LOCUS       sequence-261059-        4401 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression of nSaCas9 intradomain insertion of single evolved TadA 
            monomer (V82G) at the specified position (M741 ).
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 M741 miniABEMax V82G
FEATURES             Location/Qualifiers
     source          1..4401
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             2224..2244
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2260..2280
                     /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 gagctgtcca caaaggagca gatcagccgc
      421 aattccaagg ccctggagga gaagtatgtg gccgagctgc agctggagcg gctgaagaag
      481 gatggcgagg tgaggggctc catcaatcgc ttcaagacct ctgactacgt gaaggaggcc
      541 aagcagctgc tgaaggtgca gaaggcctac caccagctgg atcagtcctt tatcgataca
      601 tatatcgacc tgctggagac aaggcgcaca tactatgagg gaccaggaga gggctctccc
      661 ttcggctgga aggacatcaa ggagtggtac gagatgctga tgggccactg cacctatttt
      721 ccagaggagc tgagaagcgt gaagtacgcc tataacgccg atctgtacaa cgccctgaat
      781 gacctgaaca acctggtcat caccagggat gagaacgaga agctggagta ctatgagaag
      841 ttccagatca tcgagaacgt gttcaagcag aagaagaagc ctacactgaa gcagatcgcc
      901 aaggagatcc tggtgaacga ggaggacatc aagggctacc gcgtgacctc cacaggcaag
      961 ccagagttca ccaatctgaa ggtgtatcac gatatcaagg acatcacagc ccggaaggag
     1021 atcatcgaga acgccgagct gctggatcag atcgccaaga tcctgaccat ctatcagagc
     1081 tccgaggaca tccaggagga gctgaccaac ctgaatagcg agctgacaca ggaggagatc
     1141 gagcagatca gcaatctgaa gggctacacc ggcacacaca acctgagcct gaaggccatc
     1201 aatctgatcc tggatgagct gtggcacaca aacgacaatc agatcgccat ctttaaccgg
     1261 ctgaagctgg tgccaaagaa ggtggacctg tcccagcaga aggagatccc aaccacactg
     1321 gtggacgatt tcatcctgtc tcccgtggtg aagcggagct tcatccagag catcaaagtg
     1381 atcaacgcca tcatcaagaa gtacggcctg cccaatgata tcatcatcga gctggccagg
     1441 gagaagaact ccaaggacgc ccagaagatg atcaatgaga tgcagaagag gaaccgccag
     1501 accaatgagc ggatcgagga gatcatcaga accacaggca aggagaacgc caagtacctg
     1561 atcgagaaga tcaagctgca cgatatgcag gagggcaagt gtctgtattc tctggaggcc
     1621 atccctctgg aggacctgct gaacaatcca ttcaactacg aggtggatca catcatcccc
     1681 cggagcgtga gcttcgacaa ttcttttaac aataaggtgc tggtgaagca ggaggagaac
     1741 agcaagaagg gcaataggac ccctttccag tacctgtcta gctccgattc taagatcagc
     1801 tacgagacat tcaagaagca catcctgaat ctggccaagg gcaagggccg catcagcaag
     1861 accaagaagg agtacctgct ggaggagcgg gacatcaaca gattctccgt gcagaaggac
     1921 ttcatcaacc ggaatctggt ggacaccaga tacgccacac gcggcctgat gaatctgctg
     1981 cggtcttatt tcagagtgaa caatctggat gtgaaggtga agagcatcaa cggcggcttc
     2041 acctcctttc tgcggagaaa gtggaagttt aagaaggagc gcaacaaggg ctataagcac
     2101 cacgccgagg atgccctgat catcgccaat gccgacttca tctttaagga gtggaagaag
     2161 ctggacaagg ccaagaaagt gatggagaac cagatgttcg aggagaagca ggccgagagc
     2221 atgccgaaga agaaacgtaa ggtgggtagc agcggtagcc cgaaaaagaa gcgcaaagtg
     2281 ggcagcgacg cgctggacga tttcgatctg gacatgctgg gcagcgatgc gctggatgat
     2341 ttcggcggtg gcagctctga ggtggagttt tcccacgagt actggatgag acatgccctg
     2401 accctggcca agagggcacg cgatgagagg gaggtgcctg tgggagccgt gctggtgctg
     2461 aacaatagag tgatcggcga gggctggaac agagccatcg gcctgcacga cccaacagcc
     2521 catgccgaaa ttatggccct gagacagggc ggcctggtca tgcagaacta cagactgatt
     2581 gacgccaccc tgtacggcac attcgagcct tgcgtgatgt gcgccggcgc catgatccac
     2641 tctaggatcg gccgcgtggt gtttggcgtg aggaacgcaa aaaccggcgc cgcaggctcc
     2701 ctgatggacg tgctgcacta ccccggcatg aatcaccgcg tcgaaattac cgagggaatc
     2761 ctggcagatg aatgtgccgc cctgctgtgc tatttctttc ggatgcctag acaggtgttc
     2821 aatgctcaga agaaggccca gagctccacc gacggcggta gcggcggtag cggtggtagc
     2881 ggcggcagcg gtggtagcgg tggcagcggt ggtatgcccg 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
//
