LOCUS       sequence-384714-        5253 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses human codon-optimized SpCas9-NRCH-ABE8.17-m+V106W and 
            blasticidin resistance: EFS 
            promoter-SpCas9-NRCH-ABE8.17-m+V106W-NLS-FLAG-P2A-BSD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5253)
  AUTHORS   Kim N, Choi S, Kim S, Song M, Seo JH, Min S, Park J, Cho SR, Kim HH
  TITLE     Deep learning models to predict the editing efficiencies and 
            outcomes of diverse base editors.
  JOURNAL   Nat Biotechnol. 2024 Mar;42(3):484-497. doi: 
            10.1038/s41587-023-01792-x
  PUBMED    37188916
REFERENCE   2  (bases 1 to 5253)
  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"; doi: "10"; journalName: 
            "Nat Biotechnol"; date: "2024-03"; volume: "42"; issue: "3"; pages: 
            "484-497"
COMMENT     Sequence Label: SpCas9-NRCH-ABE8.17-m+V106W
FEATURES             Location/Qualifiers
     source          1..5253
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             4..501
                     /codon_start=1
                     /product="DNA adenosine deaminase evolved from the E. coli 
                     tRNA adenosine deaminase TadA (Gaudelli et al., 2017)"
                     /label=TadA*(7.10)
                     /translation="SEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWN
                     RAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYSTFEPCVMCAGAMIHSRIGRVVFG
                     WRNAKTGAAGSLMDVLHYPGMNHRVEITEGILADECAALLCYFFRMPRRVFNAQKKAQS
                     STD"
     CDS             598..4698
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMVKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGIIPHQIHLGELHAILRRQGDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRLRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGGH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKGNSDKLIARKKDWDPKKYGGFNSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGVLQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTINRKQYNTTKEVLD
                     ATLIRQSITGLYETRIDLSQLGGD"
     CDS             4744..4764
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4765..4788
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4798..4854
                     /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             4855..5253
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
ORIGIN
        1 atgtccgaag tcgagttttc ccatgagtac tggatgagac acgcattgac tctcgcaaag
       61 agggctcgag atgaacgcga ggtgcccgtg ggggcagtac tcgtgctcaa caatcgcgta
      121 atcggcgaag gttggaatag ggcaatcgga ctccacgacc ccactgcaca tgcggaaatc
      181 atggcccttc gacagggagg gcttgtgatg cagaattatc gacttatcga tgcgacgctg
      241 tactcgacgt ttgaaccttg cgtaatgtgc gcgggagcta tgattcactc ccgcattgga
      301 cgagttgtat tcggttggcg caacgccaag acgggtgccg caggttcact gatggacgtg
      361 ctgcattacc caggcatgaa ccaccgggta gaaatcacag aaggcatatt ggcggacgaa
      421 tgtgcggcgc tgttgtgtta cttttttcgc atgcccaggc gtgtctttaa cgcccagaaa
      481 aaagcacaat cctctactga ctctggtggt tcttctggtg gttctagcgg cagcgagact
      541 cccgggacct cagagtccgc cacacccgaa agttctggtg gttcttctgg tggttctgac
      601 aagaagtaca gcatcggcct ggccatcggc accaactctg tgggctgggc cgtgatcacc
      661 gacgagtaca aggtgcccag caagaaattc aaggtgctgg gcaacaccga ccggcacagc
      721 atcaagaaga acctgatcgg agccctgctg ttcgacagcg gcgaaacagc cgaggccacc
      781 cggctgaaga gaaccgccag aagaagatac accagacgga agaaccggat ctgctatctg
      841 caagagatct tcagcaacga gatggccaag gtggacgaca gcttcttcca cagactggaa
      901 gagtccttcc tggtggaaga ggataagaag cacgagcggc accccatctt cggcaacatc
      961 gtggacgagg tggcctacca cgagaagtac cccaccatct accacctgag aaagaaactg
     1021 gtggacagca ccgacaaggc cgacctgcgg ctgatctatc tggccctggc ccacatgatc
     1081 aagttccggg gccacttcct gatcgagggc gacctgaacc ccgacaacag cgacgtggac
     1141 aagctgttca tccagctggt gcagacctac aaccagctgt tcgaggaaaa ccccatcaac
     1201 gccagcggcg tggacgccaa ggccatcctg tctgccagac tgagcaagag cagacggctg
     1261 gaaaatctga tcgcccagct gcccggcgag aagaagaatg gcctgttcgg aaacctgatt
     1321 gccctgagcc tgggcctgac ccccaacttc aagagcaact tcgacctggc cgaggatgcc
     1381 aaactgcagc tgagcaagga cacctacgac gacgacctgg acaacctgct ggcccagatc
     1441 ggcgaccagt acgccgacct gtttctggcc gccaagaacc tgtccgacgc catcctgctg
     1501 agcgacatcc tgagagtgaa caccgagatc accaaggccc ccctgagcgc ctctatggtg
     1561 aagagatacg acgagcacca ccaggacctg accctgctga aagctctcgt gcggcagcag
     1621 ctgcctgaga agtacaaaga gattttcttc gaccagagca agaacggcta cgccggctac
