LOCUS       sequence-261023-        5661 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression of intradomain insertion of truncated human cytosine 
            deaminase, hAIDx, at position 1058 of nSpyCas9 with uracil DNA 
            glycosylase inhibitor (UGI) at the C terminus .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5661)
  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 5661)
  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: AIDx-ID-1058
FEATURES             Location/Qualifiers
     source          1..5661
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      7..16
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3187..3207
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3223..3243
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4870..5118
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5119..5166
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5167..5190
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             5200..5256
                     /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"
ORIGIN
        1 aagcttgcca ccatggataa aaaatatagc attggtctgg cgattggcac caactccgtg
       61 ggctgggctg tgattaccga tgaatacaag gttccgagca agaagtttaa ggtgctgggt
      121 aacaccgacc gtcacagcat caagaagaac ctgattggcg ctctgctgtt tgacagcggc
      181 gagaccgcgg aagccacccg cctgaagcgc accgcgcgtc gccgttatac ccgtcgtaag
      241 aaccgcattt gctatctgca agaaatcttc agcaacgaga tggcgaaagt ggatgacagc
      301 tttttccacc gcctggaaga gagctttctg gtggaggaag ataagaagca cgagcgtcac
      361 ccgatcttcg gtaacattgt ggacgaggtg gcctatcacg aaaagtatcc gaccatctac
      421 cacctgcgta aaaaactggt ggatagcacc gacaaagctg atctgcgcct gatctatctg
      481 gctctggcgc acatgatcaa attccgcggc cacttcctga ttgagggtga cctgaacccg
      541 gacaacagcg atgtggataa actgtttatt cagctggtgc agacctataa ccagctgttt
      601 gaagagaacc cgatcaacgc gagcggcgtg gacgcgaaag ctattctgag cgcgcgtctg
      661 agcaagagcc gtcgtctgga aaacctgatc gcgcagctgc cgggcgagaa gaaaaacggt
      721 ctgtttggca acctgattgc cctgagcctg ggtctgaccc cgaactttaa gagcaacttc
      781 gacctggcgg aggacgcgaa actgcaactg agcaaagata cctatgacga cgacctggat
      841 aacctgctgg cgcagatcgg cgatcagtat gctgacctgt ttctggcggc caaaaacctg
      901 agcgacgcga ttctgctgag cgacatcctg cgtgtgaaca ccgagattac caaagcgccg
      961 ctgagcgcca gcatgattaa gcgttacgac gaacaccacc aggacctgac cctgctgaaa
     1021 gccctggtgc gccagcagct gccggagaag tacaaagaaa tctttttcga tcagagcaaa
     1081 aacggttatg ccggttatat tgacggtggc gcgagccagg aggagtttta taagttcatt
     1141 aagccgattc tggaaaagat ggacggcacc gaggagctgc tggtgaagct gaaccgcgaa
     1201 gacctgctgc gcaaacagcg tacctttgat aacggtagca ttccgcacca gattcacctg
     1261 ggcgaactgc acgcgattct gcgtcgccag gaggacttct atccgttcct gaaggataac
