LOCUS       sequence-315178-        6980 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express human YTHDF3(1-385) 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 6980)
  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 6980)
  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-N
FEATURES             Location/Qualifiers
     source          1..6980
                     /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             1488..2033
                     /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          2144..2373
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     2356..2375
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    2446..3017
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2611..2628)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2838..2857
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3053..3856
                     /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(3116..3135)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3726..3745
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3986..4107
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4023..4042)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4209..4225)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4209..4225)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4222..4244)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4233..4249
                     /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(4257..4263,4264..4281,4282..4287))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4302..4323
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4440..4457)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4611..5199)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4691..4710)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5370..6161,6162..6230))
                     /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     5993..6012
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6231..6335)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6410..6429)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6601..6980
                     /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 acctcaaact 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 gtgcttcaga attcaccatg gacaaagact
     1501 gcgaaatgaa gcgcaccacc ctggatagcc ctctgggcaa gctggaactg tctgggtgcg
     1561 aacagggcct gcaccgtatc atcttcctgg gcaaaggaac atctgccgcc gacgccgtgg
     1621 aagtgcctgc cccagccgcc gtgctgggcg gaccagagcc actgatgcag gccaccgcct
     1681 ggctcaacgc ctactttcac cagcctgagg ccatcgagga gttccctgtg ccagccctgc
     1741 accacccagt gttccagcag gagagcttta cccgccaggt gctgtggaaa ctgctgaaag
     1801 tggtgaagtt cggagaggtc atcagctaca gccacctggc cgccctggcc ggcaatcccg
     1861 ccgccaccgc cgccgtgaaa accgccctga gcggaaatcc cgtgcccatt ctgatcccct
     1921 gccaccgggt ggtgcagggc gacctggacg tggggggcta cgagggcggg ctcgccgtga
     1981 aagagtggct gctggcccac gagggccaca gactgggcaa gcctgggctg ggtcctgcag
     2041 gcggatccgc gtttaaactc gaggttaatt aatgagcggc cgcatagata actgatccag
     2101 tgtgctggaa ttaattcgct gtctgcgagg gccagctgtt ggggtgagta ctccctctca
     2161 aaagcgggca tgacttctgc gctaagattg tcagtttcca aaaacgagga ggatttgata
     2221 ttcacctggc ccgcggtgat gcctttgagg gtggccgcgt ccatctggtc agaaaagaca
     2281 atctttttgt tgtcaagctt gaggtgtggc aggcttgaga tctggccata cacttgagtg
     2341 acaatgacat ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgcagg
     2401 tcgagcatgc atctagggcg gccaattccg cccctctccc ccccccccct cycccccccc
     2461 cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt
     2521 tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct
     2581 tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga
     2641 atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga
     2701 ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac
     2761 gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag
     2821 ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc
     2881 agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg
     2941 tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt
     3001 gaaaaacacg atgataagct tgccacaacc cgggataatt cctgcagcca atatgggatc
     3061 ggccattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     3121 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     3181 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     3241 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     3301 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     3361 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     3421 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     3481 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     3541 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     3601 cggcgatgat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     3661 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     3721 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     3781 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     3841 tgacgagttc ttctgagggg atcaattctc tagataactg atcataatca gccataccac
     3901 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     3961 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     4021 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4081 tttgtccaaa ctcatcaatg tatcttaacg cgtcgagtgc attctagttg tggtttgtcc
     4141 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     4201 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     4261 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     4321 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     4381 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     4441 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4501 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4561 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4621 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4681 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4741 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4801 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4861 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4921 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4981 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5041 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5101 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     5161 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     5221 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     5281 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     5341 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     5401 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     5461 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     5521 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     5581 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     5641 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     5701 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     5761 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     5821 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     5881 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     5941 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     6001 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     6061 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     6121 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     6181 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     6241 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     6301 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     6361 gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct
     6421 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
     6481 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
     6541 atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
     6601 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
     6661 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     6721 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     6781 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     6841 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     6901 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     6961 tagtcatcgc tattaccatg
//
