LOCUS       sequence-422672-        6967 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ExpressesTALED_Right-Cox3.1-1397C-AD in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6967)
  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 6967)
  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-Cox3.1-1397C-AD.
FEATURES             Location/Qualifiers
     source          1..6967
                     /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             885..962
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MASVLTPLLLRGLTGSARRLPVPRAK"
     CDS             975..1040
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(4179..4196)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4185..4409
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    4640..4761
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4677..4696)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4731..4750
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4810..4826)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4810..4826)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4823..4845)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4834..4850
                     /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(4858..4864,4865..4882,4883..4888))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4903..4924
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5041..5058)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5212..5800)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5292..5311)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5971..6762,6763..6831))
                     /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     6594..6613
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6832..6936)
                     /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 atgggtggac 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 ggagatggcc tctgtcttga
      901 caccactcct gctcaggggc ctgactggat cagctaggcg gctccccgtt cctcgcgcaa
      961 aaatacacag cctggactat aaggaccatg acggtgatta caaagatcac gacatagatt
     1021 ataaggatga tgatgataag gggatccgaa ttcaagatct acgcacgctc ggctacagcc
     1081 agcagcaaca ggagaagatc aaaccgaagg ttcgttcgac agtggcgcag caccacgagg
     1141 cactggtcgg ccatgggttt acacacgcgc acatcgttgc gctcagccaa cacccggcag
     1201 cgttagggac cgtcgctgtc aagtatcagg acatgatcgc agcgttgcca gaggcgacac
     1261 acgaagcgat cgttggcgtc ggcaaacagt ggtccggcgc acgcgctctg gaggccttgc
     1321 tcacggtggc gggagagttg agaggtccac cgttacagtt ggacacaggc caacttctca
     1381 agattgcaaa acgtggcggc gtgaccgcag tggaggcagt gcatgcatgg cgcaatgcac
     1441 tgacgggtgc ccccctgaac cttacccccg atcaggtggt ggcaattgcc agtaacattg
     1501 gaggtaaaca agctttggaa accgtgcaaa ggctcctgcc agtcctctgt caggaccacg
     1561 ggctcacacc agctcaggtc gtcgccattg cttctaataa tggggggaaa caagccctcg
     1621 agactgtcca gcgcctcctt cctgttctgt gccaggacca tggcttgact cctgcccaag
     1681 tggtcgctat cgcatctaac ataggcggaa agcaggcatt ggagacagtc cagcgccttc
     1741 tgccagtgct ttgtcaagcc cacgggttga ccccagcaca agtggtcgcc atcgcatcca
     1801 acggtggtgg gaaacaggca ttggagacag ttcaacggct gcttcctgtg ctgtgccagg
     1861 ctcacggact gactcctgac caagttgttg caatcgccag caataatggt ggcaaacaag
     1921 cccttgagac agtccagcgg ctcctccccg tcctttgtca agcacatggc ctcacaccag
     1981 ctcaagtcgt tgctattgct tcacatgatg gaggtaaaca agcacttgaa accgtgcaaa
     2041 gactcttgcc cgtgctttgt caggctcacg ggttgacacc agatcaagtt gtggccatcg
     2101 catctcatga cggcgggaag caggctctgg aaacagttca acgcttgctt cctgtcttgt
     2161 gccaagacca tgggctcacc cctgaccagg tcgttgcaat cgcttcaaat aatgggggga
     2221 aacaagcact cgaaaccgtg cagcgcctct tgccagtttt gtgtcaggat cacggcctta
     2281 cccctgctca ggtggtcgca atcgcaagca acgggggcgg gaagcaggca cttgagactg
     2341 ttcagagact cctccctgtc ctctgtcaag ctcatggcct gactcccgaa caggtggtcg
     2401 caattgcatc acacgacgga gggaagcagg ccttggaaac agtgcaaaga ctcttgcctg
     2461 ttctctgcca agctcacggg ctcactcctg ctcaagttgt ggccatcgct agcaacaatg
     2521 gtggtaaaca ggcattggaa acagtccaga ggcttctccc agttctttgc caggatcatg
     2581 ggttgacccc cgaccaagtg gttgccattg cctccaataa cgggggaaaa caggctctgg
     2641 aaaccgttca gcgccttctc cctgtgttgt gtcaagatca cggcctgact cctgatcagg
     2701 tcgtcgccat cgctagtaac attggcggta aacaggcctt ggagacagtc cagaggttgc
     2761 tccccgtgct ctgccaagca catggcctga ctcccgctca ggttgtggct attgcctcca
     2821 atatcggggg gaagcaagca cttgaaaccg tccagaggct gctccctgtg ttgtgtcaag
