LOCUS       sequence-422671-        6865 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TALED_Right-ND6-1397C-AD 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-ND6-1397C-AD
FEATURES             Location/Qualifiers
     source          1..6865
                     /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(4077..4094)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4083..4307
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    4538..4659
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4575..4594)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4629..4648
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4708..4724)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4708..4724)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4721..4743)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4732..4748
                     /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(4756..4762,4763..4780,4781..4786))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4801..4822
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4939..4956)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5110..5698)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5190..5209)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5869..6660,6661..6729))
                     /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     6492..6511
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6730..6834)
                     /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 ttgacccctg agcaagtggt tgccattgcc agtaatatcg
     1501 gcggcaagca ggctttggaa actgttcaga gattgctgcc cgttctttgc caagcacatg
     1561 gcttgacacc cgatcaagtt gttgctatcg ctagccatga tggagggaaa caagcccttg
     1621 agactgtgca acggctgctt ccagtgttgt gccaagctca tggacttact cccgatcagg
     1681 tcgtggctat tgcatcaaat ggtggtggca aacaagcact ggaaaccgtt caaaggttgc
     1741 ttcctgttct gtgtcaggac cacgggttga cacctgatca ggtggtcgca atcgccagca
     1801 acattggagg aaagcaggct cttgaaacag tgcagcgcct gcttcccgtg ttgtgccagg
     1861 atcatggcct tactcctgat caagttgttg caatcgcctc acatgacggc gggaagcaag
     1921 ctttggagac tgttcaaaga ctcctccctg tcctgtgcca agaccatggg ctcacacccg
     1981 cacaggtggt ggctattgct tccaacatag gagggaaaca agcccttgaa acagtccaaa
     2041 gattgctccc tgtcctttgc caggctcacg gcttgacacc agctcaggtc gtcgctattg
     2101 caagtaacaa cggaggcaaa caggccttgg agacagtcca aagactcctc ccagttcttt
     2161 gccaggccca cggtcttact cccgatcaag tggtggcaat cgcttcccat gatggtggaa
     2221 agcaggcctt ggaaactgtg cagaggctgc tccccgtttt gtgccaagac cacggattga
     2281 cccctgcaca ggtggttgct atcgcttcca ataatggagg gaagcaagca cttgagacag
     2341 tgcagcgctt gctgcctgtt ctgtgtcaag ctcacggttt gacccctgaa caagtcgtgg
     2401 ctattgctag taacatcggt ggtaaacagg ctcttgagac agttcaaaga ttgctgccag
     2461 tgttgtgtca ggctcacggt ttgactccag ctcaagttgt tgcaatcgcc agtaacggtg
     2521 gggggaagca ggcactcgaa accgtgcaac ggcttctccc cgttctgtgc caggcacacg
     2581 gactgacacc agcccaagtt gtcgccatcg cttcaaataa cgggggtaaa caagctctcg
     2641 agacagtgca acgcttgttg cctgtgcttt gccaagatca tgggttgact ccagatcagg
     2701 tcgtcgcaat cgctagcaat aacggaggca aacaagccct ggaaactgtc cagcgcttgc
     2761 tccctgttct ttgccaagcc catggactga cccccgcaca agttgtcgct attgcatcac
     2821 acgatggagg taaacaggca ttggaaacag tgcaaagatt gcttcccgtt ctgtgtcaag
     2881 atcatggctt gacacccgac caggttgtgg ccatcgcaag caatggtggt ggtaagcaag
     2941 ctctcgagac agttcaaagg ctgctgccag ttttgtgcca ggctcatgga ttgacccctg
     3001 agcaagtggt tgccattgcc agtaatatcg gcggcaagca ggctttggaa actgttcaga
     3061 gattgctgcc cgttctttgc caagcacatg gccttactcc cgatcaggtc gtggctattg
     3121 catcaaatgg tggtggcaaa caagcactgg aaaccgttca aaggttgctt cctgttctgt
     3181 gtcaggacca cggtctgact ccggaacagg tggtggcgat tgcaagcaac ggcggcggca
     3241 aacaggctct agagagcatt gttgcccagc tctccagacc tgatccggcg ctagccgcgt
     3301 tgacaaatga tcatcttgtt gctcttgctt gtcttggcgg acgacctgcc ctggatgctg
     3361 tgaaaaaggg gttgctagtc ggcagtgcca tacctgtgaa gcggggagca acaggggaga
     3421 caaaggtgtt cacaggcaac tctaacagtc caaagagccc caccaaaggc gggtgttcag
     3481 gaagcgaaac tcctggtacc tcagagtccg ctactcccga atcctctgag gtggagtttt
     3541 cccacgagta ctggatgaga catgccctga ccctggccaa gagggcacgg gatgagaggg
     3601 aggtgcctgt gggagccgtg ctggtgctga acaatagagt gatcggcgag ggctggaaca
     3661 gagccatcgg cctgcacgac ccaacagccc atgccgaaat tatggccctg agacagggcg
     3721 gcctggtcat gcagaactac agactgattg acgccaccct gtacgtgaca ttcgagcctt
     3781 gcgtgatgtg cgccggcgcc atgatccact ctaggatcgg ccgcgtggtg tttggcgtga
     3841 ggaactcaaa aagaggcgcc gcaggctccc tgatgaacgt gctgaactac cccggcatga
     3901 atcaccgcgt cgaaattacc gagggaatcc tggcagatga atgtgccgcc ctgctgtgcg
     3961 atttctatcg gatgcctaga caggtgttca atgctcagaa gaaggcccag agctccatca
     4021 actagtcagg gccctattct ataggtgtca cctaaatgct agagctcgct gatcagcctc
     4081 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     4141 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     4201 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     4261 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     4321 aagaaccagc tggggctcta gggggtatcc ccacgctgca gtgagcggga ctctggggtt
     4381 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     4441 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     4501 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     4561 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     4621 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     4681 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     4741 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     4801 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     4861 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     4921 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     4981 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5041 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     5101 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     5161 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5221 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5281 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     5341 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     5401 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5461 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5521 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     5581 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     5641 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     5701 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     5761 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     5821 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     5881 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     5941 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     6001 gataccgcga gatccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     6061 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     6121 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     6181 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     6241 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     6301 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     6361 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     6421 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     6481 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     6541 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     6601 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     6661 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     6721 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     6781 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     6841 tccccgaaaa gtgccacctg acgtc
//
