LOCUS       sequence-305067-        7887 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 7887)
  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 7887)
  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 left TALE–G1333 DddAtox C
FEATURES             Location/Qualifiers
     source          1..7887
                     /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             997..1023
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1027..1053
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1057..1083
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3616..3867
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     rep_origin      4118..4546
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4205..4224)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4415..4436
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4555..4575)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4560..4890
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4741..4876
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4803..4822
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4938..4985
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5004..5402
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    5560..5681
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5597..5616)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5651..5670
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5730..5746)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5730..5746)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5743..5765)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5754..5770
                     /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(5778..5784,5785..5802,5803..5808))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5823..5844
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5961..5978)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6132..6720)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6212..6231)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6891..7682,7683..7751))
                     /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     7514..7533
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7752..7856)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 accatggccc tgtcccgtgc ggtttgtggc acctcccgtc aactggctcc ggttctgggt
      961 tatctgggtt cccgtcaaaa acactccctg ccggactacc cgtatgatgt tccggattac
     1021 gctggctacc catacgacgt cccagactac gctggctacc catacgacgt cccagactac
     1081 gctatggaca tcgcggatct gcgtaccctg ggttacagcc agcagcagca ggagaagatc
     1141 aagccgaagg tgcgcagcac cgtggctcag caccacgaag ccctggtggg ccacggtttc
     1201 acccacgctc acatcgtggc cctgagccag cacccagccg cgctgggcac cgtggccgtg
     1261 aaatatcagg acatgattgc tgccctgcca gaggccaccc atgaagctat tgtgggcgtg
     1321 ggcaagcagt ggagcggtgc tcgtgcgctg gaggcgctgc tgaccgtggc tggtgaactg
     1381 cgtggtccgc cgctgcagct ggataccggt cagctgctga aaatcgcgaa acgtggcggt
     1441 gtgaccgcgg tggaagccgt gcatgcttgg cgtaatgctc tgaccggtgc gccgctgaac
     1501 ctgaccccgg aacaggtggt ggcaatcgca agcaataatg gtggtaaaca ggctctggaa
     1561 accgtgcaag ctctgctgcc agttctgtgc caggctcatg gtctgacccc gcagcaggtg
     1621 gtggctattg ccagccatga tggcggcaag caggccctgg agaccgtgca gcgtctgctg
     1681 ccggtgctgt gccaggccca tggcctgacc ccgcagcaag ttgtggctat tgccagcaac
     1741 ggcggtggca aacaggctct ggagaccgtg cagcgcctgt taccggtgct gtgccaagcc
     1801 catggtctga ccccggagca ggtggtggcg atcgctagca acattggtgg caagcaggcc
     1861 ctggaaaccg tgcaggcgct gttgccggtg ctgtgccaag cccatgggct gaccccggaa
     1921 caagttgttg ccattgccag caacaatggt ggcaaacagg ctctggaaac tgtgcaggcc
     1981 ctgcttccgg tgctgtgcca ggcccatgga ttaaccccgg aacaagttgt ggctattgcg
     2041 agcaatggcg gcggcaagca ggcgctggaa accgtgcagg ctctgttacc ggtgctgtgc
     2101 caggcgcatg gcctgacccc ggagcaagtg gtggccatcg ctagcaacat tggcggtaaa
     2161 caggcgctgg agaccgttca gcgtctgtta ccggtgctgt gccaggcaca tggccttacc
     2221 ccggaacaag ttgtggccat tgccagcaac atcggcggca agcaggccct ggaaacggtg
     2281 caggcgctgc tcccggtgct gtgccaggcc catgggttaa ccccggaaca agtggttgct
     2341 attgctagcc atgatggcgg taaacaggcc ctggagaccg ttcagcgcct gctaccggtg
     2401 ctgtgccagg cccatggttt aaccccggaa caggttgttg cgattgctag ccacgatggc
     2461 ggcaagcagg ctctggagac cgttcaggcc ctgctcccgg tgctgtgcca ggcccatggg
     2521 cttaccccgg agcaagttgt tgctattgcc tccaatattg gcggtaaaca ggcgctggaa
     2581 accgttcagg ctctgcttcc ggtgctgtgc caggctcatg gcctcacccc ggaacaagtt
     2641 gtggcgattg cgtcccatga tggcggcaag caggccctgg aaactgtgca ggcgctgcta
     2701 ccggtgctgt gccaggccca tgggctaacc ccggaacagg tggttgcgat tgctagcaac
     2761 aacggcggta aacaggctct ggagactgtt cagcgtctgc ttccggtgct gtgccaggct
     2821 catgggctga ccccgcagca agtggttgct attgccagca atggcggtgg caagcaggcg
     2881 ctggagactg ttcagcgcct gctcccggtg ctgtgccagg ctcatggttt aaccccggag
     2941 caggttgtgg cgatcgccag caatggtggc ggtaaacagg ctctggaaac ggtgcaggcc
     3001 ctgctcccgg tgctgtgcca ggctcatgga ctgaccccgg agcaagttgt tgccattgcg
     3061 tcccacgacg gcggcaagca ggcgctggag acggtgcagg ctctgctccc ggtgctgtgc
     3121 caggctcacg gtctgacccc gcaacaggtg gtggcaatcg caagcaacgg tggtggtcgt
     3181 ccggcactgg agagcattgt ggcgcagctg agccgtccag acccggccct ggcggctctg
