LOCUS       sequence-261041-        4392 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression of SaCas9 nickase (D10A) with single evolved TadA monomer
            (ABEMax) with a V82G substitution.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4392)
  AUTHORS   Nguyen Tran MT, Mohd Khalid MKN, Wang Q, Walker JKR, Lidgerwood GE, 
            Dilworth KL, Lisowski L, Pebay A, Hewitt AW
  TITLE     Engineering domain-inlaid SaCas9 adenine base editors with reduced 
            RNA off-targets and increased on-target DNA editing.
  JOURNAL   Nat Commun. 2020 Sep 25;11(1):4871. doi: 10.1038/s41467-020-18715-y.
  PUBMED    32978399
REFERENCE   2  (bases 1 to 4392)
  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: "2020-09-25"; volume: "11(1):4871. doi"; pages: " 
            10.1038/s41467-020-18715-y"
COMMENT     Sequence Label: miniABEMax V82G nSaCas9
FEATURES             Location/Qualifiers
     source          1..4392
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             37..57
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3823..3843
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3856..3903
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             3904..3927
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3937..3993
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             3994..4392
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
ORIGIN
        1 atgaaacgga cagccgacgg aagcgagttc gagtcaccaa agaagaagcg gaaagtctct
       61 gaggtggagt tttcccacga gtactggatg agacatgccc tgaccctggc caagagggca
      121 cgcgatgaga gggaggtgcc tgtgggagcc gtgctggtgc tgaacaatag agtgatcggc
      181 gagggctgga acagagccat cggcctgcac gacccaacag cccatgccga aattatggcc
      241 ctgagacagg gcggcctggt catgcagaac tacagactga ttgacgccac cctgtacggc
      301 acattcgagc cttgcgtgat gtgcgccggc gccatgatcc actctaggat cggccgcgtg
      361 gtgtttggcg tgaggaacgc aaaaaccggc gccgcaggct ccctgatgga cgtgctgcac
      421 taccccggca tgaatcaccg cgtcgaaatt accgagggaa tcctggcaga tgaatgtgcc
      481 gccctgctgt gctatttctt tcggatgcct agacaggtgt tcaatgctca gaagaaggcc
      541 cagagctcca ccgactccgg aggatctagc ggaggctcct ctggctctga gacacctggc
      601 acaagcgaga gcgcaacacc tgaaagcagc gggggcagca gcggggggtc agggaagcga
      661 aattacattc tggggctggc cattggcatt acatcagtgg gctatggcat cattgactac
      721 gagacaaggg acgtgatcga cgccggcgtg agactgttca aggaggccaa cgtggagaac
      781 aatgagggcc ggagatccaa gaggggagca aggcgcctga agcggagaag gcgccacaga
      841 atccagagag tgaagaagct gctgttcgat tacaacctgc tgaccgacca ctccgagctg
      901 tctggcatca atccttatga ggccagagtg aagggcctgt cccagaagct gtctgaggag
      961 gagtttagcg ccgccctgct gcacctggca aagaggagag gcgtgcacaa cgtgaatgag
     1021 gtggaggagg acaccggcaa cgagctgtcc acaaaggagc agatcagccg caattccaag
     1081 gccctggagg agaagtatgt ggccgagctg cagctggagc ggctgaagaa ggatggcgag
     1141 gtgaggggct ccatcaatcg cttcaagacc tctgactacg tgaaggaggc caagcagctg
     1201 ctgaaggtgc agaaggccta ccaccagctg gatcagtcct ttatcgatac atatatcgac
     1261 ctgctggaga caaggcgcac atactatgag ggaccaggag agggctctcc cttcggctgg
     1321 aaggacatca aggagtggta cgagatgctg atgggccact gcacctattt tccagaggag
     1381 ctgagaagcg tgaagtacgc ctataacgcc gatctgtaca acgccctgaa tgacctgaac
     1441 aacctggtca tcaccaggga tgagaacgag aagctggagt actatgagaa gttccagatc
     1501 atcgagaacg tgttcaagca gaagaagaag cctacactga agcagatcgc caaggagatc
     1561 ctggtgaacg aggaggacat caagggctac cgcgtgacct ccacaggcaa gccagagttc
