LOCUS       sequence-230080-        4111 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  MoClo golden gate assembly CD part for Cas9 (S. pyogenes 
            gRNA-targeted endonuclease Cas9, with bbsI cut sites removed 
            synonymously). Please see Supplemental Documents for annotated 
            Genbank file..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4111)
  TITLE     CIDAR MoClo Extension
REFERENCE   2  (bases 1 to 4111)
  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: C114m
FEATURES             Location/Qualifiers
     source          1..4111
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             join(2..28,29..31,32..2518,2519..2521,2522..4108)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
ORIGIN
        1 aatggataag aaatactcaa taggcttagc tatcggcaca aatagcgtcg gatgggcggt
       61 gatcactgat gaatataagg ttccgtctaa aaagttcaag gttctgggaa atacagaccg
      121 ccacagtatc aaaaaaaatc ttataggggc tcttttattt gacagtggag agacagcgga
      181 agcgactcgt ctcaaacgga cagctcgtag aaggtataca cgtcggaaga atcgtatttg
      241 ttatctacag gagatttttt caaatgagat ggcgaaagta gatgatagtt tctttcatcg
      301 acttgaagag tcttttttgg tggaagaaga taagaagcat gaacgtcatc ctatttttgg
      361 aaatatagta gatgaagttg cttatcatga gaaatatcca actatctatc atctgcgaaa
      421 aaaattggta gattctactg ataaagcgga tttgcgctta atctatttgg ccttagcgca
      481 tatgattaag tttcgtggtc attttttgat tgagggagat ttaaatcctg ataatagtga
      541 tgtggacaaa ctatttatcc agttggtaca aacctacaat caattatttg aagaaaaccc
      601 tattaacgca agtggagtag atgctaaagc gattctttct gcacgattga gtaaatcaag
      661 acgattagaa aatctcattg ctcagctccc cggtgagaag aaaaatggct tatttgggaa
      721 tctcattgct ttgtcattgg gtttgacccc taattttaaa tcaaattttg atttggcaga
      781 agatgctaaa ttacagcttt caaaagatac ttacgatgat gatttagata atttattggc
      841 gcaaattgga gatcaatatg ctgatttgtt tttggcagct aagaatttat cagatgctat
      901 tttactttca gatatcctaa gagtaaatac tgaaataact aaggctcccc tatcagcttc
      961 aatgattaaa cgctacgatg aacatcatca agacttgact cttttaaaag ctttagttcg
     1021 acaacaactt ccagaaaagt ataaagaaat cttttttgat caatcaaaaa acggatatgc
     1081 aggttatatt gatgggggag ctagccaaga agaattttat aaatttatca aaccaatttt
     1141 agaaaaaatg gatggtactg aggaattatt ggtgaaacta aatcgtgaag atttgctgcg
     1201 caagcaacgg acctttgaca acggctctat tccccatcaa attcacttgg gtgagctgca
     1261 tgctattttg agaaggcaag aggactttta tccattttta aaagacaatc gtgagaagat
     1321 tgaaaaaatc ttgacttttc gaattcctta ttatgttggt ccattggcgc gtggcaatag
     1381 tcgttttgca tggatgactc ggaagtctga agaaacaatt accccatgga attttgaaga
     1441 agttgtcgat aaaggtgctt cagctcaatc atttattgaa cgcatgacaa actttgataa
     1501 aaatcttcca aatgaaaaag tactaccaaa acatagtttg ctttatgagt attttacggt
     1561 ttataacgaa ttgacaaagg tcaaatatgt tactgaagga atgcgaaaac cagcatttct
     1621 ttcaggtgaa cagaagaaag ccattgttga tttactcttc aaaacaaatc gaaaagtaac
     1681 cgttaagcaa ttaaaagaag attatttcaa aaaaatagaa tgttttgata gtgttgaaat
