LOCUS       sequence-396616-        5605 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N terminus half of DddAtox in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5605)
  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 5605)
  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-N
FEATURES             Location/Qualifiers
     source          1..5605
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(143..952)
                     /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     866..885
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     944..963
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     misc_feature    join(1017..1028,1029..1216)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     promoter        complement(1239..1257)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        1343..1646
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1648..1924
                     /label=chicken beta-actin promoter
     primer_bind     2180..2199
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             2260..2337
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MASVLTPLLLRGLTGSARRLPVPRAK"
     primer_bind     2907..2929
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2921..2938
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2922..2938
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2948..2966
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3014..3032)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3053..3069)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3053..3069)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        complement(join(3101..3107,3108..3125,3126..3131))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3146..3167
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             3689..3940
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     polyA_signal    4130..4211
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    4335..4416
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        4604..4622
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    join(4642..4804,4805..4816)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(4846..4863)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5017..5605)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5097..5116)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
       61 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt tgccagtgtt
      121 acaaccaatt aaccaattct gattagaaaa actcatcgag catcaaatga aactgcaatt
      181 tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag
      241 aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga
      301 ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg
      361 agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagttta tgcatttctt
      421 tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca
      481 aaccgttatt cattcgtgat tgcgcctgag caagacgaaa tacgcgatcg ctgttaaaag
      541 gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa
      601 tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttt ccggggatcg
      661 cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag
      721 gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc
      781 tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac aagcgataga
      841 ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat
      901 ccatgttgga atttaatcgc ggcctcgacg tttcccgttg aatatggctc ataacacccc
      961 ttgtattact gtttatgtaa gcagacagtt ttattgttca cgatgcagct ggctcccaga
     1021 ggtgtaaagt acttgagtaa ttttacttga ttactgtact taagtattat ttttggggat
     1081 ttttacttta cttgagtaca attaaaaatc aatactttta cttttactta attacatttt
     1141 tttagaaaaa aaagtacttt ttactcctta caattttatt tacagtcaaa aagtacttat
     1201 tttttggaga tcacttcatt ctattttccc ttggtctccc tatagtgagt cgtattagat
     1261 cccgggatta ctagtttaaa cctagttatt aatagtaatc aattacgggg tcattagttc
     1321 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     1381 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     1441 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     1501 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     1561 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     1621 acgtattagt catcgctatt accatggtcg aggtgagccc cacgttctgc ttcactctcc
     1681 ccatctcccc cccctcccca cccccaattt tgtatttatt tattttttaa ttattttgtg
     1741 cagcgatggg ggcggggggg ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg
     1801 gcggggcggg gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa
     1861 gtttcctttt atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg
     1921 ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg
     1981 cccgccccgg ctctgactga ccgcgttact cccacaggtg agcgggcggg acggcccttc
     2041 tcctccgggc tgtaattagc gcttggttta atgacggctt gtttcttttc tgtggctgcg
     2101 tgaaagcctt gaggggctcc gggagctaga gcctctgcta accatgttca tgccttcttc
     2161 tttttcctac agctcctggg caacgtgctg gttattgtgc tgtctcatca ttttggcaaa
     2221 gaattttaag ctaggatcta actcgagaga tctagcacca tggcctccgt tctgaccccg
     2281 ctgctgctgc gtggcctgac cggctccgct cgtcgtctgc cagttccgcg tgcgaaaatc
     2341 cattccctgg tggatctacg cacgctcggc tacagtcagc agcagcaaga gaagatcaaa
     2401 ccgaaggtgc gttcgacagt ggcgcagcac cacgaggcac tggtgggcca tgggtttaca
     2461 cacgcgcaca tcgttgcgct cagccaacac ccggcagcgt tagggaccgt cgctgtcacg
     2521 tatcagcaca taatcacggc gttgccagag gcgacacacg aagacatcgt tggcgtcggc
     2581 aaacagtggt ccggcgcacg cgccctggag gccttgctca cggatgcggg ggagttgaga
     2641 ggtccgccgt tacagttgga cacaggccaa cttgtgaaga ttgcaaaacg tggcggcgtg
     2701 accgcaatgg aggcagtgca tgcatcgcgc aatgcactga cgggtgcccc cctggagacg
     2761 ggcgccgcta cagggcgcgt cccatttgcc attcaggctg cgcaactgtt gggaagggcg
     2821 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     2881 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga
     2941 gcgcgcgtaa tacgactcac tatagggcga attgggtacc gggccccccc tcgaggtcct
     3001 ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc
     3061 tgtttcctgt gtgaaattgt tacccgctca caattccaca caacatacga gccggaagca
     3121 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     3181 cactgcccgc tttccaccgg tcgtctccaa cgaccacctc gtcgccttgg cctgcctcgg
     3241 cggacgtcct gccatggatg cagtgaaaaa gggattgccg cacgcgccgg aattgatcag
     3301 aagagtcaat cgccgtattg gcgaacgcac gtcccatcgc gttgcctcta gaggcagcta
     3361 cgccctgggt ccgtatcaga ttagcgcccc gcagctgcca gcttacaatg gtcagaccgt
     3421 gggtaccttc tactatgtga acgacgcggg cggtctggag agcaaggtgt ttagcagcgg
     3481 cggtccaacc ccgtacccaa actatgccaa tgccggtcat gtggagggtc agagcgccct
     3541 gttcatgcgt gataacggca tcagcgaggg tctggtgttc cacaacaacc cggaaggcac
     3601 ctgcggtttt tgcgtgaaca tgaccgagac cctgctgccg gaaaacgcga aaatgaccgt
     3661 ggtgccgccg gaaggttctg gcggatccac taatctgagc gacatcattg agaaggagac
     3721 tgggaaacag ctggtcattc aggagtccat cctgatgctg cctgaggagg tggaggaagt
     3781 gatcggcaac aagccagagt ctgacatcct ggtgcacacc gcctacgacg agtccacaga
     3841 tgagaatgtg atgctgctga cctctgacgc ccccgagtat aagccttggg ccctggtcat
     3901 ccaggattct aacggcgaga ataagatcaa gatgctgtga taaggggtct ttgtcctctg
     3961 tactgtctct ctccttgccc ctaacccaaa aagcttcatt tttctgtgta ggctgcacaa
     4021 gagccttgat tgaagatata ttctttctga acagtattta aggtttccaa taaaatgtac
     4081 acccctcaga agaattcatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     4141 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     4201 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt
     4261 gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tcattgatga gtttggacaa
     4321 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     4381 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     4441 atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa
     4501 tgtggtcatt cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     4561 tgcgcagcct gaatggcgaa tggaaattgt aagcgttgat atcaattaac cctcactaaa
     4621 gggagaccac tctagttttc tttcttgctt ttacttttac ttccttaata ctcaagtaca
     4681 attttaatgg agtacttttt tacttttact caagtaagat tctagccaga tacttttact
     4741 tttaattgag taaaattttc cctaagtact tgtactttca cttgagtaaa atttttgagt
     4801 actttttaca cctctgctcg gtaccattta aatctgttcc gcttcctcgc tcactgactc
     4861 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4921 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4981 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5041 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5101 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5161 taccggatac ctgtccgcct ttctcccttc gggaaccgtg gcgctttctc atagctcacg
     5221 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5281 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5341 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5401 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     5461 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5521 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5581 tacgcgcaga aaaaaaggat ctcaa
//
