LOCUS       sequence-297678-        5131 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Cas9 and gRNA expression plasmid for P. falciparum with yDHODH 
            selectable marker..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5131)
  AUTHORS   Nessel T, Beck JM, Rayatpisheh S, Jami-Alahmadi Y, Wohlschlegel JA, 
            Goldberg DE, Beck JR
  TITLE     EXP1 is required for organisation of EXP2 in the intraerythrocytic 
            malaria parasite vacuole.
  JOURNAL   Cell Microbiol. 2020 Jan 28:e13168. doi: 10.1111/cmi.13168.
  PUBMED    31990132
REFERENCE   2  (bases 1 to 5131)
  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: "Cell 
            Microbiol. 2020 Jan 28:e13168. doi: 10.1111/cmi.13168."
COMMENT     Sequence Label: pUF-Cas9-pre-sgRNA
FEATURES             Location/Qualifiers
     source          1..5131
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(947..970)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(971..991)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(998..1018)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(1019..5122)
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(1019..5122)
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
ORIGIN
        1 tttaatatat tatgtgtgat ataataataa atccattgtc cataaataga tgaaatagcg
       61 gccgcctacc ctgaagaaga aaagtccgat gatgttgtgg acataaagaa ttgagctgtc
      121 tcaaaaaatg agaaaacaaa caaacaaaca aaataccatt atgcgcatat cctctgctca
      181 atatcaattt ttctttatgc cagttttgtt tcgacactat ttgagcaaca ccttcccttt
      241 actatctact actgttacaa tgttatttta ttcacctttt tacaatttta tcaaaatttt
      301 tttgtatgca tataaattat acattgtaaa gccaaagtgt agataaagaa caccttgttt
      361 atttttcccc aatttttttt ttttaccatt tttttattaa ccttgctcat ttacctgcta
      421 atacgattgc tatatgatat cgaaattgaa ggaaaaaaca tcatttgtgt ttatatgttt
      481 ttttttaatt tttcaacttt ttttatttaa ctatgctaat catattataa acattttaaa
      541 aatcaagtcc aactatttat gaatcattga agagacaaca taattataac agtaaaaata
      601 ataataataa taataatttt gtcttattgt ttcaaacata ttatgttcca taaatatcta
      661 aatttttaat atggtcataa attcatagga tttaatttat ataaaatatg gctattcata
      721 ctagccattt tatgtgtgta aagttttttt tttctgaaca aaaaataaaa tgttcataat
      781 tttagctatt tacatgcatg tgcatgcaca aaaaaaaata tgcacacaac atacacattt
      841 ttacagttat aaatacaatc aattggtata aatatataaa taagaaaaac gaacattaag
      901 ctgccatatc cttaattaag tcgacctgca gggtaccccc gggttacttg tcgtcgtcgt
      961 ccttgtagtc cactttccgc tttttcttag gatcttccac cttgcgtttt ttcttggggt
     1021 cgcctcccag ctgagacagg tcgatccgtg tctcgtacag gccggtgatg ctctggtgga
     1081 tcagggtggc gtccagcacc tctttggtgc tggtgtacct cttccggtcg atggtggtgt
     1141 caaagtactt gaaggcggca ggggctccca gattggtcag ggtaaacagg tggatgatat
     1201 tctcggcctg ctctctgata ggcttgtctc tgtgcttgtt gtaggcgctc agcaccttgt
     1261 ccagattagc gtcggccagg atcactctct tggagaactc gctgatctgc tcgatgatct
     1321 cgtccaggta gtgtttgtgc tgttccacaa acagctgttt ctgctcatta tcctcggggg
     1381 agcccttcag cttctcatag tggctggcca ggtacaggaa gttcacatat ttggagggca
     1441 gggccagttc gtttcccttc tgcagttcgc cggcagaggc cagcattctc ttccggccgt
     1501 tttccagctc gaacagggag tacttaggca gcttgatgat caggtccttt ttcacttctt
     1561 tgtagccctt ggcttccaga aagtcgatgg gattcttctc gaagctgctt ctttccatga
     1621 tggtgatccc cagcagctct ttcacactct tcagtttctt ggacttgccc ttttccactt
     1681 tggccaccac cagcacagaa taggccacgg tggggctgtc gaagccgccg tacttcttag
     1741 ggtcccagtc cttctttctg gcgatcagct tgtcgctgtt cctcttgggc aggatagact
     1801 ctttgctgaa gccgcctgtc tgcacctcgg tctttttcac gatattcact tggggcatag
     1861 acagcacttt ccgcacggtg gcaaagtccc ggcccttatc ccacacgatc tcgcctgttt
     1921 cgccgtttgt ctcgatcaga ggccgcttcc ggatctcgcc gttggccagg gtaatctcgg
     1981 tcttgaaaaa gttcatgatg ttgctgtaga agaagtactt ggcggtagcc ttgccgattt
     2041 cctgctcgct cttggcgatc atcttccgca cgtcgtacac cttgtagtcg ccgtacacga
