LOCUS       sequence-375488-        5909 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses the left-side half ND6-UGI-NES in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5909)
  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 5909)
  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_left
FEATURES             Location/Qualifiers
     source          1..5909
                     /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..2697
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             2710..2736
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     primer_bind     complement(2964..2981)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2970..3194
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3265..3281)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3265..3281)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3278..3300)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3289..3305
                     /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(3313..3319,3320..3337,3338..3343))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3358..3379
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3496..3513)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3667..4255)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3747..3766)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4426..5217,5218..5286))
                     /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     5049..5068
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5287..5391)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5470..5489)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(5661..5909,1..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 gccagaccgt 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 gatgctggga
     2701 tctctgcagc tgcctccact tgaaagactg acactgtaat agaccacgat cgttatgctg
     2761 atcataccct aatgatccca gcaagataat gtcctgtctt ctaagatgtg catcaagcct
     2821 ggtacatact gaaaacccta taaggtcctg gataattttt gtttgattat tcattgaaga
     2881 aacatttatt ttccaattgt gtgaagtttt tgactgttaa taaaagaatc tgtcaaccat
     2941 caaagtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     3001 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     3061 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     3121 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     3181 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctcaatac cgtcgacctc
     3241 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     3301 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctagg gtgcctaatg
     3361 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     3421 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     3481 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     3541 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     3601 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3661 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3721 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3781 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3841 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3901 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3961 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4021 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4081 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     4141 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4201 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4261 tcctttgatc ttttctacgg ggtctgacac tcagtggaac gaaaactcac gttaagggat
     4321 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     4381 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     4441 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     4501 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     4561 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     4621 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     4681 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     4741 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     4801 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     4861 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     4921 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     4981 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     5041 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     5101 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     5161 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     5221 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     5281 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     5341 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     5401 ccgaaaagtg ccacctgacg tcgacggatc gggagatcga tctcccgatc ccctagggtc
     5461 gactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg
     5521 tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt
     5581 gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt
     5641 acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac
     5701 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     5761 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     5821 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     5881 tgcccacttg gcagtacatc aagtgtatc
//
