LOCUS       sequence-271545-        7082 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  full length YTHDF2 with N-terminal flag tag for mammalian cell 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7082)
  AUTHORS   Wang X, Lu Z, Gomez A, Hon GC, Yue Y, Han D, Fu Y, Parisien M, Dai 
            Q, Jia G, Ren B, Pan T, He C
  TITLE     N6-methyladenosine-dependent regulation of messenger RNA stability.
  JOURNAL   Nature. 2014 Jan 2;505(7481):117-20. doi: 10.1038/nature12730. Epub 
            2013 Nov 27.
  PUBMED    24284625
REFERENCE   2  (bases 1 to 7082)
  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/nature12730"; journalName: "Nature"; date: "2014-01-2- 2"; 
            volume: "505"; issue: "7481"; pages: "117-20"
COMMENT     Sequence Label: pcDNA3-flag-YTHDF2
FEATURES             Location/Qualifiers
     source          1..7082
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(66..82)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(66..82)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(79..101)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    90..106
                     /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)."
     promoter        complement(join(114..120,121..138,139..144))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    159..180
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(297..314)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(468..1056)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(548..567)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1227..2018,2019..2087))
                     /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     1850..1869
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2088..2192)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2267..2286)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2458..2837
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2838..3041
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2992..3012
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3086..3105
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3086..3104
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3139..3162
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(4930..4948)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4931..4948)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4968..4985)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4974..5198
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5244..5672
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5331..5350)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5541..5562
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5681..5701)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5725..5935
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        5747..5943
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5794..5929
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5856..5875
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6010..6804
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6064..6083)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6674..6693
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    join(6978..7082,1..17)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7015..7034)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     join(7069..7082,1..6)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
       61 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
      121 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
      181 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
      241 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
      301 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
      361 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
      421 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
      481 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
      541 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
      601 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
      661 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
      721 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
      781 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
      841 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
      901 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
      961 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     1021 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     1081 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     1141 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     1201 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     1261 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     1321 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     1381 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     1441 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     1501 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     1561 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     1621 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     1681 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     1741 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     1801 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     1861 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     1921 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     1981 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     2041 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     2101 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     2161 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     2221 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
     2281 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
     2341 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
     2401 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
     2461 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
     2521 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     2581 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     2641 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     2701 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     2761 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     2821 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     2881 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     2941 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     3001 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta actagagaac
     3061 ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc caagcttggt
     3121 atcgagctcg gatccatgga ttacaaggac gacgatgaca agatgtcggc cagcagcctc
     3181 ttggagcaga gaccaaaagg tcaaggaaac aaagtacaaa atggatctgt acatcaaaag
     3241 gatggattaa acgatgatga ttttgaacct tacttgagtc cacaggcaag gcccaataat
     3301 gcatatactg ccatgtcaga ttcctactta cccagttact acagtccctc cattggcttc
     3361 tcctattctt tgggtgaagc tgcttggtct acggggggtg acacagccat gccctactta
     3421 acttcttatg gacagctgag caacggagag ccccacttcc taccagatgc aatgtttggg
     3481 caaccaggag ccctaggtag cactccattt cttggtcagc atggttttaa tttctttccc
     3541 agtgggattg acttctcagc atggggaaat aacagttctc agggacagtc tactcagagc
     3601 tctggatata gtagcaatta tgcttatgca cctagctcct taggtggagc catgattgat
     3661 ggacagtcag cttttgccaa tgagaccctc aataaggctc ctggcatgaa tactatagac
     3721 caagggatgg cagcactgaa gttgggtagc acagaagttg caagcaatgt tccaaaagtt
     3781 gtaggttctg ctgttggtag cgggtccatt actagtaaca tcgtggcttc caatagtttg
     3841 cctccagcca ccattgctcc tccaaaacca gcatcttggg ctgatattgc tagcaagcct
     3901 gcaaaacagc aacctaaact gaagaccaag aatggcattg cagggtcaag tcttccgcca
     3961 cccccgataa agcataacat ggatattgga acttgggata acaagggtcc cgttgcaaaa
     4021 gccccctcac aggctttggt tcagaatata ggtcagccaa cccaggggtc tcctcagcct
     4081 gtaggtcagc aggctaacaa tagcccacca gtggctcagg catcagtagg gcaacagaca
     4141 cagccattgc ctccacctcc accacagcct gcccagcttt cagtccagca acaggcagct
     4201 cagccaaccc gctgggtagc acctcggaac cgtggcagtg ggttcggtca taatggggtg
     4261 gatggtaatg gagtaggaca gtctcaggct ggttctggat ctactccttc agaaccccac
     4321 ccagtgttgg agaagcttcg gtccattaat aactataacc ccaaagattt tgactggaat
     4381 ctgaaacatg gccgggtttt catcattaag agctactctg aggacgatat tcaccgttcc
     4441 attaagtata atatttggtg cagcacagag catggtaaca agagactgga tgctgcttat
     4501 cgttccatga acgggaaagg ccccgtttac ttacttttca gtgtcaacgg cagtggacac
     4561 ttctgtggcg tggcagaaat gaaatctgct gtggactaca acacatgtgc aggtgtgtgg
     4621 tcccaggaca aatggaaggg tcgttttgat gtcaggtgga tttttgtgaa ggacgttccc
     4681 aatagccaac tgcgacacat tcgcctagag aacaacgaga ataaaccagt gaccaactct
     4741 agggacactc aggaagtgcc tctggaaaag gctaagcagg tgttgaaaat tatagccagc
     4801 tacaagcaca ccacttccat ttttgatgac ttctcacact atgagaaacg ccaagaggaa
     4861 gaagaaagtg ttaaaaagga acgtcaaggt cgtgggaaat gactcgagca tgcatctaga
     4921 gggccctatt ctatagtgtc acctaaatgc tagagctcgc tgatcagcct cgactgtgcc
     4981 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     5041 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     5101 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     5161 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     5221 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     5281 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     5341 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     5401 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     5461 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     5521 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     5581 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     5641 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     5701 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     5761 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     5821 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     5881 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     5941 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     6001 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     6061 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     6121 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     6181 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     6241 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     6301 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     6361 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     6421 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     6481 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     6541 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     6601 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     6661 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     6721 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     6781 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     6841 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     6901 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     6961 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     7021 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     7081 aa
//
