LOCUS       sequence-307664-        7004 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses right-side TALE–DddAtox half of ND6-DdCBE in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7004)
  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 7004)
  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: ND6-DdCBE-right side TALE
FEATURES             Location/Qualifiers
     source          1..7004
                     /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             904..981
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MASVLTPLLLRGLTGSARRLPVPRAK"
     CDS             994..1059
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2788..3039
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     rep_origin      3236..3664
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3323..3342)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3533..3554
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3673..3693)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3678..4007
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3858..3993
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3920..3939
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4055..4102
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4121..4519
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4677..4798
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4714..4733)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4768..4787
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4847..4863)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4847..4863)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4860..4882)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4871..4887
                     /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(4895..4901,4902..4919,4920..4925))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4940..4961
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5078..5095)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5249..5837)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5329..5348)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6008..6799,6800..6868))
                     /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     6631..6650
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6869..6973)
                     /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 accatggcct ccgttctgac cccgctgctg ctgcgtggcc tgaccggctc cgctcgtcgt
      961 ctgccagttc cgcgtgcgaa aatccattcc ctggactaca aagaccatga cggtgattat
     1021 aaagatcatg acatcgatta caaggatgac gatgacaaga tggacatcgc ggatctgcgt
     1081 accctgggtt acagccagca gcagcaggag aagatcaagc caaaggtgcg cagcaccgtg
     1141 gcccagcacc atgaagctct ggtgggtcac ggcttcaccc acgcgcacat cgtggctctg
     1201 agccagcacc cagccgcgct gggtaccgtg gccgtgaaat atcaggacat gattgctgcc
     1261 ctgccagagg ctacccatga agcgattgtg ggtgtgggca agagaggagc cggtgctcgt
     1321 gcgctggagg ccctgctgac cgtggccggt gaactgcgtg gcccgccgct gcagctggat
     1381 accggccagc tgctgaaaat cgcgaaacgt ggcggtgtga ccgctgtgga agctgtgcat
     1441 gcctggcgta atgctctgac cggtgccccg ctgaacctga ccccgcagca ggtggtggct
     1501 attgccagca acaacggcgg taaacaggct ctggaaaccg tgcagcgcct gctgccggtg
     1561 ctgtgccagg ctcatggtct gaccccggag caggtggtgg cgatcgctag caacatcggc
     1621 ggcaagcagg ctctggagac cgttcaggcc ctgttaccgg tgctgtgcca agcccatggt
     1681 ctgaccccgc agcaagttgt ggctattgcc agcaatggcg gtggcaaaca ggcgctggag
     1741 accgtgcagc gtctgttgcc ggtgctgtgc caagcccatg ggctgacccc gcagcaagtg
     1801 gttgccatcg ccagcaacaa cggtggcaag caggccctgg agaccgttca gcgcctgtta
     1861 ccggtgctgt gccaggccca tggcttaacc ccgcagcaag ttgtggccat cgctagcaac
     1921 aacggtggca aacaggctct ggagactgtt cagcgtctgc ttccggtgct gtgccaagcg
     1981 catggcctga ccccggaaca agttgttgct attgccagcc atgatggtgg caagcaggcg
     2041 ctggaaaccg ttcagcgcct gcttccggtg ctgtgccagg cgcatggatt aaccccgcag
     2101 caagtggtgg ccatcgccag caatggtggc ggtaaacagg ccctggaaac cgttcagcgt
     2161 ctgttaccgg tgctgtgcca ggcccatgga ttaaccccgg aacaagttgt ggctattgcg
     2221 tccaatatcg gcggcaagca ggcgctggaa actgtgcagg ctctgctccc ggtgctgtgc
     2281 caggcccatg ggttaacccc gcagcaggtt gttgccattg cgagcaacgg cggtggcaaa
     2341 caggctctgg agacggttca gcgcctgctc ccggtgctgt gccaggccca tggtttaacc
     2401 ccgcagcagg tggttgctat tgctagcaat ggcggcggca agcaggcgct ggaaacggtg
     2461 cagcgtctgc taccggtgct gtgccaggca catggcctta ccccgcagca agttgtggcc
     2521 attgctagca atggcggtgg ccgtccggcc ctggaaagca ttgtggcgca gctgagccgt
     2581 ccagacccgg ccctggcggc tctgaccaat gatcacctgg tggccctggc ctgcctgggt
     2641 ggccgtccgg ctctggatgc cgtgaagaaa ggtctgggcg gatccgccat tccagtgaag
     2701 cgcggcgcta ccggtgaaac caaagtgttt accggtaaca gcaacagccc gaagagcccg
     2761 accaaaggcg gttgctctgg cggctcaact aatctgagcg acatcattga gaaggagact
     2821 gggaaacagc tggtcattca ggagtccatc ctgatgctgc ctgaggaggt ggaggaagtg
     2881 atcggcaaca agccagagtc tgacatcctg gtgcacaccg cctacgacga gtccacagat
     2941 gagaatgtga tgctgctgac ctctgacgcc cccgagtata agccttgggc cctggtcatc
     3001 caggattcta acggcgagaa taagatcaag atgctgtgat aaggggtctt tgtcctctgt
     3061 actgtctctc tccttgcccc taacccaaaa agcttcattt ttctgtgtag gctgcacaag
     3121 agccttgatt gaagatatat tctttctgaa cagtatttaa ggtttccaat aaaatgtaca
     3181 cccctcagaa cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     3241 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3301 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3361 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3421 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     3481 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3541 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     3601 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3661 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3721 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3781 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3841 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3901 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     3961 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     4021 cttgtatatc cattttcgga tctgatcagc acgtgttgac aattaatcat cggcatagta
     4081 tatcggcata gtataatacg acaaggtgag gaactaaacc atggccaagc ctttgtctca
     4141 agaagaatcc accctcattg aaagagcaac ggctacaatc aacagcatcc ccatctctga
     4201 agactacagc gtcgccagcg cagctctctc tagcgacggc cgcatcttca ctggtgtcaa
     4261 tgtatatcat tttactgggg gaccttgtgc agaactcgtg gtgctgggca ctgctgctgc
     4321 tgcggcagct ggcaacctga cttgtatcgt cgcgatcgga aatgagaaca ggggcatctt
     4381 gagcccctgc ggacggtgcc gacaggtgct tctcgatctg catcctggga tcaaagccat
     4441 agtgaaggac agtgatggac agccgacggc agttgggatt cgtgaattgc tgccctctgg
     4501 ttatgtgtgg gagggctaag cacttcgtgg ccgaggagca ggactgacac gtgctacgag
     4561 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     4621 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     4681 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     4741 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     4801 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     4861 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     4921 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     4981 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     5041 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     5101 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     5161 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     5221 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5281 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5341 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5401 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5461 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5521 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5581 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5641 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     5701 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5761 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5821 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5881 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5941 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     6001 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6061 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6121 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6181 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6241 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6301 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6361 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6421 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6481 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6541 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6601 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6661 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6721 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6781 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6841 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6901 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6961 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
