LOCUS       sequence-276193-        5001 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AsCpf1 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 5001)
  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 5001)
  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: pAIO-AsCpf1
FEATURES             Location/Qualifiers
     source          1..5001
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(898..921)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(928..975)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(976..4896)
                     /codon_start=1
                     /product="CRISPR-associated protein from Acidaminococcus 
                     sp. BV3L6 (Zetsche et al., 2015)"
                     /label=AsCpf1
                     /translation="MTQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARND
                     HYKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRN
                     AIHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFD
                     KFTTYFSGFYENRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHF
                     ENVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVL
                     NLAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLL
                     RNENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTG
                     KITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTL
                     KKQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKA
                     RNYATKKPYSVEKFKLNFQMPTLASGWDVNKEKNNGAILFVKNGLYYLGIMPKQKGRYK
                     ALSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEP
                     LEITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSID
                     LSSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKG
                     HHGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLK
                     DQKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFF
                     HVPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQR
                     SLNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAV
                     VVLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQ
                     FTSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYD
                     VKTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIEN
                     HRFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQM
                     RNSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKES
                     KDLKLQNGISNQDWLAYIQELRN"
     regulatory      1247..1256
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 ataataaatc cattgtccat aaatagatga aatagcggcc gcctaccctg aagaagaaaa
       61 gtccgatgat gttgtggaca taaagaattg agctgtctca aaaaatgaga aaacaaacaa
      121 acaaacaaaa taccattatg cgcatatcct ctgctcaata tcaatttttc tttatgccag
      181 ttttgtttcg acactatttg agcaacacct tccctttact atctactact gttacaatgt
      241 tattttattc acctttttac aattttatca aaattttttt gtatgcatat aaattataca
      301 ttgtaaagcc aaagtgtaga taaagaacac cttgtttatt tttccccaat tttttttttt
      361 ttaccatttt tttattaacc ttgctcattt acctgctaat acgattgcta tatgatatcg
      421 aaattgaagg aaaaaacatc atttgtgttt atatgttttt ttttaatttt tcaacttttt
      481 ttatttaact atgctaatca tattataaac attttaaaaa tcaagtccaa ctatttatga
      541 atcattgaag agacaacata attataacag taaaaataat aataataata ataattttgt
      601 cttattgttt caaacatatt atgttccata aatatctaaa tttttaatat ggtcataaat
      661 tcataggatt taatttatat aaaatatggc tattcatact agccatttta tgtgtgtaaa
      721 gttttttttt ctgaacaaaa aataaaatgt tcataatttt agctatttac atgcatgtgc
      781 atgcacaaaa aaaaatatgc acacaacata cacattttta cagttataaa tacaatcaat
      841 tggtataaat atataaataa gaaaaacgaa cattaagctg ccatatccct cgagttactt
      901 gtcgtcgtcg tccttgtagt cggatccctt tttctttttt gcctggccgg cctttttcgt
      961 ggccgccggc cttttgttgc gcagctcctg gatgtaggcc agccagtcct gattggagat
     1021 gccgttctgc agcttcagat ccttgctctc cttcaggtga ttcagcagca gctggccctt
     1081 cagggcgatg tggtaggcgc cattggcatc ggcgtccatg ggccactctg ggttctgaaa
     1141 ccgggagtcg aagcacacgc cattcagatc gcgcacgggg ctgttgatat agtcctcgcc
     1201 tgtggcggca ttggagttcc gcatctgcag cacgctgcgg atcagggcca ccatggtgtc
     1261 gatggcgtga gaatcgtcat tctccagcag ctttggcagg atgttggagc catccctgaa
     1321 cacgatgccc ttctcctcca gcagggcgat cagctcgttg gcaggataca ggtcccggta
     1381 tctgccggtg aatctgtgat tctcgatcac tggcacgatt ctcttgccgg cgatgaaagg
