LOCUS       sequence-375493-        5753 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses the right-side half ND6-UGI-NES in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5753)
  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 5753)
  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_UGI-NES_right
FEATURES             Location/Qualifiers
     source          1..5753
                     /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..2541
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             2554..2580
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     primer_bind     complement(2808..2825)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2814..3038
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3109..3125)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3109..3125)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3122..3144)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3133..3149
                     /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(3157..3163,3164..3181,3182..3187))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3202..3223
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3340..3357)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3511..4099)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3591..3610)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4270..5061,5062..5130))
                     /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     4893..4912
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5131..5235)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5314..5333)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(5505..5753,1..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 gggatctctg cagctgcctc cacttgaaag actgacactg
     2581 taatagacca cgatcgttat gctgatcata ccctaatgat cccagcaaga taatgtcctg
     2641 tcttctaaga tgtgcatcaa gcctggtaca tactgaaaac cctataaggt cctggataat
     2701 ttttgtttga ttattcattg aagaaacatt tattttccaa ttgtgtgaag tttttgactg
     2761 ttaataaaag aatctgtcaa ccatcaaagt ttaaacccgc tgatcagcct cgactgtgcc
     2821 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     2881 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     2941 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     3001 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     3061 ctggggctca ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     3121 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     3181 gtgtaaagcc tagggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     3241 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     3301 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     3361 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     3421 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3481 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3541 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3601 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3661 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3721 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3781 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3841 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3901 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3961 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4021 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4081 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acactcagtg
     4141 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4201 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4261 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4321 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4381 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4441 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4501 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4561 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4621 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     4681 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     4741 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     4801 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     4861 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     4921 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4981 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5041 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5101 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     5161 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     5221 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     5281 tcgatctccc gatcccctag ggtcgactct cagtacaatc tgctctgatg ccgcatagtt
     5341 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat
     5401 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag
     5461 gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac
     5521 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
     5581 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     5641 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     5701 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atc
//
