LOCUS       sequence-307656-        7694 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses left-side TALE–DddAtox half ND4-DdCBE in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7694)
  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 7694)
  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-DdCBE- left side TALE
FEATURES             Location/Qualifiers
     source          1..7694
                     /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             3424..3675
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     rep_origin      3926..4354
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4013..4032)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4223..4244
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4363..4383)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4368..4697
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4548..4683
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4610..4629
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4745..4792
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4811..5209
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    5367..5488
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5404..5423)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5458..5477
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5537..5553)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5537..5553)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5550..5572)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5561..5577
                     /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(5585..5591,5592..5609,5610..5615))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5630..5651
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5768..5785)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5939..6527)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6019..6038)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6698..7489,7490..7558))
                     /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     7321..7340
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7559..7663)
                     /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 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 cgccattcca gtgaagcgcg gcgctaccgg tgaaaccaaa
     3361 gtgtttaccg gtaacagcaa cagcccgaag agcccgacca aaggcggttg ctctggcggc
     3421 tcaactaatc tgagcgacat cattgagaag gagactggga aacagctggt cattcaggag
     3481 tccatcctga tgctgcctga ggaggtggag gaagtgatcg gcaacaagcc agagtctgac
     3541 atcctggtgc acaccgccta cgacgagtcc acagatgaga atgtgatgct gctgacctct
     3601 gacgcccccg agtataagcc ttgggccctg gtcatccagg attctaacgg cgagaataag
     3661 atcaagatgc tgtgataaac cacgatcgtt atgctgatca taccctaatg atcccagcaa
     3721 gataatgtcc tgtcttctaa gatgtgcatc aagcctggta catactgaaa accctataag
     3781 gtcctggata atttttgttt gattattcat tgaagaaaca tttattttcc aattgtgtga
     3841 agtttttgac tgttaataaa agaatctgtc aaccatcaaa cttctgaggc ggaaagaacc
     3901 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3961 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     4021 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     4081 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     4141 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     4201 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     4261 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     4321 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     4381 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4441 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4501 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4561 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4621 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4681 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc
     4741 acgtgttgac aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag
     4801 gaactaaacc atggccaagc ctttgtctca agaagaatcc accctcattg aaagagcaac
     4861 ggctacaatc aacagcatcc ccatctctga agactacagc gtcgccagcg cagctctctc
     4921 tagcgacggc cgcatcttca ctggtgtcaa tgtatatcat tttactgggg gaccttgtgc
     4981 agaactcgtg gtgctgggca ctgctgctgc tgcggcagct ggcaacctga cttgtatcgt
     5041 cgcgatcgga aatgagaaca ggggcatctt gagcccctgc ggacggtgcc gacaggtgct
     5101 tctcgatctg catcctggga tcaaagccat agtgaaggac agtgatggac agccgacggc
     5161 agttgggatt cgtgaattgc tgccctctgg ttatgtgtgg gagggctaag cacttcgtgg
     5221 ccgaggagca ggactgacac gtgctacgag atttcgattc caccgccgcc ttctatgaaa
     5281 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     5341 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     5401 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     5461 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     5521 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     5581 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     5641 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     5701 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     5761 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     5821 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     5881 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5941 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     6001 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     6061 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     6121 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     6181 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     6241 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     6301 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     6361 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     6421 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     6481 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     6541 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     6601 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     6661 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     6721 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     6781 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     6841 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     6901 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6961 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     7021 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     7081 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     7141 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     7201 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     7261 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     7321 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     7381 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     7441 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     7501 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     7561 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     7621 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     7681 tgccacctga cgtc
//
