LOCUS       sequence-384713-        6171 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses human codon-optimized SpCas9-NRCH-SsAPOBEC3B and 
            blasticidin resistance: EFS 
            promoter-SpCas9-NRCH-SsAPOBEC3B-NLS-FLAG-P2A-BSD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6171)
  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 6171)
  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-SsAPOBEC3B
FEATURES             Location/Qualifiers
     source          1..6171
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             937..5037
                     /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             5068..5316
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5347..5595
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5596..5616
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5662..5682
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5683..5706
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             5716..5772
                     /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             5773..6171
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
ORIGIN
        1 atggacccac agaggctgcg ccagtggccc ggccctggcc cagcaagcag gggcggctac
       61 ggccagcggc caagaatcag gaaccccgag gagtggtttc acgagctgtc tccccggacc
      121 ttcagctttc acttccgcaa cctgaggttc gcatccggcc gcaatcggtc ttatatctgc
      181 tgtcaggtgg agggcaagaa ctgcttcttt cagggcatct ttcagaatca ggtgccacct
      241 gacccaccat gccacgcaga gctgtgcttc ctgtcttggt tccagagctg gggcctgtcc
      301 cccgatgagc actactatgt gacatggttt atctcttgga gcccttgctg tgagtgtgcc
      361 gccaaggtgg cccagttcct ggaggagaac cgcaacgtga gcctgtctct gagcgccgca
      421 aggctgtact atttctggaa gtccgagtct agagagggac tgcggagact gagcgacctg
      481 ggagcacaag tgggaatcat gtcctttcag gatttccagc actgctggaa caattttgtg
      541 cacaacctgg gcatgccctt ccagccttgg aagaagctgc acaagaatta ccagaggctg
      601 gtgaccgagc tgaagcagat cctgcgcgag gagcctgcca catatggctc tccacaggcc
      661 cagggcaagg tgagaatcgg aagcaccgca gcaggactga ggcacagcca ctcccacaca
      721 cgctccgagg cacacctgag gcctaaccac agctccagac agcacaggat cctgaatcct
      781 ccacgggagg ccagagccag gacctgcgtg ctggtggatg cctcttggat ctgttacaga
      841 agcggaggat ctagcggcgg ctctagcgga tctgagacac ctggcacaag cgagtctgcc
      901 acacctgaga gtagcggcgg atcttctggt ggctctgaca agaagtacag catcggcctg
      961 gccatcggca ccaactctgt gggctgggcc gtgatcaccg acgagtacaa ggtgcccagc
     1021 aagaaattca aggtgctggg caacaccgac cggcacagca tcaagaagaa cctgatcgga
     1081 gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc ggctgaagag aaccgccaga
     1141 agaagataca ccagacggaa gaaccggatc tgctatctgc aagagatctt cagcaacgag
     1201 atggccaagg tggacgacag cttcttccac agactggaag agtccttcct ggtggaagag
     1261 gataagaagc acgagcggca ccccatcttc ggcaacatcg tggacgaggt ggcctaccac
     1321 gagaagtacc ccaccatcta ccacctgaga aagaaactgg tggacagcac cgacaaggcc
     1381 gacctgcggc tgatctatct ggccctggcc cacatgatca agttccgggg ccacttcctg
