LOCUS       sequence-322563-        4101 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  This plasmid encodes a Cas9 protein as well as a sgRNA that targets 
            the C-terminus of Whi5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4101)
  TITLE     Heinemann Lab Plasmids
REFERENCE   2  (bases 1 to 4101)
  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"
COMMENT     Sequence Label: HCP8
FEATURES             Location/Qualifiers
     source          1..4101
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             1..4101
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             1..4101
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
ORIGIN
        1 gacaagaagt attctatcgg actggatatc gggactaata gcgtcgggtg ggccgtgatc
       61 actgacgagt acaaggtgcc ctctaagaag ttcaaggtgc tcgggaacac cgaccggcat
      121 tccatcaaga aaaatctgat cggagctctc ctctttgatt caggggaaac cgctgaagca
      181 acccgcctca agcggactgc tagacggcgg tacaccagga ggaagaaccg gatttgttac
      241 cttcaagaga tattctccaa cgaaatggca aaggtcgacg acagcttctt ccataggctg
      301 gaagaatcat tcctcgtgga agaggataag aagcatgaac ggcatcccat cttcggtaat
      361 atcgtcgacg aggtggccta tcacgagaaa tacccaacca tctaccatct tcgcaaaaag
      421 ctggtggact caaccgacaa ggcagacctc cggcttatct acctggccct ggcccacatg
      481 atcaagttca gaggccactt cctgatcgag ggcgacctca atcctgacaa tagcgatgtg
      541 gataaactgt tcatccagct ggtgcagact tacaaccagc tctttgaaga gaaccccatc
      601 aatgcaagcg gagtcgatgc caaggccatt ctgtcagccc ggctgtcaaa gagccgcaga
      661 cttgagaatc ttatcgctca gctgccgggt gaaaagaaaa atggactgtt cgggaacctg
      721 attgctcttt cacttgggct gactcccaat ttcaagtcta atttcgacct ggcagaggat
      781 gccaagctgc aactgtccaa ggacacctat gatgacgatc tcgacaacct cctggcccag
      841 atcggtgacc aatacgccga ccttttcctt gctgctaaga atctttctga cgccatcctg
      901 ctgtctgaca ttctccgcgt gaacactgaa atcaccaagg cccctctttc agcttcaatg
      961 attaagcggt atgatgagca ccaccaggac ctgaccctgc ttaaggcact cgtccggcag
     1021 cagcttccgg agaagtacaa ggaaatcttc tttgaccagt caaagaatgg atacgccggc
     1081 tacatcgacg gaggtgcctc ccaagaggaa ttttataagt ttatcaaacc tatccttgag
     1141 aagatggacg gcaccgaaga gctcctcgtg aaactgaatc gggaggatct gctgcggaag
     1201 cagcgcactt tcgacaatgg gagcattccc caccagatcc atcttgggga gcttcacgcc
     1261 atccttcggc gccaagagga cttctacccc tttcttaagg acaacaggga gaagattgag
     1321 aaaattctca ctttccgcat cccctactac gtgggacccc tcgccagagg aaatagccgg
     1381 tttgcttgga tgaccagaaa gtcagaagaa actatcactc cctggaactt cgaagaggtg
     1441 gtggacaagg gagccagcgc tcagtcattc atcgaacgga tgactaactt cgataagaac
     1501 ctccccaatg agaaggtcct gccgaaacat tccctgctct acgagtactt taccgtgtac
     1561 aacgagctga ccaaggtgaa atatgtcacc gaagggatga ggaagcccgc attcctgtca
     1621 ggcgaacaaa agaaggcaat tgtggacctt ctgttcaaga ccaatagaaa ggtgaccgtg
     1681 aagcagctga aggaggacta tttcaagaaa attgaatgct tcgactctgt ggagattagc
     1741 ggggtcgaag atcggttcaa cgcaagcctg ggtacttacc atgatctgct taagatcatc
