LOCUS       sequence-308712-        7808 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses the right-side ND4 TALE–DddAtox half  in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7808)
  AUTHORS   Mok BY, de Moraes MH, Zeng J, Bosch DE, Kotrys AV, Raguram A, Hsu F,
            Radey MC, Peterson SB, Mootha VK, Mougous JD, Liu DR
  TITLE     A bacterial cytidine deaminase toxin enables CRISPR-free 
            mitochondrial base editing.
  JOURNAL   Nature. 2020 Jul 8. pii: 10.1038/s41586-020-2477-4. doi: 
            10.1038/s41586-020-2477-4.
  PUBMED    32641830
REFERENCE   2  (bases 1 to 7808)
  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. 
            2020 Jul 8. pii: 10.1038/s41586-020-2477-4. doi: 
            10.1038/s41586-020-2477-4."
COMMENT     Sequence Label: ND4 right TALE–G1333 DddAtox C
FEATURES             Location/Qualifiers
     source          1..7808
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        127..506
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        507..710
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     661..681
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     755..774
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        755..773
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             796..873
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MASVLTPLLLRGLTGSARRLPVPRAK"
     CDS             886..951
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3484..3735
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     rep_origin      3932..4360
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4019..4038)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4229..4250
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4369..4389)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4374..4703
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4554..4689
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4616..4635
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4751..4798
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4817..5215
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    5373..5494
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5410..5429)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5464..5483
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5543..5559)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5543..5559)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5556..5578)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5567..5583
                     /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(5591..5597,5598..5615,5616..5621))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5636..5657
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5774..5791)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5945..6533)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6025..6044)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6704..7495,7496..7564))
                     /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     7327..7346
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7565..7669)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7744..7763)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
       61 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
      121 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
      181 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      241 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      301 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      361 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      421 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      481 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      541 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      601 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      661 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa
      721 ctagagaacc cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc
      781 aagctggcta gcaccatggc ctccgttctg accccgctgc tgctgcgtgg cctgaccggc
      841 tccgctcgtc gtctgccagt tccgcgtgcg aaaatccatt ccctggacta caaagaccat
      901 gacggtgatt ataaagatca tgacatcgat tacaaggatg acgatgacaa gatggacatc
      961 gcggatctgc gtaccctggg ttacagccag cagcagcagg agaagatcaa gccaaaggtg
     1021 cgcagcaccg tggcccagca ccatgaagct ctggtgggtc acggcttcac ccacgcgcac
     1081 atcgtggctc tgagccagca cccagccgcg ctgggtaccg tggccgtgaa atatcaggac
     1141 atgattgctg ccctgccaga ggctacccat gaagcgattg tgggtgtggg caagcagtgg
     1201 agcggtgctc gtgcgctgga ggccctgctg accgtggccg gtgaactgcg tggcccgccg
     1261 ctgcagctgg ataccggcca gctgctgaaa atcgcgaaac gtggcggtgt gaccgctgtg
     1321 gaagctgtgc atgcctggcg taatgctctg accggtgccc cgctgaacct gaccccggaa
     1381 caagtggttg cgattgcgtc ccatgatggt ggtaaacaag ccctggagac cgttcaagct
     1441 ctgctgccag tgctgtgcca ggctcatggt ctgaccccgc agcaggtggt ggctattgcc
     1501 agccatgatg gcggcaagca ggctctggag accgtgcagc gtctgctgcc ggtgctgtgc
     1561 caagcccatg gcctgacccc gcagcaagtt gtggctattg ccagcaacgg cggtggcaaa
     1621 caggcgctgg aaaccgtgca gcgcctgtta ccggtgctgt gccaagcgca tggtctgacc
     1681 ccggagcagg tggtggccat cgctagcaac aacggtggca agcaggccct ggaaaccgtg
     1741 caggcgctgt tgccggtgct gtgccaggcc catggcttaa ccccggaaca agtggtggcc
     1801 attgcgagca atggtggcgg taaacaggct ctggaaaccg tgcaggccct gcttccggtg
     1861 ctgtgccagg cccatgggct gaccccggaa caagttgtgg ctatcgccag caacatcggc
     1921 ggcaagcagg cgctggagac cgttcaggct ctgttaccgg tgctgtgcca ggcgcatggc
     1981 ctgaccccgg aacaggtggt ggcgatcgct agcaacattg gcggtaaaca ggccctggag
     2041 accgttcagc gcctgctccc ggtgctgtgc caggcccatg gtctgacccc ggaacaggtt
     2101 gttgctattg ccagcaacaa cggcggcaag caggccctgg agaccgtgca ggcgctgcta
     2161 ccggtgctgt gccaggccca tggactgacc ccggagcagg ttgtggccat cgcgtccaat
     2221 ggcggtggca aacaggctct ggagaccgtt cagcgtctgt taccggtgct gtgccaggca
     2281 catggcctga ccccggagca agttgttgcc atcgctagca acattggtgg caagcaggcg
