LOCUS       sequence-384719-        5304 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses human codon-optimized SpCas9-NRCH-ABE8e(V106W) and 
            blasticidin resistance: EFS 
            promoter-SpCas9-NRCH-ABE8e(V106W)-NLS-FLAG-P2A-BSD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5304)
  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 5304)
  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-ABE8e(V106W)
FEATURES             Location/Qualifiers
     source          1..5304
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             37..57
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             652..4752
                     /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             4795..4815
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4816..4839
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4849..4905
                     /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             4906..5304
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
ORIGIN
        1 atgaaacgga cagccgacgg aagcgagttc gagtcaccaa agaagaagcg gaaagtctct
       61 gaggtggagt tttcccacga gtactggatg agacatgccc tgaccctggc caagagggca
      121 cgggatgaga gggaggtgcc tgtgggagcc gtgctggtgc tgaacaatag agtgatcggc
      181 gagggctgga acagagccat cggcctgcac gacccaacag cccatgccga aattatggcc
      241 ctgagacagg gcggcctggt catgcagaac tacagactga ttgacgccac cctgtacgtg
      301 acattcgagc cttgcgtgat gtgcgccggc gccatgatcc actctaggat cggccgcgtg
      361 gtgtttggat ggagaaattc taaaagaggc gccgcaggct ccctgatgaa cgtgctgaac
      421 taccccggca tgaatcaccg cgtcgaaatt accgagggaa tcctggcaga tgaatgtgcc
      481 gccctgctgt gcgatttcta tcggatgcct agacaggtgt tcaatgctca gaagaaggcc
      541 cagagctcca tcaactccgg aggatctagc ggaggctcct ctggctctga gacacctggc
      601 acaagcgaga gcgcaacacc tgaaagcagc gggggcagca gcggggggtc agacaagaag
      661 tacagcatcg gcctggccat cggcaccaac tctgtgggct gggccgtgat caccgacgag
      721 tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca ccgaccggca cagcatcaag
      781 aagaacctga tcggagccct gctgttcgac agcggcgaaa cagccgaggc cacccggctg
      841 aagagaaccg ccagaagaag atacaccaga cggaagaacc ggatctgcta tctgcaagag
      901 atcttcagca acgagatggc caaggtggac gacagcttct tccacagact ggaagagtcc
      961 ttcctggtgg aagaggataa gaagcacgag cggcacccca tcttcggcaa catcgtggac
     1021 gaggtggcct accacgagaa gtaccccacc atctaccacc tgagaaagaa actggtggac
     1081 agcaccgaca aggccgacct gcggctgatc tatctggccc tggcccacat gatcaagttc
     1141 cggggccact tcctgatcga gggcgacctg aaccccgaca acagcgacgt ggacaagctg
     1201 ttcatccagc tggtgcagac ctacaaccag ctgttcgagg aaaaccccat caacgccagc
     1261 ggcgtggacg ccaaggccat cctgtctgcc agactgagca agagcagacg gctggaaaat
     1321 ctgatcgccc agctgcccgg cgagaagaag aatggcctgt tcggaaacct gattgccctg
     1381 agcctgggcc tgacccccaa cttcaagagc aacttcgacc tggccgagga tgccaaactg
     1441 cagctgagca aggacaccta cgacgacgac ctggacaacc tgctggccca gatcggcgac
     1501 cagtacgccg acctgtttct ggccgccaag aacctgtccg acgccatcct gctgagcgac
     1561 atcctgagag tgaacaccga gatcaccaag gcccccctga gcgcctctat ggtgaagaga
     1621 tacgacgagc accaccagga cctgaccctg ctgaaagctc tcgtgcggca gcagctgcct
     1681 gagaagtaca aagagatttt cttcgaccag agcaagaacg gctacgccgg ctacattgac
     1741 ggcggagcca gccaggaaga gttctacaag ttcatcaagc ccatcctgga aaagatggac
     1801 ggcaccgagg aactgctcgt gaagctgaac agagaggacc tgctgcggaa gcagcggacc
     1861 ttcgacaacg gcatcatccc ccaccagatc cacctgggag agctgcacgc cattctgcgg
     1921 cggcagggcg atttttaccc attcctgaag gacaaccggg aaaagatcga gaagatcctg
     1981 accttccgca tcccctacta cgtgggccct ctggccaggg gaaacagcag attcgcctgg
     2041 atgaccagaa agagcgagga aaccatcacc ccctggaact tcgaggaagt ggtggacaag
     2101 ggcgcttccg cccagagctt catcgagcgg atgaccaact tcgataagaa cctgcccaac
     2161 gagaaggtgc tgcccaagca cagcctgctg tacgagtact tcaccgtgta taacgagctg