     1801 aaggacaagg attttctgga caatgaggag aacgaggaca tccttgagga cattgtcctg
     1861 actctcactc tgttcgagga ccgggaaatg atcgaggaga ggcttaagac ctacgcccat
     1921 ctgttcgacg ataaagtgat gaagcaactt aaacggagaa gatataccgg atggggacgc
     1981 cttagccgca aactcatcaa cggaatccgg gacaaacaga gcggaaagac cattcttgat
     2041 ttccttaaga gcgacggatt cgctaatcgc aacttcatgc aacttatcca tgatgattcc
     2101 ctgaccttta aggaggacat ccagaaggcc caagtgtctg gacaaggtga ctcactgcac
     2161 gagcatatcg caaatctggc tggttcaccc gctattaaga agggtattct ccagaccgtg
     2221 aaagtcgtgg acgagctggt caaggtgatg ggtcgccata aaccagagaa cattgtcatc
     2281 gagatggcca gggaaaacca gactacccag aagggacaga agaacagcag ggagcggatg
     2341 aaaagaattg aggaagggat taaggagctc gggtcacaga tccttaaaga gcacccggtg
     2401 gaaaacaccc agcttcagaa tgagaagctc tatctgtact accttcaaaa tggacgcgat
     2461 atgtatgtgg accaagagct tgatatcaac aggctctcag actacgacgt ggaccatatc
     2521 gtccctcaga gcttcctcaa agacgactca attgacaata aggtgctgac tcgctcagac
     2581 aagaaccggg gaaagtcaga taacgtgccc tcagaggaag tcgtgaaaaa gatgaagaac
     2641 tattggcgcc agcttctgaa cgcaaagctg atcactcagc ggaagttcga caatctcact
     2701 aaggctgaga ggggcggact gagcgaactg gacaaagcag gattcattaa acggcaactt
     2761 gtggagactc ggcagattac taaacatgtc gcccaaatcc ttgactcacg catgaatacc
     2821 aagtacgacg aaaacgacaa acttatccgc gaggtgaagg tgattaccct gaagtccaag
     2881 ctggtcagcg atttcagaaa ggactttcaa ttctacaaag tgcgggagat caataactat
     2941 catcatgctc atgacgcata tctgaatgcc gtggtgggaa ccgccctgat caagaagtac
     3001 ccaaagctgg aaagcgagtt cgtgtacgga gactacaagg tctacgacgt gcgcaagatg
     3061 attgccaaat ctgagcagga gatcggaaag gccaccgcaa agtacttctt ctacagcaac
     3121 atcatgaatt tcttcaagac cgaaatcacc cttgcaaacg gtgagatccg gaagaggccg
     3181 ctcatcgaga ctaatgggga gactggcgaa atcgtgtggg acaagggcag agatttcgct
     3241 accgtgcgca aagtgctttc tatgcctcaa gtgaacatcg tgaagaaaac cgaggtgcaa
     3301 accggaggct tttctaagga atcaatcctc cccaagcgca actccgacaa gctcattgca
     3361 aggaagaagg attgggaccc taagaagtac ggcggattcg attcaccaac tgtggcttat
     3421 tctgtcctgg tcgtggctaa ggtggaaaaa ggaaagtcta agaagctcaa gagcgtgaag
     3481 gaactgctgg gtatcaccat tatggagcgc agctccttcg agaagaaccc aattgacttt
     3541 ctcgaagcca aaggttacaa ggaagtcaag aaggacctta tcatcaagct cccaaagtat
     3601 agcctgttcg aactggagaa tgggcggaag cggatgctcg cctccgctgg cgaacttcag
     3661 aagggtaatg agctggctct cccctccaag tacgtgaatt tcctctacct tgcaagccat
     3721 tacgagaagc tgaaggggag ccccgaggac aacgagcaaa agcaactgtt tgtggagcag
     3781 cataagcatt atctggacga gatcattgag cagatttccg agttttctaa acgcgtcatt
     3841 ctcgctgatg ccaacctcga taaagtcctt agcgcataca ataagcacag agacaaacca
     3901 attcgggagc aggctgagaa tatcatccac ctgttcaccc tcaccaatct tggtgcccct
     3961 gccgcattca agtacttcga caccaccatc gaccggaaac gctatacctc caccaaagaa
     4021 gtgctggacg ccaccctcat ccaccagagc atcaccggac tttacgaaac tcggattgac
     4081 ctctcacagc tcggagggga t
//
