LOCUS       sequence-385975-        6692 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Right-side TALE-DddAtox Nterm half ND4-FusXTBE in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6692)
  AUTHORS   Sabharwal A, Kar B, Restrepo-Castillo S, Holmberg SR, Mathew ND, 
            Kendall BL, Cotter RP, WareJoncas Z, Seiler C, Nakamaru-Ogiso E, 
            Clark KJ, Ekker SC
  TITLE     The FusX TALE Base Editor (FusXTBE) for Rapid Mitochondrial DNA 
            Programming of Human Cells In Vitro and Zebrafish Disease Models In 
            Vivo.
  JOURNAL   CRISPR J. 2021 Dec;4(6):799-821. doi: 10.1089/crispr.2021.0061. Epub
            2021 Nov 30.
  PUBMED    34847747
REFERENCE   2  (bases 1 to 6692)
  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: 
            "10.1089/crispr.2021.0061"; journalName: "CRISPR J"; date: 
            "2021-12"; volume: "4"; issue: "6"; pages: "799-821"
COMMENT     Sequence Label: ND4-FusXTBE-Right TALE
FEATURES             Location/Qualifiers
     source          1..6692
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    join(19..30,31..218)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(240..259)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(241..259)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        345..648
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        650..925
                     /label=chicken beta-actin promoter
     primer_bind     1181..1200
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1261..1338
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MASVLTPLLLRGLTGSARRLPVPRAK"
     CDS             3778..4029
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     polyA_signal    4219..4300
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    4424..4505
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        4693..4711
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    join(4731..4893,4894..4905)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(4935..4952)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5106..5694)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5186..5205)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5837..6646)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     6560..6579
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6638..6657
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 cacgatgcag ctggctccca gaggtgtaaa gtacttgagt aattttactt gattactgta
       61 cttaagtatt atttttgggg atttttactt tacttgagta caattaaaaa tcaatacttt
      121 tacttttact taattacatt tttttagaaa aaaaagtact ttttactcct tacaatttta
      181 tttacagtca aaaagtactt attttttgga gatcacttca ttctattttc ccttggtctc
      241 cctatagtga gtcgtattag atcccgggat tactagttta aacctagtta ttaatagtaa
      301 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
      361 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
      421 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
      481 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
      541 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
      601 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc
      661 cccacgttct gcttcactct ccccatctcc cccccctccc cacccccaat tttgtattta
      721 tttatttttt aattattttg tgcagcgatg ggggcggggg gggggggggc gcgcgccagg
      781 cggggcgggg cggggcgagg ggcggggcgg ggcgaggcgg agaggtgcgg cggcagccaa
      841 tcagagcggc gcgctccgaa agtttccttt tatggcgagg cggcggcggc ggcggcccta
      901 taaaaagcga agcgcgcggc gggcgggagt cgctgcgcgc tgccttcgcc ccgtgccccg
      961 ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg accgcgttac tcccacaggt
     1021 gagcgggcgg gacggccctt ctcctccggg ctgtaattag cgcttggttt aatgacggct
     1081 tgtttctttt ctgtggctgc gtgaaagcct tgaggggctc cgggagctag agcctctgct
     1141 aaccatgttc atgccttctt ctttttccta cagctcctgg gcaacgtgct ggttattgtg
     1201 ctgtctcatc attttggcaa agaattttaa gctaggatct aactcgagag atctagcacc
     1261 atggcctccg ttctgacccc gctgctgctg cgtggcctga ccggctccgc tcgtcgtctg
     1321 ccagttccgc gtgcgaaaat ccattccctg gtggatctac gcacgctcgg ctacagtcag
     1381 cagcagcaag agaagatcaa accgaaggtg cgttcgacag tggcgcagca ccacgaggca
     1441 ctggtgggcc atgggtttac acacgcgcac atcgttgcgc tcagccaaca cccggcagcg
     1501 ttagggaccg tcgctgtcac gtatcagcac ataatcacgg cgttgccaga ggcgacacac
     1561 gaagacatcg ttggcgtcgg caaacagtgg tccggcgcac gcgccctgga ggccttgctc
     1621 acggatgcgg gggagttgag aggtccgccg ttacagttgg acacaggcca acttgtgaag
     1681 attgcaaaac gtggcggcgt gaccgcaatg gaggcagtgc atgcatcgcg caatgcactg
     1741 acgggtgccc ccctgaacct gaccccggac caagtggtgg ctatcgccag caacaatggc
     1801 ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc
     1861 ctgaccccgg accaagtggt ggctatcgcc agcaacggtg gcggcaagca agcgctcgaa
     1921 acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg gcctgacccc ggaccaagtg
     1981 gtggctatcg ccagcaacat tggcggcaag caagcgctcg aaacggtgca gcggctgttg
     2041 ccggtgctgt gccaggacca tggcctgacc ccggaccaag tggtggctat cgccagcaac
     2101 attggcggca agcaagcgct cgaaacggtg cagcggctgt tgccggtgct gtgccaggac
     2161 catggcctga ccccggacca agtggtggct atcgccagca acaatggcgg caagcaagcg
     2221 ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg accatggcct gaccccggac
