LOCUS       sequence-311017-        7658 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 7658)
  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 7658)
  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 N
FEATURES             Location/Qualifiers
     source          1..7658
                     /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             3334..3585
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     rep_origin      3782..4210
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3869..3888)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4079..4100
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4219..4239)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4224..4553
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4404..4539
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4466..4485
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4601..4648
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4667..5065
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    5223..5344
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5260..5279)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5314..5333
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5393..5409)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5393..5409)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5406..5428)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5417..5433
                     /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(5441..5447,5448..5465,5466..5471))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5486..5507
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5624..5641)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5795..6383)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5875..5894)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6554..7345,7346..7414))
                     /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     7177..7196
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7415..7519)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7594..7613)
                     /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 ggcggatccg gcagctacgc cctgggtccg tatcagatta gcgccccgca gctgccagct
     3241 tacaatggtc agaccgtggg taccttctac tatgtgaacg acgcgggcgg tctggagagc
     3301 aaggtgttta gcagcggcgg ttctggcggc tcaactaatc tgagcgacat cattgagaag
     3361 gagactggga aacagctggt cattcaggag tccatcctga tgctgcctga ggaggtggag
     3421 gaagtgatcg gcaacaagcc agagtctgac atcctggtgc acaccgccta cgacgagtcc
     3481 acagatgaga atgtgatgct gctgacctct gacgcccccg agtataagcc ttgggccctg
     3541 gtcatccagg attctaacgg cgagaataag atcaagatgc tgtgataagg ggtctttgtc
     3601 ctctgtactg tctctctcct tgcccctaac ccaaaaagct tcatttttct gtgtaggctg
     3661 cacaagagcc ttgattgaag atatattctt tctgaacagt atttaaggtt tccaataaaa
     3721 tgtacacccc tcagaacttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     3781 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3841 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3901 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     3961 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     4021 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     4081 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     4141 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     4201 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     4261 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     4321 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4381 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4441 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     4501 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     4561 cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc
     4621 atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatgg ccaagccttt
     4681 gtctcaagaa gaatccaccc tcattgaaag agcaacggct acaatcaaca gcatccccat
     4741 ctctgaagac tacagcgtcg ccagcgcagc tctctctagc gacggccgca tcttcactgg
     4801 tgtcaatgta tatcatttta ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc
     4861 tgctgctgcg gcagctggca acctgacttg tatcgtcgcg atcggaaatg agaacagggg
     4921 catcttgagc ccctgcggac ggtgccgaca ggtgcttctc gatctgcatc ctgggatcaa
     4981 agccatagtg aaggacagtg atggacagcc gacggcagtt gggattcgtg aattgctgcc
     5041 ctctggttat gtgtgggagg gctaagcact tcgtggccga ggagcaggac tgacacgtgc
     5101 tacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     5161 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     5221 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     5281 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     5341 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     5401 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     5461 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5521 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5581 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5641 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5701 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5761 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5821 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5881 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5941 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     6001 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     6061 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     6121 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     6181 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     6241 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     6301 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     6361 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6421 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     6481 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     6541 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6601 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6661 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6721 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6781 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6841 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     6901 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6961 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     7021 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     7081 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     7141 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     7201 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     7261 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     7321 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     7381 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     7441 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     7501 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg
     7561 gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     7621 aagccagtat ctgctccctg cttgtgtgtt ggaggtcg
//
