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     3601 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     3661 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     3721 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     3781 tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc
     3841 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     3901 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
     3961 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt
     4021 tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
     4081 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     4141 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     4201 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     4261 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     4321 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     4381 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
     4441 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
     4501 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
     4561 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctctg gctaactaga
     4621 gaacccactg cttactggct tatcgaaatt aatacgactc actataggga gacccaagct
     4681 tatggattac aaggatgacg atgacaagtc ggacacgtgg agctctatcc aggcccacaa
     4741 gaagcagctg gactctctgc gggagaggct gcagcggagg cggaagcagg actcggggca
     4801 cttggatcta cggaatccag aggcagcatt gtctccaacc ttccgtagtg acagcccagt
     4861 gcctactgca cccacctctg gtggccctaa gcccagcaca gcttcagcag ttcctgaatt
     4921 agctacagat cctgagttag agaagaagtt gctacaccac ctctctgatc tggccttaac
     4981 attgcccact gatgctgtgt ccatctgtct tgccatctcc acgccagatg ctcctgccac
     5041 tcaagatggg gtagaaagcc tcctgcagaa gtttgcagct caggagttga ttgaggtaaa
     5101 gcgaggtctc ctacaagatg atgcacatcc tactcttgta acctatgctg accattccaa
     5161 gctctctgcc atgatgggtg ctgtggcaga aaagaagggc cctggggagg tagcagggac
     5221 tgtcacaggg cagaagcggc gtgcagaaca ggactcgact acagtagctg cctttgccag
     5281 ttcgttagtc tctggtctga actcttcagc atcggaacca gcaaaggagc cagccaagaa
     5341 atcaaggaaa catgctgcct cagatgttga tctggagata gagagccttc tgaaccaaca
     5401 gtccactaag gaacaacaga gcaagaaggt cagtcaggag atcctagagc tattaaatac
     5461 tacaacagcc aaggaacaat ccattgttga aaaatttcgc tctcgaggtc gggcccaagt
     5521 gcaagaattc tgtgactatg gaaccaagga ggagtgcatg aaagccagtg atgctgatcg
     5581 accctgtcgc aagctgcact tcagacgaat tatcaataaa cacactgatg agtctttagg
     5641 tgactgctct ttccttaata catgtttcca catggatacc tgcaagtatg ttcactatga
     5701 aattgatgct tgcatggatt ctgaggcccc tggcagcaaa gaccacacgc caagccagga
     5761 gcttgctctt acacagagtg tcggaggtga ttccagtgca gaccgactct tcccacctca
     5821 gtggatctgt tgtgatatcc gctacctgga cgtcagtatc ttgggcaagt ttgcagttgt
     5881 gatggctgac ccaccctggg atattcacat ggaactgccc tatgggaccc tgacagatga
     5941 tgagatgcgc aggctcaaca tacccgtact acaggatgat ggctttctct tcctctgggt
     6001 cacaggcagg gccatggagt tggggagaga atgtctaaat ctctgggggt atgaacgggt
     6061 agatgaaatt atttgggtga agacaaatca actgcaacgc atcattcgga caggccgtac
     6121 aggtcactgg ttgaaccatg ggaaggaaca ctgcttggtt ggtgtcaaag gaaatcccca
     6181 aggcttcaac cagggtctgg attgtgatgt gatcgtagct gaggttcgtt ccaccagtca
     6241 taaaccagat gaaatctatg gcatgattga aagactatct cctggcactc gcaagattga
     6301 gttatttgga cgaccacaca atgtgcaacc caactggatc acccttggaa accaactgga
     6361 tgggatccac ctactagacc cagatgtggt tgcacggttc aagcaaaggt acccagatgg
     6421 tatcatctct aaacctaaga atttatagtc tagagggccc tattctatag tgtcacctaa
     6481 atgctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     6541 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     6601 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     6661 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     6721 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     6781 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     6841 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     6901 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     6961 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     7021 cgccctgata gacggttttt cgccctttga cg
//
