LOCUS       sequence-227043-        7875 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cmv d0 dPspCas13b-GGS-NYTHDF1 fusion protein (full N-terminal domain
            of YTHDF1, C-terminal YTH domain omitted).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7875)
  AUTHORS   Rauch S, He C, Dickinson BC
  TITLE     Targeted m(6)A Reader Proteins To Study Epitranscriptomic Regulation
            of Single RNAs.
  JOURNAL   J Am Chem Soc. 2018 Sep 26;140(38):11974-11981. doi: 
            10.1021/jacs.8b05012. Epub 2018 Sep 18.
  PUBMED    30183280
REFERENCE   2  (bases 1 to 7875)
  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"; doi: 
            "10.1021/jacs.8b05012"; journalName: "J Am Chem Soc"; date: 
            "2018-09-26- 26"; volume: "140"; issue: "38"; pages: "11974-11981"
COMMENT     Sequence Label: Cmv d0 dPspCas13b-GGS-NYTHDF1
FEATURES             Location/Qualifiers
     source          1..7875
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             5153..5176
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(5294..5312)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    5349..5470
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5360..5379)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5414..5433
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             5869..6663
                     /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="MLEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5923..5942)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6533..6552
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(6835..7423)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6915..6934)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7539..7822
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggcagatca ctagaagctt acgctaccac catgaacatc cccgctctgg tggaaaacca
     1081 gaagaagtac tttggcacct acagcgtgat ggccatgctg aacgctcaga ccgtgctgga
     1141 ccacatccag aaggtggccg atattgaggg cgagcagaac gagaacaacg agaatctgtg
     1201 gtttcacccc gtgatgagcc acctgtacaa cgccaagaac ggctacgaca agcagcccga
     1261 gaaaaccatg ttcatcatcg agcggctgca gagctacttc ccattcctga agatcatggc
     1321 cgagaaccag agagagtaca gcaacggcaa gtacaagcag aaccgcgtgg aagtgaacag
     1381 caacgacatc ttcgaggtgc tgaagcgcgc cttcggcgtg ctgaagatgt acagggacct
     1441 gaccaacgca tacaagacct acgaggaaaa gctgaacgac ggctgcgagt tcctgaccag
     1501 cacagagcaa cctctgagcg gcatgatcaa caactactac acagtggccc tgcggaacat
     1561 gaacgagaga tacggctaca agacagagga cctggccttc atccaggaca agcggttcaa
     1621 gttcgtgaag gacgcctacg gcaagaaaaa gtcccaagtg aataccggat tcttcctgag
     1681 cctgcaggac tacaacggcg acacacagaa gaagctgcac ctgagcggag tgggaatcgc
     1741 cctgctgatc tgcctgttcc tggacaagca gtacatcaac atctttctga gcaggctgcc
     1801 catcttctcc agctacaatg cccagagcga ggaacggcgg atcatcatca gatccttcgg
     1861 catcaacagc atcaagctgc ccaaggaccg gatccacagc gagaagtcca acaagagcgt
     1921 ggccatggat atgctcaacg aagtgaagcg gtgccccgac gagctgttca caacactgtc
     1981 tgccgagaag cagtcccggt tcagaatcat cagcgacgac cacaatgaag tgctgatgaa
     2041 gcggagcagc gacagattcg tgcctctgct gctgcagtat atcgattacg gcaagctgtt
     2101 cgaccacatc aggttccacg tgaacatggg caagctgaga tacctgctga aggccgacaa
     2161 gacctgcatc gacggccaga ccagagtcag agtgatcgag cagcccctga acggcttcgg
     2221 cagactggaa gaggccgaga caatgcggaa gcaagagaac ggcaccttcg gcaacagcgg
     2281 catccggatc agagacttcg agaacatgaa gcgggacgac gccaatcctg ccaactatcc
     2341 ctacatcgtg gacacctaca cacactacat cctggaaaac aacaaggtcg agatgtttat
     2401 caacgacaaa gaggacagcg ccccactgct gcccgtgatc gaggatgata gatacgtggt
     2461 caagacaatc cccagctgcc ggatgagcac cctggaaatt ccagccatgg ccttccacat
     2521 gtttctgttc ggcagcaaga aaaccgagaa gctgatcgtg gacgtgcaca accggtacaa
     2581 gagactgttc caggccatgc agaaagaaga agtgaccgcc gagaatatcg ccagcttcgg
     2641 aatcgccgag agcgacctgc ctcagaagat cctggatctg atcagcggca atgcccacgg
     2701 caaggatgtg gacgccttca tcagactgac cgtggacgac atgctgaccg acaccgagcg
     2761 gagaatcaag agattcaagg acgaccggaa gtccattcgg agcgccgaca acaagatggg
     2821 aaagagaggc ttcaagcaga tctccacagg caagctggcc gacttcctgg ccaaggacat
     2881 cgtgctgttt cagcccagcg tgaacgatgg cgagaacaag atcaccggcc tgaactaccg
     2941 gatcatgcag agcgccattg ccgtgtacga tagcggcgac gattacgagg ccaagcagca
     3001 gttcaagctg atgttcgaga aggcccggct gatcggcaag ggcacaacag agcctcatcc
     3061 atttctgtac aaggtgttcg cccgcagcat ccccgccaat gccgtcgagt tctacgagcg
     3121 ctacctgatc gagcggaagt tctacctgac cggcctgtcc aacgagatca agaaaggcaa
     3181 cagagtggat gtgcccttca tccggcggga ccagaacaag tggaaaacac ccgccatgaa
     3241 gaccctgggc agaatctaca gcgaggatct gcccgtggaa ctgcccagac agatgttcga
     3301 caatgagatc aagtcccacc tgaagtccct gccacagatg gaaggcatcg acttcaacaa
     3361 tgccaacgtg acctatctga tcgccgagta catgaagaga gtgctggacg acgacttcca
     3421 gaccttctac cagtggaacc gcaactaccg gtacatggac atgcttaagg gcgagtacga
     3481 cagaaagggc tccctgcagc actgcttcac cagcgtggaa gagagagaag gcctctggaa
     3541 agagcgggcc tccagaacag agcggtacag aaagcaggcc agcaacaaga tccgcagcaa
