LOCUS       sequence-308876-        7504 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express full-length human YTHDF1 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 7504)
  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 7504)
  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-hYTHDF1
FEATURES             Location/Qualifiers
     source          1..7504
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2390..2935
                     /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          3046..3275
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     3258..3277
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    3354..3921
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3515..3532)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3742..3761
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3957..4760
                     /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(4020..4039)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4630..4649
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4890..5011
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4927..4946)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5113..5129)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5113..5129)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5126..5148)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5137..5153
                     /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(5161..5167,5168..5185,5186..5191))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5206..5227
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5344..5361)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5515..6103)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5595..5614)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6274..7065,7066..7134))
                     /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     6897..6916
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7135..7239)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7314..7333)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ggtaccgagc tcggatcgtc gctagcgata tcggcgcgcc accatgtcgg ccaccagcgt
      721 ggacacccag agaacaaaag gacaagataa taaagtacaa aatggttcgt tacatcagaa
      781 ggatacagtt catgacaatg actttgagcc ctaccttact ggacagtcaa atcagagtaa
      841 cagttacccc tcaatgagcg acccctacct gtccagctat tacccgccgt ccattggatt
      901 tccttactcc ctcaatgagg ctccgtggtc tactgcaggg gaccctccga ttccatacct
      961 caccacctac ggacagctca gtaacggaga ccatcatttt atgcacgatg ctgtttttgg
     1021 gcagcctggg ggcctgggga acaacatcta tcagcacagg ttcaattttt tccctgaaaa
     1081 ccctgcgttc tcagcatggg ggacaagtgg gtctcaaggt cagcagaccc agagctccgc
     1141 gtatgggagc agctacacct accccccgag ctccctgggt ggcacggtgg ttgatgggca
     1201 gccaggcttt cacagcgaca ccctcagcaa ggcccccggg atgaacagcc tggagcaggg
     1261 catggttggc ctgaagattg gggacgtcag ctcctccgcc gtcaagacgg tgggctctgt
     1321 cgtcagcagc gtggcactga ctggtgtcct ttctggcaac ggtgggacaa atgtgaacat
     1381 gccagtttca aagccgacct cgtgggctgc cattgccagc aagcctgcaa aaccacagcc
     1441 taaaatgaaa acaaagagcg ggcctgtcat ggggggtggg ctgccccctc cacccataaa
     1501 gcataacatg gacattggca cctgggataa caaggggcct gtgccgaagg ccccagtccc
     1561 ccagcaggca ccctctccac aggctgcccc acagccccag caggtggctc agcctctccc
     1621 agcacagccc ccagctttgg ctcaaccgca gtatcagagc cctcagcagc caccccagac
     1681 ccgctgggtt gccccacgca acagaaacgc ggcgtttggg cagagcggag gggctggcag
     1741 cgatagcaac tctcctggaa acgtccagcc taattctgcc cccagcgtcg aatcccaccc
     1801 cgtccttgaa aaactgaagg ctgctcacag ctacaacccg aaagagtttg agtggaatct
     1861 gaaaagcggg cgtgtgttca tcatcaagag ctactctgag gacgacatcc accgctccat
     1921 taagtactcc atctggtgta gcacagagca cggcaacaag cgcctggaca gcgccttccg
     1981 ctgcatgagc agcaaggggc ccgtctacct gctcttcagc gtcaatggga gtgggcattt
     2041 ttgtggggtg gccgagatga agtcccccgt ggactacggc accagtgccg gggtctggtc
     2101 tcaggacaag tggaagggga agtttgatgt ccagtggatt tttgttaagg atgtacccaa
     2161 taaccagctc cggcacatca ggctggagaa taacgacaac aaaccggtca caaactcccg
     2221 ggacacccag gaggtgccct tagaaaaagc caagcaagtg ctgaaaatta tcagttccta
     2281 caagcacaca acctccatct tcgacgactt tgctcactac gagaagcgcc aggaggagga
     2341 ggaggtggtg cgcaaggaac ggcagagtcg aaacaaacaa gaattcacca tggacaaaga
     2401 ctgcgaaatg aagcgcacca ccctggatag ccctctgggc aagctggaac tgtctgggtg
     2461 cgaacagggc ctgcaccgta tcatcttcct gggcaaagga acatctgccg ccgacgccgt
     2521 ggaagtgcct gccccagccg ccgtgctggg cggaccagag ccactgatgc aggccaccgc
     2581 ctggctcaac gcctactttc accagcctga ggccatcgag gagttccctg tgccagccct
     2641 gcaccaccca gtgttccagc aggagagctt tacccgccag gtgctgtgga aactgctgaa
     2701 agtggtgaag ttcggagagg tcatcagcta cagccacctg gccgccctgg ccggcaatcc
     2761 cgccgccacc gccgccgtga aaaccgccct gagcggaaat cccgtgccca ttctgatccc
     2821 ctgccaccgg gtggtgcagg gcgacctgga cgtggggggc tacgagggcg ggctcgccgt
     2881 gaaagagtgg ctgctggccc acgagggcca cagactgggc aagcctgggc tgggtcctgc
     2941 aggcggatcc gcgtttaaac tcgaggttaa ttaatgagcg gccgcataga taactgatcc
     3001 agtgtgctgg aattaattcg ctgtctgcga gggccagctg ttggggtgag tactccctct
     3061 caaaagcggg catgacttct gcgctaagat tgtcagtttc caaaaacgag gaggatttga
     3121 tattcacctg gcccgcggtg atgcctttga gggtggccgc gtccatctgg tcagaaaaga
     3181 caatcttttt gttgtcaagc ttgaggtgtg gcaggcttga gatctggcca tacacttgag
     3241 tgacaatgac atccactttg cctttctctc cacaggtgtc cactcccagg tccaactgca
     3301 ggtcgagcat gcatctaggg cggccaattc cgcccctctc cccccccccn ctctccctcc
     3361 ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat
     3421 atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct
     3481 gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg
     3541 ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta
     3601 gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag
     3661 ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg
     3721 atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat
     3781 gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca
     3841 tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc
     3901 ctttgaaaaa cacgatgata agcttgccac aacccgggat aattcctgca gccaatatgg
     3961 gatcggccat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     4021 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     4081 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     4141 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     4201 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     4261 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     4321 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     4381 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     4441 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     4501 ccgacggcga tgatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     4561 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     4621 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     4681 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     4741 ttcttgacga gttcttctga ggggatcaat tctctagata actgatcata atcagccata
     4801 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     4861 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     4921 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     4981 gtggtttgtc caaactcatc aatgtatctt aacgcgtcga gtgcattcta gttgtggttt
     5041 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     5101 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     5161 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     5221 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     5281 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5341 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5401 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5461 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5521 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5581 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     5641 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     5701 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     5761 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     5821 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     5881 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     5941 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6001 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     6061 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6121 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6181 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6241 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     6301 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     6361 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     6421 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     6481 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     6541 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     6601 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     6661 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     6721 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     6781 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     6841 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     6901 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     6961 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     7021 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     7081 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     7141 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     7201 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     7261 acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc
     7321 tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct
     7381 gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg
     7441 aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg
     7501 cgtt
//
