LOCUS       sequence-362135-        7764 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of SuperFi-Cas9 ABE7 base editor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7764)
  AUTHORS   Kulcsar PI, Talas A, Ligeti Z, Krausz SL, Welker E
  TITLE     SuperFi-Cas9 exhibits remarkable fidelity but severely reduced 
            activity yet works effectively with ABE8e.
  JOURNAL   Nat Commun. 2022 Nov 11;13(1):6858. doi: 10.1038/s41467-022-34527-8.
  PUBMED    36369279
REFERENCE   2  (bases 1 to 7764)
  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: "2022-11-11"; volume: "13(1):6858. doi"; pages: " 
            10.1038/s41467-022-34527-8"
COMMENT     Sequence Label: pAT15544_nABE-SuperFi-Cas9
FEATURES             Location/Qualifiers
     source          1..7764
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(387..1178,1179..1247))
                     /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     1010..1029
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1248..1352)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1435..1646
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      1666..1675
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1675..1698
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1705..1725
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2938..7038
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVDGDDKVDDDDKMIAKSEDEIGDATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7039..7086
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(7114..7131)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7120..7327
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(join(7392..7764,1..216))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7472..7491)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
       61 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
      121 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
      181 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
      241 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
      301 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
      361 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
      421 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
      481 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
      541 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
      601 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
      661 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
      721 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
      781 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
      841 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
      901 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
      961 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     1021 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     1081 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     1141 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     1201 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     1261 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     1321 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     1381 gtccagcaga gatccacttt ggcgccggct cgagtggctc cggtgcccgt cagtgggcag
     1441 agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaaccggtg
     1501 cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt
     1561 ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc
     1621 gcaacgggtt tgccgccaga acacaggtgt cgtgacgcgg gatccgccac catggattac
     1681 aaagacgatg acgataagat ggccccaaag aagaagcgga aggtcggtat ccacggagtc
     1741 ccagcagcca gtgaggtcga atttagtcat gagtattgga tgagacacgc cctgaccctt
