LOCUS       sequence-386140-        6794 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Right-side TALE-DddAtox Nterm half ND2-FusXTBE in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6794)
  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 6794)
  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: ND2-FusXTBE-Right TALE
FEATURES             Location/Qualifiers
     source          1..6794
                     /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             3880..4131
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     polyA_signal    4321..4402
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    4526..4607
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        4795..4813
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    join(4833..4995,4996..5007)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(5037..5054)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5208..5796)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5288..5307)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5939..6748)
                     /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     6662..6681
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6740..6759
                     /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 caacattggc
     1801 ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc
     1861 ctgaccccgg accaagtggt ggctatcgcc agcaacaatg gcggcaagca agcgctcgaa
     1921 acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg gcctgactcc ggaccaagtg
     1981 gtggctatcg ccagccacga tggcggcaag caagcgctcg aaacggtgca gcggctgttg
     2041 ccggtgctgt gccaggacca tggcctgacc ccggaccaag tggtggctat cgccagcaac
     2101 ggtggcggca agcaagcgct cgaaacggtg cagcggctgt tgccggtgct gtgccaggac
     2161 catggcctga ccccggacca agtggtggct atcgccagca acaatggcgg caagcaagcg
     2221 ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg accatggcct gaccccggac
     2281 caagtggtgg ctatcgccag caacaatggc ggcaagcaag cgctcgaaac ggtgcagcgg
     2341 ctgttgccgg tgctgtgcca ggaccatggc ctgaccccgg accaagtggt ggctatcgcc
     2401 agcaacaatg gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc
     2461 caggaccatg gcctcacacc cgatcaggtc gtggctatcg ccagcaacgg tggcggcaag
     2521 caagcgctcg aaacggtgca gcggctgttg ccggtgctgt gccaggacca tggcctgacc
     2581 ccggaccaag tggtggctat cgccagcaac ggtggcggca agcaagcgct cgaaacggtg
     2641 cagcggctgt tgccggtgct gtgccaggac catggcctga ccccggacca agtggtggct
     2701 atcgccagca acggtggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg
     2761 ctgtgccagg accatggcct gaccccggac caagtggtgg ctatcgccag caacaatggc
     2821 ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc
     2881 ctgaccccgg accaagtggt ggctatcgcc agcaacaatg gcggcaagca agcgctcgaa
     2941 acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg gcctgacccc ggaccaagtg
     3001 gtggctatcg ccagcaacgg 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 caacattggc 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 agaggcagct acgccctggg tccgtatcag attagcgccc cgcagctgcc agcttacaat
     3601 ggtcagaccg tgggtacctt ctactatgtg aacgacgcgg gcggtctgga gagcaaggtg
     3661 tttagcagcg gcggtccaac cccgtaccca aactatgcca atgccggtca tgtggagggt
     3721 cagagcgccc tgttcatgcg tgataacggc atcagcgagg gtctggtgtt ccacaacaac
     3781 ccggaaggca cctgcggttt ttgcgtgaac atgaccgaga ccctgctgcc ggaaaacgcg
     3841 aaaatgaccg tggtgccgcc ggaaggttct ggcggatcca ctaatctgag cgacatcatt
     3901 gagaaggaga ctgggaaaca gctggtcatt caggagtcca tcctgatgct gcctgaggag
     3961 gtggaggaag tgatcggcaa caagccagag tctgacatcc tggtgcacac cgcctacgac
     4021 gagtccacag atgagaatgt gatgctgctg acctctgacg cccccgagta taagccttgg
     4081 gccctggtca tccaggattc taacggcgag aataagatca agatgctgtg ataaggggtc
     4141 tttgtcctct gtactgtctc tctccttgcc cctaacccaa aaagcttcat ttttctgtgt
     4201 aggctgcaca agagccttga ttgaagatat attctttctg aacagtattt aaggtttcca
     4261 ataaaatgta cacccctcag aagaattcat tgatgagttt ggacaaacca caactagaat
     4321 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     4381 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     4441 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtcattgatg
     4501 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     4561 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     4621 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa
     4681 acctctacaa atgtggtcat tcagctggcg taatagcgaa gaggcccgca ccgatcgccc
     4741 ttcccaacag ttgcgcagcc tgaatggcga atggaaattg taagcgttga tatcaattaa
     4801 ccctcactaa agggagacca ctctagtttt ctttcttgct tttactttta cttccttaat
     4861 actcaagtac aattttaatg gagtactttt ttacttttac tcaagtaaga ttctagccag
     4921 atacttttac ttttaattga gtaaaatttt ccctaagtac ttgtactttc acttgagtaa
     4981 aatttttgag tactttttac acctctgctc ggtaccattt aaatctgttc cgcttcctcg
     5041 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     5101 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5161 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5221 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5281 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5341 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaaccgt ggcgctttct
     5401 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5461 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5521 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5581 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5641 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5701 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5761 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5821 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5881 aaaaggatct tcacctagat cctttttgcc agtgttacaa ccaattaacc aattctgatt
     5941 agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac
     6001 catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata
     6061 ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta
     6121 ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg
     6181 aatccggtga gaatggcaaa agtttatgca tttctttcca gacttgttca acaggccagc
     6241 cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg
     6301 cctgagcaag acgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgaat
     6361 gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt
     6421 cttctaatac ctggaatgct gtttttccgg ggatcgcagt ggtgagtaac catgcatcat
     6481 caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta
     6541 gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca
     6601 actctggcgc atcgggcttc ccatacaagc gatagattgt cgcacctgat tgcccgacat
     6661 tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc
     6721 tcgacgtttc ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag
     6781 acagttttat tgtt
//
