LOCUS       sequence-362133-        7764 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of nuclease inactive 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: pAT15543_dABE-SuperFi-Cas9
FEATURES             Location/Qualifiers
     source          1..7764
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(123..711)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(203..222)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(882..1673,1674..1742))
                     /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     1505..1524
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1743..1847)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1930..2141
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2161..2170
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2170..2193
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2200..2220
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(3433..3456,3457..3459,3460..6015,6016..6018,
                     6019..7533)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the N863A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVDGDDKVDDDDKMIAKSEDEIGDATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7534..7581
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(7609..7626)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    join(7615..7764,1..58)
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 gggtggggca ggacagcaag ggggaggatt gggaagagaa tagcaggcat gctggggagc
       61 ggccgctgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
      121 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
      181 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
      241 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
      301 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
      361 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
      421 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
      481 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
      541 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
      601 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
      661 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
      721 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
      781 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
      841 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
      901 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
      961 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     1021 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     1081 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     1141 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     1201 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     1261 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     1321 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     1381 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     1441 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     1501 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     1561 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     1621 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     1681 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     1741 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     1801 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     1861 aaagtgccac ctgacgtcca gcagagatcc actttggcgc cggctcgagt ggctccggtg
     1921 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
     1981 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
     2041 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
     2101 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtgtcgtga cgcgggatcc
     2161 gccaccatgg attacaaaga cgatgacgat aagatggccc caaagaagaa gcggaaggtc
     2221 ggtatccacg gagtcccagc agccagtgag gtcgaattta gtcatgagta ttggatgaga
     2281 cacgccctga cccttgcaaa acgcgcctgg gatgaaaggg aagtccctgt gggggccgtc
     2341 cttgtccata ataatcgagt gattggagag ggctggaatc gccctattgg aaggcacgac
     2401 cccactgcac acgcagagat tatggctctc cgacagggtg gactggtaat gcagaattac
     2461 cggctgatcg acgccaccct ctatgtcact cttgaaccct gtgtaatgtg cgctggcgcc
     2521 atgatccaca gcagaatagg aagagtcgtc ttcggcgcta gagatgctaa aactggagct
     2581 gcagggagtt tgatggatgt actccaccac cccgggatga atcatcgggt ggagataacc
     2641 gaaggaatcc tggctgatga atgcgctgct ctgttgagcg atttctttag gatgaggagg
     2701 caggagatta aggcacaaaa gaaagctcag agctctactg acagtggggg gagttccggt
     2761 ggatctagtg gtagcgagac acccgggact tccgaaagtg ctaccccaga atcatccggg
     2821 gggagttcag gcggaagttc tgaagtagag ttctctcacg agtattggat gcgccacgca
     2881 ctgacactgg ctaagcgggc aagggacgaa cgagaagtcc cagtcggggc tgtcctcgtc
     2941 ttgaataata gagttattgg ggaggggtgg aaccgagcta ttggactgca tgacccaact
     3001 gcacacgctg aaattatggc cttgagacag ggcggtctcg taatgcagaa ttatagattg
     3061 atagatgcta ctttgtatgt gactttcgag ccatgcgtca tgtgtgccgg ggcaatgatc
     3121 cacagcagaa ttggaagggt tgtattcggc gtccgaaacg ctaagaccgg ggctgccggg
     3181 tctctcatgg acgtccttca ctatcctggt atgaatcacc gagtggaaat taccgaagga
     3241 atcctcgctg acgaatgcgc agccctcctc tgttatttct ttcggatgcc aagacaggtc
     3301 tttaatgctc agaagaaagc tcagtcctcc actgactcag gtggctccag cggtggaagc
     3361 tcaggatctg agaccccagg aacatctgag tcagccactc ctgaatcctc aggtggtagc
     3421 tctggggggt ctgacaagaa gtacagcatc ggcctggcca tcggcaccaa ctctgtgggc
     3481 tgggccgtga tcaccgacga gtacaaggtg cccagcaaga aattcaaggt gctgggcaac
     3541 accgaccggc acagcatcaa gaagaacctg atcggagccc tgctgttcga cagcggcgaa
     3601 acagccgagg ccacccggct gaagagaacc gccagaagaa gatacaccag acggaagaac
     3661 cggatctgct atctgcaaga gatcttcagc aacgagatgg ccaaggtgga cgacagcttc
     3721 ttccacagac tggaagagtc cttcctggtg gaagaggata agaagcacga gcggcacccc
     3781 atcttcggca acatcgtgga cgaggtggcc taccacgaga agtaccccac catctaccac
     3841 ctgagaaaga aactggtgga cagcaccgac aaggccgacc tgcggctgat ctatctggcc
     3901 ctggcccaca tgatcaagtt ccggggccac ttcctgatcg agggcgacct gaaccccgac
     3961 aacagcgacg tggacaagct gttcatccag ctggtgcaga cctacaacca gctgttcgag
     4021 gaaaacccca tcaacgccag cggcgtggac gccaaggcca tcctgtctgc cagactgagc
     4081 aagagcagac ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg
     4141 ttcggcaacc tgattgccct gagcctgggc ctgaccccca acttcaagag caacttcgac
     4201 ctggccgagg atgccaaact gcagctgagc aaggacacct acgacgacga cctggacaac
     4261 ctgctggccc agatcggcga ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc
     4321 gacgccatcc tgctgagcga catcctgaga gtgaacaccg agatcaccaa ggcccccctg
     4381 agcgcctcta tgatcaagag atacgacgag caccaccagg acctgaccct gctgaaagct
     4441 ctcgtgcggc agcagctgcc tgagaagtac aaagagattt tcttcgacca gagcaagaac
     4501 ggctacgccg gctacattga cggcggagcc agccaggaag agttctacaa gttcatcaag
     4561 cccatcctgg aaaagatgga cggcaccgag gaactgctcg tgaagctgaa cagagaggac
     4621 ctgctgcgga agcagcggac cttcgacaac ggcagcatcc cccaccagat ccacctggga
     4681 gagctgcacg ccattctgcg gcggcaggaa gatttttacc cattcctgaa ggacaaccgg
     4741 gaaaagatcg agaagatcct gaccttccgc atcccctact acgtgggccc tctggccagg
     4801 ggaaacagca gattcgcctg gatgaccaga aagagcgagg aaaccatcac cccctggaac
     4861 ttcgaggaag tggtggacaa gggcgcttcc gcccagagct tcatcgagcg gatgaccaac
