LOCUS       sequence-315180-        6423 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express human YTHDF1(360-559) 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 6423)
  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 6423)
  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-C2
FEATURES             Location/Qualifiers
     source          1..6423
                     /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             936..1481
                     /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          1592..1821
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     1804..1823
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    1998..2460
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2054..2071)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2281..2300
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2496..3299
                     /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(2559..2578)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3169..3188
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3429..3550
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3466..3485)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3652..3668)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3652..3668)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3665..3687)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3676..3692
                     /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(3700..3706,3707..3724,3725..3730))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3745..3766
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3883..3900)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4054..4642)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4134..4153)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4813..5604,5605..5673))
                     /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     5436..5455
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5674..5778)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5853..5872)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6044..6423
                     /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 accatgccca gcgtcgaatc ccaccccgtc cttgaaaaac
      361 tgaaggctgc tcacagctac aacccgaaag agtttgagtg gaatctgaaa agcgggcgtg
      421 tgttcatcat caagagctac tctgaggacg acatccaccg ctccattaag tactccatct
      481 ggtgtagcac agagcacggc aacaagcgcc tggacagcgc cttccgctgc atgagcagca
      541 aggggcccgt ctacctgctc ttcagcgtca atgggagtgg gcatttttgt ggggtggccg
      601 agatgaagtc ccccgtggac tacggcacca gtgccggggt ctggtctcag gacaagtgga
      661 aggggaagtt tgatgtccag tggatttttg ttaaggatgt acccaataac cagctccggc
      721 acatcaggct ggagaataac gacaacaaac cggtcacaaa ctcccgggac acccaggagg
      781 tgcccttaga aaaagccaag caagtgctga aaattatcag ttcctacaag cacacaacct
      841 ccatcttcga cgactttgct cactacgaga agcgccagga ggaggaggag gtggtgcgca
      901 aggaacggca gagtcgaaac aaacaagaat tcaccatgga caaagactgc gaaatgaagc
      961 gcaccaccct ggatagccct ctgggcaagc tggaactgtc tgggtgcgaa cagggcctgc
     1021 accgtatcat cttcctgggc aaaggaacat ctgccgccga cgccgtggaa gtgcctgccc
     1081 cagccgccgt gctgggcgga ccagagccac tgatgcaggc caccgcctgg ctcaacgcct
     1141 actttcacca gcctgaggcc atcgaggagt tccctgtgcc agccctgcac cacccagtgt
     1201 tccagcagga gagctttacc cgccaggtgc tgtggaaact gctgaaagtg gtgaagttcg
     1261 gagaggtcat cagctacagc cacctggccg ccctggccgg caatcccgcc gccaccgccg
     1321 ccgtgaaaac cgccctgagc ggaaatcccg tgcccattct gatcccctgc caccgggtgg
     1381 tgcagggcga cctggacgtg gggggctacg agggcgggct cgccgtgaaa gagtggctgc
     1441 tggcccacga gggccacaga ctgggcaagc ctgggctggg tcctgcaggc ggatccgcgt
     1501 ttaaactcga ggttaattaa tgagcggccg catagataac tgatccagtg tgctggaatt
     1561 aattcgctgt ctgcgagggc cagctgttgg ggtgagtact ccctctcaaa agcgggcatg
     1621 acttctgcgc taagattgtc agtttccaaa aacgaggagg atttgatatt cacctggccc
     1681 gcggtgatgc ctttgagggt ggccgcgtcc atctggtcag aaaagacaat ctttttgttg
     1741 tcaagcttga ggtgtggcag gcttgagatc tggccataca cttgagtgac aatgacatcc
     1801 actttgcctt tctctccaca ggtgtccact cccaggtcca actgcaggtc gagcatgcat
     1861 ctagggcggc caattccgcc cctctccccc cccccctccc ccccccctaa cgttactggc
     1921 cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg
     1981 ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct
     2041 aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca
     2101 gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg
     2161 aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct
     2221 gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa
     2281 tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt
     2341 atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa
     2401 aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa
     2461 gcttgccaca acccgggata attcctgcag ccaatatggg atcggccatt gaacaagatg
     2521 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     2581 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     2641 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     2701 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     2761 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     2821 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     2881 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     2941 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     3001 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgat gatctcgtcg
     3061 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     3121 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     3181 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     3241 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     3301 gggatcaatt ctctagataa ctgatcataa tcagccatac cacatttgta gaggttttac
     3361 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     3421 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     3481 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     3541 atgtatctta acgcgtcgag tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     3601 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     3661 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     3721 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     3781 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     3841 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     3901 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     3961 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4021 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4081 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4141 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4201 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4261 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4321 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4381 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4441 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     4501 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4561 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4621 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4681 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     4741 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     4801 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4861 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     4921 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     4981 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5041 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5101 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5161 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5221 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5281 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5341 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5401 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5461 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     5521 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5581 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     5641 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     5701 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     5761 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     5821 agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     5881 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
     5941 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg
     6001 cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact
     6061 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     6121 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     6181 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     6241 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     6301 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     6361 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     6421 atg
//
