LOCUS       sequence-360286-        6816 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TALED_Right-ND1-1397N-UGI in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6816)
  AUTHORS   Cho SI, Lee S, Mok YG, Lim K, Lee J, Lee JM, Chung E, Kim JS
  TITLE     Targeted A-to-G base editing in human mitochondrial DNA with 
            programmable deaminases.
  JOURNAL   Cell. 2022 May 12;185(10):1764-1776.e12. doi: 
            10.1016/j.cell.2022.03.039. Epub 2022 Apr 25.
  PUBMED    35472302
REFERENCE   2  (bases 1 to 6816)
  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.2022.03.039"; journalName: "Cell"; date: 
            "2022-05-12- 12"; volume: "185"; issue: "10"; pages: "1764-1776.e12"
COMMENT     Sequence Label: TALED_Right-ND1-1397N-UGI
FEATURES             Location/Qualifiers
     source          1..6816
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(64..81)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(235..823)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(315..334)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(994..1785,1786..1854))
                     /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     1617..1636
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1855..1959)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2034..2053)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2225..2604
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2605..2808
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2759..2779
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2853..2872
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2853..2871
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2875..2952
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MASVLTPLLLRGLTGSARRLPVPRAK"
     CDS             2965..3030
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             5716..5967
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     promoter        complement(5980..5998)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5981..5998)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(6018..6035)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6024..6248
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    6479..6600
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6516..6535)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6570..6589
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6649..6665)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6649..6665)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6662..6684)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6673..6689
                     /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(6697..6703,6704..6721,6722..6727))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6742..6763
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
       61 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
      121 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
      181 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
      241 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
      301 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
      361 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
      421 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
      481 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
      541 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
      601 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
      661 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
      721 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
      781 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
      841 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
      901 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
      961 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     1021 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     1081 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagatcc
     1141 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     1201 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     1261 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     1321 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     1381 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     1441 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     1501 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     1561 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     1621 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     1681 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     1741 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     1801 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     1861 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     1921 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     1981 acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc
     2041 tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct
     2101 gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg
     2161 aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg
     2221 cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
     2281 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
     2341 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