     4921 ttcgataaga acctgcccaa cgagaaggtg ctgcccaagc acagcctgct gtacgagtac
     4981 ttcaccgtgt ataacgagct gaccaaagtg aaatacgtga ccgagggaat gagaaagccc
     5041 gccttcctga gcggcgagca gaaaaaggcc atcgtggacc tgctgttcaa gaccaaccgg
     5101 aaagtgaccg tgaagcagct gaaagaggac tacttcaaga aaatcgagtg cttcgactcc
     5161 gtggaaatct ccggcgtgga agatcggttc aacgcctccc tgggcacata ccacgatctg
     5221 ctgaaaatta tcaaggacaa ggacttcctg gacaatgagg aaaacgagga cattctggaa
     5281 gatatcgtgc tgaccctgac actgtttgag gacagagaga tgatcgagga acggctgaaa
     5341 acctatgccc acctgttcga cgacaaagtg atgaagcagc tgaagcggcg gagatacacc
     5401 ggctggggca ggctgagccg gaagctgatc aacggcatcc gggacaagca gtccggcaag
     5461 acaatcctgg atttcctgaa gtccgacggc ttcgccaaca gaaacttcat gcagctgatc
     5521 cacgacgaca gcctgacctt taaagaggac atccagaaag cccaggtgtc cggccagggc
     5581 gatagcctgc acgagcacat tgccaatctg gccggcagcc ccgccattaa gaagggcatc
     5641 ctgcagacag tgaaggtggt ggacgagctc gtgaaagtga tgggccggca caagcccgag
     5701 aacatcgtga tcgaaatggc cagagagaac cagaccaccc agaagggaca gaagaacagc
     5761 cgcgagagaa tgaagcggat cgaagagggc atcaaagagc tgggcagcca gatcctgaaa
     5821 gaacacccag tggaaaacac ccagctgcag aacgagaagc tgtacctgta ctacctgcag
     5881 aatgggcggg atatgtacgt ggaccaggaa ctggacatca accggctgtc cgactacgat
     5941 gtggacgcca tcgtgcctca gagctttctg aaggacgact ccatcgacaa caaggtgctg
     6001 accagaagcg acaagaaccg gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag
     6061 aagatgaaga actactggcg gcagctgctg aacgccaagc tgattaccca gagaaagttc
     6121 gacaatctga ccaaggccga gagaggcggc ctgagcgaac tggataaggc cggcttcatc
     6181 aagagacagc tggtggaaac ccggcagatc acaaagcacg tggcacagat cctggactcc
     6241 cggatgaaca ctaagtacga cgagaatgac aagctgatcc gggaagtgaa agtgatcacc
     6301 ctgaagtcca agctggtgtc cgatttccgg aaggatttcc agttttacaa agtgcgcgag
     6361 atcaacaact accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg
     6421 atcaaaaagt accctaagct ggaaagcgag ttcgtggacg gcgacgacaa ggtggacgac
     6481 gacgacaaga tgatcgccaa gagcgaagac gaaatcggcg acgctaccgc caagtacttc
     6541 ttctacagca acatcatgaa ctttttcaag accgagatta ccctggccaa cggcgagatc
     6601 cggaagcggc ctctgatcga gacaaacggc gaaaccgggg agatcgtgtg ggataagggc
     6661 cgggattttg ccaccgtgcg gaaagtgctg agcatgcccc aagtgaatat cgtgaaaaag
     6721 accgaggtgc agacaggcgg cttcagcaaa gagtctatcc tgcccaagag gaacagcgat
     6781 aagctgatcg ccagaaagaa ggactgggac cctaagaagt acggcggctt cgacagcccc
     6841 accgtggcct attctgtgct ggtggtggcc aaagtggaaa agggcaagtc caagaaactg
     6901 aagagtgtga aagagctgct ggggatcacc atcatggaaa gaagcagctt cgagaagaat
     6961 cccatcgact ttctggaagc caagggctac aaagaagtga aaaaggacct gatcatcaag
     7021 ctgcctaagt actccctgtt cgagctggaa aacggccgga agagaatgct ggcctctgcc
     7081 ggcgaactgc agaagggaaa cgaactggcc ctgccctcca aatatgtgaa cttcctgtac
     7141 ctggccagcc actatgagaa gctgaagggc tcccccgagg ataatgagca gaaacagctg
     7201 tttgtggaac agcacaagca ctacctggac gagatcatcg agcagatcag cgagttctcc
     7261 aagagagtga tcctggccga cgctaatctg gacaaagtgc tgtccgccta caacaagcac
     7321 cgggataagc ccatcagaga gcaggccgag aatatcatcc acctgtttac cctgaccaat
     7381 ctgggagccc ctgccgcctt caagtacttt gacaccacca tcgaccggaa gaggtacacc
     7441 agcaccaaag aggtgctgga cgccaccctg atccaccaga gcatcaccgg cctgtacgag
     7501 acacggatcg acctgtctca gctgggaggc gacaagcgtc ctgctgctac taagaaagct
     7561 ggtcaagcta agaaaaagaa atgacatgtc ctagagctcg ctgatcagcc tcgactgtgc
     7621 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     7681 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     7741 ggtgtcattc tattctgggg ggtg
//
