LOCUS       sequence-449146-        8343 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing fusion protein of dPspCas13b and inactive mutant of human
            FTO.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8343)
  AUTHORS   Wei J, Yu X, Yang L, Liu X, Gao B, Huang B, Dou X, Liu J, Zou Z, Cui
            XL, Zhang LS, Zhao X, Liu Q, He PC, Sepich-Poore C, Zhong N, Liu W, 
            Li Y, Kou X, Zhao Y, Wu Y, Cheng X, Chen C, An Y, Dong X, Wang H, 
            Shu Q, Hao Z, Duan T, He YY, Li X, Gao S, Gao Y, He C
  TITLE     FTO mediates LINE1 m(6)A demethylation and chromatin regulation in 
            mESCs and mouse development.
  JOURNAL   Science. 2022 May 27;376(6596):968-973. doi: 
            10.1126/science.abe9582. Epub 2022 May 5.
  PUBMED    35511947
REFERENCE   2  (bases 1 to 8343)
  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: 
            "10.1126/science.abe9582"; journalName: "Science"; date: 
            "2022-05-27- 27"; volume: "376"; issue: "6596"; pages: "968-973"
COMMENT     Sequence Label: dPspCas13b-mutFTO
FEATURES             Location/Qualifiers
     source          1..8343
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        403..782
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        783..986
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     937..957
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          1122..1254
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     CDS             5832..5879
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5886..5909
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(6027..6045)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    6082..6203
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6093..6112)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6147..6166
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             6602..7396
                     /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(6656..6675)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7266..7285
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(7568..8156)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7648..7667)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    join(8272..8343,1..212)
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 tggattaagt gggggtaatg tggcctgtac cctctggttg cataggtatt catacggtta
       61 aaatttatca ggcgcgattg cggcagtttt tcgggtggtt tgttgccatt tttacctgtc
      121 tgctgccgtg atcgcgctga acgcgtttta gcggtgcgta caattaaggg attatggtaa
      181 atccacttac tgtctgccct cgtagccatc gagataaacc gcagtactcc ggccacgatg
      241 cgtccggcgt agaggatcga gatcttcaat attggccatt agccatatta ttcattggtt
      301 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata
      361 tgtacattta tattggctca tgtccaatat gaccgccatg ttggcattga ttattgacta
      421 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      481 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      541 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      601 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      661 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      721 tgaccttacg ggactttcct aattggcagt acatctacgt attagtcatc gctattacca
      781 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
      841 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      901 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
      961 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatcact agaagcttta
     1021 ttgcggtagt ttatcacagt taaattgcta acgcagtcag tgcttctgac acaacagtct
     1081 cgaacttaag ctgcagaagt tggtcgtgag gcactgggca ggtaagtatc aaggttacaa
     1141 gacaggttta aggagaccaa tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt
     1201 tctgataggc acctattggt cttactgaca tccactttgc ctttctctcc acaggtgtcc
     1261 actcccagtt caattacagc tcttaaggca gatcactaga agcttacgct accaccatga
     1321 acatccccgc tctggtggaa aaccagaaga agtactttgg cacctacagc gtgatggcca
     1381 tgctgaacgc tcagaccgtg ctggaccaca tccagaaggt ggccgatatt gagggcgagc
     1441 agaacgagaa caacgagaat ctgtggtttc accccgtgat gagccacctg tacaacgcca
