LOCUS       sequence-261050-        4401 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression of nSaCas9 intradomain insertion of single evolved TadA 
            monomer (V82G) at the specified position (E131 ).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4401)
  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 4401)
  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: nSaCas9 E131 miniABEMax V82G
FEATURES             Location/Qualifiers
     source          1..4401
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             394..414
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             430..450
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3865..3912
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             3913..3936
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3946..4002
                     /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             4003..4401
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
ORIGIN
        1 atgaagcgaa attacattct ggggctggcc attggcatta catcagtggg ctatggcatc
       61 attgactacg agacaaggga cgtgatcgac gccggcgtga gactgttcaa ggaggccaac
      121 gtggagaaca atgagggccg gagatccaag aggggagcaa ggcgcctgaa gcggagaagg
      181 cgccacagaa tccagagagt gaagaagctg ctgttcgatt acaacctgct gaccgaccac
      241 tccgagctgt ctggcatcaa tccttatgag gccagagtga agggcctgtc ccagaagctg
      301 tctgaggagg agtttagcgc cgccctgctg cacctggcaa agaggagagg cgtgcacaac
      361 gtgaatgagg tggaggagga caccggcaac gagccgaaga agaaacgtaa ggtgggtagc
      421 agcggtagcc cgaaaaagaa gcgcaaagtg ggcagcgacg cgctggacga tttcgatctg
      481 gacatgctgg gcagcgatgc gctggatgat ttcggcggtg gcagctctga ggtggagttt
      541 tcccacgagt actggatgag acatgccctg accctggcca agagggcacg cgatgagagg
      601 gaggtgcctg tgggagccgt gctggtgctg aacaatagag tgatcggcga gggctggaac
      661 agagccatcg gcctgcacga cccaacagcc catgccgaaa ttatggccct gagacagggc
      721 ggcctggtca tgcagaacta cagactgatt gacgccaccc tgtacggcac attcgagcct
      781 tgcgtgatgt gcgccggcgc catgatccac tctaggatcg gccgcgtggt gtttggcgtg
      841 aggaacgcaa aaaccggcgc cgcaggctcc ctgatggacg tgctgcacta ccccggcatg
      901 aatcaccgcg tcgaaattac cgagggaatc ctggcagatg aatgtgccgc cctgctgtgc
      961 tatttctttc ggatgcctag acaggtgttc aatgctcaga agaaggccca gagctccacc
     1021 gacggcggta gcggcggtag cggtggtagc ggcggcagcg gtggtagcgg tggcagcggt
     1081 ggtgagctgt ccacaaagga gcagatcagc cgcaattcca aggccctgga ggagaagtat
     1141 gtggccgagc tgcagctgga gcggctgaag aaggatggcg aggtgagggg ctccatcaat
     1201 cgcttcaaga cctctgacta cgtgaaggag gccaagcagc tgctgaaggt gcagaaggcc
     1261 taccaccagc tggatcagtc ctttatcgat acatatatcg acctgctgga gacaaggcgc
     1321 acatactatg agggaccagg agagggctct cccttcggct ggaaggacat caaggagtgg
     1381 tacgagatgc tgatgggcca ctgcacctat tttccagagg agctgagaag cgtgaagtac
     1441 gcctataacg ccgatctgta caacgccctg aatgacctga acaacctggt catcaccagg
     1501 gatgagaacg agaagctgga gtactatgag aagttccaga tcatcgagaa cgtgttcaag
     1561 cagaagaaga agcctacact gaagcagatc gccaaggaga tcctggtgaa cgaggaggac
     1621 atcaagggct accgcgtgac ctccacaggc aagccagagt tcaccaatct gaaggtgtat
     1681 cacgatatca aggacatcac agcccggaag gagatcatcg agaacgccga gctgctggat
     1741 cagatcgcca agatcctgac catctatcag agctccgagg acatccagga ggagctgacc
