LOCUS       sequence-196857-        4179 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Broad-host-range Cas9/gRNA co-expression backbone plasmid (no gRNA).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4179)
  AUTHORS   Juergens H, Varela JA, Gorter de Vries AR, Perli T, Gast VJM, 
            Gyurchev NY, Rajkumar AS, Mans R, Pronk JT, Morrissey JP, Daran JG
  TITLE     Genome editing in Kluyveromyces and Ogataea yeasts using a 
            broad-host-range Cas9/gRNA co-expression plasmid.
  JOURNAL   FEMS Yeast Res. 2018 May 1;18(3). pii: 4847887. doi: 
            10.1093/femsyr/foy012.
  PUBMED    29438517
REFERENCE   2  (bases 1 to 4179)
  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"; journalName: "FEMS Yeast 
            Res."; date: "2018-05-1"; volume: "18(3). pii: 4847887. doi"; pages:
            " 10.1093/femsyr/foy012"
COMMENT     Sequence Label: pUDP002
FEATURES             Location/Qualifiers
     source          1..4179
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             4..27
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             31..51
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             52..4152
                     /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
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTYKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSITHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4156..4176
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 atggattata aagatgacga tgacaaacct ccaaaaaaga agagaaaggt cgataagaaa
       61 tactcaatag gcttagatat cggcacaaat agcgtcggat gggcggtgat cactgatgaa
      121 tataaggttc cgtctaaaaa gttcaaggtt ctgggaaata cagaccgcca cagtatcaaa
      181 aaaaatctta taggggctct tttatttgac agtggagaga cagcggaagc gactcgtctc
      241 aaacggacag ctcgtagaag gtatacacgt cggaagaatc gtatttgtta tctacaggag
      301 attttttcaa atgagatggc gaaagtagat gatagtttct ttcatcgact tgaagagtct
      361 tttttggtgg aagaagacaa gaagcatgaa cgtcatccta tttttggaaa tatagtagat
      421 gaagttgctt atcatgagaa atatccaact atctatcatc tgcgaaaaaa attggtagat
      481 tctacttata aagcggattt gcgcttaatc tatttggcct tagcgcatat gattaagttt
      541 cgtggtcatt ttttgattga gggagattta aatcctgata atagtgatgt ggacaaacta
      601 tttatccagt tggtacaaac ctacaatcaa ttatttgaag aaaaccctat taacgcaagt
      661 ggagtagatg ctaaagcgat tctttctgca cgattgagta aatcaagacg attagaaaat
      721 ctcattgctc agctccccgg tgagaagaaa aatggcttat ttgggaatct cattgctttg
      781 tcattgggtt tgacccctaa ttttaaatca aattttgatt tggcagaaga tgctaaatta
      841 cagctttcaa aagatactta cgatgatgat ttagataatt tattggcgca aattggagat
      901 caatatgctg atttgttttt ggcagctaag aatttatcag atgctatttt actttcagat
      961 atcctaagag taaatactga aataactaag gctcccctat cagcttcaat gattaaacgc
     1021 tacgatgaac atcatcaaga cttgactctt ttaaaagctt tagttcgaca acaacttcca
     1081 gaaaagtata aagaaatctt ttttgatcaa tcaaaaaacg gatatgcagg ttatattgat
     1141 gggggagcta gccaagaaga attttataaa tttatcaaac caattttaga aaaaatggat
     1201 ggtactgagg aattattggt gaaactaaat cgtgaagatt tgctgcgcaa gcaacggacc
     1261 tttgacaacg gctctattac ccatcaaatt cacttgggtg agctgcatgc tattttgaga
     1321 agacaagaag acttttatcc atttttaaaa gacaatcgtg agaagattga aaaaatcttg
     1381 acttttcgaa ttccttatta tgttggtcca ttggcgcgtg gcaatagtcg ttttgcatgg
     1441 atgactcgga agtctgaaga aacaattacc ccatggaatt ttgaagaagt tgtcgataaa
