LOCUS       sequence-386807-        6560 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses left-side TALE-DddAtox Cterm half ND4-FusXTBE in mammalian
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6560)
  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 6560)
  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-Left TALE
FEATURES             Location/Qualifiers
     source          1..6560
                     /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             3646..3897
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     polyA_signal    4087..4168
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    4292..4373
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        4561..4579
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    join(4599..4761,4762..4773)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(4803..4820)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4974..5562)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5054..5073)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5705..6514)
                     /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     6428..6447
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6506..6525
                     /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 cggggcgrgg 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 ctgactccgg accaagtggt ggctatcgcc agccacgatg gcggcaagca agcgctcgaa
     1921 acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg gcctgacccc ggaccaagtg
     1981 gtggctatcg ccagcaacgg 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 ccagcaacat tggcggcaag
     2521 caagcgctcg aaacggtgca gcggctgttg ccggtgctgt gccaggacca tggcctgact
     2581 ccggaccaag tggtggctat cgccagccac gatggcggca agcaagcgct cgaaacggtg
     2641 cagcggctgt tgccggtgct gtgccaggac catggcctga ccccggacca agtggtggct
     2701 atcgccagcc acgatggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg
     2761 ctgtgccagg accatggcct gaccccggac caagtggtgg ctatcgccag caacattggc
     2821 ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc
     2881 ctgactccgg accaagtggt ggctatcgcc agccacgatg 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 acggtggcgg caagcaagcg
     3241 ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg accatggcct gaccccggac
     3301 caagtggtgg ctatcgccag ccacgatggc ggcaagcaag cgctcgaaag cattgtggcc
     3361 cagctgagcc ggcctgatcc ggcgttggcc gcgttgacca acgaccacct cgtcgccttg
     3421 gcctgcctcg gcggacgtcc tgccatggat gcagtgaaaa agggattgcc gcacgcgccg
     3481 gaattgatca gaagagtcaa tcgccgtatt ggcgaacgca cgtcccatcg cgttgcctct
     3541 agagccattc cagtgaagcg cggcgctacc ggtgaaacca aagtgtttac cggtaacagc
     3601 aacagcccga agagcccgac caaaggcggt tgctctggcg gatccactaa tctgagcgac
     3661 atcattgaga aggagactgg gaaacagctg gtcattcagg agtccatcct gatgctgcct
     3721 gaggaggtgg aggaagtgat cggcaacaag ccagagtctg acatcctggt gcacaccgcc
     3781 tacgacgagt ccacagatga gaatgtgatg ctgctgacct ctgacgcccc cgagtataag
     3841 ccttgggccc tggtcatcca ggattctaac ggcgagaata agatcaagat gctgtgataa
     3901 ggggtctttg tcctctgtac tgtctctctc cttgccccta acccaaaaag cttcattttt
     3961 ctgtgtaggc tgcacaagag ccttgattga agatatattc tttctgaaca gtatttaagg
     4021 tttccaataa aatgtacacc cctcagaaga attcattgat gagtttggac aaaccacaac
     4081 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     4141 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     4201 ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtca
     4261 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     4321 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     4381 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca
     4441 agtaaaacct ctacaaatgt ggtcattcag ctggcgtaat agcgaagagg cccgcaccga
     4501 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg aaattgtaag cgttgatatc
     4561 aattaaccct cactaaaggg agaccactct agttttcttt cttgctttta cttttacttc
     4621 cttaatactc aagtacaatt ttaatggagt acttttttac ttttactcaa gtaagattct
     4681 agccagatac ttttactttt aattgagtaa aattttccct aagtacttgt actttcactt
     4741 gagtaaaatt tttgagtact ttttacacct ctgctcggta ccatttaaat ctgttccgct
     4801 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     4861 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     4921 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4981 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5041 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5101 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aaccgtggcg
     5161 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5221 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5281 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5341 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5401 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5461 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5521 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5581 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     5641 ttatcaaaaa ggatcttcac ctagatcctt tttgccagtg ttacaaccaa ttaaccaatt
     5701 ctgattagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat
     5761 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt
     5821 tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac
     5881 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga
     5941 cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag
     6001 gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg
     6061 attgcgcctg agcaagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa
     6121 tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag
     6181 gatattcttc taatacctgg aatgctgttt ttccggggat cgcagtggtg agtaaccatg
     6241 catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc
     6301 agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca
     6361 gaaacaactc tggcgcatcg ggcttcccat acaagcgata gattgtcgca cctgattgcc
     6421 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc
     6481 gcggcctcga cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt
     6541 aagcagacag ttttattgtt
//
