LOCUS       sequence-434573-        1459 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Backbone plasmid for FusX-based assembly of DdCBE arm with DddA6/11 
            1397C.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1459)
  AUTHORS   Castillo SR, Simone BW, Clark KJ, Devaux P, Ekker SC
  TITLE     Unconstrained Precision Mitochondrial Genome Editing with 
            alphaDdCBEs.
  JOURNAL   Hum Gene Ther. 2024 Sep 24. doi: 10.1089/hum.2024.073.
  PUBMED    39212664
REFERENCE   2  (bases 1 to 1459)
  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"; journalName: "Hum Gene 
            Ther. 2024 Sep 24. doi: 10.1089/hum.2024.073."
COMMENT     Sequence Label: FusX DdCBE DddA6-11 1397C
FEATURES             Location/Qualifiers
     source          1..1459
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             1..78
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MASVLTPLLLRGLTGSARRLPVPRAK"
     CDS             91..156
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(583..876)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCS"
     primer_bind     726..748
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     740..757
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     741..757
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     767..786
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        767..785
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(833..853)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(833..851)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(872..888)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(872..888)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(885..907)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    896..912
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(920..926,927..944,945..950))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    965..986
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             1208..1459
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
ORIGIN
        1 atggcctccg ttctgacccc gctgctgctg cgtggcctga ccggctccgc tcgtcgtctg
       61 ccagttccgc gtgcgaaaat ccattccctg gactacaaag accatgacgg tgattataaa
      121 gatcatgaca tcgattacaa ggatgacgat gacaagatgg acatcgcgga tctgcgtacc
      181 ctgggttaca gccagcagca gcaggagaag atcaagccaa aggtgcgcag caccgtggcc
      241 cagcaccatg aagctctggt gggtcacggc ttcacccacg cgcacatcgt ggctctgagc
      301 cagcacccag ccgcgctggg taccgtggcc gtgaaatatc aggacatgat tgctgccctg
      361 ccagaggcta cccatgaagc gattgtgggt gtgggcaagc agtggagcgg tgctcgtgcg
      421 ctggaggccc tgctgaccgt ggccggtgaa ctgcgtggcc cgccgctgca gctggatacc
      481 ggccagctgc tgaaaatcgc gaaacgtggc ggtgtgaccg ctgtggaagc tgtgcatgcc
      541 tggcgtaatg ctctgaccgg tgcccccctg gagacgggcg ccttagctac agggcgcgtc
      601 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
      661 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      721 gttttcccag tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat acgactcact
      781 atagggcgaa ttgggtaccg ggccccccct cgaggtcctc cagcttttgt tccctttagt
      841 gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
      901 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
      961 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccaccggt
     1021 cgtctccaac gatcacctgg tggccctggc ctgcctgggc ggtcgtccgg ccctggatgc
     1081 ggtgaagaaa ggcctgggtg gatccgccat tccagtgaag cgcggcgcta ccggtgaaac
     1141 caaagtgttt atcggtaaca gcaacagccc gaagagcccg accaaaggcg gttgctctgg
     1201 cggctcaact aatctgagcg acatcattga gaaggagact gggaaacagc tggtcattca
     1261 ggagtccatc ctgatgctgc ctgaggaggt ggaggaagtg atcggcaaca agccagagtc
     1321 tgacatcctg gtgcacaccg cctacgacga gtccacagat gagaatgtga tgctgctgac
     1381 ctctgacgcc cccgagtata agccttgggc cctggtcatc caggattcta acggcgagaa
     1441 taagatcaag atgctgtga
//