     1681 attgacggcg gagccagcca ggaagagttc tacaagttca tcaagcccat cctggaaaag
     1741 atggacggca ccgaggaact gctcgtgaag ctgaacagag aggacctgct gcggaagcag
     1801 cggaccttcg acaacggcat catcccccac cagatccacc tgggagagct gcacgccatt
     1861 ctgcggcggc agggcgattt ttacccattc ctgaaggaca accgggaaaa gatcgagaag
     1921 atcctgacct tccgcatccc ctactacgtg ggccctctgg ccaggggaaa cagcagattc
     1981 gcctggatga ccagaaagag cgaggaaacc atcaccccct ggaacttcga ggaagtggtg
     2041 gacaagggcg cttccgccca gagcttcatc gagcggatga ccaacttcga taagaacctg
     2101 cccaacgaga aggtgctgcc caagcacagc ctgctgtacg agtacttcac cgtgtataac
     2161 gagctgacca aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc
     2221 gagcagaaaa aggccatcgt ggacctgctg ttcaagacca accggaaagt gaccgtgaag
     2281 cagctgaaag aggactactt caagaaaatc gagtgcttcg actccgtgga aatctccggc
     2341 gtggaagatc ggttcaacgc ctccctgggc acataccacg atctgctgaa aattatcaag
     2401 gacaaggact tcctggacaa tgaggaaaac gaggacattc tggaagatat cgtgctgacc
     2461 ctgacactgt ttgaggacag agagatgatc gaggaacggc tgaaaaccta tgcccacctg
     2521 ttcgacgaca aagtgatgaa gcagctgaag cggctgagat acaccggctg gggcaggctg
     2581 agccggaagc tgatcaacgg catccgggac aagcagtccg gcaagacaat cctggatttc
     2641 ctgaagtccg acggcttcgc caacagaaac ttcatgcagc tgatccacga cgacagcctg
     2701 acctttaaag aggacatcca gaaagcccag gtgtccggcc agggcgatag cctgcacgag
     2761 cacattgcca atctggccgg cagccccgcc attaagaagg gcatcctgca gacagtgaag
     2821 gtggtggacg agctcgtgaa agtgatgggc ggccacaagc ccgagaacat cgtgatcgaa
     2881 atggccagag agaaccagac cacccagaag ggacagaaga acagccgcga gagaatgaag
     2941 cggatcgaag agggcatcaa agagctgggc agccagatcc tgaaagaaca ccccgtggaa
     3001 aacacccagc tgcagaacga gaagctgtac ctgtactacc tgcagaatgg gcgggatatg
     3061 tacgtggacc aggaactgga catcaaccgg ctgtccgact acgatgtgga ccatatcgtg
     3121 cctcagagct ttctgaagga cgactccatc gacaacaagg tgctgaccag aagcgacaag
     3181 aaccggggca agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat gaagaactac
     3241 tggcggcagc tgctgaacgc caagctgatt acccagagaa agttcgacaa tctgaccaag
     3301 gccgagagag gcggcctgag cgaactggat aaggccggct tcatcaagag acagctggtg
     3361 gaaacccggc agatcacaaa gcacgtggca cagatcctgg actcccggat gaacactaag
     3421 tacgacgaga atgacaagct gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg
     3481 gtgtccgatt tccggaagga tttccagttt tacaaagtgc gcgagatcaa caactaccac
     3541 cacgcccacg acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct
     3601 aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc
     3661 gccaagagcg agcaggaaat cggcaaggct accgccaagt acttcttcta cagcaacatc
     3721 atgaactttt tcaagaccga gattaccctg gccaacggcg agatccggaa gcggcctctg
     3781 atcgagacaa acggcgaaac cggggagatc gtgtgggata agggccggga ttttgccacc
     3841 gtgcggaaag tgctgagcat gccccaagtg aatatcgtga aaaagaccga ggtgcagaca
     3901 ggcggcttca gcaaagagtc tatcctgccc aagggcaaca gcgataagct gatcgccaga
     3961 aagaaggact gggaccctaa gaagtacggc ggcttcaaca gccccaccgt ggcctattct
     4021 gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga aactgaagag tgtgaaagag
     4081 ctactgggga tcaccatcat ggaaagaagc agcttcgaga agaatcccat cgactttctg
     4141 gaagccaagg gctacaaaga agtgaaaaag gacctgatca tcaagctgcc taagtactcc
     4201 ctgttcgagc tggaaaacgg ccggaagaga atgctggcct ctgccggcgt gctgcagaag
     4261 ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc tgtacctggc cagccactat
     4321 gagaagctga agggctcccc cgaggataat gagcagaaac agctgtttgt ggaacagcac
     4381 aagcactacc tggacgagat catcgagcag atcagcgagt tctccaagag agtgatcctg
     4441 gccgacgcta atctggacaa agtgctgtcc gcctacaaca agcaccggga taagcccatc
     4501 agagagcagg ccgagaatat catccacctg tttaccctga ccaatctggg agcccctgcc
     4561 gccttcaagt actttgacac caccatcaac cggaagcagt acaacaccac caaagaggtg
     4621 ctggacgcca ccctgatccg gcagagcatc accggcctgt acgagacacg gatcgacctg
     4681 tctcagctgg gaggcgacga gggagctgat aagcgcaccg ccgatggttc cgagttcgaa
     4741 agccccaaga agaagaggaa agtcgattac aaagacgatg acgataaggg atccggcgca
     4801 acaaacttct ctctgctgaa acaagccgga gatgtcgaag agaatcctgg accgatggcc
     4861 aagcctttgt ctcaagaaga atccaccctc attgaaagag caacggctac aatcaacagc
     4921 atccccatct ctgaagacta cagcgtcgcc agcgcagctc tctctagcga cggccgcatc
     4981 ttcactggtg tcaatgtata tcattttact gggggacctt gtgcagaact cgtggtgctg
     5041 ggcactgctg ctgctgcggc agctggcaac ctgacttgta tcgtcgcgat cggaaatgag
     5101 aacaggggca tcttgagccc ctgcggacgg tgccgacagg tgcttctcga tctgcatcct
     5161 gggatcaaag ccatagtgaa ggacagtgat ggacagccga cggcagttgg gattcgtgaa
     5221 ttgctgccct ctggttatgt gtgggagggc taa
//
