LOCUS       sequence-414911-        6496 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mitoBE expression vector, target mtND4 gene..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6496)
  AUTHORS   Yi Z, Zhang X, Tang W, Yu Y, Wei X, Zhang X, Wei W
  TITLE     Strand-selective base editing of human mitochondrial DNA using 
            mitoBEs.
  JOURNAL   Nat Biotechnol. 2023 May 22. doi: 10.1038/s41587-023-01791-y.
  PUBMED    37217751
REFERENCE   2  (bases 1 to 6496)
  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: "Nat 
            Biotechnol. 2023 May 22. doi: 10.1038/s41587-023-01791-y."
COMMENT     Sequence Label: 9ao-SOD2-3xHA-TALE(ND4-Left)-Nt.BspD6I(C)
FEATURES             Location/Qualifiers
     source          1..6496
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        132..335
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     286..306
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     377..396
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        377..395
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      421..430
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             520..546
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             550..576
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             580..606
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3852..3868)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3852..3868)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3865..3887)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3876..3892
                     /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(3900..3906,3907..3924,3925..3930))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3945..3966
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4083..4100)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4254..4842)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4334..4353)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5013..5804,5805..5873))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5636..5655
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5874..5978)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6057..6076)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(6248..6496,1..131)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
       61 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      121 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      181 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      241 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      301 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      361 agagatccgc ggccgctaat acgactcact atagggagac ccaagctggc tagcaccgcc
      421 gccaccatgg ccctgtcccg tgcggtttgt ggcacctccc gtcaactggc tccggttctg
      481 ggttatctgg gttcccgtca aaaacactcc ctgccggact acccgtatga tgttccggat
      541 tacgctggct acccatacga cgtcccagac tacgctggct acccatacga cgtcccagac
      601 tacgctatgg acatcgcgga cctcaggacg ctgggctaca gccagcaaca gcaagagaag
      661 atcaagccca aagtaaggag caccgtggcc cagcaccacg aggccctggt gggtcacggc
      721 ttcacccacg cgcatatcgt tgctctgagc caacatcccg cagctctggg taccgttgcg
      781 gtgaagtatc aggacatgat cgcggcactg cctgaagcta cacacgaagc catagtgggc
      841 gttggcaagc agtggagcgg tgccagagcg cttgaggcac tgttgacggt ggctggcgag
      901 ctgaggggac cgccactgca actggacacc ggccaactgc tgaagatcgc caagagggga
      961 ggcgtgacgg cggtggaggc cgtgcatgcc tggaggaacg cactgaccgg cgcgcccctg
     1021 aacctgacac cagagcaagt agtggctatt gcaagtaaca acggtggcaa acaagcgctg
     1081 gagaccgtgc agaggctcct tccggtgctc tgccaagcac acggtctcac tccggaacag
     1141 gtggtcgcaa tcgcgagcca cgacggcggc aagcaagccc ttgagacagt ccaaagactt
     1201 ttgcctgtcc tttgtcaggc gcatggcctt acgcctgagc aagtcgttgc gatcgcctcc
     1261 aacggcggcg gaaaacaggc tttggaaacc gtgcagcggt tgctgcccgt tttgtgccaa
     1321 gcccacggat tgacccccga acaggttgta gccatagctt ctaacatcgg aggtaagcag
     1381 gcactggaaa ccgtgcagcg cctgctccca gtactgtgtc aggctcatgg gctgacacca
     1441 gagcaagtag tggctattgc aagtaacaac ggtggcaaac aagcgctgga gaccgtgcag
     1501 aggctccttc cggtgctctg ccaagcacac ggtctcactc cggaacaggt ggtcgcaatc
     1561 gcgagcaacg gcggcggcaa gcaagccctt gagacagtcc aaagactttt gcctgtcctt
     1621 tgtcaggcgc atggccttac gcctgagcaa gtcgttgcga tcgcctccaa catcggcgga
     1681 aaacaggctt tggaaaccgt gcagcggttg ctgcccgttt tgtgccaagc ccacggattg
     1741 acccccgaac aggttgtagc catagcttct aacatcggag gtaagcaggc actggaaacc
     1801 gtgcagcgcc tgctcccagt actgtgtcag gctcatgggc tcactccgga acaggtggtc
     1861 gcaatcgcga gccacgacgg cggcaagcaa gcccttgaga cagtccaaag acttttgcct
     1921 gtcctttgtc aggcgcatgg ccttacgcct gagcaagtcg ttgcgatcgc ctcccacgac
     1981 ggcggaaaac aggctttgga aaccgtgcag cggttgctgc ccgttttgtg ccaagcccac
     2041 ggattgaccc ccgaacaggt tgtagccata gcttctaaca tcggaggtaa gcaggcactg
     2101 gaaaccgtgc agcgcctgct cccagtactg tgtcaggctc atgggctgac accagagcaa