     1621 accaatctga aggtgtatca cgatatcaag gacatcacag cccggaagga gatcatcgag
     1681 aacgccgagc tgctggatca gatcgccaag atcctgacca tctatcagag ctccgaggac
     1741 atccaggagg agctgaccaa cctgaatagc gagctgacac aggaggagat cgagcagatc
     1801 agcaatctga agggctacac cggcacacac aacctgagcc tgaaggccat caatctgatc
     1861 ctggatgagc tgtggcacac aaacgacaat cagatcgcca tctttaaccg gctgaagctg
     1921 gtgccaaaga aggtggacct gtcccagcag aaggagatcc caaccacact ggtggacgat
     1981 ttcatcctgt ctcccgtggt gaagcggagc ttcatccaga gcatcaaagt gatcaacgcc
     2041 atcatcaaga agtacggcct gcccaatgat atcatcatcg agctggccag ggagaagaac
     2101 tccaaggacg cccagaagat gatcaatgag atgcagaaga ggaaccgcca gaccaatgag
     2161 cggatcgagg agatcatcag aaccacaggc aaggagaacg ccaagtacct gatcgagaag
     2221 atcaagctgc acgatatgca ggagggcaag tgtctgtatt ctctggaggc catccctctg
     2281 gaggacctgc tgaacaatcc attcaactac gaggtggatc acatcatccc ccggagcgtg
     2341 agcttcgaca attcttttaa caataaggtg ctggtgaagc aggaggagaa cagcaagaag
     2401 ggcaatagga cccctttcca gtacctgtct agctccgatt ctaagatcag ctacgagaca
     2461 ttcaagaagc acatcctgaa tctggccaag ggcaagggcc gcatcagcaa gaccaagaag
     2521 gagtacctgc tggaggagcg ggacatcaac agattctccg tgcagaagga cttcatcaac
     2581 cggaatctgg tggacaccag atacgccaca cgcggcctga tgaatctgct gcggtcttat
     2641 ttcagagtga acaatctgga tgtgaaggtg aagagcatca acggcggctt cacctccttt
     2701 ctgcggagaa agtggaagtt taagaaggag cgcaacaagg gctataagca ccacgccgag
     2761 gatgccctga tcatcgccaa tgccgacttc atctttaagg agtggaagaa gctggacaag
     2821 gccaagaaag tgatggagaa ccagatgttc gaggagaagc aggccgagag catgcccgag
     2881 atcgagacag agcaggagta caaggagatt ttcatcacac ctcaccagat caagcacatc
     2941 aaggacttca aggactacaa gtattctcac agggtggata agaagcccaa ccgcgagctg
     3001 atcaatgaca ccctgtatag cacacggaag gacgataagg gcaataccct gatcgtgaac
     3061 aatctgaacg gcctgtacga caaggataat gacaagctga agaagctgat caacaagtct
     3121 cccgagaagc tgctgatgta ccaccacgat cctcagacat atcagaagct gaagctgatc
     3181 atggagcagt acggcgacga gaagaaccca ctgtataagt actatgagga gacaggcaac
     3241 tacctgacaa agtatagcaa gaaggataat ggccccgtga tcaagaagat caagtactat
     3301 ggcaacaagc tgaatgccca cctggacatc accgacgatt accctaactc tcgcaataag
     3361 gtggtgaagc tgagcctgaa gccataccgg ttcgacgtgt acctggacaa cggcgtgtat
     3421 aagtttgtga cagtgaagaa tctggatgtg atcaagaagg agaactacta tgaggtgaac
     3481 agcaagtgct acgaggaggc caagaagctg aagaagatca gcaaccaggc cgagttcatc
     3541 gcctcttttt acaacaatga cctgatcaag atcaatggcg agctgtatag agtgatcggc
     3601 gtgaacaatg atctgctgaa cagaatcgaa gtgaatatga tcgacatcac ctacagggag
     3661 tatctggaga acatgaatga taagaggccc cctcgcatca tcaagaccat cgcctctaag
     3721 acacagagca tcaagaagta cagcacagac atcctgggga acctgtatga agtcaagagc
     3781 aagaaacatc ctcagattat caagaaaggc tctggtggtt ctcccaagaa gaagaggaaa
     3841 gtctctggtg gttctaagcg acctgccgcc acaaagaagg ctggacaggc taagaagaag
     3901 aaagattaca aagacgatga cgataaggga tccggcgcaa caaacttctc tctgctgaaa
     3961 caagccggag atgtcgaaga gaatcctgga ccgatggcca agcctttgtc tcaagaagaa
     4021 tccaccctca ttgaaagagc aacggctaca atcaacagca tccccatctc tgaagactac
     4081 agcgtcgcca gcgcagctct ctctagcgac ggccgcatct tcactggtgt caatgtatat
     4141 cattttactg ggggaccttg tgcagaactc gtggtgctgg gcactgctgc tgctgcggca
     4201 gctggcaacc tgacttgtat cgtcgcgatc ggaaatgaga acaggggcat cttgagcccc
     4261 tgcggacggt gccgacaggt gcttctcgat ctgcatcctg ggatcaaagc catagtgaag
     4321 gacagtgatg gacagccgac ggcagttggg attcgtgaat tgctgccctc tggttatgtg
     4381 tgggagggct aa
//
