LOCUS       sequence-231780-        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"
     intron          143..275
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     258..277
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             4439..4462
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(4580..4598)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    4635..4756
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4646..4665)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4700..4719
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             5155..5949
                     /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(5209..5228)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5819..5838
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(6121..6709)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6201..6220)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6825..7108
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     enhancer        7299..7678
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(7679..7875,1..7)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7833..7853
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 cagagctggt ttagtgaacc gtcagatcac tagaagcttt attgcggtag tttatcacag
       61 ttaaattgct aacgcagtca gtgcttctga cacaacagtc tcgaacttaa gctgcagaag
      121 ttggtcgtga ggcactgggc aggtaagtat caaggttaca agacaggttt aaggagacca
      181 atagaaactg ggcttgtcga gacagagaag actcttgcgt ttctgatagg cacctattgg
      241 tcttactgac atccactttg cctttctctc cacaggtgtc cactcccagt tcaattacag
      301 ctcttaaggc agatcactag aagcttacgc taccaccatg aacatccccg ctctggtgga
      361 aaaccagaag aagtactttg gcacctacag cgtgatggcc atgctgaacg ctcagaccgt
      421 gctggaccac atccagaagg tggccgatat tgagggcgag cagaacgaga acaacgagaa
      481 tctgtggttt caccccgtga tgagccacct gtacaacgcc aagaacggct acgacaagca
      541 gcccgagaaa accatgttca tcatcgagcg gctgcagagc tacttcccat tcctgaagat
      601 catggccgag aaccagagag agtacagcaa cggcaagtac aagcagaacc gcgtggaagt
      661 gaacagcaac gacatcttcg aggtgctgaa gcgcgccttc ggcgtgctga agatgtacag
      721 ggacctgacc aacgcataca agacctacga ggaaaagctg aacgacggct gcgagttcct
      781 gaccagcaca gagcaacctc tgagcggcat gatcaacaac tactacacag tggccctgcg
      841 gaacatgaac gagagatacg gctacaagac agaggacctg gccttcatcc aggacaagcg
      901 gttcaagttc gtgaaggacg cctacggcaa gaaaaagtcc caagtgaata ccggattctt
      961 cctgagcctg caggactaca acggcgacac acagaagaag ctgcacctga gcggagtggg
     1021 aatcgccctg ctgatctgcc tgttcctgga caagcagtac atcaacatct ttctgagcag
     1081 gctgcccatc ttctccagct acaatgccca gagcgaggaa cggcggatca tcatcagatc
     1141 cttcggtatc aacagcatca agctgcccaa ggaccggatc cacagcgaga agtccaacaa
     1201 gagcgtggcc atggatatgc tcaacgaagt gaagcggtgc cccgacgagc tgttcacaac
     1261 actgtctgcc gagaagcagt cccggttcag aatcatcagc gacgaccaca atgaagtgct
     1321 gatgaagcgg agcagcgaca gattcgtgcc tctgctgctg cagtatatcg attacggcaa
     1381 gctgttcgac cacatcaggt tccacgtgaa catgggcaag ctgagatacc tgctgaaggc
     1441 cgacaagacc tgcatcgacg gccagaccag agtcagagtg atcgagcagc ccctgaacgg
     1501 cttcggcaga ctggaagagg ccgagacaat gcggaagcaa gagaacggca ccttcggcaa
     1561 cagcggcatc cggatcagag acttcgagaa catgaagcgg gacgacgcca atcctgccaa
     1621 ctatccctac atcgtggaca cctacacaca ctacatcctg gaaaacaaca aggtcgagat
     1681 gtttatcaac gacaaagagg acagcgcccc actgctgccc gtgatcgagg atgatagata
     1741 cgtggtcaag acaatcccca gctgccggat gagcaccctg gaaattccag ccatggcctt
     1801 ccacatgttt ctgttcggca gcaagaaaac cgagaagctg atcgtggacg tgcacaaccg
     1861 gtacaagaga ctgttccagg ccatgcagaa agaagaagtg accgccgaga atatcgccag
