LOCUS       sequence-374326-        6077 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mitochondrial v8 ZF-DdCBE in mammalian cells 
            (C-terminal DddAN, Left).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6077)
  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 6077)
  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: pB5_R8-3i-ATP8_v8_AGKS
FEATURES             Location/Qualifiers
     source          1..6077
                     /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             1807..2058
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     rep_origin      2309..2737
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2396..2415)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2606..2627
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2746..2766)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2751..3080
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2931..3066
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2993..3012
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3128..3175
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3194..3592
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    3750..3871
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3787..3806)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3841..3860
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3920..3936)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3920..3936)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3933..3955)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3944..3960
                     /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(3968..3974,3975..3992,3993..3998))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4013..4034
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4151..4168)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4322..4910)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4402..4421)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5081..5872,5873..5941))
                     /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     5704..5723
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5942..6046)
                     /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 tacctccggg tccctgagcc ggcacatccg cacccacacc
     1261 ggcgagaagc cttacgcatg tccagagtgt gggaagtcat tctctcaatc cggttccctt
     1321 acccgccata tcagaaccca cactggcgaa aagccgtacg cttgcccaga atgcggtaag
     1381 tccttctctc ggagcgacgc actctcacaa cataccaaga tacacctgcg gggatccggc
     1441 ggaggaggtt ctggcggttc tggaggaagt ggcagctacg ccctgggtcc gtatcagatt
     1501 agcgccccgc agctgccagc ttacaatggt cagaccgtgg gtaccttcta ctatgtgaac
     1561 gacgcgggcg gtctggagag caaggtgttt agcagcggcg gtccaacccc gtacccaaac
     1621 tatgccaatg ccggtcatgt ggagggtcag agcgccctgt tcatgcgtga taacggcatc
     1681 agcgagggtc tggtgttcca caacaacccg gaaggcacct gcggtttttg cgtgaacatg
     1741 atcgagaccc tgctgccgga aaacgcgaaa atgaccgtgg tgccgccgaa aggttctggc
     1801 ggctcaacta atctgagcga catcattgag aaggagactg ggaaacagct ggtcattcag
     1861 gagtccatcc tgatgctgcc tgaggaggtg gaggaagtga tcggcaacaa gccagagtct
     1921 gacatcctgg tgcacaccgc ctacgacgag tccacagatg agaatgtgat gctgctgacc
     1981 tctgacgccc ccgagtataa gccttgggcc ctggtcatcc aggattctaa cggcgagaat
     2041 aagatcaaga tgctgtgata aaccacgatc gttatgctga tcatacccta atgatcccag
     2101 caagataatg tcctgtcttc taagatgtgc atcaagcctg gtacatactg aaaaccctat
     2161 aaggtcctgg ataatttttg tttgattatt cattgaagaa acatttattt tccaattgtg
     2221 tgaagttttt gactgttaat aaaagaatct gtcaaccatc aaacttctga ggcggaaaga
     2281 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     2341 ggtgtggtgg tcacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     2401 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2461 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     2521 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     2581 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     2641 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     2701 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     2761 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2821 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2881 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     2941 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3001 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3061 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     3121 agcacgtgtt gacaattaat catcggcata gtatatcggc atagtataat acggcaaggt
     3181 gaggaactaa accatggcca agcctttgtc tcaagaagaa tccaccctca ttgaaagagc
     3241 aacggctaca atcaacagca tccccatctc tgaagactac agcgtcgcca gcgcagctct
     3301 ctctagcgac ggccgcatct tcactggtgt caatgtatat cattttactg ggggaccttg
     3361 tgcagaactc gtggtgctgg gcactgctgc tgctgcggca gctggcaacc tgacttgtat
     3421 cgtcgcgatc ggaaatgaga acaggggcat cttgagcccc tgcggacggt gccgacaggt
     3481 gcttctcgat ctgcatcctg ggatcaaagc catagtgaag gacagtgatg gacagccgac
     3541 ggcagttggg attcgtgaat tgctgccctc tggttatgtg tgggagggct aagcacttcg
     3601 tggccgagga gcaggactga cacgtgctac gagatttcga ttccaccgcc gccttctatg
     3661 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     3721 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     3781 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     3841 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     3901 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     3961 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     4021 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     4081 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     4141 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     4201 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     4261 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     4321 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4381 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4441 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4501 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4561 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4621 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4681 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4741 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     4801 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4861 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4921 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4981 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     5041 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     5101 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     5161 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     5221 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     5281 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     5341 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     5401 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     5461 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     5521 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     5581 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     5641 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     5701 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5761 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     5821 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     5881 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     5941 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6001 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     6061 aagtgccacc tgacgtc
//