     3241 accaacgatc acctggtggc gctggcttgc ctgggcggtc gtccggccct ggatgcggtg
     3301 aagaaaggcc tgggtggatc cccaaccccg tacccaaact atgccaatgc cggtcatgtg
     3361 gagggtcaga gcgccctgtt catgcgtgat aacggcatca gcgagggtct ggtgttccac
     3421 aacaacccgg aaggcacctg cggtttttgc gtgaacatga ccgagaccct gctgccggaa
     3481 aacgcgaaaa tgaccgtggt gccgccggaa ggtgccattc cagtgaagcg cggcgctacc
     3541 ggtgaaacca aagtgtttac cggtaacagc aacagcccga agagcccgac caaaggcggt
     3601 tgctctggcg gctcaactaa tctgagcgac atcattgaga aggagactgg gaaacagctg
     3661 gtcattcagg agtccatcct gatgctgcct gaggaggtgg aggaagtgat cggcaacaag
     3721 ccagagtctg acatcctggt gcacaccgcc tacgacgagt ccacagatga gaatgtgatg
     3781 ctgctgacct ctgacgcccc cgagtataag ccttgggccc tggtcatcca ggattctaac
     3841 ggcgagaata agatcaagat gctgtgataa accacgatcg ttatgctgat cataccctaa
     3901 tgatcccagc aagataatgt cctgtcttct aagatgtgca tcaagcctgg tacatactga
     3961 aaaccctata aggtcctgga taatttttgt ttgattattc attgaagaaa catttatttt
     4021 ccaattgtgt gaagtttttg actgttaata aaagaatctg tcaaccatca aacttctgag
     4081 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     4141 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     4201 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     4261 gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     4321 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4381 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     4441 acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg gatttcggcc
     4501 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     4561 tgtgtcagtt agggtgtgga aagtccccag gctccccagg caggcagaag tatgcaaagc
     4621 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     4681 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     4741 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     4801 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     4861 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     4921 ggatctgatc agcacgtgtt gacaattaat catcggcata gtatatcggc atagtataat
     4981 acgacaaggt gaggaactaa accatggcca agcctttgtc tcaagaagaa tccaccctca
     5041 ttgaaagagc aacggctaca atcaacagca tccccatctc tgaagactac agcgtcgcca
     5101 gcgcagctct ctctagcgac ggccgcatct tcactggtgt caatgtatat cattttactg
     5161 ggggaccttg tgcagaactc gtggtgctgg gcactgctgc tgctgcggca gctggcaacc
     5221 tgacttgtat cgtcgcgatc ggaaatgaga acaggggcat cttgagcccc tgcggacggt
     5281 gtcgacaggt gcttctcgat ctgcatcctg ggatcaaagc gatagtgaag gacagtgatg
     5341 gacagccgac ggcagttggg attcgtgaat tgctgccctc tggttatgtg tgggagggct
     5401 aagcacttcg tggccgagga gcaggactga cacgtgctac gagatttcga ttccaccgcc
     5461 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     5521 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     5581 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     5641 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     5701 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     5761 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     5821 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     5881 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     5941 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     6001 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     6061 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     6121 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     6181 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     6241 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     6301 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag
     6361 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     6421 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     6481 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     6541 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     6601 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     6661 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     6721 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     6781 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     6841 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     6901 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     6961 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     7021 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     7081 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     7141 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     7201 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     7261 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     7321 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     7381 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     7441 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     7501 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     7561 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     7621 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     7681 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     7741 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     7801 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     7861 tttccccgaa aagtgccacc tgacgtc
//