     2161 gtagtggcta ttgcaagtca cgacggtggc aaacaagcgc tggagaccgt gcagaggctc
     2221 cttccggtgc tctgccaagc acacggtctc actccggaac aggtggtcgc aatcgcgagc
     2281 aacaacggcg gcaagcaagc ccttgagaca gtccaaagac ttttgcctgt cctttgtcag
     2341 gcgcatggcc ttacgcctga gcaagtcgtt gcgatcgcct ccaacggcgg cggaaaacag
     2401 gctttggaaa ccgtgcagcg gttgctgccc gttttgtgcc aagcccacgg attgaccccc
     2461 gaacaggttg tagccatagc ttctaacggc ggaggtaagc aggcactgga aaccgtgcag
     2521 cgcctgctcc cagtactgtg tcaggctcat gggctgacac cagagcaagt agtggctatt
     2581 gcaagtcacg acggtggcaa acaagcgctg gagaccgtgc agaggctcct tccggtgctc
     2641 tgccaagcac acggtctgac accagagcaa gtagtggcta ttgcaagtaa cggcggtggc
     2701 aaacaagcgc tggagagcat cgtggcccag ctgagcagac ccgaccctgc cctcgccgct
     2761 ctgaccaacg accacctggt ggccctggct tgcctcgggg gcaggccagc tcttgacgcc
     2821 gtgaagaagg gccttggtgg atcccggcag ctggaggagg tgattgatct gctggaggtg
     2881 taccatgaga agaagaacgt gatcgaggag aagatcaagg cccggtttat cgccaacaag
     2941 aacaccgtgt tcgagtggct gacatggaac ggcttcatta ttctgggcaa tgccctcgag
     3001 tacaagaata atttcgtgat cgatgaagag ctgcagcccg tgacacacgc tgccggcaac
     3061 cagcctgata tggagattat ctacgaggac tttattgtgc tgggcgaagt gaccacatca
     3121 aagggggcca cacagtttaa gatggagtct gagccagtca ctcgccacta cctgaacaaa
     3181 aagaaggagc tggaaaaaca gggcgtggag aaggaactgt attgcctgtt catcgcccct
     3241 gaaatcaaca aaaacacctt tgaggagttt atgaagtaca acatcgtgca gaacaccagg
     3301 atcatccctc tgtctctgaa gcagttcaac atgctgctga tggtgcagaa gaagctgatc
     3361 gaaaaaggcc gcagactgtc cagctacgac attaagaacc tgatggtgag cctgtaccgg
     3421 accacaatcg agtgcgaaag aaagtacacc cagattaaag ccggcctgga ggaaaccctg
     3481 aataactggg tggtggacaa ggaggtgaga ttttagtgat aaaccacgat cgttatgctg
     3541 atcataccct aatgatccca gcaagataat gtcctgtctt ctaagatgtg catcaagcct
     3601 ggtacatact gaaaacccta taaggtcctg gataattttt gtttgattat tcattgaaga
     3661 aacatttatt ttccaattgt gtgaagtttt tgactgttaa taaaagaatc tgtcaaccat
     3721 caaaaccggt gtttaaacga agacaatagc aggcatgctg gggatgcggt gggctctatg
     3781 gcttctgagg cggaaagaac cagctggggc tcgataccgt cgacctctag ctagagcttg
     3841 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     3901 aacatacgag ccggaagcat aaagtgtaaa gcctagggtg cctaatgagt gagctaactc
     3961 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     4021 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     4081 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     4141 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     4201 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4261 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4321 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4381 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4441 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4501 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4561 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4621 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     4681 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4741 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     4801 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     4861 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     4921 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     4981 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     5041 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     5101 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     5161 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     5221 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     5281 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     5341 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     5401 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     5461 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     5521 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     5581 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     5641 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     5701 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     5761 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     5821 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     5881 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     5941 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     6001 cctgacgtcg acggatcggg agatcgatct cccgatcccc tagggtcgac tctcagtaca
     6061 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
     6121 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
     6181 atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat
     6241 acgcgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
     6301 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     6361 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     6421 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     6481 gtacatcaag tgtatc
//
