LOCUS       sequence-216920-        8574 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expressing ABE7.10 (TadA-TadA*7.10-nCas9-NLS-HA), human codon 
            optimized.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8574)
  AUTHORS   Ryu SM, Koo T, Kim K, Lim K, Baek G, Kim ST, Kim HS, Kim DE, Lee H, 
            Chung E, Kim JS
  TITLE     Adenine base editing in mouse embryos and an adult mouse model of 
            Duchenne muscular dystrophy.
  JOURNAL   Nat Biotechnol. 2018 Jul;36(6):536-539. doi: 10.1038/nbt.4148. Epub 
            2018 Apr 27.
  PUBMED    29702637
REFERENCE   2  (bases 1 to 8574)
  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: "10.1038/nbt.4148"; 
            journalName: "Nat Biotechnol"; date: "2018-07"; volume: "36"; issue:
            "6"; pages: "536-539"
COMMENT     Sequence Label: p3s-ABE7.10
FEATURES             Location/Qualifiers
     source          1..8574
                     /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             join(2092..2118,2119..2121,2122..6195)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             6211..6231
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6232..6258
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             6232..6258
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(6292..6310)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6293..6310)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(6330..6347)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6336..6560
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6648..6665)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6819..7407)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6899..6918)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7578..8369,8370..8438))
                     /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     8201..8220
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8439..8543)
                     /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 gcttgccacc
      901 atgagtgaag tggaattctc ccacgagtat tggatgagac acgcattgac gctcgcaaaa
      961 agagcgtggg acgagaggga agtacccgtc ggagcagtcc tcgtgcataa caatcgggtg
     1021 ataggcgagg gctggaatcg acctatcggc cgacatgacc cgaccgccca tgcagaaatt
     1081 atggctctgc gacagggagg gttggtgatg cagaactatc ggcttatcga tgccacactc
     1141 tacgttacgc ttgaaccgtg cgttatgtgt gcgggggcga tgatacatag tagaataggg
     1201 agggtcgtgt tcggggcgcg agatgcgaaa accggggccg caggctcatt gatggacgtc
     1261 ctgcaccacc ccggcatgaa ccatcgggtc gagataaccg aaggcatatt ggcagacgaa
     1321 tgtgctgcgc tcctttccga tttctttagg atgcgacgcc aggagatcaa ggcgcaaaaa
     1381 aaagcccagt ctagtaccga ctccgggggc agcagtgggg ggtctagtgg cagtgagacg
     1441 cccggtacga gcgagagcgc cacgcctgaa tcatcaggtg gaagcagtgg tggctcttct
     1501 gaagtcgaat tttcccacga gtactggatg agacacgcac ttacgcttgc caagagggca
     1561 agagatgaac gcgaggtccc ggtcggagca gtgcttgtcc ttaataatag agtgataggc
     1621 gaaggctgga atcgagctat cggcctccac gatcctaccg cgcatgctga gatcatggcg
     1681 ctgcgacaag gggggctggt aatgcaaaac tatagactca tcgatgcgac gctgtatgtg
     1741 acctttgagc cctgtgttat gtgtgctggg gctatgatac atagcagaat tggcagggta
     1801 gtcttcggcg tccgcaatgc caaaaccggg gcagctggat ccttgatgga tgttttgcat
     1861 taccctggaa tgaatcatag ggttgagatt acagaaggta tcttggcgga tgaatgcgcc
     1921 gcacttctct gttatttttt tcggatgccg aggcaagtct tcaatgccca gaaaaaagcg
     1981 cagtctagta cagacagtgg aggctcaagt gggggcagct caggaagcga aactcctggt
     2041 acctcagagt ccgctactcc cgaatcctct gggggtagct cagggggcag tatggacaag
     2101 aagtacagca tcggcctggc catcggtacc aacagcgtgg gctgggccgt gatcaccgac
     2161 gagtacaagg tgcccagcaa gaagttcaag gtgctgggca acaccgaccg ccacagcatc
     2221 aagaagaacc tgatcggcgc cctgctgttc gacagcggcg agaccgccga ggccacccgc