     3601 ccggcagatg agaaacgcca gcagcgaaga gatcgagaca atcctggata agcggctgag
     3661 caacagccgg aacgagtacc agaaaagcga gaaagtgatc cggcgctaca gagtgcagga
     3721 tgccctgctg tttctgctgg ccaaaaagac cctgaccgaa ctggccgatt tcgacggcga
     3781 gaggttcaaa ctgaaagaaa tcatgcccga cgccgagaag ggaatcctga gcgagatcat
     3841 gcccatgagc ttcaccttcg agaaaggcgg caagaagtac accatcacca gcgagggcat
     3901 gaagctgaag aactacggcg acttctttgt gctggctagc gacaagagga tcggcaacct
     3961 gctggaactc gtgggcagcg acatcgtgtc caaagaggat ggcggtagtg gaggaagcgg
     4021 ggggagcgga gggagtggtg gatccggcgg ctccatgtcg gccaccagcg tggacaccca
     4081 gagaacaaaa ggacaagata ataaagtaca aaatggttcg ttacatcaga aggatacagt
     4141 tcatgacaat gactttgagc cctaccttac tggacagtca aatcagagta acagttaccc
     4201 ctcaatgagc gacccctacc tgtccagcta ttacccgccg tccattggat ttccttactc
     4261 cctcaatgag gctccgtggt ctactgcagg ggaccctccg attccatacc tcaccaccta
     4321 cggacagctc agtaacggag accatcattt tatgcacgat gctgtttttg ggcagcctgg
     4381 gggcctgggg aacaacatct atcagcacag gttcaatttt ttccctgaaa accctgcgtt
     4441 ctcagcatgg gggacaagtg ggtctcaagg tcagcagacc cagagctccg cgtatgggag
     4501 cagctacacc taccccccga gctccctggg tggcacggtg gttgatgggc agccaggctt
     4561 tcacagcgac accctcagca aggcccccgg gatgaacagc ctggagcagg gcatggttgg
     4621 cctgaagatt ggggacgtca gctcctccgc cgtcaagacg gtgggctctg tcgtcagcag
     4681 cgtggcactg actggtgtcc tttctggcaa cggtgggaca aatgtgaaca tgccagtttc
     4741 aaagccgacc tcgtgggctg ccattgccag caagcctgca aaaccacagc ctaaaatgaa
     4801 aacaaagagc gggcctgtca tggggggtgg gctgccccct ccacccataa agcataacat
     4861 ggacattggc acctgggata acaaggggcc tgtgccgaag gccccagtcc cccagcaggc
     4921 accctctcca caggctgccc cacagcccca gcaggtggct cagcctctcc cagcacagcc
     4981 cccagctttg gctcaaccgc agtatcagag ccctcagcag ccaccccaga cccgctgggt
     5041 tgccccacgc aacagaaacg cggcgtttgg gcagagcgga ggggctggca gcgatagcaa
     5101 ctctcctgga aacgtccagc ctaattctgc ccccagcgtc gaatccggat ccgattacaa
     5161 ggacgacgat gacaagtagt gtttaaacga attcgggctc ggtacacggg gatcctctag
     5221 agtcgacctg caggcatgca agctgatccg gctgctaaca aagcccgaaa ggaagctgag
     5281 ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcggc cgcttcgagc
     5341 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     5401 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     5461 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggagatgtg
     5521 ggaggttttt ttaagcaagt aaaacctcta caaatgtggt aaaatcgaat tctaatggat
     5581 cctctttgcg cttgcgtttt cccttgtcca gatagcccag tagctgacat tcatccgggg
     5641 tcagcaccgt ttctgcggac tggctttcta cgtgttccgc ttcctttagc agcccttgcg
     5701 ccctgagtgc ttgcggcagc gtgagcttca aaagaattgc cagctggggc gccctctggt
     5761 aaggttggga agccctgcaa agtaaactgg atggctttct tgccgccaag gatctgatgg
     5821 cgcaggggat caagatctga tcaagagaca ggatgacggt cgtttcgcat gcttgaacaa
     5881 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     5941 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     6001 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     6061 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     6121 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     6181 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     6241 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     6301 cgcactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     6361 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgta tgccggatgg tgaggatctc
     6421 gtcgtgactc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     6481 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     6541 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     6601 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     6661 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaaccgg tatcagctca
     6721 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     6781 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     6841 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     6901 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6961 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     7021 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     7081 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     7141 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     7201 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     7261 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     7321 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     7381 tgtttgcaag cagcagatta cgcgcagaaa aaaaggattt caagaagatc ctttgatctt
     7441 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     7501 attatcaaaa aggatcttca cctagatcct tttatagtcc ggaaatacag gaacgcacgc
     7561 tggatggccc ttcgctggga tggtgaaacc atgaaaaatg gcagcttcag tggattaagt
     7621 gggggtaatg tggcctgtac cctctggttg cataggtatt catacggtta aaatttatca
     7681 ggcgcgattg cggcagtttt tcgggtggtt tgttgccatt tttacctgtc tgctgccgtg
     7741 atcgcgctga acgcgtttta gcggtgcgta caattaaggg attatggtaa atccacttac
     7801 tgtctgccct cgtagccatc gagataaacc gcagtactcc ggccacgatg cgtccggcgt
     7861 agaggatcga gatct
//
