LOCUS       sequence-308877-        6955 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express human YTHDF2(1-381) 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 6955)
  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 6955)
  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-N
FEATURES             Location/Qualifiers
     source          1..6955
                     /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             1856..2401
                     /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          2512..2741
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     2724..2743
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    2799..3372
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2966..2983)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3193..3212
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3408..4211
                     /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(3471..3490)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4081..4100
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4341..4462
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4378..4397)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4564..4580)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4564..4580)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4577..4599)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4588..4604
                     /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(4612..4618,4619..4636,4637..4642))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4657..4678
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4795..4812)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4966..5554)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5046..5065)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5725..6516,6517..6585))
                     /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     6348..6367
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6586..6690)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6765..6784)
                     /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 tctactgaat tcaccatgga
     1861 caaagactgc gaaatgaagc gcaccaccct ggatagccct ctgggcaagc tggaactgtc
     1921 tgggtgcgaa cagggcctgc accgtatcat cttcctgggc aaaggaacat ctgccgccga
     1981 cgccgtggaa gtgcctgccc cagccgccgt gctgggcgga ccagagccac tgatgcaggc
     2041 caccgcctgg ctcaacgcct actttcacca gcctgaggcc atcgaggagt tccctgtgcc
     2101 agccctgcac cacccagtgt tccagcagga gagctttacc cgccaggtgc tgtggaaact
     2161 gctgaaagtg gtgaagttcg gagaggtcat cagctacagc cacctggccg ccctggccgg
     2221 caatcccgcc gccaccgccg ccgtgaaaac cgccctgagc ggaaatcccg tgcccattct
     2281 gatcccctgc caccgggtgg tgcagggcga cctggacgtg gggggctacg agggcgggct
     2341 cgccgtgaaa gagtggctgc tggcccacga gggccacaga ctgggcaagc ctgggctggg
     2401 tcctgcaggc ggatccgcgt ttaaactcga ggttaattaa tgagcggccg catagataac
     2461 tgatccagtg tgctggaatt aattcgctgt ctgcgagggc cagctgttgg ggtgagtact
     2521 ccctctcaaa agcgggcatg acttctgcgc taagattgtc agtttccaaa aacgaggagg
     2581 atttgatatt cacctggccc gcggtgatgc ctttgagggt ggccgcgtcc atctggtcag
     2641 aaaagacaat ctttttgttg tcaagcttga ggtgtggcag gcttgagatc tggccataca
     2701 cttgagtgac aatgacatcc actttgcctt tctctccaca ggtgtccact cccaggtcca
     2761 actgcaggtc gagcatgcat ctagggcggc caattccgcc cctctcccyc ccccccccct
     2821 aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt
     2881 tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg
     2941 acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc
     3001 gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt
     3061 tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta
     3121 taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg
     3181 gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag
     3241 gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag
     3301 tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa
     3361 acacgatgat aagcttgcca caacccggga taattcctgc agccaatatg ggatcggcca
     3421 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     3481 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     3541 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     3601 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     3661 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     3721 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     3781 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     3841 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     3901 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     3961 atgatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     4021 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     4081 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     4141 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     4201 agttcttctg aggggatcaa ttctctagat aactgatcat aatcagccat accacatttg
     4261 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     4321 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     4381 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     4441 ccaaactcat caatgtatct taacgcgtcg agtgcattct agttgtggtt tgtccaaact
     4501 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     4561 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     4621 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     4681 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     4741 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     4801 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     4861 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     4921 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     4981 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5041 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5101 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5161 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5221 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5281 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5341 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5401 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5461 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5521 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     5581 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     5641 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     5701 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     5761 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     5821 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     5881 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     5941 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6001 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     6061 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6121 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6181 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6241 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6301 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     6361 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     6421 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     6481 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     6541 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     6601 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     6661 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     6721 cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat
     6781 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
     6841 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
     6901 cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgtt
//