     1501 agaacggcta cgacaagcag cccgagaaaa ccatgttcat catcgagcgg ctgcagagct
     1561 acttcccatt cctgaagatc atggccgaga accagagaga gtacagcaac ggcaagtaca
     1621 agcagaaccg cgtggaagtg aacagcaacg acatcttcga ggtgctgaag cgcgccttcg
     1681 gcgtgctgaa gatgtacagg gacctgacca acgcatacaa gacctacgag gaaaagctga
     1741 acgacggctg cgagttcctg accagcacag agcaacctct gagcggcatg atcaacaact
     1801 actacacagt ggccctgcgg aacatgaacg agagatacgg ctacaagaca gaggacctgg
     1861 ccttcatcca ggacaagcgg ttcaagttcg tgaaggacgc ctacggcaag aaaaagtccc
     1921 aagtgaatac cggattcttc ctgagcctgc aggactacaa cggcgacaca cagaagaagc
     1981 tgcacctgag cggagtggga atcgccctgc tgatctgcct gttcctggac aagcagtaca
     2041 tcaacatctt tctgagcagg ctgcccatct tctccagcta caatgcccag agcgaggaac
     2101 ggcggatcat catcagatcc ttcggtatca acagcatcaa gctgcccaag gaccggatcc
     2161 acagcgagaa gtccaacaag agcgtggcca tggatatgct caacgaagtg aagcggtgcc
     2221 ccgacgagct gttcacaaca ctgtctgccg agaagcagtc ccggttcaga atcatcagcg
     2281 acgaccacaa tgaagtgctg atgaagcgga gcagcgacag attcgtgcct ctgctgctgc
     2341 agtatatcga ttacggcaag ctgttcgacc acatcaggtt ccacgtgaac atgggcaagc
     2401 tgagatacct gctgaaggcc gacaagacct gcatcgacgg ccagaccaga gtcagagtga
     2461 tcgagcagcc cctgaacggc ttcggcagac tggaagaggc cgagacaatg cggaagcaag
     2521 agaacggcac cttcggcaac agcggcatcc ggatcagaga cttcgagaac atgaagcggg
     2581 acgacgccaa tcctgccaac tatccctaca tcgtggacac ctacacacac tacatcctgg
     2641 aaaacaacaa ggtcgagatg tttatcaacg acaaagagga cagcgcccca ctgctgcccg
     2701 tgatcgagga tgatagatac gtggtcaaga caatccccag ctgccggatg agcaccctgg
     2761 aaattccagc catggccttc cacatgtttc tgttcggcag caagaaaacc gagaagctga
     2821 tcgtggacgt gcacaaccgg tacaagagac tgttccaggc catgcagaaa gaagaagtga
     2881 ccgccgagaa tatcgccagc ttcggaatcg ccgagagcga cctgcctcag aagatcctgg
     2941 atctgatcag cggcaatgcc cacggcaagg atgtggacgc cttcatcaga ctgaccgtgg
     3001 acgacatgct gaccgacacc gagcggagaa tcaagagatt caaggacgac cggaagtcca
     3061 ttcggagcgc cgacaacaag atgggaaaga gaggcttcaa gcagatctcc acaggcaagc
     3121 tggccgactt cctggccaag gacatcgtgc tgtttcagcc cagcgtgaac gatggcgaga
     3181 acaagatcac cggcctgaac taccggatca tgcagagcgc cattgccgtg tacgatagcg
     3241 gcgacgatta cgaggccaag cagcagttca agctgatgtt cgagaaggcc cggctgatcg
     3301 gcaagggcac aacagagcct catccatttc tgtacaaggt gttcgcccgc agcatccccg
     3361 ccaatgccgt cgagttctac gagcgctacc tgatcgagcg gaagttctac ctgaccggcc
     3421 tgtccaacga gatcaagaaa ggcaacagag tggatgtgcc cttcatccgg cgggaccaga
     3481 acaagtggaa aacacccgcc atgaagaccc tgggcagaat ctacagcgag gatctgcccg
     3541 tggaactgcc cagacagatg ttcgacaatg agatcaagtc ccacctgaag tccctgccac
     3601 agatggaagg catcgacttc aacaatgcca acgtgaccta tctgatcgcc gagtacatga
     3661 agagagtgct ggacgacgac ttccagacct tctaccagtg gaaccgcaac taccggtaca
     3721 tggacatgct taagggcgag tacgacagaa agggctccct gcagcactgc ttcaccagcg
     3781 tggaagagag agaaggcctc tggaaagagc gggcctccag aacagagcgg tacagaaagc
     3841 aggccagcaa caagatccgc agcaaccggc agatgagaaa cgccagcagc gaagagatcg
     3901 agacaatcct ggataagcgg ctgagcaaca gccggaacga gtaccagaaa agcgagaaag
     3961 tgatccggcg ctacagagtg caggatgccc tgctgtttct gctggccaaa aagaccctga
     4021 ccgaactggc cgatttcgac ggcgagaggt tcaaactgaa agaaatcatg cccgacgccg
     4081 agaagggaat cctgagcgag atcatgccca tgagcttcac cttcgagaaa ggcggcaaga
     4141 agtacaccat caccagcgag ggcatgaagc tgaagaacta cggcgacttc tttgtgctgg
     4201 ctagcgacaa gaggatcggc aacctgctgg aactcgtggg cagcgacatc gtgtccaaag
     4261 aggatggcgg tagtggagga agcgggggga gcggagggag tggtggatcc ggcggctcca
     4321 agcgcacccc gactgccgag gaacgagagc gcgaagctaa gaaactgagg cttcttgaag
     4381 agcttgaaga cacttggctc ccttatctga cccccaaaga tgatgaattc tatcagcagt
     4441 ggcagctgaa atatcctaaa ctaattctcc gagaagccag cagtgtatct gaggagctcc
     4501 ataaagaggt tcaagaagcc tttctcacac tgcacaagca tggctgctta tttcgggacc
     4561 tggttaggat ccaaggcaaa gatctgctca ctccggtatc tcgcatcctc attggtaatc
     4621 caggctgcac ctacaagtac ctgaacacca ggctctttac ggtcccctgg ccagtgaaag
     4681 ggtctaatat aaaacacacc gaggctgaaa tagccgctgc ttgtgagacc ttcctcaagc
     4741 tcaatgacta cctgcagata gaaaccatcc aggctttgga agaacttgct gccaaagaga
