LOCUS       sequence-442770-        9681 bp    DNA     circular SYN 17-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 9681)
  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 9681)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 17, 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..9681
                     /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             7170..7217
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7224..7247
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(7365..7383)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    7420..7541
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7431..7450)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7485..7504
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             7940..8734
                     /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(7994..8013)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8604..8623
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(8906..9494)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8986..9005)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    join(9610..9681,1..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 ctgatgaatc ccctaatgat tttggtaaaa atcattaagt
     5341 taaggtggat acacatcttg tcatatgatc aaatggtttc gcgaaaaatc aataatcaga
     5401 caacaagatg tgcgaactcg atattttaca cgactctctt taccaattct gccccgaatt
     5461 acacttaaaa cgactcaaca gcttaacgtt ggcttgccac gcattacttg actgtaaaac
     5521 tctcactctt accgaacttg gccgtaacct gccaaccaaa gcgagaacaa aacataacat
     5581 caaacgaatc gaccgattgt taggtaatcg tcacctccac aaagagcgac tcgctgtata
     5641 ccgttggcat gctagcttta tctgttcggg caatacgatg cccattgtac ttgttgactg
     5701 gtctgatatt cgtgagcaaa aacgacttat ggtattgcga gcttcagtcg cactacacgg
     5761 tcgttctgtt actctttatg agaaagcgtt cccgctttca gagcaatgtt caaagaaagc
     5821 tcatgaccaa tttctagccg accttgcgag cattctaccg agtaacacca caccgctcat
     5881 tgtcagtgat gctggcttta aagtgccatg gtataaatcc gttgagaagc tgggttggta
     5941 ctggttaagt cgagtaagag gaaaagtaca atatgcagac ctaggagcgg aaaactggaa
     6001 acctatcagc aacttacatg atatgtcatc tagtcactca aagactttag gctataagag
     6061 gctgactaaa agcaatccaa tctcatgcca aattctattg tataaatctc gctctaaagg
     6121 ccgaaaaaat cagcgctcga cacggactca ttgtcaccac ccgtcaccta aaatctactc
     6181 agcgtcggca aaggagccat gggttctagc aactaactta cctgttgaaa ttcgaacacc
     6241 caaacaactt gttaatatct attcgaagcg aatgcagatt gaagaaacct tccgagactt
     6301 gaaaagtcct gcctacggac taggcctacg ccatagccga acgagcagct cagagcgttt
     6361 tgatatcatg ctgctaatcg ccctgatgct tcaactaaca tgttggcttg cgggcgttca
     6421 tgctcagaaa caaggttggg acaagcactt ccaggctaac acagtcagaa atcgaaacgt
     6481 actctcaaca gttcgcttag gcatggaagt tttgcggcat tctggctaca caataacaag
     6541 ggaagactta ctcgtggctg caaccctact agctcaaaat ttattcacac atggttacgc
     6601 tttggggaaa ttatgagggg atctctcagt gctcaacagg aaccttggat tatattttac
     6661 aacgctgtca gttggctctg cagaatgtct gtgacgatgt ggacaatgat gatgtctctt
     6721 tgaaatcctt tgagcctgca gttttgaaac aaggagaaga aattcataat gaggtcgagt
     6781 ttgagtggct gaggcagttt tggtttcaag gcaatcgata cagaaagtgc actgactggt
     6841 ggtgtcaacc catggctcaa ctggaagcac tgtggaagaa gatggagggt gtgacaaatg
     6901 ctgtgcttca tgaagttaaa agagaggggc tccccgtgga acaaaggaat gaaatcttga
     6961 ctgccatcct tgcctcgctc actgcacgcc agaacctgag gagagaatgg catgccaggt
     7021 gccagtcacg aattgcccga acattacctg ctgatcagaa gccagaatgt cggccatact
     7081 gggaaaagga tgatgcttcg atgcctctgc cgtttgacct cacagacatc gtttcagaac
     7141 tcagaggtca gcttctggaa gcaaaaccca aaaggccggc ggccacgaaa aaggccggcc
     7201 aggcaaaaaa gaaaaaggga tccgattaca aggacgacga tgacaagtag tgtttaaacg
     7261 aattcgggct cggtacacgg ggatcctcta gagtcgacct gcaggcatgc aagctgatcc
     7321 ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact
     7381 agcataaccc cttggggcgg ccgcttcgag cagacatgat aagatacatt gatgagtttg
     7441 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     7501 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     7561 attttatgtt tcaggttcag ggggagatgt gggaggtttt tttaagcaag taaaacctct
     7621 acaaatgtgg taaaatcgaa ttctaatgga tcctctttgc gcttgcgttt tcccttgtcc
     7681 agatagccca gtagctgaca ttcatccggg gtcagcaccg tttctgcgga ctggctttct
     7741 acgtgttccg cttcctttag cagcccttgc gccctgagtg cttgcggcag cgtgagcttc
     7801 aaaagaattg ccagctgggg cgccctctgg taaggttggg aagccctgca aagtaaactg
     7861 gatggctttc ttgccgccaa ggatctgatg gcgcagggga tcaagatctg atcaagagac
     7921 aggatgacgg tcgtttcgca tgcttgaaca agatggattg cacgcaggtt ctccggccgc
     7981 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     8041 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     8101 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     8161 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     8221 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     8281 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     8341 ccaccaagcg aaacatcgca tcgagcgagc acgcactcgg atggaagccg gtcttgtcga
     8401 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     8461 caaggcgcgt atgccggatg gtgaggatct cgtcgtgact catggcgatg cctgcttgcc
     8521 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     8581 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     8641 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     8701 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     8761 gaccaagcga cgcccaaccg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     8821 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     8881 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     8941 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     9001 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     9061 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     9121 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     9181 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     9241 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     9301 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     9361 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     9421 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     9481 aaaaaggatt tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     9541 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     9601 ttttatagtc cggaaataca ggaacgcacg ctggatggcc cttcgctggg atggtgaaac
     9661 catgaaaaat ggcagcttca g
//
