LOCUS       sequence-315179-        7563 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express full-length human YTHDF3 with a C-terminal SNAP-tag in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7563)
  AUTHORS   Fu Y, Zhuang X
  TITLE     m(6)A-binding YTHDF proteins promote stress granule formation.
  JOURNAL   Nat Chem Biol. 2020 May 25. pii: 10.1038/s41589-020-0524-y. doi: 
            10.1038/s41589-020-0524-y.
  PUBMED    32451507
REFERENCE   2  (bases 1 to 7563)
  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: "Nat Chem 
            Biol. 2020 May 25. pii: 10.1038/s41589-020-0524-y. doi: 
            10.1038/s41589-020-0524-y."
COMMENT     Sequence Label: pSNAPf-hYTHDF3
FEATURES             Location/Qualifiers
     source          1..7563
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     249..268
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        249..267
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2088..2633
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     intron          2744..2973
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     2956..2975
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    3138..3600
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3194..3211)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3421..3440
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3636..4439
                     /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3699..3718)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4309..4328
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4569..4690
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4606..4625)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4792..4808)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4792..4808)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4805..4827)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4816..4832
                     /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(4840..4846,4847..4864,4865..4870))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4885..4906
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5023..5040)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5194..5782)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5274..5293)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5953..6744,6745..6813))
                     /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     6576..6595
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6814..6918)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6993..7012)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        7184..7563
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc ttactggctt
      241 atcgaaatta atacgactca ctatagggag acccaagctt ggtaccgagc tcggatcgtc
      301 gctagcgata tcggcgcgcc accatgtcag ccactagcgt ggatcagaga cctaaagggc
      361 aaggaaataa agtttcagta caaaacggtt cgattcatca aaaagatgct gtaaatgatg
      421 atgattttga gccatactta agtagccaga caaatcagag taacagctat ccaccaatgt
      481 cagatccata catgcctagt tactatgctc catccattgg atttccatat tctcttgggg
      541 aagcagcgtg gtccacagct ggagaccagc ctatgccata tctgacaacc tatggacaaa
      601 tgagtaatgg agaacatcac tatataccag atggtgtatt tagtcaacct ggggcattag
      661 gaaatacccc tccatttctt ggtcaacatg gatttaactt ttttcctggt aatgctgatt
      721 tctctacatg ggggacaagt ggatctcagg gacaatcaac acaaagttct gcttatagta
      781 gcagttatgg ctatccacct agttctcttg ggagagctat tactgatgga caggctggat
      841 ttggcaatga tactttgagt aaggtgcctg gcattagcag tattgagcaa ggcatgactg
      901 gactgaaaat tggtggtgac ctgacagctg cagtgacaaa aactgtaggt acagctttga
      961 gcagcagtgg tatgactagc attgcaacca atagtgtgcc cccagttagc agtgcagcac
     1021 ctaaaccaac ctcctgggct gccattgcca gaaagcctgc caaacctcaa ccgaaactta
     1081 aacccaaggg caatgtggga attgggggtt ctgctgtacc accacctcct ataaaacaca
     1141 acatgaatat tggaacttgg gatgaaaaag ggtcagtggt aaaggctcca ccaacccaac
     1201 cagttctgcc ccctcaaact ataatccagc agcctcagcc attaattcaa ccaccaccat
     1261 tggtgcaaag ccaactgcct caacagcagc ctcaaccacc acaaccacag cagcaacaag
     1321 gacctcagcc acaggcccag cctcaccaag tgcagcctca acagcagcag ctgcagaatc
     1381 gctgggtagc tcctcgtaac aggggagcag gcttcaacca gaacaatgga gcgggcagtg
     1441 aaaactttgg tttaggtgtt gtacctgtca gtgcttcacc ttctagtgta gaagtgcatc
     1501 ccgtgctgga aaagctaaag gccataaaca actataatcc caaagacttt gattggaatc
     1561 tgaagaatgg acgtgtgttt ataattaaaa gctactctga ggatgacata catcgttcca
     1621 ttaaatactc tatctggtgt agtactgagc atggtaataa gcgtttggat gcagcttacc
     1681 gttccctgaa tgggaaaggc ccactctatt tactcttcag tgtgaatggc agtggacatt
     1741 tttgtggagt ggctgaaatg aagtctgttg tggactataa tgcgtatgct ggtgtctggt
     1801 ctcaggataa gtggaagggc aaatttgaag ttaaatggat ctttgtcaaa gatgttccca
     1861 ataaccaatt acggcatatt cgcttagaaa ataatgacaa caaaccggtt accaattcaa
     1921 gggacactca agaggtaccc ctagaaaaag ctaagcaagt gcttaaaata attgctactt
     1981 tcaagcatac cacctcaatc tttgatgact ttgcacatta tgaaaagcgt caagaagagg
     2041 aggaagccat gcgtagggag agaaatagaa acaaacaaga attcaccatg gacaaagact
     2101 gcgaaatgaa gcgcaccacc ctggatagcc ctctgggcaa gctggaactg tctgggtgcg
     2161 aacagggcct gcaccgtatc atcttcctgg gcaaaggaac atctgccgcc gacgccgtgg
     2221 aagtgcctgc cccagccgcc gtgctgggcg gaccagagcc actgatgcag gccaccgcct
     2281 ggctcaacgc ctactttcac cagcctgagg ccatcgagga gttccctgtg ccagccctgc
     2341 accacccagt gttccagcag gagagcttta cccgccaggt gctgtggaaa ctgctgaaag
     2401 tggtgaagtt cggagaggtc atcagctaca gccacctggc cgccctggcc ggcaatcccg