     1801 gcaaaacgcg cctgggatga aagggaagtc cctgtggggg ccgtccttgt ccataataat
     1861 cgagtgattg gagagggctg gaatcgccct attggaaggc acgaccccac tgcacacgca
     1921 gagattatgg ctctccgaca gggtggactg gtaatgcaga attaccggct gatcgacgcc
     1981 accctctatg tcactcttga accctgtgta atgtgcgctg gcgccatgat ccacagcaga
     2041 ataggaagag tcgtcttcgg cgctagagat gctaaaactg gagctgcagg gagtttgatg
     2101 gatgtactcc accaccccgg gatgaatcat cgggtggaga taaccgaagg aatcctggct
     2161 gatgaatgcg ctgctctgtt gagcgatttc tttaggatga ggaggcagga gattaaggca
     2221 caaaagaaag ctcagagctc tactgacagt ggggggagtt ccggtggatc tagtggtagc
     2281 gagacacccg ggacttccga aagtgctacc ccagaatcat ccggggggag ttcaggcgga
     2341 agttctgaag tagagttctc tcacgagtat tggatgcgcc acgcactgac actggctaag
     2401 cgggcaaggg acgaacgaga agtcccagtc ggggctgtcc tcgtcttgaa taatagagtt
     2461 attggggagg ggtggaaccg agctattgga ctgcatgacc caactgcaca cgctgaaatt
     2521 atggccttga gacagggcgg tctcgtaatg cagaattata gattgataga tgctactttg
     2581 tatgtgactt tcgagccatg cgtcatgtgt gccggggcaa tgatccacag cagaattgga
     2641 agggttgtat tcggcgtccg aaacgctaag accggggctg ccgggtctct catggacgtc
     2701 cttcactatc ctggtatgaa tcaccgagtg gaaattaccg aaggaatcct cgctgacgaa
     2761 tgcgcagccc tcctctgtta tttctttcgg atgccaagac aggtctttaa tgctcagaag
     2821 aaagctcagt cctccactga ctcaggtggc tccagcggtg gaagctcagg atctgagacc
     2881 ccaggaacat ctgagtcagc cactcctgaa tcctcaggtg gtagctctgg ggggtctgac
     2941 aagaagtaca gcatcggcct ggccatcggc accaactctg tgggctgggc cgtgatcacc
     3001 gacgagtaca aggtgcccag caagaaattc aaggtgctgg gcaacaccga ccggcacagc
     3061 atcaagaaga acctgatcgg agccctgctg ttcgacagcg gcgaaacagc cgaggccacc
     3121 cggctgaaga gaaccgccag aagaagatac accagacgga agaaccggat ctgctatctg
     3181 caagagatct tcagcaacga gatggccaag gtggacgaca gcttcttcca cagactggaa
     3241 gagtccttcc tggtggaaga ggataagaag cacgagcggc accccatctt cggcaacatc
     3301 gtggacgagg tggcctacca cgagaagtac cccaccatct accacctgag aaagaaactg
     3361 gtggacagca ccgacaaggc cgacctgcgg ctgatctatc tggccctggc ccacatgatc
     3421 aagttccggg gccacttcct gatcgagggc gacctgaacc ccgacaacag cgacgtggac
     3481 aagctgttca tccagctggt gcagacctac aaccagctgt tcgaggaaaa ccccatcaac
     3541 gccagcggcg tggacgccaa ggccatcctg tctgccagac tgagcaagag cagacggctg
     3601 gaaaatctga tcgcccagct gcccggcgag aagaagaatg gcctgttcgg caacctgatt
     3661 gccctgagcc tgggcctgac ccccaacttc aagagcaact tcgacctggc cgaggatgcc
     3721 aaactgcagc tgagcaagga cacctacgac gacgacctgg acaacctgct ggcccagatc
     3781 ggcgaccagt acgccgacct gtttctggcc gccaagaacc tgtccgacgc catcctgctg
     3841 agcgacatcc tgagagtgaa caccgagatc accaaggccc ccctgagcgc ctctatgatc
     3901 aagagatacg acgagcacca ccaggacctg accctgctga aagctctcgt gcggcagcag
     3961 ctgcctgaga agtacaaaga gattttcttc gaccagagca agaacggcta cgccggctac
     4021 attgacggcg gagccagcca ggaagagttc tacaagttca tcaagcccat cctggaaaag
     4081 atggacggca ccgaggaact gctcgtgaag ctgaacagag aggacctgct gcggaagcag
     4141 cggaccttcg acaacggcag catcccccac cagatccacc tgggagagct gcacgccatt
     4201 ctgcggcggc aggaagattt ttacccattc ctgaaggaca accgggaaaa gatcgagaag
     4261 atcctgacct tccgcatccc ctactacgtg ggccctctgg ccaggggaaa cagcagattc
     4321 gcctggatga ccagaaagag cgaggaaacc atcaccccct ggaacttcga ggaagtggtg
     4381 gacaagggcg cttccgccca gagcttcatc gagcggatga ccaacttcga taagaacctg
     4441 cccaacgaga aggtgctgcc caagcacagc ctgctgtacg agtacttcac cgtgtataac
     4501 gagctgacca aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc
     4561 gagcagaaaa aggccatcgt ggacctgctg ttcaagacca accggaaagt gaccgtgaag
     4621 cagctgaaag aggactactt caagaaaatc gagtgcttcg actccgtgga aatctccggc
     4681 gtggaagatc ggttcaacgc ctccctgggc acataccacg atctgctgaa aattatcaag
     4741 gacaaggact tcctggacaa tgaggaaaac gaggacattc tggaagatat cgtgctgacc
