LOCUS       sequence-437187-        6865 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TALED_Right-mouse-ATP6-1397C-AD(V28R) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6865)
  AUTHORS   Cho SI, Lim K, Hong S, Lee J, Kim A, Lim CJ, Ryou S, Lee JM, Mok YG,
            Chung E, Kim S, Han S, Cho SM, Kim J, Kim EK, Nam KH, Oh Y, Choi M, 
            An TH, Oh KJ, Lee S, Lee H, Kim JS
  TITLE     Engineering TALE-linked deaminases to facilitate precision adenine 
            base editing in mitochondrial DNA.
  JOURNAL   Cell. 2024 Jan 4;187(1):95-109.e26. doi: 10.1016/j.cell.2023.11.035.
  PUBMED    38181745
REFERENCE   2  (bases 1 to 6865)
  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"; doi: 
            "10.1016/j.cell.2023.11"; journalName: "Cell"; date: "2024-01-4- 4";
            volume: "187"; issue: "1"; pages: "95-109.e26"
COMMENT     Sequence Label: TALED_Right-mouse-ATP6-1397C-AD(V28R)
FEATURES             Location/Qualifiers
     source          1..6865
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1765..1782)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1771..1995
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    2226..2347
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2263..2282)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2317..2336
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2396..2412)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2396..2412)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2409..2431)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2420..2436
                     /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(2444..2450,2451..2468,2469..2474))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2489..2510
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2627..2644)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2798..3386)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2878..2897)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3557..4348,4349..4417))
                     /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     4180..4199
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4418..4522)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4597..4616)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4788..5167
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5168..5371
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5322..5342
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5416..5435
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5416..5434
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5438..5515
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MASVLTPLLLRGLTGSARRLPVPRAK"
     CDS             5528..5593
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
ORIGIN
        1 ataggaggaa aacaagcatt ggagacagtg caaaggcttt tgccagtttt gtgtcaagca
       61 catggcctta cacctgcaca agttgttgca attgcaagca acggcggcgg aaaacaggca
      121 ctggaaactg tgcaaagact cctgccagtt ttgtgccagg ctcatggcct gactccagac
      181 caggtcgttg ctattgccag caataacgga ggtaagcaag cattggagac tgttcagcgg
      241 ctcctccctg tcctctgtca ggcccacggg ctgactcctg cacaggtggt ggccatcgct
      301 tcaaacgggg gggggaagca agccctcgaa actgttcaac ggttgttgcc tgttttgtgt
      361 caggatcatg ggctgactcc agcccaggtt gttgcaattg catctaatat cggtggaaag
      421 caggctttgg aaaccgtcca gaggcttctc ccagtgctgt gtcaggcaca tgggttgaca
      481 cccgaccaag tggtcgcaat tgcttcaaat ggagggggaa aacaagccct tgaaacagtg
      541 cagaggttgc ttcctgttct ttgtcaagct cacggtctta ctcctgatca ggtggtggct
      601 atcgcaagca acggcggggg gaaacaggca ctcgaaactg tgcaacgcct cctgcctgtg
      661 ttgtgccagg ctcatggatt gacccctgag caagtggttg ccattgccag taatatcggc
      721 ggcaagcagg ctttggaaac tgttcagaga ttgctgcccg ttctttgcca agcacatggc
      781 cttactcccg atcaggtcgt ggctattgca tcaaatggtg gtggcaaaca agcactggaa
      841 accgttcaaa ggttgcttcc tgttctgtgt caggaccacg gtctgactcc ggaacaggtg
      901 gtggcgattg caagcaacgg cggcggcaaa caggctctag agagcattgt tgcccagctc
      961 tccagacctg atccggcgct agccgcgttg acaaatgatc atcttgttgc tcttgcttgt
     1021 cttggcggac gacctgccct ggatgctgtg aaaaaggggt tgctagtcgg cagtgccata
     1081 cctgtgaagc ggggagcaac aggggagaca aaggtgttca caggcaactc taacagtcca
     1141 aagagcccca ccaaaggcgg gtgttcagga agcgaaactc ctggtacctc agagtccgct
     1201 actcccgaat cctctgaggt ggagttttcc cacgagtact ggatgagaca tgccctgacc
     1261 ctggccaaga gggcacggga tgagagggag aggcctgtgg gagccgtgct ggtgctgaac
     1321 aatagagtga tcggcgaggg ctggaacaga gccatcggcc tgcacgaccc aacagcccat
     1381 gccgaaatta tggccctgag acagggcggc ctggtcatgc agaactacag actgattgac
     1441 gccaccctgt acgtgacatt cgagccttgc gtgatgtgcg ccggcgccat gatccactct
     1501 aggatcggcc gcgtggtgtt tggcgtgagg aactcaaaaa gaggcgccgc aggctccctg
     1561 atgaacgtgc tgaactaccc cggcatgaat caccgcgtcg aaattaccga gggaatcctg
     1621 gcagatgaat gtgccgccct gctgtgcgat ttctatcgga tgcctagaca ggtgttcaat
     1681 gctcagaaga aggcccagag ctccatcaac tagtcagggc cctattctat aggtgtcacc
     1741 taaatgctag agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     1801 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     1861 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     1921 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     1981 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
