LOCUS       sequence-360282-        6991 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TALED_Left-RNR2-1397C-AD in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6991)
  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 6991)
  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_Left-RNR2-1397C-AD
FEATURES             Location/Qualifiers
     source          1..6991
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(103..122)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        294..673
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        674..877
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     828..848
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     922..941
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        922..940
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1037..1063
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1067..1093
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1097..1123
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(4262..4279)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4268..4492
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    4723..4844
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4760..4779)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4814..4833
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4893..4909)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4893..4909)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4906..4928)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4917..4933
                     /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(4941..4947,4948..4965,4966..4971))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4986..5007
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5124..5141)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5295..5883)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5375..5394)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6054..6845,6846..6914))
                     /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     6677..6696
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(6915..6991,1..28))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
       61 acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc
      121 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
      181 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
      241 tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg
      301 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
      361 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
      421 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
      481 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
      541 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
      601 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
      661 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
      721 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
      781 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
      841 taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta gagaacccac
      901 tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagatggctc ttagtcgagc
      961 cgtctgtggt acttcccgac agcttgctcc agtcttgggg taccttggct cccgacaaaa
     1021 gcactctctc cctgattacc cttacgatgt acctgactac gccggctacc cttacgacgt
     1081 accagattac gctggatatc catacgatgt ccctgattat gctgggatcc gaattcaaga
     1141 tctacgcacg ctcggctaca gccagcagca acaggagaag atcaaaccga aggttcgttc
     1201 gacagtggcg cagcaccacg aggcactggt cggccatggg tttacacacg cgcacatcgt
     1261 tgcgctcagc caacacccgg cagcgttagg gaccgtcgct gtcaagtatc aggacatgat
     1321 cgcagcgttg ccagaggcga cacacgaagc gatcgttggc gtcggcaaac agtggtccgg
     1381 cgcacgcgct ctggaggcct tgctcacggt ggcgggagag ttgagaggtc caccgttaca
     1441 gttggacaca ggccaacttc tcaagattgc aaaacgtggc ggcgtgaccg cagtggaggc
     1501 agtgcatgca tggcgcaatg cactgacggg tgcccccctg aacctgaccc ctgcccaggt
     1561 ggtggccatt gcctcccacg atggaggtaa gcaggctctg gagacagttc aaagattgct
     1621 tccagttctt tgccaggatc acggtttgac acccgaccaa gtcgttgcaa tcgccagtca
     1681 tgatggtggt aagcaagcac tcgaaaccgt ccagcgcttg ctgcccgtgc tctgccaggc
     1741 tcatgggctg acacctgatc aggtcgtggc catcgcatca aatatagggg gtaaacaagc
     1801 tttggagact gtccagaggc tcctccccgt cttgtgtcaa gcccatgggt tgacacctga
     1861 ccaggtcgtg gccattgcca gcaatggagg agggaagcaa gctcttgaga ctgtgcagcg
     1921 cttgcttccc gtcctctgcc aagcacatgg gctcacaccc gatcaagttg ttgctattgc
     1981 aagcaatatc ggaggcaagc aggccttgga aactgtccaa agattgttgc cagtcttgtg
     2041 ccaggatcac ggactcaccc cagaccaagt ggttgcaatt gcttcaaatg gcgggggcaa
     2101 acaggccctg gaaaccgttc agcggcttct tcctgtcctt tgccaggccc atggtcttac
     2161 accagctcag gtcgttgcta ttgcatcaca cgatgggggc aaacaggcac tcgagacagt
     2221 gcagcggttg ctgccagtgc tgtgtcaaga tcacggtttg accccagagc aagtggtggc
     2281 catcgcttca aacataggag gaaagcaagc cctcgagact gttcaaagac tgcttcccgt
     2341 tctctgtcaa gcacatgggt tgactccagc acaggttgtt gccattgcta gtaatattgg
     2401 tggtaaacag gcattggaaa ctgtgcaaag actccttcct gtcctgtgtc aagcacatgg