     2281 caagtggtgg ctatcgccag caacggtggc ggcaagcaag cgctcgaaac ggtgcagcgg
     2341 ctgttgccgg tgctgtgcca ggaccatggc ctgaccccgg accaagtggt ggctatcgcc
     2401 agcaacattg gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc
     2461 caggaccatg gcctcacacc cgatcaggtc gtggctatcg ccagcaacaa tggcggcaag
     2521 caagcgctcg aaacggtgca gcggctgttg ccggtgctgt gccaggacca tggcctgacc
     2581 ccggaccaag tggtggctat cgccagcaac aatggcggca agcaagcgct cgaaacggtg
     2641 cagcggctgt tgccggtgct gtgccaggac catggcctga ccccggacca agtggtggct
     2701 atcgccagca acattggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg
     2761 ctgtgccagg accatggcct gaccccggac caagtggtgg ctatcgccag caacaatggc
     2821 ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc
     2881 ctgaccccgg accaagtggt ggctatcgcc agcaacattg gcggcaagca agcgctcgaa
     2941 acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg gcctgacccc ggaccaagtg
     3001 gtggctatcg ccagcaacaa tggcggcaag caagcgctcg aaacggtgca gcggctgttg
     3061 ccggtgctgt gccaggacca tggcctgacc ccggaccaag tggtggctat cgccagcaac
     3121 ggtggcggca agcaagcgct cgaaacggtg cagcggctgt tgccggtgct gtgccaggac
     3181 catggcctga ccccggacca agtggtggct atcgccagca acaatggcgg caagcaagcg
     3241 ctcgaaagca ttgtggccca gctgagccgg cctgatccgg cgttggccgc gttgaccaac
     3301 gaccacctcg tcgccttggc ctgcctcggc ggacgtcctg ccatggatgc agtgaaaaag
     3361 ggattgccgc acgcgccgga attgatcaga agagtcaatc gccgtattgg cgaacgcacg
     3421 tcccatcgcg ttgcctctag aggcagctac gccctgggtc cgtatcagat tagcgccccg
     3481 cagctgccag cttacaatgg tcagaccgtg ggtaccttct actatgtgaa cgacgcgggc
     3541 ggtctggaga gcaaggtgtt tagcagcggc ggtccaaccc cgtacccaaa ctatgccaat
     3601 gccggtcatg tggagggtca gagcgccctg ttcatgcgtg ataacggcat cagcgagggt
     3661 ctggtgttcc acaacaaccc ggaaggcacc tgcggttttt gcgtgaacat gaccgagacc
     3721 ctgctgccgg aaaacgcgaa aatgaccgtg gtgccgccgg aaggttctgg cggatccact
     3781 aatctgagcg acatcattga gaaggagact gggaaacagc tggtcattca ggagtccatc
     3841 ctgatgctgc ctgaggaggt ggaggaagtg atcggcaaca agccagagtc tgacatcctg
     3901 gtgcacaccg cctacgacga gtccacagat gagaatgtga tgctgctgac ctctgacgcc
     3961 cccgagtata agccttgggc cctggtcatc caggattcta acggcgagaa taagatcaag
     4021 atgctgtgat aaggggtctt tgtcctctgt actgtctctc tccttgcccc taacccaaaa
     4081 agcttcattt ttctgtgtag gctgcacaag agccttgatt gaagatatat tctttctgaa
     4141 cagtatttaa ggtttccaat aaaatgtaca cccctcagaa gaattcattg atgagtttgg
     4201 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     4261 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     4321 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     4381 caaatgtggt cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     4441 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     4501 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
     4561 ttttttaaag caagtaaaac ctctacaaat gtggtcattc agctggcgta atagcgaaga
     4621 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggaaattgta
     4681 agcgttgata tcaattaacc ctcactaaag ggagaccact ctagttttct ttcttgcttt
     4741 tacttttact tccttaatac tcaagtacaa ttttaatgga gtactttttt acttttactc
     4801 aagtaagatt ctagccagat acttttactt ttaattgagt aaaattttcc ctaagtactt
     4861 gtactttcac ttgagtaaaa tttttgagta ctttttacac ctctgctcgg taccatttaa
     4921 atctgttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     4981 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     5041 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     5101 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     5161 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     5221 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     5281 ggaaccgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     5341 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5401 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5461 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     5521 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     5581 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5641 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     5701 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5761 ttggtcatga gattatcaaa aaggatcttc acctagatcc tttttgccag tgttacaacc
     5821 aattaaccaa ttctgattag aaaaactcat cgagcatcaa atgaaactgc aatttattca
     5881 tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact
     5941 caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc
     6001 caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat
     6061 caccatgagt gacgactgaa tccggtgaga atggcaaaag tttatgcatt tctttccaga
     6121 cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt
     6181 tattcattcg tgattgcgcc tgagcaagac gaaatacgcg atcgctgtta aaaggacaat
     6241 tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt
     6301 cacctgaatc aggatattct tctaatacct ggaatgctgt ttttccgggg atcgcagtgg
     6361 tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa
     6421 attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt
     6481 tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaagcga tagattgtcg
     6541 cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt
     6601 tggaatttaa tcgcggcctc gacgtttccc gttgaatatg gctcataaca ccccttgtat
     6661 tactgtttat gtaagcagac agttttattg tt
//