     1501 ggtgcttcag ctcaatcatt tattgaacgc atgacaaact ttgataaaaa tcttccaaat
     1561 gaaaaagtac taccaaaaca tagtttgctt tatgagtatt ttacggttta taacgaattg
     1621 acaaaggtca aatatgttac tgaaggaatg cgaaaaccag catttctttc aggtgaacag
     1681 aagaaagcca ttgttgattt actcttcaaa acaaatcgaa aagtaaccgt taagcaatta
     1741 aaagaagatt atttcaaaaa aatagaatgt tttgatagtg ttgaaatttc aggagttgaa
     1801 gatagattta atgcttcatt aggtacctac catgatttgc taaaaattat taaagataaa
     1861 gattttttgg ataatgaaga aaatgaagat atcttagagg atattgtttt aacattgacc
     1921 ttatttgaag atagggagat gattgaggaa agacttaaaa catatgctca cctctttgat
     1981 gataaggtga tgaaacagct taaacgtcgc cgttatactg gttggggacg tttgtctcga
     2041 aaattgatta atggtattag ggataagcaa tctggcaaaa caatattaga ttttttgaaa
     2101 tcagatggtt ttgccaatcg caattttatg cagctgatcc atgatgatag tttgacattt
     2161 aaagaagaca ttcaaaaagc acaagtgtct ggacaaggcg atagtttaca tgaacatatt
     2221 gcaaatttag ctggtagccc tgctattaaa aaaggtattt tacagactgt aaaagttgtt
     2281 gatgaattgg tcaaagtaat ggggcggcat aagccagaaa atatcgttat tgaaatggca
     2341 cgtgaaaatc agacaactca aaagggccag aaaaattcgc gagagcgtat gaaacgaatc
     2401 gaagaaggta tcaaagaatt aggaagtcag attcttaaag agcatcctgt tgaaaatact
     2461 caattgcaaa atgaaaagct ctatctctat tatctccaaa atggaagaga catgtatgtg
     2521 gaccaagaat tagatattaa tcgtttaagt gattatgatg tcgatcacat tgttccacaa
     2581 agtttcctta aagacgattc aatagacaat aaggtcttaa cgcgttctga taaaaatcgt
     2641 ggtaaatcgg ataacgttcc aagtgaagaa gtagtcaaaa agatgaaaaa ctattggaga
     2701 caacttctaa acgccaagtt aatcactcaa cgtaagtttg ataatttaac gaaagctgaa
     2761 cgtggaggtt tgagtgaact tgataaagct ggttttatca aacgccaatt ggttgaaact
     2821 cgccaaatca ctaagcatgt ggcacaaatt ttggatagtc gcatgaatac taaatacgat
     2881 gaaaatgata aacttattcg agaggttaaa gtgattacct taaaatctaa attagtttct
     2941 gacttccgaa aagatttcca attctataaa gtacgtgaga ttaacaatta ccatcatgcc
     3001 catgatgcgt atctaaatgc cgtcgttgga actgctttga ttaagaaata tccaaaactt
     3061 gaatcggagt ttgtctatgg tgattataaa gtttatgatg ttcgtaaaat gattgctaag
     3121 tctgagcaag aaataggcaa agcaaccgca aaatatttct tttactctaa tatcatgaac
     3181 ttcttcaaaa cagaaattac acttgcaaat ggagagattc gcaaacgccc tctaatcgaa
     3241 actaatgggg aaactggaga aattgtctgg gataaagggc gagattttgc cacagtgcgc
     3301 aaagtattgt ccatgcccca agtcaatatt gtcaagaaaa cagaagtaca gacaggcgga
     3361 ttctccaagg agtcaatttt accaaaaaga aattcggaca agcttattgc tcgtaaaaaa
     3421 gactgggatc caaaaaaata tggtggtttt gatagtccaa cggtagctta ttcagtccta
     3481 gtggttgcta aggtggaaaa agggaaatcg aagaagttaa aatccgttaa agagttacta
     3541 gggatcacaa ttatggaaag aagttccttt gaaaaaaatc cgattgactt tttagaagct
     3601 aaaggatata aggaagttaa aaaagactta atcattaaac tacctaaata tagtcttttt
     3661 gagttagaaa acggtcgtaa acggatgctg gctagtgccg gagaattaca aaaaggaaat
     3721 gagctggctc tgccaagcaa atatgtgaat tttttatatt tagctagtca ttatgaaaag
     3781 ttgaagggta gtccagaaga taacgaacaa aaacaattgt ttgtggagca gcataagcat
     3841 tatttagatg agattattga gcaaatcagt gaattttcta agcgtgttat tttagcagat
     3901 gccaatttag ataaagttct tagtgcatat aacaaacata gagacaaacc aatacgtgaa
     3961 caagcagaaa atattattca tttatttacg ttgacgaatc ttggagctcc cgctgctttt
     4021 aaatattttg atacaacaat tgatcgtaaa cgatatacgt ctacaaaaga agttttagat
     4081 gccactctta tccatcaatc catcactggt ctttatgaaa cacgcattga tttgagtcag
     4141 ctaggaggtg accctccaaa aaagaagaga aaggtctga
//
