LOCUS       sequence-360283-        6657 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TALED_Right-RNR2-1397N in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6657)
  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 6657)
  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-RNR2-1397N
FEATURES             Location/Qualifiers
     source          1..6657
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(23..40)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(194..782)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(274..293)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(953..1744,1745..1813))
                     /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     1576..1595
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1814..1918)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1993..2012)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2184..2563
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2564..2767
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2718..2738
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2812..2831
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2812..2830
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2834..2911
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MASVLTPLLLRGLTGSARRLPVPRAK"
     CDS             2924..2989
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     promoter        complement(5780..5798)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5781..5798)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5818..5835)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5824..6048
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    6279..6400
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6316..6335)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6370..6389
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6449..6465)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6449..6465)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6462..6484)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6473..6489
                     /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(6497..6503,6504..6521,6522..6527))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6542..6563
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
       61 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
      121 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
      181 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
      241 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
      301 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
      361 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
      421 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
      481 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
      541 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
      601 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
      661 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
      721 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
      781 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
      841 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
      901 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
      961 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     1021 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     1081 caatgatacc gcgagatcca cgctcaccgg ctccagattt atcagcaata aaccagccag
     1141 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     1201 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     1261 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     1321 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     1381 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     1441 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     1501 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     1561 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     1621 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     1681 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     1741 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     1801 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     1861 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     1921 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     1981 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
     2041 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
     2101 cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga
     2161 tgtacgggcc agatatacgc gttgacattg attattgact agttattaat agtaatcaat
     2221 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
     2281 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     2341 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggact atttacggta
     2401 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
     2461 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