     1801 aacctgaata gcgagctgac acaggaggag atcgagcaga tcagcaatct gaagggctac
     1861 accggcacac acaacctgag cctgaaggcc atcaatctga tcctggatga gctgtggcac
     1921 acaaacgaca atcagatcgc catctttaac cggctgaagc tggtgccaaa gaaggtggac
     1981 ctgtcccagc agaaggagat cccaaccaca ctggtggacg atttcatcct gtctcccgtg
     2041 gtgaagcgga gcttcatcca gagcatcaaa gtgatcaacg ccatcatcaa gaagtacggc
     2101 ctgcccaatg atatcatcat cgagctggcc agggagaaga actccaagga cgcccagaag
     2161 atgatcaatg agatgcagaa gaggaaccgc cagaccaatg agcggatcga ggagatcatc
     2221 agaaccacag gcaaggagaa cgccaagtac ctgatcgaga agatcaagct gcacgatatg
     2281 caggagggca agtgtctgta ttctctggag gccatccctc tggaggacct gctgaacaat
     2341 ccattcaact acgaggtgga tcacatcatc ccccggagcg tgagcttcga caattctttt
     2401 aacaataagg tgctggtgaa gcaggaggag aacagcaaga agggcaatag gacccctttc
     2461 cagtacctgt ctagctccga ttctaagatc agctacgaga cattcaagaa gcacatcctg
     2521 aatctggcca agggcaaggg ccgcatcagc aagaccaaga aggagtacct gctggaggag
     2581 cgggacatca acagattctc cgtgcagaag gacttcatca accggaatct ggtggacacc
     2641 agatacgcca cacgcggcct gatgaatctg ctgcggtctt atttcagagt gaacaatctg
     2701 gatgtgaagg tgaagagcat caacggcggc ttcacctcct ttctgcggag aaagtggaag
     2761 tttaagaagg agcgcaacaa gggctataag caccacgccg aggatgccct gatcatcgcc
     2821 aatgccgact tcatctttaa ggagtggaag aagctggaca aggccaagaa agtgatggag
     2881 aaccagatgt tcgaggagaa gcaggccgag agcatgcccg agatcgagac agagcaggag
     2941 tacaaggaga ttttcatcac acctcaccag atcaagcaca tcaaggactt caaggactac
     3001 aagtattctc acagggtgga taagaagccc aaccgcgagc tgatcaatga caccctgtat
     3061 agcacacgga aggacgataa gggcaatacc ctgatcgtga acaatctgaa cggcctgtac
     3121 gacaaggata atgacaagct gaagaagctg atcaacaagt ctcccgagaa gctgctgatg
     3181 taccaccacg atcctcagac atatcagaag ctgaagctga tcatggagca gtacggcgac
     3241 gagaagaacc cactgtataa gtactatgag gagacaggca actacctgac aaagtatagc
     3301 aagaaggata atggccccgt gatcaagaag atcaagtact atggcaacaa gctgaatgcc
     3361 cacctggaca tcaccgacga ttaccctaac tctcgcaata aggtggtgaa gctgagcctg
     3421 aagccatacc ggttcgacgt gtacctggac aacggcgtgt ataagtttgt gacagtgaag
     3481 aatctggatg tgatcaagaa ggagaactac tatgaggtga acagcaagtg ctacgaggag
     3541 gccaagaagc tgaagaagat cagcaaccag gccgagttca tcgcctcttt ttacaacaat
     3601 gacctgatca agatcaatgg cgagctgtat agagtgatcg gcgtgaacaa tgatctgctg
     3661 aacagaatcg aagtgaatat gatcgacatc acctacaggg agtatctgga gaacatgaat
     3721 gataagaggc cccctcgcat catcaagacc atcgcctcta agacacagag catcaagaag
     3781 tacagcacag acatcctggg gaacctgtat gaagtcaaga gcaagaaaca tcctcagatt
     3841 atcaagaaag gctctggtgg ttctaagcga cctgccgcca caaagaaggc tggacaggct
     3901 aagaagaaga aagattacaa agacgatgac gataagggat ccggcgcaac aaacttctct
     3961 ctgctgaaac aagccggaga tgtcgaagag aatcctggac cgatggccaa gcctttgtct
     4021 caagaagaat ccaccctcat tgaaagagca acggctacaa tcaacagcat ccccatctct
     4081 gaagactaca gcgtcgccag cgcagctctc tctagcgacg gccgcatctt cactggtgtc
     4141 aatgtatatc attttactgg gggaccttgt gcagaactcg tggtgctggg cactgctgct
     4201 gctgcggcag ctggcaacct gacttgtatc gtcgcgatcg gaaatgagaa caggggcatc
     4261 ttgagcccct gcggacggtg ccgacaggtg cttctcgatc tgcatcctgg gatcaaagcc
     4321 atagtgaagg acagtgatgg acagccgacg gcagttggga ttcgtgaatt gctgccctct
     4381 ggttatgtgt gggagggcta a
//
