LOCUS       sequence-308097-        7928 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 7928)
  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 7928)
  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–G1397 DddAtox N
FEATURES             Location/Qualifiers
     source          1..7928
                     /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             3658..3909
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     rep_origin      4160..4588
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4247..4266)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4457..4478
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4597..4617)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4602..4931
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4782..4917
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4844..4863
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4979..5026
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5045..5443
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    5601..5722
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5638..5657)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5692..5711
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5771..5787)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5771..5787)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5784..5806)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5795..5811
                     /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(5819..5825,5826..5843,5844..5849))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5864..5885
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6002..6019)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6173..6761)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6253..6272)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6932..7723,7724..7792))
                     /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     7555..7574
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7793..7897)
                     /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 acagtgggag 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 cggcagctac gccctgggtc cgtatcagat tagcgccccg
     3361 cagctgccag cttacaatgg tcagaccgtg ggtaccttct actatgtgaa cgacgcgggc
     3421 ggtctggaga gcaaggtgtt tagcagcggc ggtccaaccc cgtacccaaa ctatgccaat
     3481 gccggtcatg tggagggtca gagcgccctg ttcatgcgtg ataacggcat cagcgagggt
     3541 ctggtgttcc acaacaaccc ggaaggcacc tgcggttttt gcgtgaacat gaccgagacc
     3601 ctgctgccgg aaaacgcgaa aatgaccgtg gtgccgccgg aaggttctgg cggctcaact
     3661 aatctgagcg acatcattga gaaggagact gggaaacagc tggtcattca ggagtccatc
     3721 ctgatgctgc ctgaggaggt ggaggaagtg atcggcaaca agccagagtc tgacatcctg
     3781 gtgcacaccg cctacgacga gtccacagat gagaatgtga tgctgctgac ctctgacgcc
     3841 cccgagtata agccttgggc cctggtcatc caggattcta acggcgagaa taagatcaag
     3901 atgctgtgat aaaccacgat cgttatgctg atcataccct aatgatccca gcaagataat
     3961 gtcctgtctt ctaagatgtg catcaagcct ggtacatact gaaaacccta taaggtcctg
     4021 gataattttt gtttgattat tcattgaaga aacatttatt ttccaattgt gtgaagtttt
     4081 tgactgttaa taaaagaatc tgtcaaccat caaacttctg aggcggaaag aaccagctgg
     4141 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     4201 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     4261 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     4321 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     4381 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     4441 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     4501 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     4561 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     4621 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4681 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4741 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     4801 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     4861 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4921 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat cagcacgtgt
     4981 tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta
     5041 aaccatggcc aagcctttgt ctcaagaaga atccaccctc attgaaagag caacggctac
     5101 aatcaacagc atccccatct ctgaagacta cagcgtcgcc agcgcagctc tctctagcga
     5161 cggccgcatc ttcactggtg tcaatgtata tcattttact gggggacctt gtgcagaact
     5221 cgtggtgctg ggcactgctg ctgctgcggc agctggcaac ctgacttgta tcgtcgcgat
     5281 cggaaatgag aacaggggca tcttgagccc ctgcggacgg tgccgacagg tgcttctcga
     5341 tctgcatcct gggatcaaag ccatagtgaa ggacagtgat ggacagccga cggcagttgg
     5401 gattcgtgaa ttgctgccct ctggttatgt gtgggagggc taagcacttc gtggccgagg
     5461 agcaggactg acacgtgcta cgagatttcg attccaccgc cgccttctat gaaaggttgg
     5521 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     5581 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     5641 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     5701 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     5761 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     5821 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     5881 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     5941 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     6001 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     6061 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     6121 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     6181 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     6241 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     6301 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     6361 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     6421 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     6481 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     6541 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     6601 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     6661 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     6721 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     6781 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     6841 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     6901 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     6961 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     7021 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     7081 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     7141 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     7201 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     7261 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     7321 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     7381 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     7441 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     7501 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     7561 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     7621 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     7681 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     7741 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     7801 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     7861 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     7921 ctgacgtc
//
