LOCUS       sequence-308713-        7616 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 7616)
  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 7616)
  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–G1397 DddAtox C
FEATURES             Location/Qualifiers
     source          1..7616
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        113..492
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        493..696
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     647..667
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     741..760
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        741..759
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             782..859
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MASVLTPLLLRGLTGSARRLPVPRAK"
     CDS             872..937
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3278..3529
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     rep_origin      3726..4154
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3813..3832)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4023..4044
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4163..4183)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4168..4497
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4348..4483
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4410..4429
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4545..4592
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4611..5009
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    5167..5288
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5204..5223)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5258..5277
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5337..5353)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5337..5353)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5350..5372)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5361..5377
                     /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(5385..5391,5392..5409,5410..5415))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5430..5451
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5568..5585)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5739..6327)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5819..5838)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6498..7289,7290..7358))
                     /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     7121..7140
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7359..7463)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7538..7557)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
       61 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga
      121 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
      181 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
      241 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      301 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      361 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      421 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      481 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      541 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      601 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      661 aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact
      721 gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc tggctagcac
      781 catggcctcc gttctgaccc cgctgctgct gcgtggcctg accggctccg ctcgtcgtct
      841 gccagttccg cgtgcgaaaa tccattccct ggactacaaa gaccatgacg gtgattataa
      901 agatcatgac atcgattaca aggatgacga tgacaagatg gacatcgcgg atctgcgtac
      961 cctgggttac agccagcagc agcaggagaa gatcaagcca aaggtgcgca gcaccgtggc
     1021 ccagcaccat gaagctctgg tgggtcacgg cttcacccac gcgcacatcg tggctctgag
     1081 ccagcaccca gccgcgctgg gtaccgtggc cgtgaaatat caggacatga ttgctgccct
     1141 gccagaggct acccatgaag cgattgtggg tgtgggcaag cagtggagcg gtgctcgtgc
     1201 gctggaggcc ctgctgaccg tggccggtga actgcgtggc ccgccgctgc agctggatac
     1261 cggccagctg ctgaaaatcg cgaaacgtgg cggtgtgacc gctgtggaag ctgtgcatgc
     1321 ctggcgtaat gctctgaccg gtgccccgct gaacctgacc ccggaacaag tggttgcgat
     1381 tgcgtcccat gatggtggta aacaagccct ggagaccgtt caagctctgc tgccagtgct
     1441 gtgccaggct catggtctga ccccgcagca ggtggtggct attgccagcc atgatggcgg
     1501 caagcaggct ctggagaccg tgcagcgtct gctgccggtg ctgtgccaag cccatggcct
     1561 gaccccgcag caagttgtgg ctattgccag caacggcggt ggcaaacagg cgctggaaac
     1621 cgtgcagcgc ctgttaccgg tgctgtgcca agcgcatggt ctgaccccgg agcaggtggt
     1681 ggccatcgct agcaacaacg gtggcaagca ggccctggaa accgtgcagg cgctgttgcc
     1741 ggtgctgtgc caggcccatg gcttaacccc ggaacaagtg gtggccattg cgagcaatgg
     1801 tggcggtaaa caggctctgg aaaccgtgca ggccctgctt ccggtgctgt gccaggccca
     1861 tgggctgacc ccggaacaag ttgtggctat cgccagcaac atcggcggca agcaggcgct
     1921 ggagaccgtt caggctctgt taccggtgct gtgccaggcg catggcctga ccccggaaca
     1981 ggtggtggcg atcgctagca acattggcgg taaacaggcc ctggagaccg ttcagcgcct
     2041 gctcccggtg ctgtgccagg cccatggtct gaccccggaa caggttgttg ctattgccag
     2101 caacaacggc ggcaagcagg ccctggagac cgtgcaggcg ctgctaccgg tgctgtgcca
     2161 ggcccatgga ctgaccccgg agcaggttgt ggccatcgcg tccaatggcg gtggcaaaca
     2221 ggctctggag accgttcagc gtctgttacc ggtgctgtgc caggcacatg gcctgacccc
     2281 ggagcaagtt gttgccatcg ctagcaacat tggtggcaag caggcgctgg aaaccgttca
     2341 gcgcctgcta ccggtgctgt gccaggctca tggcttaacc ccggagcagg ttgtcgccat
     2401 tgccagcaac aatggtggca aacaggctct ggaaactgtg caggccctgc taccggtgct
     2461 gtgccaggcc catgggttaa ccccggaaca ggttgtggcc attgcctcca ataacggtgg
