LOCUS       sequence-261040-        4986 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresion of SaCas9 nickase (D10A) with ABEMax at the N terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4986)
  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 4986)
  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: ABEMax nSaCas9
FEATURES             Location/Qualifiers
     source          1..4986
                     /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             4417..4437
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4450..4497
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4498..4521
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4531..4587
                     /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             4588..4986
                     /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 gaagtcgagt ttagccacga gtattggatg aggcacgcac tgaccctggc aaagcgagca
      121 tgggatgaaa gagaagtccc cgtgggcgcc gtgctggtgc acaacaatag agtgatcgga
      181 gagggatgga acaggccaat cggccgccac gaccctaccg cacacgcaga gatcatggca
      241 ctgaggcagg gaggcctggt catgcagaat taccgcctga tcgatgccac cctgtatgtg
      301 acactggagc catgcgtgat gtgcgcagga gcaatgatcc acagcaggat cggaagagtg
      361 gtgttcggag cacgggacgc caagaccggc gcagcaggct ccctgatgga tgtgctgcac
      421 caccccggca tgaaccaccg ggtggagatc acagagggaa tcctggcaga cgagtgcgcc
      481 gccctgctga gcgatttctt tagaatgcgg agacaggaga tcaaggccca gaagaaggca
      541 cagagctcca ccgactctgg aggatctagc ggaggatcct ctggaagcga gacaccaggc
      601 acaagcgagt ccgccacacc agagagctcc ggcggctcct ccggaggatc ctctgaggtg
      661 gagttttccc acgagtactg gatgagacat gccctgaccc tggccaagag ggcacgcgat
      721 gagagggagg tgcctgtggg agccgtgctg gtgctgaaca atagagtgat cggcgagggc
      781 tggaacagag ccatcggcct gcacgaccca acagcccatg ccgaaattat ggccctgaga
      841 cagggcggcc tggtcatgca gaactacaga ctgattgacg ccaccctgta cgtgacattc
      901 gagccttgcg tgatgtgcgc cggcgccatg atccactcta ggatcggccg cgtggtgttt
      961 ggcgtgagga acgcaaaaac cggcgccgca ggctccctga tggacgtgct gcactacccc
     1021 ggcatgaatc accgcgtcga aattaccgag ggaatcctgg cagatgaatg tgccgccctg
     1081 ctgtgctatt tctttcggat gcctagacag gtgttcaatg ctcagaagaa ggcccagagc
     1141 tccaccgact ccggaggatc tagcggaggc tcctctggct ctgagacacc tggcacaagc
     1201 gagagcgcaa cacctgaaag cagcgggggc agcagcgggg ggtcagggaa gcgaaattac
     1261 attctggggc tggccattgg cattacatca gtgggctatg gcatcattga ctacgagaca
     1321 agggacgtga tcgacgccgg cgtgagactg ttcaaggagg ccaacgtgga gaacaatgag
     1381 ggccggagat ccaagagggg agcaaggcgc ctgaagcgga gaaggcgcca cagaatccag
     1441 agagtgaaga agctgctgtt cgattacaac ctgctgaccg accactccga gctgtctggc
     1501 atcaatcctt atgaggccag agtgaagggc ctgtcccaga agctgtctga ggaggagttt
     1561 agcgccgccc tgctgcacct ggcaaagagg agaggcgtgc acaacgtgaa tgaggtggag
     1621 gaggacaccg gcaacgagct gtccacaaag gagcagatca gccgcaattc caaggccctg
     1681 gaggagaagt atgtggccga gctgcagctg gagcggctga agaaggatgg cgaggtgagg
     1741 ggctccatca atcgcttcaa gacctctgac tacgtgaagg aggccaagca gctgctgaag
     1801 gtgcagaagg cctaccacca gctggatcag tcctttatcg atacatatat cgacctgctg
     1861 gagacaaggc gcacatacta tgagggacca ggagagggct ctcccttcgg ctggaaggac
     1921 atcaaggagt ggtacgagat gctgatgggc cactgcacct attttccaga ggagctgaga
     1981 agcgtgaagt acgcctataa cgccgatctg tacaacgccc tgaatgacct gaacaacctg
     2041 gtcatcacca gggatgagaa cgagaagctg gagtactatg agaagttcca gatcatcgag
     2101 aacgtgttca agcagaagaa gaagcctaca ctgaagcaga tcgccaagga gatcctggtg
