LOCUS       sequence-360287-        6696 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses TALED_Right-ND1-AD in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6696)
  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 6696)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-AD
FEATURES             Location/Qualifiers
     source          1..6696
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(50..69)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        241..620
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        621..824
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     775..795
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     869..888
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        869..887
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             891..968
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MASVLTPLLLRGLTGSARRLPVPRAK"
     CDS             981..1046
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     promoter        complement(3876..3894)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3877..3894)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3914..3931)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3920..4144
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    4375..4496
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4412..4431)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4466..4485
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4545..4561)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4545..4561)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4558..4580)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4569..4585
                     /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(4593..4599,4600..4617,4618..4623))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4638..4659
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4776..4793)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4947..5535)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5027..5046)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5706..6497,6498..6566))
                     /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     6329..6348
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6567..6671)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct
       61 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
      121 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
      181 atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
      241 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
      301 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      361 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      421 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      481 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      541 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      601 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      661 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      721 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      781 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      841 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag atggcctctg
      901 tcttgacacc actcctgctc aggggcctga ctggatcagc taggcggctc cccgttcctc
      961 gcgcaaaaat acacagcctg gactataagg accatgacgg tgattacaaa gatcacgaca
     1021 tagattataa ggatgatgat gataagggga tccgaattca agatctacgc acgctcggct
     1081 acagccagca gcaacaggag aagatcaaac cgaaggttcg ttcgacagtg gcgcagcacc
     1141 acgaggcact ggtcggccat gggtttacac acgcgcacat cgttgcgctc agccaacacc
     1201 cggcagcgtt agggaccgtc gctgtcaagt atcaggacat gatcgcagcg ttgccagagg
     1261 cgacacacga agcgatcgtt ggcgtcggca aacagtggtc cggcgcacgc gctctggagg
     1321 ccttgctcac ggtggcggga gagttgagag gtccaccgtt acagttggac acaggccaac
     1381 ttctcaagat tgcaaaacgt ggcggcgtga ccgcagtgga ggcagtgcat gcatggcgca
     1441 atgcactgac gggtgccccc ctgaacctca ctcctgatca agtggttgcc attgcctcta
     1501 acaacggcgg aaaacaggct cttgagactg tccagaggct gctgcccgtg ctctgtcagg
     1561 atcatggact cacacctgct caagtggttg caatcgccag caatattggc ggtaaacagg
     1621 ccttggaaac agttcaacgc ttgcttccag tcttgtgcca agctcacggg ctgaccccag
     1681 agcaggtcgt ggctatcgca tctaacaatg gtggtaagca agctctggag acagtgcaga
     1741 gacttctccc agttctgtgc caggcccacg ggttgacacc tgatcaagtc gttgcaatcg
     1801 catcaaatgg cggagggaag caggctcttg agactgtgca gaggttgctc cctgttctct
     1861 gccaggccca cggtttgaca cccgcacagg ttgtcgctat cgctagtaac ggtgggggta
     1921 agcaagcctt ggagactgtt caaagattgc tgcctgtgct gtgccaggcc catgggctca
     1981 ctcctgaaca ggtcgttgct attgctagca acggaggcgg taaacaagcc ctcgaaactg
     2041 tccaacgcct tttgcctgtc ctctgtcaag cacacggact tacaccagca caggttgtcg
     2101 caattgcctc caataacggt ggtaaacagg ctctggaaac agtccagaga ctcctccccg
     2161 ttttgtgtca agaccacggt ctgacccccg cccaagtggt tgctattgcc agtaacattg
     2221 gtggaaaaca agccctcgaa actgttcaga ggcttttgcc agtcctgtgc caggcccatg
     2281 ggcttacacc tgagcaagtg gttgcaatcg cttctaacgg agggggcaag caggctcttg
     2341 agacagttca acgcctgttg ccagttttgt gccaagccca cggccttact ccagatcagg
     2401 tcgtcgcaat cgctagcaat aacggaggca aacaagccct ggaaactgtc cagcgcttgc
     2461 tccctgttct ttgccaagcc catggactga cccccgcaca agttgtcgct attgcatcac
     2521 acgatggagg taaacaggca ttggaaacag tgcaaagatt gcttcccgtt ctgtgtcaag
     2581 atcatggctt gacacccgac caggttgtgg ccatcgcaag caatggtggt ggtaagcaag
     2641 ctctcgagac agttcaaagg ctgctgccag ttttgtgcca ggcacacgga ctgactcccg
     2701 atcaggtcgt tgcaattgca agccatgatg gcggcaagca ggctttggaa acagtccaga
     2761 ggttgcttcc tgtcctttgt caggctcacg ggttgactcc agaccaagtg gtggcaatcg
     2821 cctctaatat tggcggtaag caagccctcg agacagttca acggttgctc ccagtgcttt
     2881 gccaggcaca tggacttact ccagctcaag ttgttgctat tgcttctcac gatggtggta
     2941 agcaagcatt ggaaaccgtc cagcgcctgt tgccagtttt gtgccaggct catggcctga
     3001 cacctgcaca ggtggttgca atcgcaagcc atgatggcgg taagcaagct ttggagacag
     3061 tccagcgctt gcttcccgtg ctctgtcaag accacggtct gacccccgct caagttgtgg
     3121 ctattgcttc ccatgacggg ggcaagcagg ctttggagac tgtgcaaaga ttgcttcctg
     3181 tgttgtgcca ggcacacggt ctgactccgg aacaggtggt ggcgattgca agcaacggcg
     3241 gcggcaaaca ggctctagag agcattgttg cccagctctc cagacctgat ccggcgctag
     3301 ccgcgttgct agtctcagga agcgaaactc ctggtacctc agagtccgct actcccgaat
     3361 cctctgaggt ggagttttcc cacgagtact ggatgagaca tgccctgacc ctggccaaga
     3421 gggcacggga tgagagggag gtgcctgtgg gagccgtgct ggtgctgaac aatagagtga
     3481 tcggcgaggg ctggaacaga gccatcggcc tgcacgaccc aacagcccat gccgaaatta
     3541 tggccctgag acagggcggc ctggtcatgc agaactacag actgattgac gccaccctgt
     3601 acgtgacatt cgagccttgc gtgatgtgcg ccggcgccat gatccactct aggatcggcc
     3661 gcgtggtgtt tggcgtgagg aactcaaaaa gaggcgccgc aggctccctg atgaacgtgc
     3721 tgaactaccc cggcatgaat caccgcgtcg aaattaccga gggaatcctg gcagatgaat
     3781 gtgccgccct gctgtgcgat ttctatcgga tgcctagaca ggtgttcaat gctcagaaga
     3841 aggcccagag ctccatcaac tagtcagggc cctattctat agtgtcacct aaatgctaga
     3901 gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     3961 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     4021 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     4081 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     4141 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgctgcagtg
     4201 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     4261 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     4321 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     4381 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     4441 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     4501 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     4561 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     4621 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     4681 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     4741 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     4801 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     4861 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     4921 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4981 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5041 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5101 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     5161 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     5221 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     5281 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     5341 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     5401 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     5461 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     5521 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     5581 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     5641 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     5701 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     5761 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     5821 ggccccagtg ctgcaatgat accgcgagat ccacgctcac cggctccaga tttatcagca
     5881 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     5941 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6001 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     6061 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     6121 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     6181 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     6241 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     6301 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     6361 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     6421 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     6481 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     6541 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     6601 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     6661 ggggttccgc gcacatttcc ccgaaaagtg ccacct
//