     1441 atcgagggcg acctgaaccc cgacaacagc gacgtggaca agctgttcat ccagctggtg
     1501 cagacctaca accagctgtt cgaggaaaac cccatcaacg ccagcggcgt ggacgccaag
     1561 gccatcctgt ctgccagact gagcaagagc agacggctgg aaaatctgat cgcccagctg
     1621 cccggcgaga agaagaatgg cctgttcgga aacctgattg ccctgagcct gggcctgacc
     1681 cccaacttca agagcaactt cgacctggcc gaggatgcca aactgcagct gagcaaggac
     1741 acctacgacg acgacctgga caacctgctg gcccagatcg gcgaccagta cgccgacctg
     1801 tttctggccg ccaagaacct gtccgacgcc atcctgctga gcgacatcct gagagtgaac
     1861 accgagatca ccaaggcccc cctgagcgcc tctatggtga agagatacga cgagcaccac
     1921 caggacctga ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa gtacaaagag
     1981 attttcttcg accagagcaa gaacggctac gccggctaca ttgacggcgg agccagccag
     2041 gaagagttct acaagttcat caagcccatc ctggaaaaga tggacggcac cgaggaactg
     2101 ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc ggaccttcga caacggcatc
     2161 atcccccacc agatccacct gggagagctg cacgccattc tgcggcggca gggcgatttt
     2221 tacccattcc tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt ccgcatcccc
     2281 tactacgtgg gccctctggc caggggaaac agcagattcg cctggatgac cagaaagagc
     2341 gaggaaacca tcaccccctg gaacttcgag gaagtggtgg acaagggcgc ttccgcccag
     2401 agcttcatcg agcggatgac caacttcgat aagaacctgc ccaacgagaa ggtgctgccc
     2461 aagcacagcc tgctgtacga gtacttcacc gtgtataacg agctgaccaa agtgaaatac
     2521 gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg agcagaaaaa ggccatcgtg
     2581 gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga ggactacttc
     2641 aagaaaatcg agtgcttcga ctccgtggaa atctccggcg tggaagatcg gttcaacgcc
     2701 tccctgggca cataccacga tctgctgaaa attatcaagg acaaggactt cctggacaat
     2761 gaggaaaacg aggacattct ggaagatatc gtgctgaccc tgacactgtt tgaggacaga
     2821 gagatgatcg aggaacggct gaaaacctat gcccacctgt tcgacgacaa agtgatgaag
     2881 cagctgaagc ggctgagata caccggctgg ggcaggctga gccggaagct gatcaacggc
     2941 atccgggaca agcagtccgg caagacaatc ctggatttcc tgaagtccga cggcttcgcc
     3001 aacagaaact tcatgcagct gatccacgac gacagcctga cctttaaaga ggacatccag
     3061 aaagcccagg tgtccggcca gggcgatagc ctgcacgagc acattgccaa tctggccggc
     3121 agccccgcca ttaagaaggg catcctgcag acagtgaagg tggtggacga gctcgtgaaa
     3181 gtgatgggcg gccacaagcc cgagaacatc gtgatcgaaa tggccagaga gaaccagacc
     3241 acccagaagg gacagaagaa cagccgcgag agaatgaagc ggatcgaaga gggcatcaaa
     3301 gagctgggca gccagatcct gaaagaacac cccgtggaaa acacccagct gcagaacgag
     3361 aagctgtacc tgtactacct gcagaatggg cgggatatgt acgtggacca ggaactggac
     3421 atcaaccggc tgtccgacta cgatgtggac catatcgtgc ctcagagctt tctgaaggac
     3481 gactccatcg acaacaaggt gctgaccaga agcgacaaga accggggcaa gagcgacaac
     3541 gtgccctccg aagaggtcgt gaagaagatg aagaactact ggcggcagct gctgaacgcc
     3601 aagctgatta cccagagaaa gttcgacaat ctgaccaagg ccgagagagg cggcctgagc
     3661 gaactggata aggccggctt catcaagaga cagctggtgg aaacccggca gatcacaaag
     3721 cacgtggcac agatcctgga ctcccggatg aacactaagt acgacgagaa tgacaagctg