     2041 acgctgcagt gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     2101 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     2161 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     2221 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     2281 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     2341 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     2401 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     2461 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     2521 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     2581 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     2641 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     2701 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     2761 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     2821 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     2881 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     2941 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     3001 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3061 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3121 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     3181 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     3241 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     3301 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     3361 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     3421 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     3481 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     3541 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     3601 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     3661 gcttaccatc tggccccagt gctgcaatga taccgcgaga tccacgctca ccggctccag
     3721 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     3781 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     3841 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     3901 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     3961 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     4021 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     4081 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     4141 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     4201 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     4261 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     4321 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     4381 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     4441 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     4501 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
     4561 cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata
     4621 gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa
     4681 aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt
     4741 taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac attgattatt
     4801 gactagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
     4861 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
     4921 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
     4981 tcaatgggtg gactatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
     5041 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
     5101 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
     5161 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
     5221 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
     5281 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
     5341 tgtacggtgg gaggtctata taagcagagc tctctggcta actagagaac ccactgctta
     5401 ctggcttatc gaaattaata cgactcacta tagggagatg gcctctgtct tgacaccact
     5461 cctgctcagg ggcctgactg gatcagctag gcggctcccc gttcctcgcg caaaaataca
     5521 cagcctggac tataaggacc atgacggtga ttacaaagat cacgacatag attataagga
     5581 tgatgatgat aaggggatcc gaattcaaga tctacgcacg ctcggctaca gccagcagca
     5641 acaggagaag atcaaaccga aggttcgttc gacagtggcg cagcaccacg aggcactggt
     5701 cggccatggg tttacacacg cgcacatcgt tgcgctcagc caacacccgg cagcgttagg
     5761 gaccgtcgct gtcaagtatc aggacatgat cgcagcgttg ccagaggcga cacacgaagc
     5821 gatcgttggc gtcggcaaac agtggtccgg cgcacgcgct ctggaggcct tgctcacggt
     5881 ggcgggagag ttgagaggtc caccgttaca gttggacaca ggccaacttc tcaagattgc
     5941 aaaacgtggc ggcgtgaccg cagtggaggc agtgcatgca tggcgcaatg cactgacggg
     6001 tgcccccctg aacctcaccc ccgctcaggt tgtggctatc gcaagtaaca acggtggaaa
     6061 acaggccctt gaaactgtcc aacgccttct tcccgttctc tgccaggctc acggcttgac
     6121 acccgaccag gtcgttgcta ttgcctcaaa caatggagga aaacaggcac tcgaaaccgt
     6181 ccagcggctg ctccccgtct tgtgccagga ccatggactg acaccagagc aagtggttgc
     6241 catcgcctcc aacaacggtg gtaagcaagc acttgagact gtgcagaggc ttttgcctgt
     6301 gctttgtcag gcccatgggt tgactcccgc tcaggtggtg gctattgcaa gtaatggagg
     6361 agggaagcag gccttggaga cagtccaacg cttgctgccc gttctttgtc aggatcatgg
     6421 gttgacaccc gagcaagttg tggcaattgc ttcacacgat ggtggcaaac aggctttgga
     6481 aacagttcaa agattgttgc ctgtcctttg ccaagctcat ggacttactc cagcacaggt
     6541 ggtggccatc gcatccaaca taggaggtaa acaagcactg gaaaccgtcc agaggctttt
     6601 gcctgtcctc tgccaggatc acggtctgac accagaacaa gtggtcgcta ttgcatccaa
     6661 cggagggggc aagcaggctc ttgaaacagt gcaacgcctg ctgccagtcc tttgccaagc
     6721 ccatggcctt actcctgacc aagtcgtggc aattgctagt aacggtggcg ggaaacaagc
     6781 acttgaaact gttcagcgct tgttgcccgt tctctgtcag gatcacggtc ttacaccaga
     6841 tcaggttgtg gccattgcct caaac
//