     2881 atcacggtct taccccagct caagtggtcg ctattgcaag caacataggg ggaaaacaag
     2941 cactggaaac agtgcaacgc cttcttcctg tcctttgtca ggctcatggg cttacccctg
     3001 aacaagttgt ggccatcgcc agcaacgggg gaggaaaaca ggctttggaa accgtgcaaa
     3061 ggctgcttcc tgttctttgc caagcacacg ggttgacacc cgcacaggtt gtcgccatcg
     3121 ccagtaacaa cggagggaag caagcactcg agacagttca gcgccttctg ccagtcctgt
     3181 gccaagatca tggtctgacc cctgaccaag tggtcgcaat tgcttccaat aatggtggta
     3241 aacaagccct ggaaactgtc caacgcttgt tgccagtgtt gtgtcaagac cacggtctga
     3301 ctccggaaca ggtggtggcg attgcaagca acggcggcgg caaacaggct ctagagagca
     3361 ttgttgccca gctctccaga cctgatccgg cgctagccgc gttgacaaat gatcatcttg
     3421 ttgctcttgc ttgtcttggc ggacgacctg ccctggatgc tgtgaaaaag gggttgctag
     3481 tcggcagtgc catacctgtg aagcggggag caacagggga gacaaaggtg ttcacaggca
     3541 actctaacag tccaaagagc cccaccaaag gcgggtgttc aggaagcgaa actcctggta
     3601 cctcagagtc cgctactccc gaatcctctg aggtggagtt ttcccacgag tactggatga
     3661 gacatgccct gaccctggcc aagagggcac gggatgagag ggaggtgcct gtgggagccg
     3721 tgctggtgct gaacaataga gtgatcggcg agggctggaa cagagccatc ggcctgcacg
     3781 acccaacagc ccatgccgaa attatggccc tgagacaggg cggcctggtc atgcagaact
     3841 acagactgat tgacgccacc ctgtacgtga cattcgagcc ttgcgtgatg tgcgccggcg
     3901 ccatgatcca ctctaggatc ggccgcgtgg tgtttggcgt gaggaactca aaaagaggcg
     3961 ccgcaggctc cctgatgaac gtgctgaact accccggcat gaatcaccgc gtcgaaatta
     4021 ccgagggaat cctggcagat gaatgtgccg ccctgctgtg cgatttctat cggatgccta
     4081 gacaggtgtt caatgctcag aagaaggccc agagctccat caactagtca gggccctatt
     4141 ctataggtgt cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc cttctagttg
     4201 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     4261 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     4321 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     4381 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     4441 tagggggtat ccccacgctg cagtgagcgg gactctgggg ttcgaaatga ccgaccaagc
     4501 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     4561 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     4621 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     4681 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     4741 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     4801 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     4861 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     4921 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     4981 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5041 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5101 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5161 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5221 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5281 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5341 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5401 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5461 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5521 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5581 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5641 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5701 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5761 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5821 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     5881 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     5941 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6001 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6061 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagatccacg
     6121 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6181 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6241 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6301 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6361 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6421 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6481 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6541 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6601 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6661 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6721 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6781 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     6841 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     6901 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     6961 tgacgtc
//