     2281 ctgaagcgca ccgcccgccg ccgctacacc cgccgcaaga accgcatctg ctacctgcag
     2341 gagatcttca gcaacgagat ggccaaggtg gacgacagct tcttccaccg cctggaggag
     2401 agcttcctgg tggaggagga caagaagcac gagcgccacc ccatcttcgg caacatcgtg
     2461 gacgaggtgg cctaccacga gaagtacccc accatctacc acctgcgcaa gaagctggtg
     2521 gacagcaccg acaaggccga cctgcgcctg atctacctgg ccctggccca catgatcaag
     2581 ttccgcggcc acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag
     2641 ctgttcatcc agctggtgca gacctacaac cagctgttcg aggagaaccc catcaacgcc
     2701 agcggcgtgg acgccaaggc catcctgagc gcccgcctga gcaagagccg ccgcctggag
     2761 aacctgatcg cccagctgcc cggcgagaag aagaacggcc tgttcggcaa cctgatcgcc
     2821 ctgagcctgg gcctgacccc caacttcaag agcaacttcg acctggccga ggacgccaag
     2881 ctgcagctga gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc
     2941 gaccagtacg ccgacctgtt cctggccgcc aagaacctga gcgacgccat cctgctgagc
     3001 gacatcctgc gcgtgaacac cgagatcacc aaggcccccc tgagcgccag catgatcaag
     3061 cgctacgacg agcaccacca ggacctgacc ctgctgaagg ccctggtgcg ccagcagctg
     3121 cccgagaagt acaaggagat cttcttcgac cagagcaaga acggctacgc cggctacatc
     3181 gacggcggcg ccagccagga ggagttctac aagttcatca agcccatcct ggagaagatg
     3241 gacggcaccg aggagctgct ggtgaagctg aaccgcgagg acctgctgcg caagcagcgc
     3301 accttcgaca acggcagcat cccccaccag atccacctgg gcgagctgca cgccatcctg
     3361 cgccgccagg aggacttcta ccccttcctg aaggacaacc gcgagaagat cgagaagatc
     3421 ctgaccttcc gcatccccta ctacgtgggc cccctggccc gcggcaacag ccgcttcgcc
     3481 tggatgaccc gcaagagcga ggagaccatc accccctgga acttcgagga ggtggtggac
     3541 aagggcgcca gcgcccagag cttcatcgag cgcatgacca acttcgacaa gaacctgccc
     3601 aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtacaacgag
     3661 ctgaccaagg tgaagtacgt gaccgagggc atgcgcaagc ccgccttcct gagcggcgag
     3721 cagaagaagg ccatcgtgga cctgctgttc aagaccaacc gcaaggtgac cgtgaagcag
     3781 ctgaaggagg actacttcaa gaagatcgag tgcttcgaca gcgtggagat cagcggcgtg
     3841 gaggaccgct tcaacgccag cctgggcacc taccacgacc tgctgaagat catcaaggac
     3901 aaggacttcc tggacaacga ggagaacgag gacatcctgg aggacatcgt gctgaccctg
     3961 accctgttcg aggaccgcga gatgatcgag gagcgcctga agacctacgc ccacctgttc
     4021 gacgacaagg tgatgaagca gctgaagcgc cgccgctaca ccggctgggg ccgcctgagc
     4081 cgcaagctta tcaacggcat ccgcgacaag cagagcggca agaccatcct ggacttcctg
     4141 aagagcgacg gcttcgccaa ccgcaacttc atgcagctga tccacgacga cagcctgacc
     4201 ttcaaggagg acatccagaa ggcccaggtg agcggccagg gcgacagcct gcacgagcac
     4261 atcgccaacc tggccggcag ccccgccatc aagaagggca tcctgcagac cgtgaaggtg
     4321 gtggacgagc tggtgaaggt gatgggccgc cacaagcccg agaacatcgt gatcgagatg
     4381 gcccgcgaga accagaccac ccagaagggc cagaagaaca gccgcgagcg catgaagcgc
     4441 atcgaggagg gcatcaagga gctgggcagc cagatcctga aggagcaccc cgtggagaac
     4501 acccagctgc agaacgagaa gctgtacctg tactacctgc agaacggccg cgacatgtac
     4561 gtggaccagg agctggacat caaccgcctg agcgactacg acgtggacca catcgtgccc
     4621 cagagcttcc tgaaggacga cagcatcgac aacaaggtgc tgacccgcag cgacaagaac
     4681 cgcggcaaga gcgacaacgt gcccagcgag gaggtggtga agaagatgaa gaactactgg
     4741 cgccagctgc tgaacgccaa gctgatcacc cagcgcaagt tcgacaacct gaccaaggcc
     4801 gagcgcggcg gcctgagcga gctggacaag gccggcttca tcaagcgcca gctggtggag
     4861 acccgccaga tcaccaagca cgtggcccag atcctggaca gccgcatgaa caccaagtac
     4921 gacgagaacg acaagctgat ccgcgaggtg aaggtgatca ccctgaagag caagctggtg
     4981 agcgacttcc gcaaggactt ccagttctac aaggtgcgcg agatcaacaa ctaccaccac
     5041 gcccacgacg cctacctgaa cgccgtggtg ggcaccgccc tgatcaagaa gtaccccaag
     5101 ctggagagcg agttcgtgta cggcgactac aaggtgtacg acgtgcgcaa gatgatcgcc
     5161 aagagcgagc aggagatcgg caaggccacc gccaagtact tcttctacag caacatcatg
