LOCUS       sequence-261026-        5553 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression of intradomain insertion of rat cytosine deaminase, 
            rAPOBEC3, at position 1058 of nSpyCas9 lacking uracil DNA 
            glycosylase inhibitor..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5553)
  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 5553)
  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: S2 1058 APOBEC3 no UGI
FEATURES             Location/Qualifiers
     source          1..5553
                     /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             3319..4005
                     /codon_start=1
                     /product="cytidine deaminase (C to U editing enzyme) from 
                     rat"
                     /label=APOBEC-1
                     /note="can use ssDNA as a substrate (Komor et al., 2016)"
                     /translation="MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWG
                     GRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFL
                     SRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSN
                     EAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHI
                     LWATGLK"
     CDS             5011..5058
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5059..5082
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             5092..5148
                     /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 gagctcagag actggcccag tggctgtgga ccccacattg
     3361 agacggcgga tcgagcccca tgagtttgag gtattcttcg atccgagaga gctccgcaag
     3421 gagacctgcc tgctttacga aattaattgg gggggccggc actccatttg gcgacataca
     3481 tcacagaaca ctaacaagca cgtcgaagtc aacttcatcg agaagttcac gacagaaaga
     3541 tatttctgtc cgaacacaag gtgcagcatt acctggtttc tcagctggag cccatgcggc
     3601 gaatgtagta gggccatcac tgaattcctg tcaaggtatc cccacgtcac tctgtttatt
     3661 tacatcgcaa ggctgtacca ccacgctgac ccccgcaatc gacaaggcct gcgggatttg
     3721 atctcttcag gtgtgactat ccaaattatg actgagcagg agtcaggata ctgctggaga
     3781 aactttgtga attatagccc gagtaatgaa gcccactggc ctaggtatcc ccatctgtgg
     3841 gtacgactgt acgttcttga actgtactgc atcatactgg gcctgcctcc ttgtctcaac
     3901 attctgagaa ggaagcagcc acagctgaca ttctttacca tcgctcttca gtcttgtcat
     3961 taccagcgac tgcccccaca cattctctgg gccaccgggt tgaaaggcgg tagcggcggt
     4021 agcggtggta gcggcggcag cggtggtagc ggtggcagcg gtggtcgcaa acgcccgctg
     4081 atcgaaacca acggcgaaac cggtgaaatc gtgtgggata agggtcgcga tttcgcgacc
     4141 gtgcgcaagg tgctgagcat gccgcaggtg aacatcgtga agaagaccga agtgcagacc
     4201 ggcggtttta gcaaggaaag catcctgccg aagcgcaaca gcgataagct gatcgcgcgt
     4261 aagaaggact gggacccgaa aaagtatggt ggcttcgaca gcccgaccgt ggcgtacagc
     4321 gtgctggtgg tggccaaggt ggaaaagggc aaaagcaaaa aactgaaaag cgtgaaggag
     4381 ctgctgggta ttaccattat ggaacgtagc agctttgaga agaacccgat cgacttcctg
     4441 gaggcgaagg gttacaagga agtgaagaaa gatctgatca tcaaactgcc gaagtacagc
     4501 ctgtttgagc tggaaaacgg ccgtaagcgt atgctggcga gcgcgggtga gctgcaaaaa
     4561 ggtaacgagc tggcgctgcc gagcaagtac gtgaactttc tgtacctggc gagccactat
     4621 gaaaaactga aaggcagccc ggaggacaac gagcagaagc agctgtttgt ggagcagcac
     4681 aagcactatc tggacgaaat catcgaacag atcagcgagt tcagcaaacg cgtgattctg
     4741 gccgacgcca acctggacaa agtgctgagc gcctacaaca agcaccgcga taaaccgatt
     4801 cgcgaacagg ccgagaacat cattcacctg tttaccctga ccaacctggg cgcgccggcg
     4861 gcgttcaaat actttgatac caccattgat cgcaaacgct acaccagcac caaggaagtg
     4921 ctggatgcga ccctgatcca ccagagcatt accggtctgt acgaaacccg catcgatctg
     4981 agccagctgg gcggcgatag cggtggtagc aagcgtccgg cggcgaccaa aaaggccggc
     5041 caggcgaaga agaagaagga ctacaaggat gacgatgaca aaggcagcgg cgccaccaac
     5101 ttcagcctgc tgaagcaggc cggtgacgtg gaagaaaacc ccggcccgat ggcgaagccg
     5161 ctgagccagg aggaaagcac cctgattgag cgcgccaccg cgaccatcaa cagcattccg
     5221 attagcgaag actacagcgt ggcgagcgcc gctctgagca gcgatggccg tatcttcacc
     5281 ggcgtgaacg tgtaccactt taccggcggt ccgtgcgcgg aactggtggt gctgggcacc
     5341 gcggcggctg cggctgcggg taacctgacc tgcatcgtgg ccattggtaa cgaaaaccgc
     5401 ggcattctga gcccgtgcgg ccgttgccgc caggtgctgc tggacctgca ccccggtatc
     5461 aaagcgattg tgaaagatag cgatggtcaa ccgaccgctg tgggtatccg tgaactgctg
     5521 ccatccggct atgtttggga gggctaaggt acc
//
