LOCUS       sequence-374335-        5789 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mitochondrial v8 ZF-DdCBE in mammalian cells 
            (C-terminal DddAC, Right).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5789)
  AUTHORS   Willis JCW, Silva-Pinheiro P, Widdup L, Minczuk M, Liu DR
  TITLE     Compact zinc finger base editors that edit mitochondrial or nuclear 
            DNA in vitro and in vivo.
  JOURNAL   Nat Commun. 2022 Nov 23;13(1):7204. doi: 10.1038/s41467-022-34784-7.
  PUBMED    36418298
REFERENCE   2  (bases 1 to 5789)
  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 
            Commun."; date: "2022-11-23"; volume: "13(1):7204. doi"; pages: " 
            10.1038/s41467-022-34784-7"
COMMENT     Sequence Label: pB8_4-3i-ATP8_v8_AGKS
FEATURES             Location/Qualifiers
     source          1..5789
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1051..1074
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1573..1824
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     rep_origin      2021..2449
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2108..2127)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2318..2339
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2458..2478)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2463..2792
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2643..2778
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2705..2724
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2840..2887
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2906..3304
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    3462..3583
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3499..3518)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3553..3572
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3632..3648)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3632..3648)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3645..3667)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3656..3672
                     /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(3680..3686,3687..3704,3705..3710))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3725..3746
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3863..3880)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4034..4622)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4114..4133)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4793..5584,5585..5653))
                     /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     5416..5435
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5654..5758)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 accatgttgg ggtttgtggg tcgggtggcc gctgctccgg cctccggggc cttgcggaga
      961 ctcacccctt cagcgtcgct gcccccagct cagctcttac tgcgggccgc tccgacggcg
     1021 gtccatcctg tcagggacta tgcggcgcaa gattacaagg acgacgatga caaggtggat
     1081 gaaatgacca aaaagttcgg cacgctcacc attcacgaca ccgaaaaggg atcacttcag
     1141 aaaaagctgg aggaactgga gttggacgcc gctatggctg agaggcccta tgcttgccct
     1201 gagtgcggta aaagcttttc tcaagcctct aacttgatta gccacatccg cacccacacc
     1261 ggcgagaagc cttacgcatg tccagagtgt gggaagtcat tctctacgtc ccacagtttg
     1321 accgaacata tcagaaccca cactggcgaa aagccgtacg cttgcccaga atgcggtaag
     1381 tccttctctg aacgaagcca cctgcgcgaa cataccaaga tacacctgcg gggatccggc
     1441 ggaggaggtt ctggcggttc tggaggaagt gccattccag tgaagcgcgg cgctaccggt
     1501 gaaaccaaag tgtttatcgg taacagcaac agcccgaaga gcccgaccaa aggcggttgc
     1561 tctggcggct caactaatct gagcgacatc attgagaagg agactgggaa acagctggtc
     1621 attcaggagt ccatcctgat gctgcctgag gaggtggagg aagtgatcgg caacaagcca
     1681 gagtctgaca tcctggtgca caccgcctac gacgagtcca cagatgagaa tgtgatgctg
     1741 ctgacctctg acgcccccga gtataagcct tgggccctgg tcatccagga ttctaacggc
     1801 gagaataaga tcaagatgct gtgataaggg gtctttgtcc tctgtactgt ctctctcctt
     1861 gcccctaacc caaaaagctt catttttctg tgtaggctgc acaagagcct tgattgaaga
     1921 tatattcttt ctgaacagta tttaaggttt ccaataaaat gtacacccct cagaacttct
     1981 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     2041 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     2101 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     2161 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     2221 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     2281 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     2341 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     2401 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     2461 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2521 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     2581 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     2641 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     2701 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     2761 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     2821 tcggatctga tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata
     2881 atacgacaag gtgaggaact aaaccatggc caagcctttg tctcaagaag aatccaccct
     2941 cattgaaaga gcaacggcta caatcaacag catccccatc tctgaagact acagcgtcgc
     3001 cagcgcagct ctctctagcg acggccgcat cttcactggt gtcaatgtat atcattttac
     3061 tgggggacct tgtgcagaac tcgtggtgct gggcactgct gctgctgcgg cagctggcaa
     3121 cctgacttgt atcgtcgcga tcggaaatga gaacaggggc atcttgagcc cctgcggacg
     3181 gtgccgacag gtgcttctcg atctgcatcc tgggatcaaa gccatagtga aggacagtga
     3241 tggacagccg acggcagttg ggattcgtga attgctgccc tctggttatg tgtgggaggg
     3301 ctaagcactt cgtggccgag gagcaggact gacacgtgct acgagatttc gattccaccg
     3361 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     3421 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     3481 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     3541 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     3601 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     3661 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     3721 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     3781 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     3841 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     3901 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     3961 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4021 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4081 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4141 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4201 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     4261 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4321 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4381 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4441 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     4501 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4561 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     4621 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     4681 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     4741 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     4801 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     4861 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     4921 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     4981 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     5041 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     5101 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     5161 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     5221 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     5281 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     5341 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     5401 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     5461 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     5521 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     5581 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     5641 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     5701 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     5761 catttccccg aaaagtgcca cctgacgtc
//