     1741 ttcaggagtt gaagatagat ttaatgcttc attaggtacc taccatgatt tgctaaaaat
     1801 tattaaagat aaagattttt tggataatga agaaaatgaa gatatcttag aggatattgt
     1861 tttaacattg accttatttg aagataggga gatgattgag gaaagactta aaacatatgc
     1921 tcacctcttt gatgataagg tgatgaaaca gcttaaacgt cgccgttata ctggttgggg
     1981 acgtttgtct cgaaaattga ttaatggtat tagggataag caatctggca aaacaatatt
     2041 agattttttg aaatcagatg gttttgccaa tcgcaatttt atgcagctga tccatgatga
     2101 tagtttgaca tttaaagaag atattcaaaa agcacaagtg tctggacaag gcgatagttt
     2161 acatgaacat attgcaaatt tagctggtag ccctgctatt aaaaaaggta ttttacagac
     2221 tgtaaaagtt gttgatgaat tggtcaaagt aatggggcgg cataagccag aaaatatcgt
     2281 tattgaaatg gcacgtgaaa atcagacaac tcaaaagggc cagaaaaatt cgcgagagcg
     2341 tatgaaacga atcgaagaag gtatcaaaga attaggaagt cagattctta aagagcatcc
     2401 tgttgaaaat actcaattgc aaaatgaaaa gctctatctc tattatctcc aaaatggaag
     2461 agacatgtat gtggaccaag aattagatat taatcgttta agtgattatg atgtcgatgc
     2521 cattgttcca caaagtttcc ttaaagacga ttcaatagac aataaggtct taacgcgttc
     2581 tgataaaaat cgtggtaaat cggataacgt tccaagtgaa gaagtagtca aaaagatgaa
     2641 aaactattgg agacaacttc taaacgccaa gttaatcact caacgtaagt ttgataattt
     2701 aacgaaagct gaacgtggag gtttgagtga acttgataaa gctggtttta tcaaacgcca
     2761 attggttgaa actcgccaaa tcactaagca tgtggcacaa attttggata gtcgcatgaa
     2821 tactaaatac gatgaaaatg ataaacttat tcgagaggtt aaagtgatta ccttaaaatc
     2881 taaattagtt tctgacttcc gaaaagattt ccaattctat aaagtacgtg agattaacaa
     2941 ttaccatcat gcccatgatg cgtatctaaa tgccgtcgtt ggaactgctt tgattaagaa
     3001 atatccaaaa cttgaatcgg agtttgtcta tggtgattat aaagtttatg atgttcgtaa
     3061 aatgattgct aagtctgagc aagaaatagg caaagcaacc gcaaaatatt tcttttactc
     3121 taatatcatg aacttcttca aaacagaaat tacacttgca aatggagaga ttcgcaaacg
     3181 ccctctaatc gaaactaatg gggaaactgg agaaattgtc tgggataaag ggcgagattt
     3241 tgccacagtg cgcaaagtat tgtccatgcc ccaagtcaat attgtcaaga aaacagaagt
     3301 acagacaggc ggattctcca aggagtcaat tttaccaaaa agaaattcgg acaagcttat
     3361 tgctcgtaaa aaagactggg atccaaaaaa atatggtggt tttgatagtc caacggtagc
     3421 ttattcagtc ctagtggttg ctaaggtgga aaaagggaaa tcgaagaagt taaaatccgt
     3481 taaagagtta ctagggatca caattatgga aagaagttcc tttgaaaaaa atccgattga
     3541 ctttttagaa gctaaaggat ataaggaagt taaaaaagac ttaatcatta aactacctaa
     3601 atatagtctt tttgagttag aaaacggtcg taaacggatg ctggctagtg ccggagaatt
     3661 acaaaaagga aatgagctgg ctctgccaag caaatatgtg aattttttat atttagctag
     3721 tcattatgaa aagttgaagg gtagtccaga agataacgaa caaaaacaat tgtttgtgga
     3781 gcagcataag cattatttag atgagattat tgagcaaatc agtgaatttt ctaagcgtgt
     3841 tattttagca gatgccaatt tagataaagt tcttagtgca tataacaaac atagagacaa
     3901 accaatacgt gaacaagcag aaaatattat tcatttattt acgttgacga atcttggagc
     3961 tcccgctgct tttaaatatt ttgatacaac aattgatcgt aaacgatata cgtctacaaa
     4021 agaagtttta gatgccactc ttatccatca atccatcact ggtctttatg aaacacgcat
     4081 tgatttgagt cagctaggag gtgactaagg t
//
