LOCUS       sequence-261028-        5562 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression of circular permutant of nSpyCas9 lacking uracil DNA 
            glycosylase inhibitor at amino acid position 1029 encoding rat 
            cytosine deaminase, rAPOBEC3 at the N-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5562)
  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 5562)
  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: S1 1029 CP APOBEC3 no UGI
FEATURES             Location/Qualifiers
     source          1..5562
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             13..699
                     /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             700..720
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             736..756
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5020..5067
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5068..5091
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             5101..5157
                     /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 ggtaccgcca ccatgagctc agagactggc ccagtggctg tggaccccac attgagacgg
       61 cggatcgagc cccatgagtt tgaggtattc ttcgatccga gagagctccg caaggagacc
      121 tgcctgcttt acgaaattaa ttgggggggc cggcactcca tttggcgaca tacatcacag
      181 aacactaaca agcacgtcga agtcaacttc atcgagaagt tcacgacaga aagatatttc
      241 tgtccgaaca caaggtgcag cattacctgg tttctcagct ggagcccatg cggcgaatgt
      301 agtagggcca tcactgaatt cctgtcaagg tatccccacg tcactctgtt tatttacatc
      361 gcaaggctgt accaccacgc tgacccccgc aatcgacaag gcctgcggga tttgatctct
      421 tcaggtgtga ctatccaaat tatgactgag caggagtcag gatactgctg gagaaacttt
      481 gtgaattata gcccgagtaa tgaagcccac tggcctaggt atccccatct gtgggtacga
      541 ctgtacgttc ttgaactgta ctgcatcata ctgggcctgc ctccttgtct caacattctg
      601 agaaggaagc agccacagct gacattcttt accatcgctc ttcagtcttg tcattaccag
      661 cgactgcccc cacacattct ctgggccacc gggttgaaac cgaagaaaaa gcgtaaagtg
      721 ggtagcagcg gcagcccgaa aaagaaacgc aaagtgggta gcgacgctct ggatgacttc
      781 gacctggata tgctgggcag cgacgccctg gatgactttg gcggtggcag cataggcaag
      841 gcaaccgcta agtatttttt ttacagcaac atcatgaact tctttaagac cgagattacc
      901 ctggccaacg gtgaaatccg taaacgcccg ctgattgaaa ccaacggtga aaccggcgaa
      961 atcgtgtggg acaagggccg tgattttgcg accgtgcgca aagtgctgtc catgccgcag
     1021 gtgaacattg tgaagaaaac cgaggtgcag accggcggtt tcagcaagga aagcattctg
     1081 ccgaaacgta acagcgacaa gctgatcgcc cgcaagaaag attgggaccc gaagaaatac
     1141 ggcggtttcg atagcccgac cgtggcttat agcgtgctgg tggtggccaa agtggagaag
     1201 ggtaaaagca agaaactgaa gagcgtgaaa gaactgctgg gcatcaccat tatggagcgt
     1261 agcagcttcg aaaagaaccc gatcgacttt ctggaggcga agggttacaa ggaagtgaag
     1321 aaagatctga tcattaagct gccgaaatac agcctgtttg agctggaaaa tggtcgtaaa
     1381 cgtatgctgg ccagcgccgg cgagctgcaa aagggcaacg aactggcgct gccgagcaaa
     1441 tatgtgaact tcctgtacct ggccagccac tatgagaagc tgaaaggtag cccggaggac
     1501 aacgaacaga agcagctgtt tgtggagcag cacaaacact atctggatga gatcattgaa
     1561 cagatcagcg agttcagcaa gcgtgtgatt ctggcggacg ctaacctgga taaagtgctg
     1621 agcgcctaca acaaacaccg tgacaagccg atccgcgagc aggcggaaaa catcattcac
     1681 ctgttcaccc tgaccaacct gggtgcgccg gccgcgttca agtactttga caccaccatt
     1741 gatcgtaaac gctataccag caccaaggag gtgctggatg ctaccctgat ccaccagagc
     1801 attaccggcc tgtacgaaac ccgtattgat ctgagccagc tgggcggtga cggcggtagc
     1861 ggcggtagcg gcggtagcgg cggtagcggc ggtagcggcg gtagcggcgg tgacaagaaa
     1921 tatagcatcg gtctggcgat tggcaccaac agcgtgggtt gggctgtgat caccgatgag
     1981 tacaaggtgc cgagcaagaa gttcaaggtg ctgggcaaca ccgaccgcca cagcatcaag
     2041 aaaaacctga ttggtgccct gctgtttgat agcggtgaaa ccgccgaggc cacccgtctg
     2101 aagcgcaccg cccgtcgccg ttacacccgt cgtaaaaacc gtatctgcta cctgcaagag
     2161 attttcagca acgaaatggc caaagtggac gatagcttct ttcaccgcct ggaggagagc
     2221 ttcctggtgg aggaagacaa gaaacacgag cgtcacccga tctttggcaa cattgtggat
     2281 gaggtggctt accacgaaaa gtatccgacc atctaccacc tgcgtaagaa actggtggat