     1321 cgtgagaaaa ttgagaagat tctgaccttc cgtatcccgt actatgtggg cccgctggct
     1381 cgtggtaaca gccgtttcgc ttggatgacc cgtaaaagcg aggagaccat taccccgtgg
     1441 aacttcgagg aagtggtgga caaaggtgcc agcgcgcaga gctttatcga gcgcatgacc
     1501 aacttcgaca aaaacctgcc gaacgaaaaa gtgctgccga aacacagcct gctgtatgaa
     1561 tacttcaccg tgtataacga actgaccaag gtgaagtatg tgaccgaagg tatgcgcaag
     1621 ccggcttttc tgagcggcga gcagaagaag gcgattgtgg acctgctgtt caagaccaac
     1681 cgtaaagtga ccgtgaagca gctgaaggag gactacttca agaaaattga atgctttgac
     1741 agcgtggaga tcagcggcgt ggaggaccgc ttcaacgcga gcctgggcac ctaccacgat
     1801 ctgctgaaga tcattaagga taaagatttt ctggacaacg aggaaaacga agacatcctg
     1861 gaggatattg tgctgaccct gaccctgttt gaggatcgtg aaatgattga agaacgcctg
     1921 aagacctatg ctcacctgtt cgatgataaa gtgatgaaac agctgaagcg tcgtcgttat
     1981 accggttggg gccgtctgag ccgcaaactg atcaacggca ttcgtgacaa acagagcggc
     2041 aagaccatcc tggattttct gaaaagcgat ggctttgcga accgtaactt tatgcagctg
     2101 attcacgatg atagcctgac cttcaaggag gacattcaga aggcgcaggt gagcggccag
     2161 ggtgacagcc tgcacgagca cattgcgaac ctggcgggca gcccggcgat taagaaaggc
     2221 attctgcaaa ccgtgaaagt ggtggatgaa ctggtgaaag tgatgggccg ccacaagccg
     2281 gagaacattg tgatcgagat ggcccgtgag aaccagacca cccagaaagg ccagaagaac
     2341 agccgcgagc gcatgaagcg tattgaagag ggtatcaagg agctgggcag ccagatcctg
     2401 aaggaacacc cggtggaaaa cacccagctg caaaacgaga aactgtatct gtactacctg
     2461 caaaacggcc gtgacatgta tgtggatcag gaactggaca tcaaccgcct gagcgactac
     2521 gacgtggatg cgatcgtgcc gcagagcttc ctgaaagacg acagcattga taacaaggtg
     2581 ctgacccgta gcgacaagaa ccgtggcaaa agcgataacg tgccgagcga ggaggtggtg
     2641 aaaaagatga aaaactattg gcgccagctg ctgaacgcga agctgattac ccagcgcaag
     2701 ttcgacaacc tgaccaaggc tgagcgtggt ggcctgagcg agctggacaa ggctggcttt
     2761 atcaaacgcc agctggtgga gacccgccag atcaccaagc acgtggcgca gatcctggat
     2821 agccgtatga acaccaaata cgatgagaac gataagctga tccgcgaggt gaaagtgatc
     2881 accctgaaga gcaagctggt gagcgatttt cgcaaggact tccagttcta caaggtgcgc
     2941 gaaatcaaca actaccacca cgcgcacgat gcgtatctga acgcggtggt gggcaccgct
     3001 ctgatcaaga aatatccgaa gctggagagc gagttcgtgt acggtgacta taaagtgtac
     3061 gacgtgcgca aaatgattgc caagagcgag caggagattg gtaaagctac cgctaaatat
     3121 ttcttttaca gcaacatcat gaacttcttt aagaccgaga tcaccctggc caacggtgaa
     3181 atccgtccga agaagaaacg taaggtgggt agcagcggta gcccgaaaaa gaagcgcaaa
     3241 gtgggcagcg acgcgctgga cgatttcgat ctggacatgc tgggcagcga tgcgctggat
     3301 gatttcggcg gtggcagcat ggatagcctg ctgatgaacc gtcgcaagtt cctgtaccag
     3361 ttcaaaaacg tgcgttgggc gaagggtcgt cgtgagacct acctgtgcta cgtggtgaag
     3421 cgtcgtgaca gcgcgaccag cttcagcctg gattttggct atctgcgtaa caaaaacggt
