LOCUS       sequence-276190-        5285 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cas9 and gRNA expression plasmid for P. falciparum; no selection in 
            parasites..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5285)
  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 5285)
  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: pAIO3
FEATURES             Location/Qualifiers
     source          1..5285
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(951..974)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(975..995)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(1002..1022)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(1023..5126)
                     /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(1023..5126)
                     /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 tatatatata tatatatata ttttatatca tttaatatat tatgtgtgat ataataataa
       61 atccattgtc cataaataga tgaaatagcg gccgcctacc ctgaagaaga aaagtccgat
      121 gatgttgtgg acataaagaa ttgagctgtc tcaaaaaatg agaaaacaaa caaacaaaca
      181 aaataccatt atgcgcatat cctctgctca atatcaattt ttctttatgc cagttttgtt
      241 tcgacactat ttgagcaaca ccttcccttt actatctact actgttacaa tgttatttta
      301 ttcacctttt tacaatttta tcaaaatttt tttgtatgca tataaattat acattgtaaa
      361 gccaaagtgt agataaagaa caccttgttt atttttcccc aatttttttt tttttaccat
      421 ttttttatta accttgctca tttacctgct aatacgattg ctatatgata tcgaaattga
      481 aggaaaaaac atcatttgtg tttatatgtt tttttttaat ttttcaactt tttttattta
      541 actatgctaa tcatattata aacattttaa aaatcaagtc caactattta tgaatcattg
      601 aagagacaac ataattataa cagtaaaaat aataataata ataataattt tgtcttattg
      661 tttcaaacat attatgttcc ataaatatct aaatttttaa tatggtcata aattcatagg
      721 atttaattta tataaaatat ggctattcat actagccatt ttatgtgtgt aaagtttttt
      781 tttctgaaca aaaaataaaa tgttcataat tttagctatt tacatgcatg tgcatgcaca
      841 aaaaaaaata tgcacacaac atacacattt ttacagttat aaatacaatc aattggtata
      901 aatatataaa taagaaaaac gaacattaag ctgccatatc cctcgagtta cttgtcgtcg
      961 tcgtccttgt agtccacttt ccgctttttc ttaggatctt ccaccttgcg ttttttcttg
     1021 gggtcgcctc ccagctgaga caggtcgatc cgtgtctcgt acaggccggt gatgctctgg
     1081 tggatcaggg tggcgtccag cacctctttg gtgctggtgt acctcttccg gtcgatggtg
     1141 gtgtcaaagt acttgaaggc ggcaggggct cccagattgg tcagggtaaa caggtggatg
     1201 atattctcgg cctgctctct gataggcttg tctctgtgct tgttgtaggc gctcagcacc
     1261 ttgtccagat tagcgtcggc caggatcact ctcttggaga actcgctgat ctgctcgatg
     1321 atctcgtcca ggtagtgttt gtgctgttcc acaaacagct gtttctgctc attatcctcg
     1381 ggggagccct tcagcttctc atagtggctg gccaggtaca ggaagttcac atatttggag
     1441 ggcagggcca gttcgtttcc cttctgcagt tcgccggcag aggccagcat tctcttccgg
     1501 ccgttttcca gctcgaacag ggagtactta ggcagcttga tgatcaggtc ctttttcact
     1561 tctttgtagc ccttggcttc cagaaagtcg atgggattct tctcgaagct gcttctttcc
     1621 atgatggtga tccccagcag ctctttcaca ctcttcagtt tcttggactt gcccttttcc
     1681 actttggcca ccaccagcac agaataggcc acggtggggc tgtcgaagcc gccgtacttc
     1741 ttagggtccc agtccttctt tctggcgatc agcttgtcgc tgttcctctt gggcaggata
     1801 gactctttgc tgaagccgcc tgtctgcacc tcggtctttt tcacgatatt cacttggggc
     1861 atagacagca ctttccgcac ggtggcaaag tcccggccct tatcccacac gatctcgcct
     1921 gtttcgccgt ttgtctcgat cagaggccgc ttccggatct cgccgttggc cagggtaatc
     1981 tcggtcttga aaaagttcat gatgttgctg tagaagaagt acttggcggt agccttgccg
     2041 atttcctgct cgctcttggc gatcatcttc cgcacgtcgt acaccttgta gtcgccgtac
     2101 acgaactcgc tttccagctt agggtacttt ttgatcaggg cggttcccac gacggcgttc
     2161 aggtaggcgt cgtgggcgtg gtggtagttg ttgatctcgc gcactttgta aaactggaaa
     2221 tccttccgga aatcggacac cagcttggac ttcagggtga tcactttcac ttcccggatc
     2281 agtttgtcgt tctcgtcgta cttagtgttc atccgggagt ccaggatctg tgccacgtgc
