LOCUS       sequence-375376-        7927 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mitochondrial v8(HS1) ZF-DdCBE pair in mammalian cells
            (LB510-Nd1+RB54-Nd1); AAV packaging vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7927)
  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 7927)
  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: pAAV-Nd1
FEATURES             Location/Qualifiers
     source          1..7927
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    728..749
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(764..769,770..787,788..794)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    802..818
                     /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)."
     primer_bind     807..829
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     826..842
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     826..842
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   880..1009
                     /label=AAV2 ITR
     enhancer        1150..1453
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1454..1657
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1608..1628
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          1710..1842
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1825..1844
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             2007..2033
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2700..2948
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             2958..3014
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             3162..3185
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4116..4367
                     /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(4439..4459)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4857..4868)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4935..4952)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4941..5148
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5236..5365
                     /label=AAV2 ITR
     primer_bind     complement(5384..5401)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5384..5400)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5393..5415)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5542..5997
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5629..5648)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5839..5860
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6023..6126
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6127..6195,6196..6987)
                     /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     complement(6345..6364)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7158..7746
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7647..7666
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattac tgcagacgcc
       61 atgggcatta aaaaatgccc tcttgggtta tcaagagggt cattatattt gctgaggagt
      121 gccattacaa ctcgctccac gtcaactgtt caggacgttg tttggtagcg aagaattgtt
      181 tcgctgcttc aatcatttgc tctttgctca cgacacaatc cttcgataag tcgacctctc
      241 ctcctaattt gttttcgtag aacttctcgg ttcccccgga cgccgaattg cctttactta
      301 cgaagttttc accgatgtcc atatagtaga ccaccgcgat gtcgtttttt gcaaagatgt
      361 ttaactgcgg aaatccactt tcctcaaagg tcaggacgaa gttgttcgcg tcaaaagcgg
      421 ggcgacgcga caaaaattca accagagaat ccacttcgtc aatttcgatt tccccgtcga
      481 agtcgtcggc gtacattttt ttttcctcct tactctagta aaagttaaac aaaattattt
      541 gtagagggaa accgttgtag tctccctgaa tatatcatac gagccttatg catgcccgta
      601 aagttatcca gcaaccactc atagacctag ggcagcagat agggacgacg tggtgttagc
      661 tgtgctcccg aggatcctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
      721 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
      781 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
      841 accatgatta cgccagattt aattaaggcc ttaattaggc tgcgcgctcg ctcgctcact
      901 gaggccgccc gggcaaagcc cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc
      961 gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctt gtagttaatg
     1021 attaacccgc catgctactt atctacgtag ccatgctcta ggaagatcgg aattcgccct
     1081 taagctagct agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
     1141 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     1201 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     1261 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     1321 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     1381 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     1441 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
     1501 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     1561 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
     1621 taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatcct
     1681 gcagaagttg gtcgtgaggc actgggcagg taagtatcaa ggttacaaga caggtttaag
     1741 gagaccaata gaaactgggc ttgtcgagac agagaagact cttgcgtttc tgataggcac
     1801 ctattggtct tactgacatc cactttgcct ttctctccac aggtgtccag gcggccgcca
     1861 tgttgggttt tgtaggtcgg gtagcagctg ctcctgcatc aggtgcattg aggagactca
     1921 ccccttcagc gtcactgccc ccagctcagc tcttgctgcg ggccgctccg acggcggtcc
     1981 atcctgtcag ggactatgcg gcgcaatacc catatgatgt gcctgattac gctgttgacg
     2041 agatgactaa gaaatttgga acattgacaa tacatgatac agagaaaggt agcttgcaaa
     2101 agaaattgga agagttggaa cttgatgctg caatggcaga acgaccttat gcctgtcctg
     2161 aatgcggaaa gtcctttagc acgtctgggg agttggttag gcatattcgc acccatacag
     2221 gagaaaagcc ctacgcgtgt ccggaatgtg gcaagagttt ttccaggact gatacgctgc
     2281 gcgatcacat aaggacacat actggggaga aaccttatgc ttgtcccgaa tgtggaaaaa
     2341 gcttctcaac aagcggagaa ctcgtccgac acattagaac tcacacgggt gagaagccat
     2401 atgcatgccc cgagtgtgga aagagtttta gtcggagcga ccatctcact actcatatac
     2461 ggacacacac cggagagaaa ccatatgcgt gcccggaatg tgggaaaagt ttcagcacaa
     2521 gcggctctct cgttagacac acaaaaattc atttgagagg tagcggaggt ggtggaagcg
     2581 gaggcagcgg gggttctgct attccagtta aaagaggagc tactggtgaa acaaaagttt