     2461 gcttacacct gctcaggtcg ttgctattgc atcacacgat gggggcaaac aggcactcga
     2521 gacagtgcag cggttgctgc cagtgctgtg tcaagatcac ggtttgaccc cagagcaagt
     2581 ggtggccatc gcttcaaaca taggaggaaa gcaagccctc gagactgttc aaagactgct
     2641 tcccgttctc tgtcaagcac atgggttgac tccagcacag gttgttgcca ttgctagtaa
     2701 tattggtggt aaacaggcat tggaaactgt gcaaagactc cttcctgtcc tgtgtcagga
     2761 ccatggcttg actcctgcac aggttgtcgc catcgctagc aatgggggtg gcaaacaggc
     2821 acttgaaaca gtgcaacggt tgttgcccgt cttgtgtcag gaccatgggc ttacccctgc
     2881 acaggtcgtg gcaattgcct ccaacatcgg ggggaagcaa gcccttgaaa ctgtccagcg
     2941 cctgttgcca gttctttgtc aagcccatgg attgactccc gaccaggtgg tcgcaattgc
     3001 aagtaacaac ggtgggaaac aggctttgga gactgtgcag agacttcttc ccgtgttgtg
     3061 ccaagctcat gggctcaccc ccgcccaagt ggtcgccatc gcttctaata acggtggcaa
     3121 gcaagctttg gagactgtcc agaggctcct gcccgtcctc tgtcaggacc acggtttgac
     3181 acctgctcag gtggttgcca tcgcaagcaa taatggcgga aaacaggccc tggaaaccgt
     3241 ccagcgcctg ttgcctgtcc tctgccaaga tcatggattg actccagacc aagttgtggc
     3301 aattgcatct aacgggggtg gcaaacaagc ccttgagact gttcagcgcc tgttgcccgt
     3361 tttgtgccaa gcacacggtc tgactccgga acaggtggtg gcgattgcaa gcaacggcgg
     3421 cggcaaacag gctctagaga gcattgttgc ccagctctcc agacctgatc cggcgctagc
     3481 cgcgttgaca aatgatcatc ttgttgctct tgcttgtctt ggcggacgac ctgccctgga
     3541 tgctgtgaaa aaggggttgc tagtcggcag tgccatacct gtgaagcggg gagcaacagg
     3601 ggagacaaag gtgttcacag gcaactctaa cagtccaaag agccccacca aaggcgggtg
     3661 ttcaggaagc gaaactcctg gtacctcaga gtccgctact cccgaatcct ctgaggtgga
     3721 gttttcccac gagtactgga tgagacatgc cctgaccctg gccaagaggg cacgggatga
     3781 gagggaggtg cctgtgggag ccgtgctggt gctgaacaat agagtgatcg gcgagggctg
     3841 gaacagagcc atcggcctgc acgacccaac agcccatgcc gaaattatgg ccctgagaca
     3901 gggcggcctg gtcatgcaga actacagact gattgacgcc accctgtacg tgacattcga
     3961 gccttgcgtg atgtgcgccg gcgccatgat ccactctagg atcggccgcg tggtgtttgg
     4021 cgtgaggaac tcaaaaagag gcgccgcagg ctccctgatg aacgtgctga actaccccgg
     4081 catgaatcac cgcgtcgaaa ttaccgaggg aatcctggca gatgaatgtg ccgccctgct
     4141 gtgcgatttc tatcggatgc ctagacaggt gttcaatgct cagaagaagg cccagagctc
     4201 catcaactag tcagggccct attctatagg tgtcacctaa atgctagagc tcgctgatca
     4261 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     4321 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     4381 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     4441 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     4501 gcggaaagaa ccagctgggg ctctaggggg tatccccacg ctgcagtgag cgggactctg
     4561 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     4621 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     4681 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
     4741 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     4801 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     4861 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     4921 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     4981 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     5041 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     5101 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     5161 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     5221 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     5281 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     5341 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     5401 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     5461 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     5521 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     5581 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     5641 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     5701 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     5761 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     5821 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     5881 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     5941 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     6001 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     6061 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     6121 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     6181 gcaatgatac cgcgagatcc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     6241 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     6301 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     6361 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     6421 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     6481 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     6541 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     6601 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     6661 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     6721 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     6781 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     6841 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     6901 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     6961 ctcatgagcg gatacatatt tgaatgtatt t
//