     2101 actcgctttc cagcttaggg tactttttga tcagggcggt tcccacgacg gcgttcaggt
     2161 aggcgtcgtg ggcgtggtgg tagttgttga tctcgcgcac tttgtaaaac tggaaatcct
     2221 tccggaaatc ggacaccagc ttggacttca gggtgatcac tttcacttcc cggatcagtt
     2281 tgtcgttctc gtcgtactta gtgttcatcc gggagtccag gatctgtgcc acgtgctttg
     2341 tgatctgccg ggtttccacc agctgtctct tgatgaagcc ggccttatcc agttcgctca
     2401 ggccgcctct ctcggccttg gtcagattgt cgaacttcct ctgggtaatc agcttggcat
     2461 tcagcagctg gcgccagtag ttcttcatct tcttcacgac ctcttcggag ggcacgttgt
     2521 cgctcttgcc ccggttcttg tcgctccgag tcagcacttt gttatcgatg gagtcgtcct
     2581 tcagaaagct ctgaggcacg atatggtcca catcgtagtc ggacagccgg ttgatgtcca
     2641 gttcctggtc cacgtacata tcccgcccat tctgcaggta gtacaggtac agcttctcgt
     2701 tctgcagctg ggtgttttcc acggggtgtt ctttcaggat ctggctgccc agctctttga
     2761 tgccctcttc gatccgcttc attctctcgc ggctgttctt ctgtcccttc tgggtggtct
     2821 ggttctctct ggccatttcg atcacgatgt tctcgggctt gtgccggccc atcactttca
     2881 cgagctcgtc caccaccttc actgtctgca ggatgccctt cttaatggcg gggctgccgg
     2941 ccagattggc aatgtgctcg tgcaggctat cgccctggcc ggacacctgg gctttctgga
     3001 tgtcctcttt aaaggtcagg ctgtcgtcgt ggatcagctg catgaagttt ctgttggcga
     3061 agccgtcgga cttcaggaaa tccaggattg tcttgccgga ctgcttgtcc cggatgccgt
     3121 tgatcagctt ccggctcagc ctgccccagc cggtgtatct ccgccgcttc agctgcttca
     3181 tcactttgtc gtcgaacagg tgggcatagg ttttcagccg ttcctcgatc atctctctgt
     3241 cctcaaacag tgtcagggtc agcacgatat cttccagaat gtcctcgttt tcctcattgt
     3301 ccaggaagtc cttgtccttg ataattttca gcagatcgtg gtatgtgccc agggaggcgt
     3361 tgaaccgatc ttccacgccg gagatttcca cggagtcgaa gcactcgatt ttcttgaagt
     3421 agtcctcttt cagctgcttc acggtcactt tccggttggt cttgaacagc aggtccacga
     3481 tggctttttt ctgctcgccg ctcaggaagg cgggctttct cattccctcg gtcacgtatt
     3541 tcactttggt cagctcgttg tacacggtga agtactcgta cagcaggctg tgcttgggca
     3601 gcaccttctc gttgggcagg ttcttatcga agttggtcat ccgctcgatg aagctctggg
     3661 cgctggcgcc cttgtccacc acttcctcga agttccaggg ggtgatggtt tcctcgctct
     3721 ttctggtcat ccaggcgaat ctgctgtttc ccctggccag agggcccacg tagtagggga
     3781 tgcggaaggt caggatcttc tcgatctttt cccggttgtc cttcaggaat gggtaaaaat
     3841 cttcctgccg ccgcagaatg gcgtgcagct ctcccaggtg gatctggtgg gggatgctgc
     3901 cgttgtcgaa ggtccgctgc ttccgcagca ggtcctctct gttcagcttc acgagcagtt
     3961 cctcggtgcc gtccatcttt tccaggatgg gcttgatgaa cttgtagaac tcttcctggc
     4021 tggctccgcc atcgatgtag ccggcgtagc cgttcttgct ctggtcgaag aaaatctctt
     4081 tgtacttctc aggcagctgc tgccgcacga gagctttcag cagggtcagg tcctggtggt
     4141 gctcgtcgta tctcttgatc atagaggcgc tcaggggggc cttggtgatc tcggtgttca
     4201 ctctcaggat gtcgctcagc aggatggcgt cggacaggtt cttggcggcc agaaacaggt
     4261 cggcgtactg gtcgccgatc tgggccagca ggttgtccag gtcgtcgtcg taggtgtcct
     4321 tgctcagctg cagtttggca tcctcggcca ggtcgaagtt gctcttgaag ttgggggtca
     4381 ggcccaggct cagggcaatc aggttgccga acaggccatt cttcttctcg ccgggcagct
     4441 gggcgatcag attttccagc cgtctgctct tgctcagtct ggcagacagg atggccttgg
     4501 cgtccacgcc gctggcgttg atggggtttt cctcgaacag ctggttgtag gtctgcacca
     4561 gctggatgaa cagcttgtcc acgtcgctgt tgtcggggtt caggtcgccc tcgatcagga
     4621 agtggccccg gaacttgatc atgtgggcca gggccagata gatcagccgc aggtcggcct
     4681 tgtcggtgct gtccaccagt ttctttctca ggtggtagat ggtggggtac ttctcgtggt
     4741 aggccacctc gtccacgatg ttgccgaaga tggggtgccg ctcgtgcttc ttatcctctt
     4801 ccaccaggaa ggactcttcc agtctgtgga agaagctgtc gtccaccttg gccatctcgt
     4861 tgctgaagat ctcttgcaga tagcagatcc ggttcttccg tctggtgtat cttcttctgg
     4921 cggttctctt cagccgggtg gcctcggctg tttctccgct gtcgaacagc agggcgccga
     4981 tcaggttctt cttgatgctg tgccggtcgg tgttgcccag caccttgaat ttcttgctgg
     5041 gcaccttgta ctcgtcggtg atcacggccc agcccacaga gttggtgccg atgtccaggc
     5101 cgatgctgta cttcttgtcc atctcgagtt t
//
