LOCUS       sequence-308858-        7565 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express full-length human YTHDF2 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 7565)
  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 7565)
  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-hYTHDF2
FEATURES             Location/Qualifiers
     source          1..7565
                     /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             2450..2995
                     /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          3106..3335
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     3318..3337
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    3415..3982
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3576..3593)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3803..3822
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4018..4821
                     /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(4081..4100)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4691..4710
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4951..5072
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4988..5007)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5174..5190)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5174..5190)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5187..5209)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5198..5214
                     /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(5222..5228,5229..5246,5247..5252))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5267..5288
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5405..5422)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5576..6164)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5656..5675)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6335..7126,7127..7195))
                     /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     6958..6977
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7196..7300)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7375..7394)
                     /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 ccagcagcct
      721 cttggagcag agaccaaaag gtcaaggaaa caaagtacaa aatggatctg tacatcaaaa
      781 ggatggatta aacgatgatg attttgaacc ttacttgagt ccacaggcaa ggcccaataa
      841 tgcatatact gccatgtcag attcctactt acccagttac tacagtccct ccattggctt
      901 ctcctattct ttgggtgaag ctgcttggtc tacggggggt gacacagcca tgccctactt
      961 aacttcttat ggacagctga gcaacggaga gccccacttc ctaccagatg caatgtttgg
     1021 gcaaccagga gccctaggta gcactccatt tcttggtcag catggtttta atttctttcc
     1081 cagtgggatt gacttctcag catggggaaa taacagttct cagggacagt ctactcagag
     1141 ctctggatat agtagcaatt atgcttatgc acctagctcc ttaggtggag ccatgattga
     1201 tggacagtca gcttttgcca atgagaccct caataaggct cctggcatga atactataga
     1261 ccaagggatg gcagcactga agttgggtag cacagaagtt gcaagcaatg ttccaaaagt
     1321 tgtaggttct gctgttggta gcgggtccat tactagtaac atcgtggctt ccaatagttt
     1381 gcctccagcc accattgctc ctccaaaacc agcatcttgg gctgatattg ctagcaagcc
     1441 tgcaaaacag caacctaaac tgaagaccaa gaatggcatt gcagggtcaa gtcttccgcc
     1501 acccccgata aagcataaca tggatattgg aacttgggat aacaagggtc ccgttgcaaa
     1561 agccccctca caggctttgg ttcagaatat aggtcagcca acccaggggt ctcctcagcc
     1621 tgtaggtcag caggctaaca atagcccacc agtggctcag gcatcagtag ggcaacagac
     1681 acagccattg cctccacctc caccacagcc tgcccagctt tcagtccagc aacaggcagc
     1741 tcagccaacc cgctgggtag cacctcggaa ccgtggcagt gggttcggtc ataatggggt
     1801 ggatggtaat ggagtaggac agtctcaggc tggttctgga tctactcctt cagaacccca
     1861 cccagtgttg gagaagcttc ggtccattaa taactataac cccaaagatt ttgactggaa
     1921 tctgaaacat ggccgggttt tcatcattaa gagctactct gaggacgata ttcaccgttc
     1981 cattaagtat aatatttggt gcagcacaga gcatggtaac aagagactgg atgctgctta
     2041 tcgttccatg aacgggaaag gccccgttta cttacttttc agtgtcaacg gcagtggaca
     2101 cttctgtggc gtggcagaaa tgaaatctgc tgtggactac aacacatgtg caggtgtgtg
     2161 gtcccaggac aaatggaagg gtcgttttga tgtcaggtgg atttttgtga aggacgttcc
     2221 caatagccaa ctgcgacaca ttcgcctaga gaacaacgag aataaaccag tgaccaactc
     2281 tagggacact caggaagtgc ctctggaaaa ggctaagcag gtgttgaaaa ttatagccag
     2341 ctacaagcac accacttcca tttttgatga cttctcacac tatgagaaac gccaagagga
     2401 agaagaaagt gttaaaaagg aacgtcaagg tcgtgggaaa gaattcacca tggacaaaga
     2461 ctgcgaaatg aagcgcacca ccctggatag ccctctgggc aagctggaac tgtctgggtg
     2521 cgaacagggc ctgcaccgta tcatcttcct gggcaaagga acatctgccg ccgacgccgt
     2581 ggaagtgcct gccccagccg ccgtgctggg cggaccagag ccactgatgc aggccaccgc
     2641 ctggctcaac gcctactttc accagcctga ggccatcgag gagttccctg tgccagccct
     2701 gcaccaccca gtgttccagc aggagagctt tacccgccag gtgctgtgga aactgctgaa
     2761 agtggtgaag ttcggagagg tcatcagcta cagccacctg gccgccctgg ccggcaatcc
     2821 cgccgccacc gccgccgtga aaaccgccct gagcggaaat cccgtgccca ttctgatccc
     2881 ctgccaccgg gtggtgcagg gcgacctgga cgtggggggc tacgagggcg ggctcgccgt
     2941 gaaagagtgg ctgctggccc acgagggcca cagactgggc aagcctgggc tgggtcctgc
     3001 aggcggatcc gcgtttaaac tcgaggttaa ttaatgagcg gccgcataga taactgatcc
     3061 agtgtgctgg aattaattcg ctgtctgcga gggccagctg ttggggtgag tactccctct
     3121 caaaagcggg catgacttct gcgctaagat tgtcagtttc caaaaacgag gaggatttga
     3181 tattcacctg gcccgcggtg atgcctttga gggtggccgc gtccatctgg tcagaaaaga
     3241 caatcttttt gttgtcaagc ttgaggtgtg gcaggcttga gatctggcca tacacttgag
     3301 tgacaatgac atccactttg cctttctctc cacaggtgtc cactcccagg tccaactgca
     3361 ggtcgagcat gcatctaggg cggccaattc cgcccctctc cccccccccc nntctccctc
     3421 ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta
     3481 tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc
     3541 tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct
     3601 gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt
     3661 agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa
     3721 gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg
     3781 gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga
     3841 tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac
     3901 atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt
     3961 cctttgaaaa acacgatgat aagcttgcca caacccggga taattcctgc agccaatatg
     4021 ggatcggcca ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     4081 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     4141 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     4201 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4261 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4321 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4381 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4441 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     4501 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     4561 cccgacggcg atgatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     4621 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     4681 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     4741 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     4801 cttcttgacg agttcttctg aggggatcaa ttctctagat aactgatcat aatcagccat
     4861 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     4921 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     4981 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     5041 tgtggtttgt ccaaactcat caatgtatct taacgcgtcg agtgcattct agttgtggtt
     5101 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     5161 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     5221 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     5281 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     5341 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     5401 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     5461 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     5521 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5581 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5641 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5701 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5761 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5821 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     5881 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     5941 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     6001 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     6061 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     6121 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     6181 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     6241 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     6301 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     6361 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     6421 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     6481 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     6541 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     6601 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     6661 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     6721 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     6781 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     6841 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     6901 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     6961 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     7021 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     7081 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     7141 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     7201 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     7261 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     7321 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat
     7381 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
     7441 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
     7501 gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac
     7561 gcgtt
//