     2641 ttataggtaa aagcaacagc cctaagtctc ctacaaaagg tggatgtagc ggaggttcta
     2701 ctaatctgag cgacatcatt gagaaggaga ctgggaaaca gctggtcatt caggagtcca
     2761 tcctgatgct gcctgaggag gtggaggaag tgatcggcaa caagccagag tctgacatcc
     2821 tggtgcacac cgcctacgac gagtccacag atgagaatgt gatgctgctg acctctgacg
     2881 cccccgagta taagccttgg gccctggtca tccaggattc taacggcgag aataagatca
     2941 agatgctggg aagcggagct actaacttca gcctgctgaa gcaggctgga gacgtggagg
     3001 agaaccctgg acctatgctc ggattcgttg gaagagttgc tgcagcacca gctagcggag
     3061 ctctcagacg gttgacacct agcgcttctt tgcctcctgc acaattgctt ttgagagcag
     3121 cacctacagc tgttcaccca gttcgagatt acgctgcaca ggattacaag gacgacgatg
     3181 acaaggtgga tgaaatgacc aaaaagttcg gcacgctcac cattcacgac accgaaaagg
     3241 gatcacttca gaaaaagctg gaggaactgg agttggacgc cgcttctggc ggctcaggat
     3301 cttacgcttt gggtccttat caaatttctg caccacaact tccagcttat aatggacaaa
     3361 cagtgggaac attctactat gttaacgacg caggaggact tgaatcaaaa gtgtttagct
     3421 ccggtggacc aacaccttac cctaactatg caaatgcagg acatgtcgag ggacagtctg
     3481 ctctctttat gcgagacaat ggtattagtg aaggtttggt ttttcacaac aacccagaag
     3541 gaacttgtgg tttctgtgtt aatatgatag agacgttgct cccagaaaac gctaaaatga
     3601 ctgtcgtccc tccaaaagga ggatccggcg gaggaggttc tggcggttct ggaggaagta
     3661 tggctgagag gccctatgct tgccctgagt gcggtaaaag cttttctaca agcggctctc
     3721 tcgttagaca catccgcacc cacaccggcg agaagcctta cgcatgtcca gagtgtggga
     3781 agtcattctc taggcgggac gaactgaacg tgcatatcag aacccacact ggcgaaaagc
     3841 cgtacgcttg cccagaatgc ggtaagtcct tctctacgtc tggggagttg gttaggcaca
     3901 tcagaaccca tacaggcgaa aaaccctacg cctgcccaga gtgcggcaaa tccttttcac
     3961 agaaatcttc tttgatagct catattagga ctcatacggg agagaagccg tacgcgtgtc
     4021 ctgagtgcgg gaagtctttc tctcaaaaga gcagtctcat tgcccatacc aagatacacc
     4081 tgcggagcgg aggatccgga ggatctggag gcagcaccaa cctttctgat attatcgaaa
     4141 aagaaacagg aaagcaactt gttatacaag aaagcattct tatgcttccc gaagaagttg
     4201 aagaggtaat tggaaataaa cccgaaagcg atattcttgt acatacagct tatgatgaaa
     4261 gcaccgacga aaacgtcatg ctccttacaa gcgatgcacc tgaatacaaa ccatgggcat
     4321 tggttattca agacagcaat ggagaaaaca aaataaaaat gttgtgataa ggtaccgatg
     4381 gatccaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     4441 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     4501 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     4561 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     4621 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     4681 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     4741 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     4801 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     4861 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     4921 gtcttcgaga tctgcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     4981 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     5041 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     5101 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggact cgagttaagg
     5161 gcgaattccc gataaggatc ttcctagagc atggctacgt agataagtag catggcgggt
     5221 taatcattaa ctacaaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     5281 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     5341 cctcagtgag cgagcgagcg cgcagcctta attaacctaa ttcactggcc gtcgttttac
     5401 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     5461 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     5521 gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     5581 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     5641 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     5701 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     5761 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     5821 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     5881 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     5941 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg
     6001 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     6061 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaagg
     6121 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     6181 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     6241 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     6301 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     6361 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     6421 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     6481 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     6541 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     6601 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     6661 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     6721 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     6781 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     6841 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     6901 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     6961 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     7021 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     7081 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     7141 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     7201 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     7261 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     7321 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     7381 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     7441 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     7501 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     7561 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     7621 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     7681 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     7741 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     7801 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     7861 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     7921 gcggaag
//
