LOCUS       sequence-307644-        7214 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses left-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 7214)
  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 7214)
  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-left side TALE
FEATURES             Location/Qualifiers
     source          1..7214
                     /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             2944..3195
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     rep_origin      3446..3874
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3533..3552)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3743..3764
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3883..3903)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3888..4217
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4068..4203
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4130..4149
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4265..4312
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4331..4729
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4887..5008
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4924..4943)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4978..4997
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5057..5073)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5057..5073)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5070..5092)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5081..5097
                     /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(5105..5111,5112..5129,5130..5135))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5150..5171
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5288..5305)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5459..6047)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5539..5558)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6218..7009,7010..7078))
                     /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     6841..6860
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7079..7183)
                     /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 acattgtggc cctgagccag cacccagccg cgctgggcac cgtggccgtg
     1261 aaatatcagg atatgattgc tgccctgcca gaggccaccc atgaagctat tgtgggcgtg
     1321 ggcaagcagt ggagcggtgc tcgtgcgctg gaggcgctgc tgaccgtggc tggtgaactg
     1381 cgtggtccgc cgctgcagct ggacaccggt cagctgctga aaatcgcgaa acgtggcggt
     1441 gtgaccgcgg tggaagccgt gcatgcttgg cgtaatgctc tgaccggtgc gccgctgaac
     1501 ctgaccccgc agcaggtggt ggctattgcc agcaacaacg gcggtaaaca ggccctggag
     1561 accgtgcagc gcctgctgcc ggtgctgtgc caggcccatg gtctgacccc ggagcaggtg
     1621 gtggcgatcg ctagcaacat cggcggcaag caggccctgg aaaccgtgca ggcgctgtta
     1681 ccggtgctgt gccaggctca tggcctgacc ccggaacaag ttgtggctat tgccagccat
     1741 gatggcggta aacaggctct ggaaaccgtg cagcgtctgt tgccggtgct gtgccaagcc
     1801 catggcctga ccccggagca agttgtggct attgcgagcc atgatggcgg caagcaggcg
     1861 ctggaaaccg ttcagcgcct gttaccggtg ctgtgccaag ctcatggtct gaccccggaa
     1921 caggtggtgg ccattgcttc ccatgatggc ggtaaacagg ccctggaaac cgttcagcgt
     1981 ctgcttccgg tgctgtgcca ggcccatggg ctgaccccgg aacaagtggt tgctattgcc
     2041 agccacgatg gcggcaagca ggctctggag accgttcagc gcctgcttcc ggtgctgtgc
     2101 caggcccatg gcttaacccc ggaacaagtt gttgctattg ctagtcatga tggcggtaaa
     2161 caggcgctgg agaccgttca gcgtctgtta ccggtgctgt gccaggcgca tggcttaacc
     2221 ccggagcagg ttgttgccat tgcctccaat atcggcggca agcaggctct ggaaaccgtt
     2281 caggccctgt tgccggtgct gtgccaggcc catggactga ccccgcagca agttgttgcc
     2341 attgccagca atggcggtgg caaacaggcg ctggaaactg ttcagcgcct gctcccggtg
     2401 ctgtgccaag cgcatggtct gaccccgcag caagtggttg ctattgctag caatggtggc
     2461 ggtcgtccgg cgctggaaag cattgtggct cagctgagcc gtccagaccc ggccctggcg
     2521 gctctgacca acgatcacct ggtggcgctg gcttgcctgg gcggtcgtcc ggccctggat
     2581 gcggtgaaga aaggcctggg tggatccggc agctacgccc tgggtccgta tcagattagc
     2641 gccccgcagc tgccagctta caatggtcag accgtgggta ccttctacta tgtgaacgac
     2701 gcgggcggtc tggagagcaa ggtgtttagc agcggcggtc caaccccgta cccaaactat
     2761 gccaatgccg gtcatgtgga gggtcagagc gccctgttca tgcgtgataa cggcatcagc
     2821 gagggtctgg tgttccacaa caacccggaa ggcacctgcg gtttttgcgt gaacatgacc
     2881 gagaccctgc tgccggaaaa cgcgaaaatg accgtggtgc cgccggaagg ttctggcggc
     2941 tcaactaatc tgagcgacat cattgagaag gagactggga aacagctggt cattcaggag
     3001 tccatcctga tgctgcctga ggaggtggag gaagtgatcg gcaacaagcc agagtctgac
     3061 atcctggtgc acaccgccta cgacgagtcc acagatgaga atgtgatgct gctgacctct
     3121 gacgcccccg agtataagcc ttgggccctg gtcatccagg attctaacgg cgagaataag
     3181 atcaagatgc tgtgataaac cacgatcgtt atgctgatca taccctaatg atcccagcaa
     3241 gataatgtcc tgtcttctaa gatgtgcatc aagcctggta catactgaaa accctataag
     3301 gtcctggata atttttgttt gattattcat tgaagaaaca tttattttcc aattgtgtga
     3361 agtttttgac tgttaataaa agaatctgtc aaccatcaaa cttctgaggc ggaaagaacc
     3421 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3481 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3541 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3601 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3661 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3721 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     3781 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     3841 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     3901 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3961 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4021 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4081 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4141 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4201 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc
     4261 acgtgttgac aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag
     4321 gaactaaacc atggccaagc ctttgtctca agaagaatcc accctcattg aaagagcaac
     4381 ggctacaatc aacagcatcc ccatctctga agactacagc gtcgccagcg cagctctctc
     4441 tagcgacggc cgcatcttca ctggtgtcaa tgtatatcat tttactgggg gaccttgtgc
     4501 agaactcgtg gtgctgggca ctgctgctgc tgcggcagct ggcaacctga cttgtatcgt
     4561 cgcgatcgga aatgagaaca ggggcatctt gagcccctgc ggacggtgcc gacaggtgct
     4621 tctcgatctg catcctggga tcaaagccat agtgaaggac agtgatggac agccgacggc
     4681 agttgggatt cgtgaattgc tgccctctgg ttatgtgtgg gagggctaag cacttcgtgg
     4741 ccgaggagca ggactgacac gtgctacgag atttcgattc caccgccgcc ttctatgaaa
     4801 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     4861 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     4921 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     4981 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     5041 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     5101 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     5161 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     5221 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     5281 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     5341 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     5401 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5461 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5521 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     5581 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5641 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5701 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5761 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5821 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5881 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5941 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     6001 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     6061 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     6121 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     6181 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     6241 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     6301 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     6361 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     6421 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6481 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     6541 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6601 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6661 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6721 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6781 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6841 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6901 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6961 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     7021 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     7081 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     7141 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     7201 tgccacctga cgtc
//