     2221 accaaagtga aatacgtgac cgagggaatg agaaagcccg ccttcctgag cggcgagcag
     2281 aaaaaggcca tcgtggacct gctgttcaag accaaccgga aagtgaccgt gaagcagctg
     2341 aaagaggact acttcaagaa aatcgagtgc ttcgactccg tggaaatctc cggcgtggaa
     2401 gatcggttca acgcctccct gggcacatac cacgatctgc tgaaaattat caaggacaag
     2461 gacttcctgg acaatgagga aaacgaggac attctggaag atatcgtgct gaccctgaca
     2521 ctgtttgagg acagagagat gatcgaggaa cggctgaaaa cctatgccca cctgttcgac
     2581 gacaaagtga tgaagcagct gaagcggctg agatacaccg gctggggcag gctgagccgg
     2641 aagctgatca acggcatccg ggacaagcag tccggcaaga caatcctgga tttcctgaag
     2701 tccgacggct tcgccaacag aaacttcatg cagctgatcc acgacgacag cctgaccttt
     2761 aaagaggaca tccagaaagc ccaggtgtcc ggccagggcg atagcctgca cgagcacatt
     2821 gccaatctgg ccggcagccc cgccattaag aagggcatcc tgcagacagt gaaggtggtg
     2881 gacgagctcg tgaaagtgat gggcggccac aagcccgaga acatcgtgat cgaaatggcc
     2941 agagagaacc agaccaccca gaagggacag aagaacagcc gcgagagaat gaagcggatc
     3001 gaagagggca tcaaagagct gggcagccag atcctgaaag aacaccccgt ggaaaacacc
     3061 cagctgcaga acgagaagct gtacctgtac tacctgcaga atgggcggga tatgtacgtg
     3121 gaccaggaac tggacatcaa ccggctgtcc gactacgatg tggaccatat cgtgcctcag
     3181 agctttctga aggacgactc catcgacaac aaggtgctga ccagaagcga caagaaccgg
     3241 ggcaagagcg acaacgtgcc ctccgaagag gtcgtgaaga agatgaagaa ctactggcgg
     3301 cagctgctga acgccaagct gattacccag agaaagttcg acaatctgac caaggccgag
     3361 agaggcggcc tgagcgaact ggataaggcc ggcttcatca agagacagct ggtggaaacc
     3421 cggcagatca caaagcacgt ggcacagatc ctggactccc ggatgaacac taagtacgac
     3481 gagaatgaca agctgatccg ggaagtgaaa gtgatcaccc tgaagtccaa gctggtgtcc
     3541 gatttccgga aggatttcca gttttacaaa gtgcgcgaga tcaacaacta ccaccacgcc
     3601 cacgacgcct acctgaacgc cgtcgtggga accgccctga tcaaaaagta ccctaagctg
     3661 gaaagcgagt tcgtgtacgg cgactacaag gtgtacgacg tgcggaagat gatcgccaag
     3721 agcgagcagg aaatcggcaa ggctaccgcc aagtacttct tctacagcaa catcatgaac
     3781 tttttcaaga ccgagattac cctggccaac ggcgagatcc ggaagcggcc tctgatcgag
     3841 acaaacggcg aaaccgggga gatcgtgtgg gataagggcc gggattttgc caccgtgcgg
     3901 aaagtgctga gcatgcccca agtgaatatc gtgaaaaaga ccgaggtgca gacaggcggc
     3961 ttcagcaaag agtctatcct gcccaagggc aacagcgata agctgatcgc cagaaagaag
     4021 gactgggacc ctaagaagta cggcggcttc aacagcccca ccgtggccta ttctgtgctg
     4081 gtggtggcca aagtggaaaa gggcaagtcc aagaaactga agagtgtgaa agagctactg
     4141 gggatcacca tcatggaaag aagcagcttc gagaagaatc ccatcgactt tctggaagcc
     4201 aagggctaca aagaagtgaa aaaggacctg atcatcaagc tgcctaagta ctccctgttc
     4261 gagctggaaa acggccggaa gagaatgctg gcctctgccg gcgtgctgca gaagggaaac
     4321 gaactggccc tgccctccaa atatgtgaac ttcctgtacc tggccagcca ctatgagaag
     4381 ctgaagggct cccccgagga taatgagcag aaacagctgt ttgtggaaca gcacaagcac
     4441 tacctggacg agatcatcga gcagatcagc gagttctcca agagagtgat cctggccgac
     4501 gctaatctgg acaaagtgct gtccgcctac aacaagcacc gggataagcc catcagagag
     4561 caggccgaga atatcatcca cctgtttacc ctgaccaatc tgggagcccc tgccgccttc
     4621 aagtactttg acaccaccat caaccggaag cagtacaaca ccaccaaaga ggtgctggac
     4681 gccaccctga tccggcagag catcaccggc ctgtacgaga cacggatcga cctgtctcag
     4741 ctgggaggcg actctggcgg ctcaaaaaga accgccgacg gcagcgaatt cgagcccaag
     4801 aagaagagga aagtcgatta caaagacgat gacgataagg gatccggcgc aacaaacttc
     4861 tctctgctga aacaagccgg agatgtcgaa gagaatcctg gaccgatggc caagcctttg
     4921 tctcaagaag aatccaccct cattgaaaga gcaacggcta caatcaacag catccccatc
     4981 tctgaagact acagcgtcgc cagcgcagct ctctctagcg acggccgcat cttcactggt
     5041 gtcaatgtat atcattttac tgggggacct tgtgcagaac tcgtggtgct gggcactgct
     5101 gctgctgcgg cagctggcaa cctgacttgt atcgtcgcga tcggaaatga gaacaggggc
     5161 atcttgagcc cctgcggacg gtgccgacag gtgcttctcg atctgcatcc tgggatcaaa
     5221 gccatagtga aggacagtga tggacagccg acggcagttg ggattcgtga attgctgccc
     5281 tctggttatg tgtgggaggg ctaa
//