     2341 agcaccgaca aggccgatct gcgcctgatc tatctggctc tggcccacat gatcaagttc
     2401 cgtggtcact ttctgatcga gggcgacctg aacccggata acagcgatgt ggacaagctg
     2461 ttcatccagc tggtgcagac ctacaaccag ctgtttgagg aaaacccgat taacgccagc
     2521 ggtgtggacg cgaaggctat cctgagcgcg cgtctgagca aaagccgccg tctggagaac
     2581 ctgatcgcgc agctgccggg tgaaaagaaa aacggtctgt ttggcaacct gattgctctg
     2641 agcctgggcc tgaccccgaa cttcaaaagc aactttgacc tggcggagga tgctaagctg
     2701 caactgagca aggataccta tgacgatgac ctggacaacc tgctggcgca gatcggcgac
     2761 cagtacgctg atctgttcct ggctgccaag aacctgagcg atgcgatcct gctgagcgac
     2821 attctgcgtg tgaacaccga gattaccaag gccccgctga gcgcgagcat gatcaaacgc
     2881 tatgatgaac accaccagga cctgaccctg ctgaaagctc tggtgcgtca gcagctgccg
     2941 gagaagtaca aagaaatctt ctttgaccag agcaagaacg gttacgccgg ctatattgat
     3001 ggcggtgcga gccaggagga gttctacaag tttatcaaac cgattctgga gaaaatggac
     3061 ggcaccgagg aactgctggt gaagctgaac cgtgaagatc tgctgcgtaa acagcgcacc
     3121 ttcgacaacg gtagcatccc gcaccagatt cacctgggtg agctgcacgc catcctgcgc
     3181 cgtcaggaag atttctaccc gtttctgaag gacaaccgtg agaagatcga aaaaattctg
     3241 accttccgca ttccgtacta tgtgggtccg ctggctcgtg gcaacagccg ctttgcctgg
     3301 atgacccgca agagcgagga aaccatcacc ccgtggaact tcgaggaagt ggtggacaaa
     3361 ggcgccagcg cgcagagctt cattgagcgt atgaccaact ttgataagaa cctgccgaac
     3421 gaaaaagtgc tgccgaagca cagcctgctg tacgagtatt tcaccgtgta caacgaactg
     3481 accaaggtga aatatgtgac cgagggtatg cgcaagccgg ccttcctgag cggcgaacag
     3541 aagaaagcga tcgtggacct gctgtttaaa accaaccgta aggtgaccgt gaagcagctg
     3601 aaagaggatt atttcaagaa aattgaatgc tttgacagcg tggagatcag cggtgtggaa
     3661 gatcgtttta acgcgagcct gggcacctac cacgacctgc tgaaaatcat taaggataaa
     3721 gacttcctgg ataacgagga aaacgaggac atcctggaag atattgtgct gaccctgacc
     3781 ctgtttgagg accgtgaaat gatcgaggaa cgcctgaaga cctatgccca cctgttcgat
     3841 gacaaagtga tgaaacagct gaagcgccgt cgctacaccg gttggggccg tctgagccgc
     3901 aaactgatca acggtattcg cgataaacag agcggcaaga ccatcctgga ttttctgaag
     3961 agcgacggtt tcgcgaaccg taactttatg cagctgattc acgatgacag cctgaccttc
     4021 aaagaggaca tccagaaggc gcaggtgagc ggtcagggcg atagcctgca cgaacatatt
     4081 gctaacctgg ccggtagccc ggccatcaag aagggcatcc tgcaaaccgt gaaggtggtg
     4141 gacgagctgg tgaaagtgat gggtcgtcac aagccggaaa acatcgtgat tgagatggcc
     4201 cgcgaaaacc agaccaccca gaaaggccag aagaacagcc gtgagcgcat gaagcgtatc
     4261 gaggaaggca tcaaggaact gggcagccag atcctgaagg agcacccggt ggaaaacacc
     4321 cagctgcaaa acgagaaact gtacctgtac tacctgcaaa acggtcgtga catgtatgtg
     4381 gatcaggaac tggacattaa ccgcctgagc gattatgacg tggatgcgat cgtgccgcag
     4441 agcttcctga aggatgacag cattgacaac aaagtgctga cccgtagcga taaaaaccgc
     4501 ggcaagagcg acaacgtgcc gagcgaggaa gtggtgaaga aaatgaaaaa ctactggcgt
     4561 cagctgctga acgccaaact gatcacccag cgtaagtttg acaacctgac caaagctgag
     4621 cgcggcggtc tgagcgaact ggataaagcc ggtttcatta agcgtcagct ggtggaaacc
     4681 cgccagatca ccaagcacgt ggcgcagatt ctggacagcc gcatgaacac caagtatgac
     4741 gagaacgata aactgatccg tgaagtgaaa gtgatcaccc tgaagagcaa actggtgagc
     4801 gattttcgta aagacttcca gttttacaag gtgcgcgaga tcaacaacta ccaccacgct
     4861 cacgacgcct atctgaacgc ggtggtgggc accgccctga tcaagaagta cccgaagctg
     4921 gagagcgagt tcgtgtacgg cgactacaag gtgtatgatg tgcgtaagat gatcgccaaa
     4981 agcgagcagg aaattggcgg cggcgatagc ggtggtagca agcgtccggc ggctaccaag
     5041 aaagcgggcc aggctaagaa aaagaaagac tacaaagatg acgatgacaa gggtagcggc
     5101 gctaccaact tcagcctgct gaagcaggcg ggcgacgtgg aggaaaatcc gggcccgatg
     5161 gctaaaccgc tgagccagga ggaaagcacc ctgatcgagc gcgctaccgc caccattaac
     5221 agcatcccga ttagcgaaga ctatagcgtg gccagcgccg cgctgagcag cgatggtcgt
     5281 attttcaccg gcgtgaacgt gtaccacttt accggtggcc cgtgcgccga gctggtggtg
     5341 ctgggcaccg ccgccgccgc cgccgccggc aacctgacct gcatcgtggc gattggtaac
     5401 gaaaatcgtg gcatcctgag cccgtgcggc cgttgccgcc aggtgctgct ggacctgcat
     5461 ccgggcatca aggcgattgt gaaggatagc gacggccaac cgaccgccgt gggtattcgt
     5521 gaactgctgc catccggcta tgtttgggag ggctaaggat cc
//