     4801 aggctaatga ggatgctgtg ccattgtgta tgtctgcaga tttccccagg gttgggatgg
     4861 gttcatccta caacggacaa gatgaagtgg acattaagag cagagcagca tacaacgtaa
     4921 ctttgctgaa tttcatggat cctcagaaaa tgccatacct gaaagaggaa ccttattttg
     4981 gcatggggaa aatggcagtg agctgggctc atgctgaaaa tctggtggac aggtcagcgg
     5041 tggcagtgta cagttatagc tgtgaaggcc ctgaagagga aagtgaggat gactctcatc
     5101 tcgaaggcag ggatcctgat atttggcatg ttggttttaa gatctcatgg gacatagaga
     5161 cacctggttt ggcgataccc cttcaccaag gagactgcta tttcatgctt gatgatctca
     5221 atgccaccca ccaacactgt gttttggccg gttcacaacc tcggtttagt tccacccacc
     5281 gagtggcaga gtgctcaaca ggaaccttgg attatatttt acaacgctgt cagttggctc
     5341 tgcagaatgt ctgtgacgat gtggacaatg atgatgtctc tttgaaatcc tttgagcctg
     5401 cagttttgaa acaaggagaa gaaattcata atgaggtcga gtttgagtgg ctgaggcagt
     5461 tttggtttca aggcaatcga tacagaaagt gcactgactg gtggtgtcaa cccatggctc
     5521 aactggaagc actgtggaag aagatggagg gtgtgacaaa tgctgtgctt catgaagtta
     5581 aaagagaggg gctccccgtg gaacaaagga atgaaatctt gactgccatc cttgcctcgc
     5641 tcactgcacg ccagaacctg aggagagaat ggcatgccag gtgccagtca cgaattgccc
     5701 gaacattacc tgctgatcag aagccagaat gtcggccata ctgggaaaag gatgatgctt
     5761 cgatgcctct gccgtttgac ctcacagaca tcgtttcaga actcagaggt cagcttctgg
     5821 aagcaaaacc caaaaggccg gcggccacga aaaaggccgg ccaggcaaaa aagaaaaagg
     5881 gatccgatta caaggacgac gatgacaagt agtgtttaaa cgaattcggg ctcggtacac
     5941 ggggatcctc tagagtcgac ctgcaggcat gcaagctgat ccggctgcta acaaagcccg
     6001 aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc
     6061 ggccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     6121 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     6181 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     6241 agggggagat gtgggaggtt tttttaagca agtaaaacct ctacaaatgt ggtaaaatcg
     6301 aattctaatg gatcctcttt gcgcttgcgt tttcccttgt ccagatagcc cagtagctga
     6361 cattcatccg gggtcagcac cgtttctgcg gactggcttt ctacgtgttc cgcttccttt
     6421 agcagccctt gcgccctgag tgcttgcggc agcgtgagct tcaaaagaat tgccagctgg
     6481 ggcgccctct ggtaaggttg ggaagccctg caaagtaaac tggatggctt tcttgccgcc
     6541 aaggatctga tggcgcaggg gatcaagatc tgatcaagag acaggatgac ggtcgtttcg
     6601 catgcttgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     6661 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     6721 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     6781 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     6841 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     6901 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     6961 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     7021 catcgagcga gcacgcactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     7081 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gtatgccgga
     7141 tggtgaggat ctcgtcgtga ctcatggcga tgcctgcttg ccgaatatca tggtggaaaa
     7201 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     7261 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     7321 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     7381 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     7441 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     7501 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     7561 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     7621 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     7681 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     7741 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     7801 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     7861 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     7921 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     7981 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     8041 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     8101 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tttcaagaag
     8161 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     8221 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttatag tccggaaata
     8281 caggaacgca cgctggatgg cccttcgctg ggatggtgaa accatgaaaa atggcagctt
     8341 cag
//