     2341 ctggaaaccg ttcagcgcct gctaccggtg ctgtgccagg ctcatggctt aaccccggag
     2401 caggttgtcg ccattgccag caacaatggt ggcaaacagg ctctggaaac tgtgcaggcc
     2461 ctgctaccgg tgctgtgcca ggcccatggg ttaaccccgg aacaggttgt ggccattgcc
     2521 tccaataacg gtggcaagca ggcgctggaa acggtgcagg ctctgcttcc ggtgctgtgc
     2581 caggctcatg ggctgacccc ggagcaagtg gttgctattg cgtccaacat tggtggcaaa
     2641 caggccctgg aaaccgttca ggcgctgctc ccggtgctgt gccaggccca tgggctaacc
     2701 ccggaacagg tggttgccat tgcctccaac aatggtggca agcaggccct ggaaacggtt
     2761 cagcgtctgc ttccggtgct gtgccaggcc catgggctta ccccgcagca agttgttgct
     2821 atcgccagca atattggtgg caaacaggct ctggaaacgg tgcagcgcct gctaccggtg
     2881 ctgtgccagg ctcatggttt aaccccgcag caggtggttg cgattgcctc caacaacggt
     2941 ggcaagcagg cgctggaaac tgttcagcgt ctgctcccgg tgctgtgcca ggctcacggc
     3001 ctgaccccgc agcaagtggt ggctatcgcc tccaacggtg gtggtcgccc ggctctggaa
     3061 agcattgtgg cgcagctgag ccgtccagac ccggccctgg cggctctgac caatgatcac
     3121 ctggtggccc tggcctgcct gggtggccgt ccggctctgg atgccgtgaa gaaaggtctg
     3181 ggcggatccc caaccccgta cccaaactat gccaatgccg gtcatgtgga gggtcagagc
     3241 gccctgttca tgcgtgataa cggcatcagc gagggtctgg tgttccacaa caacccggaa
     3301 ggcacctgcg gtttttgcgt gaacatgacc gagaccctgc tgccggaaaa cgcgaaaatg
     3361 accgtggtgc cgccggaagg tgccattcca gtgaagcgcg gcgctaccgg tgaaaccaaa
     3421 gtgtttaccg gtaacagcaa cagcccgaag agcccgacca aaggcggttg ctctggcggc
     3481 tcaactaatc tgagcgacat cattgagaag gagactggga aacagctggt cattcaggag
     3541 tccatcctga tgctgcctga ggaggtggag gaagtgatcg gcaacaagcc agagtctgac
     3601 atcctggtgc acaccgccta cgacgagtcc acagatgaga atgtgatgct gctgacctct
     3661 gacgcccccg agtataagcc ttgggccctg gtcatccagg attctaacgg cgagaataag
     3721 atcaagatgc tgtgataagg ggtctttgtc ctctgtactg tctctctcct tgcccctaac
     3781 ccaaaaagct tcatttttct gtgtaggctg cacaagagcc ttgattgaag atatattctt
     3841 tctgaacagt atttaaggtt tccaataaaa tgtacacccc tcagaacttc tgaggcggaa
     3901 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     3961 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     4021 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4081 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4141 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4201 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4261 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     4321 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     4381 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4441 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4501 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     4561 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4621 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     4681 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     4741 atcagcacgt gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa
     4801 ggtgaggaac taaaccatgg ccaagccttt gtctcaagaa gaatccaccc tcattgaaag
     4861 agcaacggct acaatcaaca gcatccccat ctctgaagac tacagcgtcg ccagcgcagc
     4921 tctctctagc gacggccgca tcttcactgg tgtcaatgta tatcatttta ctgggggacc
     4981 ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg gcagctggca acctgacttg
     5041 tatcgtcgcg atcggaaatg agaacagggg catcttgagc ccctgcggac ggtgccgaca
     5101 ggtgcttctc gatctgcatc ctgggatcaa agccatagtg aaggacagtg atggacagcc
     5161 gacggcagtt gggattcgtg aattgctgcc ctctggttat gtgtgggagg gctaagcact
     5221 tcgtggccga ggagcaggac tgacacgtgc tacgagattt cgattccacc gccgccttct
     5281 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     5341 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     5401 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5461 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     5521 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     5581 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     5641 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5701 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5761 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5821 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5881 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5941 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6001 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6061 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6121 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6181 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6241 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6301 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6361 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6421 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6481 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6541 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6601 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6661 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6721 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6781 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6841 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6901 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6961 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     7021 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7081 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7141 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7201 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7261 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7321 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7381 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7441 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7501 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7561 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7621 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7681 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct
     7741 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
     7801 ggaggtcg
//