     2521 caagcaggcg ctggaaacgg tgcaggctct gcttccggtg ctgtgccagg ctcatgggct
     2581 gaccccggag caagtggttg ctattgcgtc caacattggt ggcaaacagg ccctggaaac
     2641 cgttcaggcg ctgctcccgg tgctgtgcca ggcccatggg ctaaccccgg aacaggtggt
     2701 tgccattgcc tccaacaatg gtggcaagca ggccctggaa acggttcagc gtctgcttcc
     2761 ggtgctgtgc caggcccatg ggcttacccc gcagcaagtt gttgctatcg ccagcaatat
     2821 tggtggcaaa caggctctgg aaacggtgca gcgcctgcta ccggtgctgt gccaggctca
     2881 tggtttaacc ccgcagcagg tggttgcgat tgcctccaac aacggtggca agcaggcgct
     2941 ggaaactgtt cagcgtctgc tcccggtgct gtgccaggct cacggcctga ccccgcagca
     3001 agtggtggct atcgcctcca acggtggtgg tcgcccggct ctggaaagca ttgtggcgca
     3061 gctgagccgt ccagacccgg ccctggcggc tctgaccaat gatcacctgg tggccctggc
     3121 ctgcctgggt ggccgtccgg ctctggatgc cgtgaagaaa ggtctgggcg gatccgccat
     3181 tccagtgaag cgcggcgcta ccggtgaaac caaagtgttt accggtaaca gcaacagccc
     3241 gaagagcccg accaaaggcg gttgctctgg cggctcaact aatctgagcg acatcattga
     3301 gaaggagact gggaaacagc tggtcattca ggagtccatc ctgatgctgc ctgaggaggt
     3361 ggaggaagtg atcggcaaca agccagagtc tgacatcctg gtgcacaccg cctacgacga
     3421 gtccacagat gagaatgtga tgctgctgac ctctgacgcc cccgagtata agccttgggc
     3481 cctggtcatc caggattcta acggcgagaa taagatcaag atgctgtgat aaggggtctt
     3541 tgtcctctgt actgtctctc tccttgcccc taacccaaaa agcttcattt ttctgtgtag
     3601 gctgcacaag agccttgatt gaagatatat tctttctgaa cagtatttaa ggtttccaat
     3661 aaaatgtaca cccctcagaa cttctgaggc ggaaagaacc agctggggct ctagggggta
     3721 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3781 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     3841 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     3901 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     3961 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     4021 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     4081 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     4141 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     4201 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     4261 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4321 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     4381 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     4441 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     4501 ctcccgggag cttgtatatc cattttcgga tctgatcagc acgtgttgac aattaatcat
     4561 cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc atggccaagc
     4621 ctttgtctca agaagaatcc accctcattg aaagagcaac ggctacaatc aacagcatcc
     4681 ccatctctga agactacagc gtcgccagcg cagctctctc tagcgacggc cgcatcttca
     4741 ctggtgtcaa tgtatatcat tttactgggg gaccttgtgc agaactcgtg gtgctgggca
     4801 ctgctgctgc tgcggcagct ggcaacctga cttgtatcgt cgcgatcgga aatgagaaca
     4861 ggggcatctt gagcccctgc ggacggtgcc gacaggtgct tctcgatctg catcctggga
     4921 tcaaagccat agtgaaggac agtgatggac agccgacggc agttgggatt cgtgaattgc
     4981 tgccctctgg ttatgtgtgg gagggctaag cacttcgtgg ccgaggagca ggactgacac
     5041 gtgctacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     5101 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     5161 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     5221 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     5281 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     5341 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     5401 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     5461 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     5521 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     5581 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5641 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5701 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5761 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5821 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5881 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     5941 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     6001 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     6061 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6121 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     6181 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6241 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6301 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6361 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6421 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6481 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     6541 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6601 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6661 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6721 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6781 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6841 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6901 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6961 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     7021 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     7081 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     7141 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     7201 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     7261 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     7321 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     7381 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     7441 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
     7501 tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat
     7561 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcg
//