     3481 tgccacgtgg aactgctgtt tctgcgctac attagcgact gggacctgga ccccggtcgc
     3541 tgctaccgcg tgacctggtt taccagctgg agcccgtgct atgattgcgc gcgtcacgtg
     3601 gccgatttcc tgcgcggtaa cccgaacctg agcctgcgca ttttcaccgc gcgcctgtat
     3661 ttctgcgagg atcgcaaggc ggagccggag ggcctgcgtc gtctgcaccg tgctggtgtg
     3721 cagattgcga ttatgacctt taaggactac ttttattgct ggaacacctt tgtggagaac
     3781 cacgagcgca ccttcaaagc gtgggagggt ctgcacgaga acagcgtgcg tctgagccgc
     3841 cagctgcgtc gcatcctgct gccgggcggt agcggcggta gcggtggtag cggcggcagc
     3901 ggtggtagcg gtggcagcgg tggtcgcaaa cgcccgctga tcgaaaccaa cggcgaaacc
     3961 ggtgaaatcg tgtgggataa gggtcgcgat ttcgcgaccg tgcgcaaggt gctgagcatg
     4021 ccgcaggtga acatcgtgaa gaagaccgaa gtgcagaccg gcggttttag caaggaaagc
     4081 atcctgccga agcgcaacag cgataagctg atcgcgcgta agaaggactg ggacccgaaa
     4141 aagtatggtg gcttcgacag cccgaccgtg gcgtacagcg tgctggtggt ggccaaggtg
     4201 gaaaagggca aaagcaaaaa actgaaaagc gtgaaggagc tgctgggtat taccattatg
     4261 gaacgtagca gctttgagaa gaacccgatc gacttcctgg aggcgaaggg ttacaaggaa
     4321 gtgaagaaag atctgatcat caaactgccg aagtacagcc tgtttgagct ggaaaacggc
     4381 cgtaagcgta tgctggcgag cgcgggtgag ctgcaaaaag gtaacgagct ggcgctgccg
     4441 agcaagtacg tgaactttct gtacctggcg agccactatg aaaaactgaa aggcagcccg
     4501 gaggacaacg agcagaagca gctgtttgtg gagcagcaca agcactatct ggacgaaatc
     4561 atcgaacaga tcagcgagtt cagcaaacgc gtgattctgg ccgacgccaa cctggacaaa
     4621 gtgctgagcg cctacaacaa gcaccgcgat aaaccgattc gcgaacaggc cgagaacatc
     4681 attcacctgt ttaccctgac caacctgggc gcgccggcgg cgttcaaata ctttgatacc
     4741 accattgatc gcaaacgcta caccagcacc aaggaagtgc tggatgcgac cctgatccac
     4801 cagagcatta ccggtctgta cgaaacccgc atcgatctga gccagctggg cggcgatagc
     4861 ggtggtagca ccaacctgag cgacatcatt gagaaagaga ccggcaagca gctggtaatc
     4921 caggaaagca ttctgatgct gccggaagaa gtggaggaag tgattggtaa caaaccggaa
     4981 agcgacatcc tggtgcacac cgcttatgac gagagcaccg acgaaaacgt gatgctgctg
     5041 accagcgacg cgccggagta taagccgtgg gcgctggtaa ttcaggatag caacggcgaa
     5101 aacaaaatca agatgctgaa gcgtccggcg gcgaccaaaa aggccggcca ggcgaagaag
     5161 aagaaggact acaaggatga cgatgacaaa ggcagcggcg ccaccaactt cagcctgctg
     5221 aagcaggccg gtgacgtgga agaaaacccc ggcccgatgg cgaagccgct gagccaggag
     5281 gaaagcaccc tgattgagcg cgccaccgcg accatcaaca gcattccgat tagcgaagac
     5341 tacagcgtgg cgagcgccgc tctgagcagc gatggccgta tcttcaccgg cgtgaacgtg
     5401 taccacttta ccggcggtcc gtgcgcggaa ctggtggtgc tgggcaccgc ggcggctgcg
     5461 gctgcgggta acctgacctg catcgtggcc attggtaacg aaaaccgcgg cattctgagc
     5521 ccgtgcggcc gttgccgcca ggtgctgctg gacctgcacc ccggtatcaa agcgattgtg
     5581 aaagatagcg atggtcaacc gaccgctgtg ggtatccgtg aactgctgcc atccggctat
     5641 gtttgggagg gctaaggtac c
//
