LOCUS       sequence-360284-        6555 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TALED_Right-ND1-1397N in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6555)
  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 6555)
  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
FEATURES             Location/Qualifiers
     source          1..6555
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(11..28)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(941..1732,1733..1801))
                     /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     1564..1583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1981..2000)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2172..2551
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2552..2755
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2706..2726
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2800..2819
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2800..2818
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2822..2899
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MASVLTPLLLRGLTGSARRLPVPRAK"
     CDS             2912..2977
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     promoter        complement(5666..5684)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5667..5684)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5704..5721)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5710..5934
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    6165..6286
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6202..6221)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6256..6275
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6335..6351)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6335..6351)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6348..6370)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6359..6375
                     /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(6383..6389,6390..6407,6408..6413))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6428..6449
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagatccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag
     1981 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
     2041 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
     2101 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag
     2161 atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
     2221 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     2281 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     2341 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     2401 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     2461 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     2521 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
     2581 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     2641 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     2701 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctctg
     2761 gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc actataggga
     2821 gatggcctct gtcttgacac cactcctgct caggggcctg actggatcag ctaggcggct
     2881 ccccgttcct cgcgcaaaaa tacacagcct ggactataag gaccatgacg gtgattacaa
     2941 agatcacgac atagattata aggatgatga tgataagggg atccgaattc aagatctacg
     3001 cacgctcggc tacagccagc agcaacagga gaagatcaaa ccgaaggttc gttcgacagt
     3061 ggcgcagcac cacgaggcac tggtcggcca tgggtttaca cacgcgcaca tcgttgcgct
     3121 cagccaacac ccggcagcgt tagggaccgt cgctgtcaag tatcaggaca tgatcgcagc
     3181 gttgccagag gcgacacacg aagcgatcgt tggcgtcggc aaacagtggt ccggcgcacg
     3241 cgctctggag gccttgctca cggtggcggg agagttgaga ggtccaccgt tacagttgga
     3301 cacaggccaa cttctcaaga ttgcaaaacg tggcggcgtg accgcagtgg aggcagtgca
     3361 tgcatggcgc aatgcactga cgggtgcccc cctgaacctc actcctgatc aagtggttgc
     3421 cattgcctct aacaacggcg gaaaacaggc tcttgagact gtccagaggc tgctgcccgt
     3481 gctctgtcag gatcatggac tcacacctgc tcaagtggtt gcaatcgcca gcaatattgg
     3541 cggtaaacag gccttggaaa cagttcaacg cttgcttcca gtcttgtgcc aagctcacgg
     3601 gctgacccca gagcaggtcg tggctatcgc atctaacaat ggtggtaagc aagctctgga
     3661 gacagtgcag agacttctcc cagttctgtg ccaggcccac gggttgacac ctgatcaagt
     3721 cgttgcaatc gcatcaaatg gcggagggaa gcaggctctt gagactgtgc agaggttgct
     3781 ccctgttctc tgccaggccc acggtttgac acccgcacag gttgtcgcta tcgctagtaa
     3841 cggtgggggt aagcaagcct tggagactgt tcaaagattg ctgcctgtgc tgtgccaggc
     3901 ccatgggctc actcctgaac aggtcgttgc tattgctagc aacggaggcg gtaaacaagc
     3961 cctcgaaact gtccaacgcc ttttgcctgt cctctgtcaa gcacacggac ttacaccagc
     4021 acaggttgtc gcaattgcct ccaataacgg tggtaaacag gctctggaaa cagtccagag
     4081 actcctcccc gttttgtgtc aagaccacgg tctgaccccc gcccaagtgg ttgctattgc
     4141 cagtaacatt ggtggaaaac aagccctcga aactgttcag aggcttttgc cagtcctgtg
     4201 ccaggcccat gggcttacac ctgagcaagt ggttgcaatc gcttctaacg gagggggcaa
     4261 gcaggctctt gagacagttc aacgcctgtt gccagttttg tgccaagccc acggccttac
     4321 tccagatcag gtcgtcgcaa tcgctagcaa taacggaggc aaacaagccc tggaaactgt
     4381 ccagcgcttg ctccctgttc tttgccaagc ccatggactg acccccgcac aagttgtcgc
     4441 tattgcatca cacgatggag gtaaacaggc attggaaaca gtgcaaagat tgcttcccgt
     4501 tctgtgtcaa gatcatggct tgacacccga ccaggttgtg gccatcgcaa gcaatggtgg
     4561 tggtaagcaa gctctcgaga cagttcaaag gctgctgcca gttttgtgcc aggcacacgg
     4621 actgactccc gatcaggtcg ttgcaattgc aagccatgat ggcggcaagc aggctttgga
     4681 aacagtccag aggttgcttc ctgtcctttg tcaggctcac gggttgactc cagaccaagt
     4741 ggtggcaatc gcctctaata ttggcggtaa gcaagccctc gagacagttc aacggttgct
     4801 cccagtgctt tgccaggcac atggacttac tccagctcaa gttgttgcta ttgcttctca
     4861 cgatggtggt aagcaagcat tggaaaccgt ccagcgcctg ttgccagttt tgtgccaggc
     4921 tcatggcctg acacctgcac aggtggttgc aatcgcaagc catgatggcg gtaagcaagc
     4981 tttggagaca gtccagcgct tgcttcccgt gctctgtcaa gaccacggtc tgacccccgc
     5041 tcaagttgtg gctattgctt cccatgacgg gggcaagcag gctttggaga ctgtgcaaag
     5101 attgcttcct gtgttgtgcc aggcacacgg tctgactccg gaacaggtgg tggcgattgc
     5161 aagcaacggc ggcggcaaac aggctctaga gagcattgtt gcccagctct ccagacctga
     5221 tccggcgcta gccgcgttga caaatgatca tcttgttgct cttgcttgtc ttggcggacg
     5281 acctgccctg gatgctgtga aaaaggggtt ggggggctct ggatcaggta gctacgcact
     5341 tggtccttac cagattagcg caccccaact ccccgcctat aatggtcaaa ccgtcgggac
     5401 cttttactac gtaaacgatg ctggtgggct ggaatccaaa gtattctcct cagggggccc
     5461 tacaccctac cccaactacg ccaatgctgg tcatgtagaa gggcagtcag cactgtttat
     5521 gcgcgataat ggtataagcg aggggttggt cttccataac aacccagagg gtacttgtgg
     5581 cttctgtgtg aatatgactg aaacccttct gcccgaaaat gccaagatga ctgtcgtccc
     5641 acctgaaggc tagtcagggc cctattctat agtgtcacct aaatgctaga gctcgctgat
     5701 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     5761 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     5821 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     5881 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     5941 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgctgcagtg agcgggactc
     6001 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     6061 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     6121 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     6181 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     6241 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     6301 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     6361 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     6421 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     6481 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     6541 tgcgtattgg gcgct
//
