LOCUS       sequence-368596-        8041 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  human mitochondrial DNA ND1 target TALE.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8041)
  AUTHORS   Mok YG, Lee JM, Chung E, Lee J, Lim K, Cho SI, Kim JS
  TITLE     Base editing in human cells with monomeric DddA-TALE fusion 
            deaminases.
  JOURNAL   Nat Commun. 2022 Jul 12;13(1):4038. doi: 10.1038/s41467-022-31745-y.
  PUBMED    35821233
REFERENCE   2  (bases 1 to 8041)
  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"; journalName: "Nat 
            Commun."; date: "2022-07-12"; volume: "13(1):4038. doi"; pages: " 
            10.1038/s41467-022-31745-y"
COMMENT     Sequence Label: pCMV-ND1 Right-GSVG-UGI
FEATURES             Location/Qualifiers
     source          1..8041
                     /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             904..981
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MASVLTPLLLRGLTGSARRLPVPRAK"
     CDS             994..1059
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3826..4077
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     rep_origin      4274..4702
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4361..4380)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4571..4592
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4711..4731)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4716..5045
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4896..5031
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4958..4977
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        5092..5139
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5158..5556
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    5714..5835
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5751..5770)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5805..5824
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5884..5900)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5884..5900)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5897..5919)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5908..5924
                     /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(5932..5938,5939..5956,5957..5962))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5977..5998
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6115..6132)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6286..6874)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6366..6385)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7045..7836,7837..7905))
                     /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     7668..7687
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7906..8010)
                     /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 atgggtggag 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 ggagacccaa gctggctagc
      901 accatggcct ccgttctgac cccgctgctg ctgcgtggcc tgaccggctc cgctcgtcgt
      961 ctgccagttc cgcgtgcgaa aatccattcc ctggactaca aagaccatga cggtgattat
     1021 aaagatcatg acatcgatta caaggatgac gatgacaaga tggacatcgc ggatctacgc
     1081 acgctcggct acagccagca gcaacaggag aagatcaaac cgaaggttcg ttcgacagtg
     1141 gcgcagcacc acgaggcact ggtcggccat gggtttacac acgcgcacat cgttgcgctc
     1201 agccaacacc cggcagcgtt agggaccgtc gctgtcaagt atcaggacat gatcgcagcg
     1261 ttgccagagg cgacacacga agcgatcgtt ggcgtcggca aacagtggtc cggcgcacgc
     1321 gctctggagg ccttgctcac ggtggcggga gagttgagag gtccaccgtt acagttggac
     1381 acaggccaac ttctcaagat tgcaaaacgt ggcggcgtga ccgcagtgga ggcagtgcat
     1441 gcatggcgca atgcactgac gggtgccccc ctgaacctca ctcctgatca agtggttgcc
     1501 attgcctcta acaacggcgg aaaacaggct cttgagactg tccagaggct gctgcccgtg
     1561 ctctgtcagg atcatggact cacacctgct caagtggttg caatcgccag caatattggc
     1621 ggtaaacagg ccttggaaac agttcaacgc ttgcttccag tcttgtgcca agctcacggg
     1681 ctgaccccag agcaggtcgt ggctatcgca tctaacaatg gtggtaagca agctctggag
     1741 acagtgcaga gacttctccc agttctgtgc caggcccacg ggttgacacc tgatcaagtc
     1801 gttgcaatcg catcaaatgg cggagggaag caggctcttg agactgtgca gaggttgctc
     1861 cctgttctct gccaggccca cggtttgaca cccgcacagg ttgtcgctat cgctagtaac
     1921 ggtgggggta agcaagcctt ggagactgtt caaagattgc tgcctgtgct gtgccaggcc
     1981 catgggctca ctcctgaaca ggtcgttgct attgctagca acggaggcgg taaacaagcc
     2041 ctcgaaactg tccaacgcct tttgcctgtc ctctgtcaag cacacggact tacaccagca
     2101 caggttgtcg caattgcctc caataacggt ggtaaacagg ctctggaaac agtccagaga
     2161 ctcctccccg ttttgtgtca agaccacggt ctgacccccg cccaagtggt tgctattgcc
     2221 agtaacattg gtggaaaaca agccctcgaa actgttcaga ggcttttgcc agtcctgtgc
     2281 caggcccatg ggcttacacc tgagcaagtg gttgcaatcg cttctaacgg agggggcaag
     2341 caggctcttg agacagttca acgcctgttg ccagttttgt gccaagccca cggccttact
     2401 ccagatcagg tcgtcgcaat cgctagcaat aacggaggca aacaagccct ggaaactgtc
     2461 cagcgcttgc tccctgttct ttgccaagcc catggactga cccccgcaca agttgtcgct
     2521 attgcatcac acgatggagg taaacaggca ttggaaacag tgcaaagatt gcttcccgtt
     2581 ctgtgtcaag atcatggctt gacacccgac caggttgtgg ccatcgcaag caatggtggt
     2641 ggtaagcaag ctctcgagac agttcaaagg ctgctgccag ttttgtgcca ggcacacgga
     2701 ctgactcccg atcaggtcgt tgcaattgca agccatgatg gcggcaagca ggctttggaa
     2761 acagtccaga ggttgcttcc tgtcctttgt caggctcacg ggttgactcc agaccaagtg
     2821 gtggcaatcg cctctaatat tggcggtaag caagccctcg agacagttca acggttgctc
     2881 ccagtgcttt gccaggcaca tggacttact ccagctcaag ttgttgctat tgcttctcac
     2941 gatggtggta agcaagcatt ggaaaccgtc cagcgcctgt tgccagtttt gtgccaggct
     3001 catggcctga cacctgcaca ggtggttgca atcgcaagcc atgatggcgg taagcaagct