     2521 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
     2581 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
     2641 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
     2701 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
     2761 cagagctctc tggctaacta gagaacccac tgcttactgg cttatcgaaa ttaatacgac
     2821 tcactatagg gagatggcct ctgtcttgac accactcctg ctcaggggcc tgactggatc
     2881 agctaggcgg ctccccgttc ctcgcgcaaa aatacacagc ctggactata aggaccatga
     2941 cggtgattac aaagatcacg acatagatta taaggatgat gatgataagg ggatccgaat
     3001 tcaagatcta cgcacgctcg gctacagcca gcagcaacag gagaagatca aaccgaaggt
     3061 tcgttcgaca gtggcgcagc accacgaggc actggtcggc catgggttta cacacgcgca
     3121 catcgttgcg ctcagccaac acccggcagc gttagggacc gtcgctgtca agtatcagga
     3181 catgatcgca gcgttgccag aggcgacaca cgaagcgatc gttggcgtcg gcaaacagtg
     3241 gtccggcgca cgcgctctgg aggccttgct cacggtggcg ggagagttga gaggtccacc
     3301 gttacagttg gacacaggcc aacttctcaa gattgcaaaa cgtggcggcg tgaccgcagt
     3361 ggaggcagtg catgcatggc gcaatgcact gacgggtgcc cccctgaacc tgacccctgc
     3421 tcaggttgtc gcaattgcta gcaacaatgg aggaaaacaa gctttggaga cagttcagag
     3481 gttgttgccc gtcctctgtc aggctcatgg cttgactcct gaccaggtgg tcgctattgc
     3541 tagtcacgac ggcggtaaac aagccctcga aacagtgcag agacttcttc ctgttctgtg
     3601 ccaagaccat ggtcttacac cagctcaggt cgttgccatc gcctctaata ttggtggaaa
     3661 acaggcactc gagactgtgc aaaggcttct gccagtcctt tgccaagcac atgggttgac
     3721 ccctgcccag gtggtggcca ttgcctccca cgatggaggt aagcaggctc tggagacagt
     3781 tcaaagattg cttccagttc tttgccagga tcacggtttg acacccgacc aagtcgttgc
     3841 aatcgccagt catgatggtg gtaagcaagc actcgaaacc gtccagcgct tgctgcccgt
     3901 gctctgccag gctcatgggc tgacacctga tcaggtcgtg gccatcgcat caaatatagg
     3961 gggtaaacaa gctttggaga ctgtccagag gctcctcccc gtcttgtgtc aagcccatgg
     4021 actgacacca gagcaggtcg tcgccattgc tagtaatggg ggagggaagc aggctttgga
     4081 aactgtccag agactcttgc cagttttgtg tcaggcacac gggcttacac ctgagcaagt
     4141 ggtggccatt gctagccacg acggggggaa acaggccctc gaaaccgttc aacggttgct
     4201 ccccgtcctc tgccaagcac acggtcttac ccctgctcaa gttgttgcaa tcgcttccaa
     4261 taacggcggt aaacaagccc ttgagacagt gcaaaggctc ttgccagtgc tctgtcaagc
     4321 tcatggactt acccctgcac aggttgtcgc aatcgcatcc aacaatggag gaaaacaagc
     4381 actcgaaacc gttcagcgcc tgcttcctgt cttgtgtcag gcacacggac tgactccaga
     4441 ccaggtggtt gctatcgcca gcaataacgg aggtaaacaa gccttggaga ctgtccaaag
     4501 gttgttgccc gttctctgcc aagaccatgg tttgacacca gctcaagtgg ttgccatcgc
     4561 ttctaatatc gggggtaagc aggcactcga gacagtccag agactcctgc ctgttctctg
     4621 ccaggctcac gggttgactc ctgcacaagt tgttgcaatt gcaagcaacg gcggcggaaa
     4681 acaggcactg gaaactgtgc aaagactcct gccagttttg tgccaggctc atggcctgac
     4741 tccagaccag gtcgttgcta ttgccagcaa taacggaggt aagcaagcat tggagactgt
     4801 tcagcggctc ctccctgtcc tctgtcaggc ccacgggctg actcctgcac aggtggtggc
     4861 catcgcttca aacggggggg ggaagcaagc cctcgaaact gttcaacggt tgttgcctgt
     4921 tttgtgtcag gatcatgggc tgactcctga ccaggtcgtt gccattgcta gtcatgacgg
     4981 gggtaaacag gctctcgaaa ctgtgcaacg cttgttgcca gttttgtgcc aagctcatgg
     5041 cttgacacct gcacaagtgg ttgcaatcgc ttcacacgac ggaggaaagc aagccctcga
     5101 aactgtgcag cggctcctcc ccgtgctgtg tcaagatcac ggccttacac ccgctcaagt
     5161 cgtcgcaatt gccagcaatg gaggcggtaa gcaggccttg gagactgttc agaggttgtt
     5221 gcccgtgctt tgccaagacc acggtctgac tccggaacag gtggtggcga ttgcaagcaa
     5281 cggcggcggc aaacaggctc tagagagcat tgttgcccag ctctccagac ctgatccggc
     5341 gctagccgcg ttgacaaatg atcatcttgt tgctcttgct tgtcttggcg gacgacctgc
     5401 cctggatgct gtgaaaaagg ggttgggggg ctctggatca ggtagctacg cacttggtcc
     5461 ttaccagatt agcgcacccc aactccccgc ctataatggt caaaccgtcg ggacctttta
     5521 ctacgtaaac gatgctggtg ggctggaatc caaagtattc tcctcagggg gccctacacc
     5581 ctaccccaac tacgccaatg ctggtcatgt agaagggcag tcagcactgt ttatgcgcga
     5641 taatggtata agcgaggggt tggtcttcca taacaaccca gagggtactt gtggcttctg
     5701 tgtgaatatg actgaaaccc ttctgcccga aaatgccaag atgactgtcg tcccacctga
     5761 aggctagtca gggccctatt ctatagtgtc acctaaatgc tagagctcgc tgatcagcct
     5821 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     5881 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     5941 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     6001 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     6061 aaagaaccag ctggggctct agggggtatc cccacgctgc agtgagcggg actctggggt
     6121 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     6181 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     6241 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     6301 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     6361 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     6421 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     6481 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     6541 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     6601 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgc
//