     3781 atccgggaag tgaaagtgat caccctgaag tccaagctgg tgtccgattt ccggaaggat
     3841 ttccagtttt acaaagtgcg cgagatcaac aactaccacc acgcccacga cgcctacctg
     3901 aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctggaaag cgagttcgtg
     3961 tacggcgact acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga gcaggaaatc
     4021 ggcaaggcta ccgccaagta cttcttctac agcaacatca tgaacttttt caagaccgag
     4081 attaccctgg ccaacggcga gatccggaag cggcctctga tcgagacaaa cggcgaaacc
     4141 ggggagatcg tgtgggataa gggccgggat tttgccaccg tgcggaaagt gctgagcatg
     4201 ccccaagtga atatcgtgaa aaagaccgag gtgcagacag gcggcttcag caaagagtct
     4261 atcctgccca agggcaacag cgataagctg atcgccagaa agaaggactg ggaccctaag
     4321 aagtacggcg gcttcaacag ccccaccgtg gcctattctg tgctggtggt ggccaaagtg
     4381 gaaaagggca agtccaagaa actgaagagt gtgaaagagc tactggggat caccatcatg
     4441 gaaagaagca gcttcgagaa gaatcccatc gactttctgg aagccaaggg ctacaaagaa
     4501 gtgaaaaagg acctgatcat caagctgcct aagtactccc tgttcgagct ggaaaacggc
     4561 cggaagagaa tgctggcctc tgccggcgtg ctgcagaagg gaaacgaact ggccctgccc
     4621 tccaaatatg tgaacttcct gtacctggcc agccactatg agaagctgaa gggctccccc
     4681 gaggataatg agcagaaaca gctgtttgtg gaacagcaca agcactacct ggacgagatc
     4741 atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa tctggacaaa
     4801 gtgctgtccg cctacaacaa gcaccgggat aagcccatca gagagcaggc cgagaatatc
     4861 atccacctgt ttaccctgac caatctggga gcccctgccg ccttcaagta ctttgacacc
     4921 accatcaacc ggaagcagta caacaccacc aaagaggtgc tggacgccac cctgatccgg
     4981 cagagcatca ccggcctgta cgagacacgg atcgacctgt ctcagctggg aggcgactct
     5041 ggtggaagcg gaggatctgg cggcagcacc aatctgagcg acatcatcga gaaagagaca
     5101 ggcaagcagc tggtcatcca agagtccatc ctgatgctgc ctgaagaggt ggaagaagtg
     5161 atcggcaaca agcccgagtc cgacatcctg gtgcacaccg cctacgatga gagcaccgac
     5221 gagaacgtga tgctgctgac ctctgacgcc cctgagtaca agccttgggc tctcgtgatc
     5281 caggacagca acggcgagaa caagatcaag atgctgagcg gcggctctgg tggctctggc
     5341 ggatctacaa acctgtccga tattattgag aaagaaaccg ggaaacagct cgtgattcaa
     5401 gagtctattc tcatgctccc ggaagaagtc gaggaagtca ttggaaacaa gcctgagagc
     5461 gatattctgg tccatacagc ctacgacgag tctaccgatg agaatgtcat gctcctcacc
     5521 agcgacgctc ccgagtataa gccatgggca cttgtcattc aggactccaa tggggaaaac
     5581 aaaatcaaaa tgctcccaaa gaaaaaacgc aaggtggagg gagctgataa gcgcaccgcc
     5641 gatggttccg agttcgaaag ccccaagaag aagaggaaag tcgattacaa agacgatgac
     5701 gataagggat ccggcgcaac aaacttctct ctgctgaaac aagccggaga tgtcgaagag
     5761 aatcctggac cgatggccaa gcctttgtct caagaagaat ccaccctcat tgaaagagca
     5821 acggctacaa tcaacagcat ccccatctct gaagactaca gcgtcgccag cgcagctctc
     5881 tctagcgacg gccgcatctt cactggtgtc aatgtatatc attttactgg gggaccttgt
     5941 gcagaactcg tggtgctggg cactgctgct gctgcggcag ctggcaacct gacttgtatc
     6001 gtcgcgatcg gaaatgagaa caggggcatc ttgagcccct gcggacggtg ccgacaggtg
     6061 cttctcgatc tgcatcctgg gatcaaagcc atagtgaagg acagtgatgg acagccgacg
     6121 gcagttggga ttcgtgaatt gctgccctct ggttatgtgt gggagggcta a
//
