LOCUS       sequence-386805-        6458 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses left-side TALE-DddAtox Cterm half ND2-FusXTBE in mammalian
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6458)
  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 6458)
  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-Left TALE
FEATURES             Location/Qualifiers
     source          1..6458
                     /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             3544..3795
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     polyA_signal    3985..4066
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    4190..4271
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        4459..4477
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    join(4497..4659,4660..4671)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(4701..4718)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4872..5460)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4952..4971)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5603..6412)
                     /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     6326..6345
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6404..6423
                     /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 agcaacggtg gcggcaagca agcgctcgaa
     1921 acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg gcctgactcc ggaccaagtg
     1981 gtggctatcg ccagccacga tggcggcaag caagcgctcg aaacggtgca gcggctgttg
     2041 ccggtgctgt gccaggacca tggcctgacc ccggaccaag tggtggctat cgccagccac
     2101 gatggcggca agcaagcgct cgaaacggtg cagcggctgt tgccggtgct gtgccaggac
     2161 catggcctga ccccggacca agtggtggct atcgccagca acattggcgg caagcaagcg
     2221 ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg accatggcct gaccccggac
     2281 caagtggtgg ctatcgccag caacggtggc ggcaagcaag cgctcgaaac ggtgcagcgg
     2341 ctgttgccgg tgctgtgcca ggaccatggc ctgaccccgg accaagtggt ggctatcgcc
     2401 agccacgatg gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc
     2461 caggaccatg gcctcacacc cgatcaggtc gtggctatcg ccagcaacat tggcggcaag
     2521 caagcgctcg aaacggtgca gcggctgttg ccggtgctgt gccaggacca tggcctgacc
     2581 ccggaccaag tggtggctat cgccagcaac ggtggcggca 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 ctgactccgg accaagtggt ggctatcgcc agccacgatg gcggcaagca agcgctcgaa
     2941 acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg gcctgacccc ggaccaagtg
     3001 gtggctatcg ccagcaacat tggcggcaag caagcgctcg aaacggtgca gcggctgttg
     3061 ccggtgctgt gccaggacca tggcctgacc ccggaccaag tggtggctat cgccagcaac
     3121 aatggcggca 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 agccattcca gtgaagcgcg gcgctaccgg tgaaaccaaa
     3481 gtgtttaccg gtaacagcaa cagcccgaag agcccgacca aaggcggttg ctctggcgga
     3541 tccactaatc tgagcgacat cattgagaag gagactggga aacagctggt cattcaggag
     3601 tccatcctga tgctgcctga ggaggtggag gaagtgatcg gcaacaagcc agagtctgac
     3661 atcctggtgc acaccgccta cgacgagtcc acagatgaga atgtgatgct gctgacctct
     3721 gacgcccccg agtataagcc ttgggccctg gtcatccagg attctaacgg cgagaataag
     3781 atcaagatgc tgtgataagg ggtctttgtc ctctgtactg tctctctcct tgcccctaac
     3841 ccaaaaagct tcatttttct gtgtaggctg cacaagagcc ttgattgaag atatattctt
     3901 tctgaacagt atttaaggtt tccaataaaa tgtacacccc tcagaagaat tcattgatga
     3961 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     4021 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     4081 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     4141 cctctacaaa tgtggtcatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     4201 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     4261 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt
     4321 gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tcattcagct ggcgtaatag
     4381 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggaa
     4441 attgtaagcg ttgatatcaa ttaaccctca ctaaagggag accactctag ttttctttct
     4501 tgcttttact tttacttcct taatactcaa gtacaatttt aatggagtac ttttttactt
     4561 ttactcaagt aagattctag ccagatactt ttacttttaa ttgagtaaaa ttttccctaa
     4621 gtacttgtac tttcacttga gtaaaatttt tgagtacttt ttacacctct gctcggtacc
     4681 atttaaatct gttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4741 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4801 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4861 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4921 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4981 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5041 ccttcgggaa ccgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     5101 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5161 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5221 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5281 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     5341 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5401 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5461 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5521 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt tgccagtgtt
     5581 acaaccaatt aaccaattct gattagaaaa actcatcgag catcaaatga aactgcaatt
     5641 tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag
     5701 aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga
     5761 ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg
     5821 agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagttta tgcatttctt
     5881 tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca
     5941 aaccgttatt cattcgtgat tgcgcctgag caagacgaaa tacgcgatcg ctgttaaaag
     6001 gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa
     6061 tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttt ccggggatcg
     6121 cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag
     6181 gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc
     6241 tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac aagcgataga
     6301 ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat
     6361 ccatgttgga atttaatcgc ggcctcgacg tttcccgttg aatatggctc ataacacccc
     6421 ttgtattact gtttatgtaa gcagacagtt ttattgtt
//