     1441 ggtgcccttg gcgtcaaact gtgtctcgtt cttctcgaac acgatatccc atgcaggcat
     1501 aaagccgggc aggcccctct ggaaggacag atttctgttc atcttaaagt gcaggatgaa
     1561 gtcgccggtt ttcacgtcgt agtgcagaaa gtcgaagccc tccaggaagt gcttgcggct
     1621 ctcgtgattc ttgatggttt tccacacgaa ggggtccacg aagccggtca ggggatcgat
     1681 cttagatgta tatggggcag gcacgtaaaa caggaagcca gactgggtgc ccatcttggc
     1741 aaaggaggtg aactggtctg tcagctggta tgggttcagc acgcctccca ctttctctgc
     1801 tggatagtcc ttcagcacca ggcaattcag cttatcgatc agcatcttct cgaactgctg
     1861 gtacacggcc ttctcggcga tgccggtcct cttgctctta aagccgaaat tcaggttctc
     1921 cagcaccacc acggcctggt agtggatcat caggtccacg atctcgtgga tgacctggct
     1981 cagatagccc tgcttcagat ccttgattgt gcccaccaca gaccaggcct gccttgctgc
     2041 caccctctcc ttctccctgt tgtccagctt cttctggtaa tcaaactgct ggatggtgtt
     2101 caggctccgc tgctccagga tcttgccggt ggagtcgatc actgtgatat agatcaggtt
     2161 tctctcgccc cgatcgatgc cgatgatagg tgtctcgggg tgctccttca ggtaggcatt
     2221 caccctctgg ttgaacttag atggggaatt ggcggcctga tagttcagtg tgataggcac
     2281 gtggaaaaag aacttgtcgc tggtaaagcg cctatccttg atgatctcgt gagacacctc
     2341 cttggtgatc acgttgggca gcagggccct ggcctcatca gacaggtcgt gggacagtct
     2401 gtgattcaca tagtcgtaca gctcctggta cagggtgtcg gggattgggg ttttctgatc
     2461 cttcagcttc ttgttcagca tcttctctcc cagccggtgt gccatcctct tcatcctgga
     2521 cttagggcgg tagaacagct cggcctggcc attcagcttg atgcttgtct tggccaggtt
     2581 ctctggagaa aacaggccgg tccaatacag tgtgtgcaga ttaggcttgc cgtggtggcc
     2641 cttggcaaag tccttgttat agatctggaa caggtacagc ttgcctgtct ccacggcatc
     2701 catgatctcc ttctcggcga ttctctggaa gctgatgtgg tacagcaggg gattcagctc
     2761 ggcatagtac tcgcccaggt ccttatactg agaggatggc cgcaggctag acagatcgat
     2821 agaggttgtc ttggtatact tggacagaaa atcccttgtg aagtcgatcc acttgcacag
     2881 ggcctctctg tagcccttct ggtcgccggt tttcttggcg taggctgtct gaaacttctt
     2941 tggctccttc tcaggattgt tcaggtcgta gatctccttt gtgatctcca gaggctcgat
     3001 gaaattgttg gacagcagga tgggggttgt gtgggtctga aagtgggctg tcacggcctt
     3061 cagctgggtg ctgcactttg ggatcatctt ggcggcatca gggaagtagt catagtacat
     3121 cttatcaaag ccctcgctgg ttttctctgt gggctcgaag ctcagggcct tatacctgcc
     3181 cttctgcttt ggcatgatgc ccagatagta caggccgttc ttcacaaaca ggatggcgcc
     3241 attgttcttc tccttattca cgtcccagcc agaggccagt gtaggcatct gaaagttcag
     3301 cttgaacttc tccacggagt agggcttctt ggtggcataa tttctggcct tgttgtagaa
     3361 gctcagagaa ggctccatct ccagcttgat gccggtcagc cgggcagaga actcggggtc
     3421 cacctcgttg gactcatcca cggcaaacca gtccagcagg tggtacaggc ccagcaggct
     3481 gtccagctga gacttcagga tctccttctc ctcctgcttc ttcagggttg taggcagtgg
     3541 ctgatccagg gcggcgtgtg cgtgggacag gatctcgctg gttttctgct tgaaggcctc
     3601 gctcagctcc ttgcctgcgg cagagatgat ctcctgcagg ttgatatcct cgtgcttcag
     3661 gctgcgctgc accttctcct tggcagactt ggtgatcttg cctgtcagct cggagattct
     3721 ccgctcatac agggcattcc tcagtgtatc ccagtggtcg cacagggcgc tgctgattgt
     3781 ctccagcttc ttgtggctga tgaagatgtg tgtcaggtcg atgctgttca gctcgttaaa
     3841 cagggcctcg gctgtctcca gcacgttctc gtttctcagc agtgtcttgt acttgcagaa
     3901 ggactggatc acttcctcgt cgctcttaaa ctcctccagg atgaaagaca gggtgttcct
     3961 atcggacagg atctgcttaa acagggggat gaatctgtgt ggcagggagg cgatgatgtg
     4021 ggctgtctca tcattcttct ggatggccag attcagcacc tcgttcaggc ccttgatctt
     4081 ctcggtgcct gcctcccgag agattcctcc cagcagctgg ttatacaggt cgatctgggt
     4141 ctgtgtcagc agctggttat aaaaagggaa ggaaaacacc tcctcgatgg aggtgctcac
     4201 gaagatgccg atggccttct tcacgttctc aaagtgctcc cgcaggctgg gcacggcggt
     4261 gatcaggcgt gtgaagatgt gacaattctc cttaaacttg gggaagttgt cctgcacgat
     4321 gcggtgtggg atggctgtgc tgatatcctc ggcgctgaac acgttcttcc tgttctcata
     4381 aaagccggag aagtaggttg taaacttgtc gaagctccgc agcagggcgt tctcgtgctc
     4441 ggttgtggtc acggtgccca gctgcttcag caccttgcca ttaaacagct cggccttgaa
     4501 caggcccttg tagatctcgg cgtgtctctt attgatggca tcggtcaggt tgtctgtccg
     4561 gccgatgaag tagtcgtgga tggcattgcg atatgtggcc tgctcctcga tcagggcgtt
     4621 ccttgtctcc tcggttttct cctttctata ggagtcgatg gcggcgctca ggttctccca
     4681 atccagctgc accagctgca ggcactggtc ggcataggtc ttgtagatcc gatcgatgat
     4741 gggcttcagc tccttgtagt gatcattgcg ggccttgtcc tcctcgatga agccctgctc
     4801 ctggatgtgc ttcagggtct tgccctgtgg gatcagctca aaccgcagtg tcttgctcac
     4861 ctgatacagg ttggtaaagc cctcgaactg tgtcatggat cctgatatat ttctattagg
     4921 tatttattat tataaaatat aaatcttgaa tgataataaa taaaatatta gttattcctt
     4981 ttctagttta aaatatacat a
//