     1921 cttcggaatc gccgagagcg acctgcctca gaagatcctg gatctgatca gcggcaatgc
     1981 ccacggcaag gatgtggacg ccttcatcag actgaccgtg gacgacatgc tgaccgacac
     2041 cgagcggaga atcaagagat tcaaggacga ccggaagtcc attcggagcg ccgacaacaa
     2101 gatgggaaag agaggcttca agcagatctc cacaggcaag ctggccgact tcctggccaa
     2161 ggacatcgtg ctgtttcagc ccagcgtgaa cgatggcgag aacaagatca ccggcctgaa
     2221 ctaccggatc atgcagagcg ccattgccgt gtacgatagc ggcgacgatt acgaggccaa
     2281 gcagcagttc aagctgatgt tcgagaaggc ccggctgatc ggcaagggca caacagagcc
     2341 tcatccattt ctgtacaagg tgttcgcccg cagcatcccc gccaatgccg tcgagttcta
     2401 cgagcgctac ctgatcgagc ggaagttcta cctgaccggc ctgtccaacg agatcaagaa
     2461 aggcaacaga gtggatgtgc ccttcatccg gcgggaccag aacaagtgga aaacacccgc
     2521 catgaagacc ctgggcagaa tctacagcga ggatctgccc gtggaactgc ccagacagat
     2581 gttcgacaat gagatcaagt cccacctgaa gtccctgcca cagatggaag gcatcgactt
     2641 caacaatgcc aacgtgacct atctgatcgc cgagtacatg aagagagtgc tggacgacga
     2701 cttccagacc ttctaccagt ggaaccgcaa ctaccggtac atggacatgc ttaagggcga
     2761 gtacgacaga aagggctccc tgcagcactg cttcaccagc gtggaagaga gagaaggcct
     2821 ctggaaagag cgggcctcca gaacagagcg gtacagaaag caggccagca acaagatccg
     2881 cagcaaccgg cagatgagaa acgccagcag cgaagagatc gagacaatcc tggataagcg
     2941 gctgagcaac agccggaacg agtaccagaa aagcgagaaa gtgatccggc gctacagagt
     3001 gcaggatgcc ctgctgtttc tgctggccaa aaagaccctg accgaactgg ccgatttcga
     3061 cggcgagagg ttcaaactga aagaaatcat gcccgacgcc gagaagggaa tcctgagcga
     3121 gatcatgccc atgagcttca ccttcgagaa aggcggcaag aagtacacca tcaccagcga
     3181 gggcatgaag ctgaagaact acggcgactt ctttgtgctg gctagcgaca agaggatcgg
     3241 caacctgctg gaactcgtgg gcagcgacat cgtgtccaaa gaggatggcg gtagtggagg
     3301 aagcgggggg agcggaggga gtggtggatc cggcggctcc atgtcggcca ccagcgtgga
     3361 cacccagaga acaaaaggac aagataataa agtacaaaat ggttcgttac atcagaagga
     3421 tacagttcat gacaatgact ttgagcccta ccttactgga cagtcaaatc agagtaacag
     3481 ttacccctca atgagcgacc cctacctgtc cagctattac ccgccgtcca ttggatttcc
     3541 ttactccctc aatgaggctc cgtggtctac tgcaggggac cctccgattc catacctcac
     3601 cacctacgga cagctcagta acggagacca tcattttatg cacgatgctg tttttgggca
     3661 gcctgggggc ctggggaaca acatctatca gcacaggttc aattttttcc ctgaaaaccc
     3721 tgcgttctca gcatggggga caagtgggtc tcaaggtcag cagacccaga gctccgcgta
     3781 tgggagcagc tacacctacc ccccgagctc cctgggtggc acggtggttg atgggcagcc
     3841 aggctttcac agcgacaccc tcagcaaggc ccccgggatg aacagcctgg agcagggcat
     3901 ggttggcctg aagattgggg acgtcagctc ctccgccgtc aagacggtgg gctctgtcgt
     3961 cagcagcgtg gcactgactg gtgtcctttc tggcaacggt gggacaaatg tgaacatgcc
     4021 agtttcaaag ccgacctcgt gggctgccat tgccagcaag cctgcaaaac cacagcctaa
     4081 aatgaaaaca aagagcgggc ctgtcatggg gggtgggctg ccccctccac ccataaagca
     4141 taacatggac attggcacct gggataacaa ggggcctgtg ccgaaggccc cagtccccca
     4201 gcaggcaccc tctccacagg ctgccccaca gccccagcag gtggctcagc ctctcccagc
     4261 acagccccca gctttggctc aaccgcagta tcagagccct cagcagccac cccagacccg
     4321 ctgggttgcc ccacgcaaca gaaacgcggc gtttgggcag agcggagggg ctggcagcga
     4381 tagcaactct cctggaaacg tccagcctaa ttctgccccc agcgtcgaat ccggatccga
     4441 ttacaaggac gacgatgaca agtagtgttt aaacgaattc gggctcggta cacggggatc
     4501 ctctagagtc gacctgcagg catgcaagct gatccggctg ctaacaaagc ccgaaaggaa
     4561 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcggccgct
     4621 tcgagcagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     4681 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     4741 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     4801 gatgtgggag gtttttttaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgaattcta
     4861 atggatcctc tttgcgcttg cgttttccct tgtccagata gcccagtagc tgacattcat
     4921 ccggggtcag caccgtttct gcggactggc tttctacgtg ttccgcttcc tttagcagcc
     4981 cttgcgccct gagtgcttgc ggcagcgtga gcttcaaaag aattgccagc tggggcgccc
     5041 tctggtaagg ttgggaagcc ctgcaaagta aactggatgg ctttcttgcc gccaaggatc
     5101 tgatggcgca ggggatcaag atctgatcaa gagacaggat gacggtcgtt tcgcatgctt
     5161 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     5221 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     5281 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     5341 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     5401 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     5461 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     5521 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     5581 cgagcacgca ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     5641 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgtatgcc ggatggtgag
     5701 gatctcgtcg tgactcatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     5761 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     5821 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     5881 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     5941 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aaccggtatc
     6001 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     6061 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     6121 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     6181 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     6241 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     6301 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     6361 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     6421 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     6481 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     6541 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     6601 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     6661 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatttcaag aagatccttt
     6721 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     6781 catgagatta tcaaaaagga tcttcaccta gatcctttta tagtccggaa atacaggaac
     6841 gcacgctgga tggcccttcg ctgggatggt gaaaccatga aaaatggcag cttcagtgga
     6901 ttaagtgggg gtaatgtggc ctgtaccctc tggttgcata ggtattcata cggttaaaat
     6961 ttatcaggcg cgattgcggc agtttttcgg gtggtttgtt gccattttta cctgtctgct
     7021 gccgtgatcg cgctgaacgc gttttagcgg tgcgtacaat taagggatta tggtaaatcc
     7081 acttactgtc tgccctcgta gccatcgaga taaaccgcag tactccggcc acgatgcgtc
     7141 cggcgtagag gatcgagatc ttcaatattg gccattagcc atattattca ttggttatat
     7201 agcataaatc aatattggct attggccatt gcatacgttg tatctatatc ataatatgta
     7261 catttatatt ggctcatgtc caatatgacc gccatgttgg cattgattat tgactagtta
     7321 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
     7381 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
     7441 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
     7501 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtcc
     7561 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
     7621 cttacgggac tttcctaatt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
     7681 gatgcggttt tggcagtaca ccaatgggcg tggatagcgg tttgactcac ggggatttcc
     7741 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     7801 tccaaaatgt cgtaataacc ccgccccgtt gacgcaaatg ggcggtaggc gtgtacggtg
     7861 ggaggtctat ataag
//
