LOCUS       sequence-375503-        5714 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses the right-side half ND6-Q1310A in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5714)
  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 5714)
  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_right
FEATURES             Location/Qualifiers
     source          1..5714
                     /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             409..486
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MASVLTPLLLRGLTGSARRLPVPRAK"
     CDS             499..564
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2293..2544
                     /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(2769..2786)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2775..2999
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3070..3086)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3070..3086)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3083..3105)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3094..3110
                     /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(3118..3124,3125..3142,3143..3148))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3163..3184
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3301..3318)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3472..4060)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3552..3571)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4231..5022,5023..5091))
                     /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     4854..4873
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5092..5196)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5275..5294)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(5466..5714,1..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 ggccagcgtg
      421 ctcacaccac tgctgctgag aggcctgaca ggaagcgcgc ggagactgcc tgtccccaga
      481 gccaagatcc acagcctgga ttataaggac cacgacggag attacaaaga ccacgatatc
      541 gactacaagg atgacgacga taagatggac atcgcggacc tcaggacgct gggctacagc
      601 cagcaacagc aagagaagat caagcccaaa gtaaggagca ccgtggccca gcaccacgag
      661 gccctggtgg gtcacggctt cacccacgcg catatcgttg ctctgagcca acatcccgca
      721 gctctgggta ccgttgcggt gaagtatcag gacatgatcg cggcactgcc tgaagctaca
      781 cacgaagcca tagtgggcgt tggcaagaga ggtgctggtg ccagagcgct tgaggcactg
      841 ttgacggtgg ctggcgagct gaggggaccg ccactgcaac tggacaccgg ccaactgctg
      901 aagatcgcca agaggggagg cgtgacggcg gtggaggccg tgcatgcctg gaggaacgca
      961 ctgaccggcg cgcccctgaa cctgacacca gagcaagtag tggctattgc aagtaacaac
     1021 ggtggcaaac aagcgctgga gaccgtgcag aggctccttc cggtgctctg ccaagcacac
     1081 ggtctcactc cggaacaggt ggtcgcaatc gcgagcaaca tcggcggcaa gcaagccctt
     1141 gagacagtcc aaagactttt gcctgtcctt tgtcaggcgc atggccttac gcctgagcaa
     1201 gtcgttgcga tcgcctccaa cggcggcgga aaacaggctt tggaaaccgt gcagcggttg
     1261 ctgcccgttt tgtgccaagc ccacggattg acccccgaac aggttgtagc catagcttct
     1321 aacaacggag gtaagcaggc actggaaacc gtgcagcgcc tgctcccagt actgtgtcag
     1381 gctcatgggc tgacaccaga gcaagtagtg gctattgcaa gtaacaacgg tggcaaacaa
     1441 gcgctggaga ccgtgcagag gctccttccg gtgctctgcc aagcacacgg tctcactccg
     1501 gaacaggtgg tcgcaatcgc gagccacgac ggcggcaagc aagcccttga gacagtccaa
     1561 agacttttgc ctgtcctttg tcaggcgcat ggccttacgc ctgagcaagt cgttgcgatc
     1621 gcctccaacg gcggcggaaa acaggctttg gaaaccgtgc agcggttgct gcccgttttg
     1681 tgccaagccc acggattgac ccccgaacag gttgtagcca tagcttctaa catcggaggt
     1741 aagcaggcac tggaaaccgt gcagcgcctg ctcccagtac tgtgtcaggc tcatgggctc
     1801 actccggaac aggtggtcgc aatcgcgagc aacggcggcg gcaagcaagc ccttgagaca
     1861 gtccaaagac ttttgcctgt cctttgtcag gcgcatggcc ttacgcctga gcaagtcgtt
     1921 gcgatcgcct ccaacggcgg cggaaaacag gctttggaaa ccgtgcagcg gttgctgccc
     1981 gttttgtgcc aagcccacgg attgaccccc gaacaggttg tagccatagc ttctaacggc
     2041 ggaggtagac ctgcactgga aagcatcgtg gcccagctga gcagacccga ccctgccctc
     2101 gccgctctga ccaacgacca cctggtggcc ctggcttgcc tcgggggcag gccagctctt
     2161 gacgccgtga agaagggcct tggcggatcc gccatccccg tgaagcgggg cgctacaggc
     2221 gagacaaaag ttttcaccgg caactccaac agccccaagt ccccaaccaa gggcggttgt
     2281 agcggcggca gcaccaacct gagcgacatc atcgagaagg aaaccggcaa gcagctggtg
     2341 atccaggaga gcatcctgat gctgcctgaa gaggtggaag aagtgatcgg caacaagcct
     2401 gagagcgaca tcctggtgca caccgcctac gacgagtcta ccgacgagaa cgtgatgctg
     2461 ctgacaagcg atgcccctga gtacaagccc tgggccctgg tgatccaaga ctccaatggc
     2521 gagaacaaga tcaagatgct gtaatagacc acgatcgtta tgctgatcat accctaatga
     2581 tcccagcaag ataatgtcct gtcttctaag atgtgcatca agcctggtac atactgaaaa
     2641 ccctataagg tcctggataa tttttgtttg attattcatt gaagaaacat ttattttcca
     2701 attgtgtgaa gtttttgact gttaataaaa gaatctgtca accatcaaag tttaaacccg
     2761 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     2821 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     2881 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     2941 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     3001 ttctgaggcg gaaagaacca gctggggctc aataccgtcg acctctagct agagcttggc
     3061 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     3121 catacgagcc ggaagcataa agtgtaaagc ctagggtgcc taatgagtga gctaactcac
     3181 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     3241 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     3301 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     3361 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     3421 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3481 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3541 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3601 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3661 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3721 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3781 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3841 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3901 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     3961 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4021 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4081 tacggggtct gacactcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4141 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     4201 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     4261 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4321 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     4381 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     4441 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4501 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     4561 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     4621 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     4681 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     4741 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     4801 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     4861 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     4921 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     4981 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5041 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     5101 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     5161 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     5221 tgacgtcgac ggatcgggag atcgatctcc cgatccccta gggtcgactc tcagtacaat
     5281 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
     5341 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
     5401 gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac
     5461 gcgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
     5521 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     5581 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     5641 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     5701 acatcaagtg tatc
//