     5221 aacttcttca agaccgagat caccctggcc aacggcgaga tccgcaagcg ccccctgatc
     5281 gagaccaacg gcgagaccgg cgagatcgtg tgggacaagg gccgcgactt cgccaccgtg
     5341 cgcaaggtgc tgagcatgcc ccaggtgaac atcgtgaaga agaccgaggt gcagaccggc
     5401 ggcttcagca aggagagcat cctgcccaag cgcaacagcg acaagctgat cgcccgcaag
     5461 aaggactggg accccaagaa gtacggcggc ttcgacagcc ccaccgtggc ctacagcgtg
     5521 ctggtggtgg ccaaggtgga gaagggcaag agcaagaagc tgaagagcgt gaaggagctg
     5581 ctgggcatca ccatcatgga gcgcagcagc ttcgagaaga accccatcga cttcctggag
     5641 gccaagggct acaaggaggt gaagaaggac ctgatcatca agctgcccaa gtacagcctg
     5701 ttcgagctgg agaacggccg caagcgcatg ctggccagcg ccggcgagct gcagaagggc
     5761 aacgagctgg ccctgcccag caagtacgtg aacttcctgt acctggccag ccactacgag
     5821 aagctgaagg gcagccccga ggacaacgag cagaagcagc tgttcgtgga gcagcacaag
     5881 cactacctgg acgagatcat cgagcagatc agcgagttca gcaagcgcgt gatcctggcc
     5941 gacgccaacc tggacaaggt gctgagcgcc tacaacaagc accgcgacaa gcccatccgc
     6001 gagcaggccg agaacatcat ccacctgttc accctgacca acctgggcgc ccccgccgcc
     6061 ttcaagtact tcgacaccac catcgaccgc aagcgctaca ccagcaccaa ggaggtgctg
     6121 gacgccaccc tgatccacca gagcatcacc ggtctgtacg agacccgcat cgacctgagc
     6181 cagctgggcg gcgacggcgg ctccggacct ccaaagaaaa agagaaaagt atacccctac
     6241 gacgtgcccg actacgccta ataactcgag catgcatcta gagggcccta ttctatagtg
     6301 tcacctaaat gctagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     6361 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     6421 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     6481 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     6541 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctgcatta atgaatcggc
     6601 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     6661 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     6721 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     6781 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     6841 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     6901 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     6961 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     7021 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     7081 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     7141 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     7201 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     7261 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     7321 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     7381 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     7441 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     7501 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     7561 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     7621 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     7681 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     7741 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     7801 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     7861 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     7921 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     7981 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     8041 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     8101 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     8161 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     8221 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     8281 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     8341 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     8401 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     8461 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     8521 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
