LOCUS       sequence-375499-        5870 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses the left-side half ND6-Q1310A in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5870)
  AUTHORS   Lei Z, Meng H, Liu L, Zhao H, Rao X, Yan Y, Wu H, Liu M, He A, Yi C
  TITLE     Mitochondrial base editor induces substantial nuclear off-target 
            mutations.
  JOURNAL   Nature. 2022 May 12. pii: 10.1038/s41586-022-04836-5. doi: 
            10.1038/s41586-022-04836-5.
  PUBMED    35551512
REFERENCE   2  (bases 1 to 5870)
  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: "Nature. 
            2022 May 12. pii: 10.1038/s41586-022-04836-5. doi: 
            10.1038/s41586-022-04836-5."
COMMENT     Sequence Label: ND6-L1397-N_Q1310A_left
FEATURES             Location/Qualifiers
     source          1..5870
                     /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      403..412
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             502..528
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             532..558
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             562..588
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2449..2700
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     primer_bind     complement(2925..2942)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2931..3155
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3226..3242)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3226..3242)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3239..3261)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3250..3266
                     /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(3274..3280,3281..3298,3299..3304))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3319..3340
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3457..3474)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3628..4216)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3708..3727)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4387..5178,5179..5247))
                     /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     5010..5029
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5248..5352)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5431..5450)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(5622..5870,1..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 atagggagag ccgccaccat ggccctgagc
      421 agagccgttt gtggaacaag ccggcagctg gctcctgtgc ttggctacct gggcagcaga
      481 cagaagcaca gcctgccaga ctacccctat gacgtgcccg actacgccgg ctacccttat
      541 gacgtgcctg attacgccgg atacccttac gacgttccag attacgccat ggacatcgcg
      601 gacctcagga cgctgggcta cagccagcaa cagcaagaga agatcaagcc caaagtaagg
      661 agcaccgtgg cccagcacca cgaggccctg gtgggtcacg gcttcaccca cgcgcatatc
      721 gttgctctga gccaacatcc cgcagctctg ggtaccgttg cggtgaagta tcaggacatg
      781 atcgcggcac tgcctgaagc tacacacgaa gccatagtgg gcgttggcaa gcagtggagc
      841 ggtgccagag cgcttgaggc actgttgacg gtggctggcg agctgagggg accgccactg
      901 caactggaca ccggccaact gctgaagatc gccaagaggg gaggcgtgac ggcggtggag
      961 gccgtgcatg cctggaggaa cgcactgacc ggcgcgcccc tgaacctgac accagagcaa
     1021 gtagtggcta ttgcaagtaa caacggtggc aaacaagcgc tggagaccgt gcagaggctc
     1081 cttccggtgc tctgccaagc acacggtctc actccggaac aggtggtcgc aatcgcgagc
     1141 aacatcggcg gcaagcaagc ccttgagaca gtccaaagac ttttgcctgt cctttgtcag
     1201 gcgcatggcc ttacgcctga gcaagtcgtt gcgatcgcct cccacgacgg cggaaaacag
     1261 gctttggaaa ccgtgcagcg gttgctgccc gttttgtgcc aagcccacgg attgaccccc
     1321 gaacaggttg tagccatagc ttctcacgac ggaggtaagc aggcactgga aaccgtgcag
     1381 cgcctgctcc cagtactgtg tcaggctcat gggctgacac cagagcaagt agtggctatt
     1441 gcaagtcacg acggtggcaa acaagcgctg gagaccgtgc agaggctcct tccggtgctc
     1501 tgccaagcac acggtctcac tccggaacag gtggtcgcaa tcgcgagcca cgacggcggc
     1561 aagcaagccc ttgagacagt ccaaagactt ttgcctgtcc tttgtcaggc gcatggcctt
     1621 acgcctgagc aagtcgttgc gatcgcctcc cacgacggcg gaaaacaggc tttggaaacc
     1681 gtgcagcggt tgctgcccgt tttgtgccaa gcccacggat tgacccccga acaggttgta
     1741 gccatagctt ctaacatcgg aggtaagcag gcactggaaa ccgtgcagcg cctgctccca
     1801 gtactgtgtc aggctcatgg gctcactccg gaacaggtgg tcgcaatcgc gagcaacggc
     1861 ggcggcaagc aagcccttga gacagtccaa agacttttgc ctgtcctttg tcaggcgcat
     1921 ggccttacgc ctgagcaagt cgttgcgatc gcctccaacg gcggcggaag acctgctttg
     1981 gaaagcatcg tggcccagct gagcagaccc gaccctgccc tcgccgctct gaccaacgac
     2041 cacctggtgg ccctggcttg cctcgggggc aggccagctc ttgacgccgt gaagaagggc
     2101 cttggtggat ccggatctta cgccctgggc ccttaccaga tctctgcccc tcagctgccc
     2161 gcctacaacg gcgctaccgt gggcacattc tactacgtga acgacgccgg cggcctggaa
     2221 agcaaggtgt tcagcagcgg cggacctacc ccttatccta actacgccaa tgctggccac
     2281 gtggaaggac aaagcgccct gttcatgaga gataacggca tcagcgaggg cctggtcttt
     2341 cacaacaacc ccgagggcac ctgcggcttc tgcgtgaata tgaccgagac actgctgcct
     2401 gaaaacgcca agatgaccgt ggtgccacct gagggaagcg gcggcagcac caacctgagc
     2461 gacatcatcg agaaggaaac cggcaagcag ctggtgatcc aggagagcat cctgatgctg
     2521 cctgaagagg tggaagaagt gatcggcaac aagcctgaga gcgacatcct ggtgcacacc
     2581 gcctacgacg agtctaccga cgagaacgtg atgctgctga caagcgatgc ccctgagtac
     2641 aagccctggg ccctggtgat ccaagactcc aatggcgaga acaagatcaa gatgctgtaa
     2701 tagaccacga tcgttatgct gatcataccc taatgatccc agcaagataa tgtcctgtct
     2761 tctaagatgt gcatcaagcc tggtacatac tgaaaaccct ataaggtcct ggataatttt
     2821 tgtttgatta ttcattgaag aaacatttat tttccaattg tgtgaagttt ttgactgtta
     2881 ataaaagaat ctgtcaacca tcaaagttta aacccgctga tcagcctcga ctgtgccttc
     2941 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     3001 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     3061 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     3121 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     3181 gggctcaata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     3241 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     3301 taaagcctag ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     3361 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     3421 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     3481 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     3541 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     3601 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     3661 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     3721 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     3781 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     3841 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3901 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     3961 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4021 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     4081 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4141 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4201 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgaca ctcagtggaa
     4261 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     4321 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     4381 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     4441 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     4501 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     4561 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     4621 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     4681 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     4741 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     4801 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     4861 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     4921 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     4981 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5041 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     5101 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5161 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5221 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     5281 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     5341 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatcg
     5401 atctcccgat cccctagggt cgactctcag tacaatctgc tctgatgccg catagttaag
     5461 ccagtagctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta
     5521 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg
     5581 ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag
     5641 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
     5701 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     5761 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     5821 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
//