     2461 ccgccaccgc cgccgtgaaa accgccctga gcggaaatcc cgtgcccatt ctgatcccct
     2521 gccaccgggt ggtgcagggc gacctggacg tggggggcta cgagggcggg ctcgccgtga
     2581 aagagtggct gctggcccac gagggccaca gactgggcaa gcctgggctg ggtcctgcag
     2641 gcggatccgc gtttaaactc gaggttaatt aatgagcggc cgcatagata actgatccag
     2701 tgtgctggaa ttaattcgct gtctgcgagg gccagctgtt ggggtgagta ctccctctca
     2761 aaagcgggca tgacttctgc gctaagattg tcagtttcca aaaacgagga ggatttgata
     2821 ttcacctggc ccgcggtgat gcctttgagg gtggccgcgt ccatctggtc agaaaagaca
     2881 atctttttgt tgtcaagctt gaggtgtggc aggcttgaga tctggccata cacttgagtg
     2941 acaatgacat ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgcagg
     3001 tcgagcatgc atctagggcg gccaattccg cccctctccc ccccccctaa cgttactggc
     3061 cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg
     3121 ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct
     3181 aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca
     3241 gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg
     3301 aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct
     3361 gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa
     3421 tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt
     3481 atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa
     3541 aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa
     3601 gcttgccaca acccgggata attcctgcag ccaatatggg atcggccatt gaacaagatg
     3661 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     3721 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     3781 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     3841 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     3901 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     3961 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     4021 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     4081 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     4141 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgat gatctcgtcg
     4201 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     4261 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     4321 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     4381 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     4441 gggatcaatt ctctagataa ctgatcataa tcagccatac cacatttgta gaggttttac
     4501 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     4561 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     4621 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     4681 atgtatctta acgcgtcgag tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     4741 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     4801 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     4861 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     4921 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     4981 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     5041 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5101 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5161 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5221 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5281 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5341 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5401 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5461 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5521 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5581 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     5641 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5701 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5761 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5821 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5881 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5941 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6001 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6061 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6121 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     6181 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6241 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6301 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6361 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6421 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6481 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6541 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     6601 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6661 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6721 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6781 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6841 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6901 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     6961 agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     7021 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
     7081 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg
     7141 cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact
     7201 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     7261 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     7321 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     7381 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     7441 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     7501 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     7561 atg
//