     3061 ttggagacag tccagcgctt gcttcccgtg ctctgtcaag accacggtct gacccccgct
     3121 caagttgtgg ctattgcttc ccatgacggg ggcaagcagg ctttggagac tgtgcaaaga
     3181 ttgcttcctg tgttgtgcca ggcacacggt ctgactccgg aacaggtggt ggcgattgca
     3241 agcaacggcg gcggcaaaca ggctctagag agcattgttg cccagctctc cagacctgat
     3301 ccggcgctag ccgcgttgac aaatgatcat cttgttgctc ttgcttgtct tggcggacga
     3361 cctgccctgg atgctgtgaa aaaggggttg gggggatccg gcagctacgc cctgggtccg
     3421 tatcagatta gcgccccgca gctgccagca tacaatggtc agaccgtggg taccttctac
     3481 tatgtgaacg acgctggcgg tctggagggc aaggtgttta gcagcggcgg tccaaccccg
     3541 tacccaaact atgccaatgc cggtcatgtg gagagtcaga gcgccctgtt catgcgtgat
     3601 aacggcatca gcgagggtct ggtgttccac aacaacccgg aaggcacctg cggtttttgc
     3661 gtgaacatga ccgagaccct gctgccggaa aacgcgaaaa tgaccgtggt gccgccggaa
     3721 ggtgtcattc cagtgaagcg cggcgctacc ggtgaaacca aagtgtttac cggtaacagc
     3781 aacggcccga agagcccgac caaaggcggt tgctctggcg gctcaactaa tctgagcgac
     3841 atcattgaga aggagactgg gaaacagctg gtcattcagg agtccatcct gatgctgcct
     3901 gaggaggtgg aggaagtgat cggcaacaag ccagagtctg acatcctggt gcacaccgcc
     3961 tacgacgagt ccacagatga gaatgtgatg ctgctgacct ctgacgcccc cgagtataag
     4021 ccttgggccc tggtcatcca ggattctaac ggcgagaata agatcaagat gctgtgataa
     4081 ggggtctttg tcctctgtac tgtctctctc cttgccccta acccaaaaag cttcattttt
     4141 ctgtgtaggc tgcacaagag ccttgattga agatatattc tttctgaaca gtatttaagg
     4201 tttccaataa aatgtacacc cctcagaact tctgaggcgg aaagaaccag ctggggctct
     4261 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     4321 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     4381 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     4441 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     4501 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     4561 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     4621 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     4681 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     4741 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4801 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4861 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4921 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4981 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     5041 gcaaaaagct cccggagctt gtatatccat tttcggatct gatcagcacg tgttgacaat
     5101 taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg
     5161 gccaagcctt tgtctcaaga agaatccacc ctcattgaaa gagcaacggc tacaatcaac
     5221 agcatcccca tctctgaaga ctacagcgtc gccagcgcag ctctctctag cgacggccgc
     5281 atcttcactg gtgtcaatgt atatcatttt actgggggac cttgtgcaga actcgtggtg
     5341 ctgggcactg ctgctgctgc ggcagctggc aacctgactt gtatcgtcgc gatcggaaat
     5401 gagaacaggg gcatcttgag cccctgcgga cggtgccgac aggtgcttct cgatctgcat
     5461 cctgggatca aagccatagt gaaggacagt gatggacagc cgacggcagt tgggattcgt
     5521 gaattgctgc cctctggtta tgtgtgggag ggctaagcac ttcgtggccg aggagcagga
     5581 ctgacacgtg ctacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     5641 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     5701 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     5761 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     5821 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     5881 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     5941 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     6001 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     6061 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     6121 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     6181 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     6241 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     6301 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     6361 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     6421 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     6481 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     6541 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     6601 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     6661 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     6721 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     6781 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     6841 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     6901 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     6961 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     7021 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     7081 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     7141 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     7201 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     7261 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     7321 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     7381 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     7441 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     7501 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     7561 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     7621 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     7681 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     7741 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     7801 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     7861 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     7921 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     7981 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     8041 c
//
