LOCUS       sequence-396619-        5370 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses C terminus half of DddAtox in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5370)
  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 5370)
  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: FusXTBE-1397-C
FEATURES             Location/Qualifiers
     source          1..5370
                     /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)"
     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"
     primer_bind     1908..1930
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1922..1939
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1923..1939
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1949..1967
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2015..2033)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2054..2070)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2054..2070)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        complement(join(2102..2108,2109..2126,2127..2132))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2147..2168
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             2456..2707
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     polyA_signal    2897..2978
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    3102..3183
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        3371..3389
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    join(3409..3571,3572..3583)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(3613..3630)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3784..4372)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3864..3883)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4515..5324)
                     /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     5238..5257
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5316..5335
                     /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 ccctggagac gggcgccgct acagggcgcg tcccatttgc cattcaggct
     1801 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     1861 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     1921 ttgtaaaacg acggccagtg agcgcgcgta atacgactca ctatagggcg aattgggtac
     1981 cgggcccccc ctcgaggtcc tccagctttt gttcccttta gtgagggtta attgcgcgct
     2041 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttacccgctc acaattccac
     2101 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     2161 tcacattaat tgcgttgcgc tcactgcccg ctttccaccg gtcgtctcca acgaccacct
     2221 cgtcgccttg gcctgcctcg gcggacgtcc tgccatggat gcagtgaaaa agggattgcc
     2281 gcacgcgccg gaattgatca gaagagtcaa tcgccgtatt ggcgaacgca cgtcccatcg
     2341 cgttgcctct agagccattc cagtgaagcg cggcgctacc ggtgaaacca aagtgtttac
     2401 cggtaacagc aacagcccga agagcccgac caaaggcggt tgctctggcg gatccactaa
     2461 tctgagcgac atcattgaga aggagactgg gaaacagctg gtcattcagg agtccatcct
     2521 gatgctgcct gaggaggtgg aggaagtgat cggcaacaag ccagagtctg acatcctggt
     2581 gcacaccgcc tacgacgagt ccacagatga gaatgtgatg ctgctgacct ctgacgcccc
     2641 cgagtataag ccttgggccc tggtcatcca ggattctaac ggcgagaata agatcaagat
     2701 gctgtgataa ggggtctttg tcctctgtac tgtctctctc cttgccccta acccaaaaag
     2761 cttcattttt ctgtgtaggc tgcacaagag ccttgattga agatatattc tttctgaaca
     2821 gtatttaagg tttccaataa aatgtacacc cctcagaaga attcattgat gagtttggac
     2881 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     2941 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     3001 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
     3061 aatgtggtca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     3121 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     3181 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     3241 ttttaaagca agtaaaacct ctacaaatgt ggtcattcag ctggcgtaat agcgaagagg
     3301 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg aaattgtaag
     3361 cgttgatatc aattaaccct cactaaaggg agaccactct agttttcttt cttgctttta
     3421 cttttacttc cttaatactc aagtacaatt ttaatggagt acttttttac ttttactcaa
     3481 gtaagattct agccagatac ttttactttt aattgagtaa aattttccct aagtacttgt
     3541 actttcactt gagtaaaatt tttgagtact ttttacacct ctgctcggta ccatttaaat
     3601 ctgttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     3661 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     3721 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3781 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3841 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3901 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3961 aaccgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     4021 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4081 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     4141 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4201 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     4261 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4321 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4381 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4441 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt tttgccagtg ttacaaccaa
     4501 ttaaccaatt ctgattagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata
     4561 tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca
     4621 ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca
     4681 acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca
     4741 ccatgagtga cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc tttccagact
     4801 tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta
     4861 ttcattcgtg attgcgcctg agcaagacga aatacgcgat cgctgttaaa aggacaatta
     4921 caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca
     4981 cctgaatcag gatattcttc taatacctgg aatgctgttt ttccggggat cgcagtggtg
     5041 agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat
     5101 tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg
     5161 ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaagcgata gattgtcgca
     5221 cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg
     5281 gaatttaatc gcggcctcga cgtttcccgt tgaatatggc tcataacacc ccttgtatta
     5341 ctgtttatgt aagcagacag ttttattgtt
//