     2341 tttgtgatct gccgggtttc caccagctgt ctcttgatga agccggcctt atccagttcg
     2401 ctcaggccgc ctctctcggc cttggtcaga ttgtcgaact tcctctgggt aatcagcttg
     2461 gcattcagca gctggcgcca gtagttcttc atcttcttca cgacctcttc ggagggcacg
     2521 ttgtcgctct tgccccggtt cttgtcgctc cgagtcagca ctttgttatc gatggagtcg
     2581 tccttcagaa agctctgagg cacgatatgg tccacatcgt agtcggacag ccggttgatg
     2641 tccagttcct ggtccacgta catatcccgc ccattctgca ggtagtacag gtacagcttc
     2701 tcgttctgca gctgggtgtt ttccacgggg tgttctttca ggatctggct gcccagctct
     2761 ttgatgccct cttcgatccg cttcattctc tcgcggctgt tcttctgtcc cttctgggtg
     2821 gtctggttct ctctggccat ttcgatcacg atgttctcgg gcttgtgccg gcccatcact
     2881 ttcacgagct cgtccaccac cttcactgtc tgcaggatgc ccttcttaat ggcggggctg
     2941 ccggccagat tggcaatgtg ctcgtgcagg ctatcgccct ggccggacac ctgggctttc
     3001 tggatgtcct ctttaaaggt caggctgtcg tcgtggatca gctgcatgaa gtttctgttg
     3061 gcgaagccgt cggacttcag gaaatccagg attgtcttgc cggactgctt gtcccggatg
     3121 ccgttgatca gcttccggct cagcctgccc cagccggtgt atctccgccg cttcagctgc
     3181 ttcatcactt tgtcgtcgaa caggtgggca taggttttca gccgttcctc gatcatctct
     3241 ctgtcctcaa acagtgtcag ggtcagcacg atatcttcca gaatgtcctc gttttcctca
     3301 ttgtccagga agtccttgtc cttgataatt ttcagcagat cgtggtatgt gcccagggag
     3361 gcgttgaacc gatcttccac gccggagatt tccacggagt cgaagcactc gattttcttg
     3421 aagtagtcct ctttcagctg cttcacggtc actttccggt tggtcttgaa cagcaggtcc
     3481 acgatggctt ttttctgctc gccgctcagg aaggcgggct ttctcattcc ctcggtcacg
     3541 tatttcactt tggtcagctc gttgtacacg gtgaagtact cgtacagcag gctgtgcttg
     3601 ggcagcacct tctcgttggg caggttctta tcgaagttgg tcatccgctc gatgaagctc
     3661 tgggcgctgg cgcccttgtc caccacttcc tcgaagttcc agggggtgat ggtttcctcg
     3721 ctctttctgg tcatccaggc gaatctgctg tttcccctgg ccagagggcc cacgtagtag
     3781 gggatgcgga aggtcaggat cttctcgatc ttttcccggt tgtccttcag gaatgggtaa
     3841 aaatcttcct gccgccgcag aatggcgtgc agctctccca ggtggatctg gtgggggatg
     3901 ctgccgttgt cgaaggtccg ctgcttccgc agcaggtcct ctctgttcag cttcacgagc
     3961 agttcctcgg tgccgtccat cttttccagg atgggcttga tgaacttgta gaactcttcc
     4021 tggctggctc cgccatcgat gtagccggcg tagccgttct tgctctggtc gaagaaaatc
     4081 tctttgtact tctcaggcag ctgctgccgc acgagagctt tcagcagggt caggtcctgg
     4141 tggtgctcgt cgtatctctt gatcatagag gcgctcaggg gggccttggt gatctcggtg
     4201 ttcactctca ggatgtcgct cagcaggatg gcgtcggaca ggttcttggc ggccagaaac
     4261 aggtcggcgt actggtcgcc gatctgggcc agcaggttgt ccaggtcgtc gtcgtaggtg
     4321 tccttgctca gctgcagttt ggcatcctcg gccaggtcga agttgctctt gaagttgggg
     4381 gtcaggccca ggctcagggc aatcaggttg ccgaacaggc cattcttctt ctcgccgggc
     4441 agctgggcga tcagattttc cagccgtctg ctcttgctca gtctggcaga caggatggcc
     4501 ttggcgtcca cgccgctggc gttgatgggg ttttcctcga acagctggtt gtaggtctgc
     4561 accagctgga tgaacagctt gtccacgtcg ctgttgtcgg ggttcaggtc gccctcgatc
     4621 aggaagtggc cccggaactt gatcatgtgg gccagggcca gatagatcag ccgcaggtcg
     4681 gccttgtcgg tgctgtccac cagtttcttt ctcaggtggt agatggtggg gtacttctcg
     4741 tggtaggcca cctcgtccac gatgttgccg aagatggggt gccgctcgtg cttcttatcc
     4801 tcttccacca ggaaggactc ttccagtctg tggaagaagc tgtcgtccac cttggccatc
     4861 tcgttgctga agatctcttg cagatagcag atccggttct tccgtctggt gtatcttctt
     4921 ctggcggttc tcttcagccg ggtggcctcg gctgtttctc cgctgtcgaa cagcagggcg
     4981 ccgatcaggt tcttcttgat gctgtgccgg tcggtgttgc ccagcacctt gaatttcttg
     5041 ctgggcacct tgtactcgtc ggtgatcacg gcccagccca cagagttggt gccgatgtcc
     5101 aggccgatgc tgtacttctt gtccatttcg aaggatcctg atatatttct attaggtatt
     5161 tattattata aaatataaat cttgaatgat aataaataaa atattagtta ttccttttct
     5221 agtttaaaat atacatatta taaatatata tatatatata tatttttatt gtgacaagaa
     5281 tatat
//