     2161 aacgaggagg acatcaaggg ctaccgcgtg acctccacag gcaagccaga gttcaccaat
     2221 ctgaaggtgt atcacgatat caaggacatc acagcccgga aggagatcat cgagaacgcc
     2281 gagctgctgg atcagatcgc caagatcctg accatctatc agagctccga ggacatccag
     2341 gaggagctga ccaacctgaa tagcgagctg acacaggagg agatcgagca gatcagcaat
     2401 ctgaagggct acaccggcac acacaacctg agcctgaagg ccatcaatct gatcctggat
     2461 gagctgtggc acacaaacga caatcagatc gccatcttta accggctgaa gctggtgcca
     2521 aagaaggtgg acctgtccca gcagaaggag atcccaacca cactggtgga cgatttcatc
     2581 ctgtctcccg tggtgaagcg gagcttcatc cagagcatca aagtgatcaa cgccatcatc
     2641 aagaagtacg gcctgcccaa tgatatcatc atcgagctgg ccagggagaa gaactccaag
     2701 gacgcccaga agatgatcaa tgagatgcag aagaggaacc gccagaccaa tgagcggatc
     2761 gaggagatca tcagaaccac aggcaaggag aacgccaagt acctgatcga gaagatcaag
     2821 ctgcacgata tgcaggaggg caagtgtctg tattctctgg aggccatccc tctggaggac
     2881 ctgctgaaca atccattcaa ctacgaggtg gatcacatca tcccccggag cgtgagcttc
     2941 gacaattctt ttaacaataa ggtgctggtg aagcaggagg agaacagcaa gaagggcaat
     3001 aggacccctt tccagtacct gtctagctcc gattctaaga tcagctacga gacattcaag
     3061 aagcacatcc tgaatctggc caagggcaag ggccgcatca gcaagaccaa gaaggagtac
     3121 ctgctggagg agcgggacat caacagattc tccgtgcaga aggacttcat caaccggaat
     3181 ctggtggaca ccagatacgc cacacgcggc ctgatgaatc tgctgcggtc ttatttcaga
     3241 gtgaacaatc tggatgtgaa ggtgaagagc atcaacggcg gcttcacctc ctttctgcgg
     3301 agaaagtgga agtttaagaa ggagcgcaac aagggctata agcaccacgc cgaggatgcc
     3361 ctgatcatcg ccaatgccga cttcatcttt aaggagtgga agaagctgga caaggccaag
     3421 aaagtgatgg agaaccagat gttcgaggag aagcaggccg agagcatgcc cgagatcgag
     3481 acagagcagg agtacaagga gattttcatc acacctcacc agatcaagca catcaaggac
     3541 ttcaaggact acaagtattc tcacagggtg gataagaagc ccaaccgcga gctgatcaat
     3601 gacaccctgt atagcacacg gaaggacgat aagggcaata ccctgatcgt gaacaatctg
     3661 aacggcctgt acgacaagga taatgacaag ctgaagaagc tgatcaacaa gtctcccgag
     3721 aagctgctga tgtaccacca cgatcctcag acatatcaga agctgaagct gatcatggag
     3781 cagtacggcg acgagaagaa cccactgtat aagtactatg aggagacagg caactacctg
     3841 acaaagtata gcaagaagga taatggcccc gtgatcaaga agatcaagta ctatggcaac
     3901 aagctgaatg cccacctgga catcaccgac gattacccta actctcgcaa taaggtggtg
     3961 aagctgagcc tgaagccata ccggttcgac gtgtacctgg acaacggcgt gtataagttt
     4021 gtgacagtga agaatctgga tgtgatcaag aaggagaact actatgaggt gaacagcaag
     4081 tgctacgagg aggccaagaa gctgaagaag atcagcaacc aggccgagtt catcgcctct
     4141 ttttacaaca atgacctgat caagatcaat ggcgagctgt atagagtgat cggcgtgaac
     4201 aatgatctgc tgaacagaat cgaagtgaat atgatcgaca tcacctacag ggagtatctg
     4261 gagaacatga atgataagag gccccctcgc atcatcaaga ccatcgcctc taagacacag
     4321 agcatcaaga agtacagcac agacatcctg gggaacctgt atgaagtcaa gagcaagaaa
     4381 catcctcaga ttatcaagaa aggctctggt ggttctccca agaagaagag gaaagtctct
     4441 ggtggttcta agcgacctgc cgccacaaag aaggctggac aggctaagaa gaagaaagat
     4501 tacaaagacg atgacgataa gggatccggc gcaacaaact tctctctgct gaaacaagcc
     4561 ggagatgtcg aagagaatcc tggaccgatg gccaagcctt tgtctcaaga agaatccacc
     4621 ctcattgaaa gagcaacggc tacaatcaac agcatcccca tctctgaaga ctacagcgtc
     4681 gccagcgcag ctctctctag cgacggccgc atcttcactg gtgtcaatgt atatcatttt
     4741 actgggggac cttgtgcaga actcgtggtg ctgggcactg ctgctgctgc ggcagctggc
     4801 aacctgactt gtatcgtcgc gatcggaaat gagaacaggg gcatcttgag cccctgcgga
     4861 cggtgccgac aggtgcttct cgatctgcat cctgggatca aagccatagt gaaggacagt
     4921 gatggacagc cgacggcagt tgggattcgt gaattgctgc cctctggtta tgtgtgggag
     4981 ggctaa
//