     4801 ctgacactgt ttgaggacag agagatgatc gaggaacggc tgaaaaccta tgcccacctg
     4861 ttcgacgaca aagtgatgaa gcagctgaag cggcggagat acaccggctg gggcaggctg
     4921 agccggaagc tgatcaacgg catccgggac aagcagtccg gcaagacaat cctggatttc
     4981 ctgaagtccg acggcttcgc caacagaaac ttcatgcagc tgatccacga cgacagcctg
     5041 acctttaaag aggacatcca gaaagcccag gtgtccggcc agggcgatag cctgcacgag
     5101 cacattgcca atctggccgg cagccccgcc attaagaagg gcatcctgca gacagtgaag
     5161 gtggtggacg agctcgtgaa agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa
     5221 atggccagag agaaccagac cacccagaag ggacagaaga acagccgcga gagaatgaag
     5281 cggatcgaag agggcatcaa agagctgggc agccagatcc tgaaagaaca ccccgtggaa
     5341 aacacccagc tgcagaacga gaagctgtac ctgtactacc tgcagaatgg gcgggatatg
     5401 tacgtggacc aggaactgga catcaaccgg ctgtccgact acgatgtgga ccatatcgtg
     5461 cctcagagct ttctgaagga cgactccatc gacaacaagg tgctgaccag aagcgacaag
     5521 aaccggggca agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat gaagaactac
     5581 tggcggcagc tgctgaacgc caagctgatt acccagagaa agttcgacaa tctgaccaag
     5641 gccgagagag gcggcctgag cgaactggat aaggccggct tcatcaagag acagctggtg
     5701 gaaacccggc agatcacaaa gcacgtggca cagatcctgg actcccggat gaacactaag
     5761 tacgacgaga atgacaagct gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg
     5821 gtgtccgatt tccggaagga tttccagttt tacaaagtgc gcgagatcaa caactaccac
     5881 cacgcccacg acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct
     5941 aagctggaaa gcgagttcgt ggacggcgac gacaaggtgg acgacgacga caagatgatc
     6001 gccaagagcg aagacgaaat cggcgacgct accgccaagt acttcttcta cagcaacatc
     6061 atgaactttt tcaagaccga gattaccctg gccaacggcg agatccggaa gcggcctctg
     6121 atcgagacaa acggcgaaac cggggagatc gtgtgggata agggccggga ttttgccacc
     6181 gtgcggaaag tgctgagcat gccccaagtg aatatcgtga aaaagaccga ggtgcagaca
     6241 ggcggcttca gcaaagagtc tatcctgccc aagaggaaca gcgataagct gatcgccaga
     6301 aagaaggact gggaccctaa gaagtacggc ggcttcgaca gccccaccgt ggcctattct
     6361 gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga aactgaagag tgtgaaagag
     6421 ctgctgggga tcaccatcat ggaaagaagc agcttcgaga agaatcccat cgactttctg
     6481 gaagccaagg gctacaaaga agtgaaaaag gacctgatca tcaagctgcc taagtactcc
     6541 ctgttcgagc tggaaaacgg ccggaagaga atgctggcct ctgccggcga actgcagaag
     6601 ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc tgtacctggc cagccactat
     6661 gagaagctga agggctcccc cgaggataat gagcagaaac agctgtttgt ggaacagcac
     6721 aagcactacc tggacgagat catcgagcag atcagcgagt tctccaagag agtgatcctg
     6781 gccgacgcta atctggacaa agtgctgtcc gcctacaaca agcaccggga taagcccatc
     6841 agagagcagg ccgagaatat catccacctg tttaccctga ccaatctggg agcccctgcc
     6901 gccttcaagt actttgacac caccatcgac cggaagaggt acaccagcac caaagaggtg
     6961 ctggacgcca ccctgatcca ccagagcatc accggcctgt acgagacacg gatcgacctg
     7021 tctcagctgg gaggcgacaa gcgtcctgct gctactaaga aagctggtca agctaagaaa
     7081 aagaaatgac atgtcctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc
     7141 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     7201 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     7261 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagagaat agcaggcatg
     7321 ctggggagcg gccgctgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     7381 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     7441 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     7501 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     7561 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     7621 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     7681 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     7741 gcagccactg gtaacaggat tagc
//