     2401 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     2461 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
     2521 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     2581 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     2641 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     2701 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     2761 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact
     2821 agagaaccca ctgcttactg gcttatcgaa attaatacga ctcactatag ggagatggcc
     2881 tctgtcttga caccactcct gctcaggggc ctgactggat cagctaggcg gctccccgtt
     2941 cctcgcgcaa aaatacacag cctggactat aaggaccatg acggtgatta caaagatcac
     3001 gacatagatt ataaggatga tgatgataag gggatccgaa ttcaagatct acgcacgctc
     3061 ggctacagcc agcagcaaca ggagaagatc aaaccgaagg ttcgttcgac agtggcgcag
     3121 caccacgagg cactggtcgg ccatgggttt acacacgcgc acatcgttgc gctcagccaa
     3181 cacccggcag cgttagggac cgtcgctgtc aagtatcagg acatgatcgc agcgttgcca
     3241 gaggcgacac acgaagcgat cgttggcgtc ggcaaacagt ggtccggcgc acgcgctctg
     3301 gaggccttgc tcacggtggc gggagagttg agaggtccac cgttacagtt ggacacaggc
     3361 caacttctca agattgcaaa acgtggcggc gtgaccgcag tggaggcagt gcatgcatgg
     3421 cgcaatgcac tgacgggtgc ccccctgaac ctcactcctg atcaagtggt tgccattgcc
     3481 tctaacaacg gcggaaaaca ggctcttgag actgtccaga ggctgctgcc cgtgctctgt
     3541 caggatcatg gactcacacc tgctcaagtg gttgcaatcg ccagcaatat tggcggtaaa
     3601 caggccttgg aaacagttca acgcttgctt ccagtcttgt gccaagctca cgggctgacc
     3661 ccagagcagg tcgtggctat cgcatctaac aatggtggta agcaagctct ggagacagtg
     3721 cagagacttc tcccagttct gtgccaggcc cacgggttga cacctgatca agtcgttgca
     3781 atcgcatcaa atggcggagg gaagcaggct cttgagactg tgcagaggtt gctccctgtt
     3841 ctctgccagg cccacggttt gacacccgca caggttgtcg ctatcgctag taacggtggg
     3901 ggtaagcaag ccttggagac tgttcaaaga ttgctgcctg tgctgtgcca ggcccatggg
     3961 ctcactcctg aacaggtcgt tgctattgct agcaacggag gcggtaaaca agccctcgaa
     4021 actgtccaac gccttttgcc tgtcctctgt caagcacacg gacttacacc agcacaggtt
     4081 gtcgcaattg cctccaataa cggtggtaaa caggctctgg aaacagtcca gagactcctc
     4141 cccgttttgt gtcaagacca cggtctgacc cccgcccaag tggttgctat tgccagtaac
     4201 attggtggaa aacaagccct cgaaactgtt cagaggcttt tgccagtcct gtgccaggcc
     4261 catgggctta cacctgagca agtggttgca atcgcttcta acggaggggg caagcaggct
     4321 cttgagacag ttcaacgcct gttgccagtt ttgtgccaag cccacggcct tactccagat
     4381 caggtcgtcg caatcgctag caataacgga ggcaaacaag ccctggaaac tgtccagcgc
     4441 ttgctccctg ttctttgcca agcccatgga ctgacccccg cacaagttgt cgctattgca
     4501 tcacacgatg gaggtaaaca ggcattggaa acagtgcaaa gattgcttcc cgttctgtgt
     4561 caagatcatg gcttgacacc cgaccaggtt gtggccatcg caagcaatgg tggtggtaag
     4621 caagctctcg agacagttca aaggctgctg ccagttttgt gccaggcaca cggactgact
     4681 cccgatcagg tcgttgcaat tgcaagccat gatggcggca agcaggcttt ggaaacagtc
     4741 cagaggttgc ttcctgtcct ttgtcaggct cacgggttga ctccagacca agtggtggca
     4801 atcgcctcta atattggcgg taagcaagcc ctcgagacag ttcaacggtt gctcccagtg
     4861 ctttgccagg cacatggact tactccagct caagttgttg ctattgcttc tcacgatggt
     4921 ggtaagcaag cattggaaac cgtccagcgc ctgttgccag ttttgtgcca ggctcatggc
     4981 ctgacacctg cacaggtggt tgcaatcgca agccatgatg gcggtaagca agctttggag
     5041 acagtccagc gcttgcttcc cgtgctctgt caagaccacg gtctgacccc cgctcaagtt
     5101 gtggctattg cttcccatga cgggggcaag caggctttgg agactgtgca aagattgctt
     5161 cctgtgttgt gccaggcaca cggtctgact ccggaacagg tggtggcgat tgcaagcaac
     5221 ggcggcggca aacaggctct agagagcatt gttgcccagc tctccagacc tgatccggcg
     5281 ctagccgcgt tgacaaatga tcatcttgtt gctcttgctt gtcttggcgg acgacctgcc
     5341 ctggatgctg tgaaaaaggg gttggggggc tctggatcag gtagctacgc acttggtcct
     5401 taccagatta gcgcacccca actccccgcc tataatggtc aaaccgtcgg gaccttttac
     5461 tacgtaaacg atgctggtgg gctggaatcc aaagtattct cctcaggggg ccctacaccc
     5521 taccccaact acgccaatgc tggtcatgta gaagggcagt cagcactgtt tatgcgcgat
     5581 aatggtataa gcgaggggtt ggtcttccat aacaacccag agggtacttg tggcttctgt
     5641 gtgaatatga ctgaaaccct tctgcccgaa aatgccaaga tgactgtcgt cccacctgaa
     5701 ggctccggtg gttcaacaaa tctgtctgac atcatagaaa aagaaactgg caaacaactg
     5761 gtgatacaag aaagtattct gatgttgcct gaggaagttg aagaggtaat aggcaacaag
     5821 cctgaaagcg atatactggt acataccgca tacgatgagt caactgacga aaacgtgatg
     5881 ctcttgacat ccgatgcccc tgaatacaag ccctgggcac ttgtaatcca agatagtaat
     5941 ggggagaaca aaatcaaaat gttgtagtca gggccctatt ctatagtgtc acctaaatgc
     6001 tagagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     6061 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     6121 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     6181 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     6241 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgctgc
     6301 agtgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     6361 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     6421 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     6481 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     6541 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     6601 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     6661 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     6721 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     6781 gcccgctttc cagtcgggaa acctgtcgtg ccagct
//
