LOCUS       sequence-276200-        5501 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  AsCpf1 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 5501)
  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 5501)
  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-AsCpf1-pre-gRNA
FEATURES             Location/Qualifiers
     source          1..5501
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(1372..1395)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(1402..1449)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(1450..5370)
                     /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      1721..1730
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 attatatatg taatgattta tataaaaaaa taaaattata caagtatata ttttgtttct
       61 ataaattgat atcttaatta tttattatta gaaatagata tttttataat aaaccaatag
      121 ataaaatttg tagagaaaaa aaaataaaaa taaaaataaa aataatataa tatataataa
      181 aataaaataa tattatataa atatatttta atttttttta caaaatgccc gggacagatc
      241 ttcggactag tcctccccgc ggaaaggggc cattggatat atatttagta ttccatagta
      301 tatagttaaa tataataata ttatagtaaa tttttgcatg aatttaaaga aaaaaaaaaa
      361 aaaaataaca aaaaggagta ttatataaat ataacaaaat aataaatact atacctatat
      421 atatatatat atatatatat attttatatc atttaatata ttatgtgtga tataataata
      481 aatccattgt ccataaatag atgaaatagc ggccgcctac cctgaagaag aaaagtccga
      541 tgatgttgtg gacataaaga attgagctgt ctcaaaaaat gagaaaacaa acaaacaaac
      601 aaaataccat tatgcgcata tcctctgctc aatatcaatt tttctttatg ccagttttgt
      661 ttcgacacta tttgagcaac accttccctt tactatctac tactgttaca atgttatttt
      721 attcaccttt ttacaatttt atcaaaattt ttttgtatgc atataaatta tacattgtaa
      781 agccaaagtg tagataaaga acaccttgtt tatttttccc caattttttt ttttttacca
      841 tttttttatt aaccttgctc atttacctgc taatacgatt gctatatgat atcgaaattg
      901 aaggaaaaaa catcatttgt gtttatatgt ttttttttaa tttttcaact ttttttattt
      961 aactatgcta atcatattat aaacatttta aaaatcaagt ccaactattt atgaatcatt
     1021 gaagagacaa cataattata acagtaaaaa taataataat aataataatt ttgtcttatt
     1081 gtttcaaaca tattatgttc cataaatatc taaattttta atatggtcat aaattcatag
     1141 gatttaattt atataaaata tggctattca tactagccat tttatgtgtg taaagttttt
     1201 ttttctgaac aaaaaataaa atgttcataa ttttagctat ttacatgcat gtgcatgcac
     1261 aaaaaaaaat atgcacacaa catacacatt tttacagtta taaatacaat caattggtat
     1321 aaatatataa ataagaaaaa cgaacattaa gctgccatat ccctcgagtt acttgtcgtc
     1381 gtcgtccttg tagtcggatc cctttttctt ttttgcctgg ccggcctttt tcgtggccgc
     1441 cggccttttg ttgcgcagct cctggatgta ggccagccag tcctgattgg agatgccgtt
     1501 ctgcagcttc agatccttgc tctccttcag gtgattcagc agcagctggc ccttcagggc
     1561 gatgtggtag gcgccattgg catcggcgtc catgggccac tctgggttct gaaaccggga
     1621 gtcgaagcac acgccattca gatcgcgcac ggggctgttg atatagtcct cgcctgtggc
     1681 ggcattggag ttccgcatct gcagcacgct gcggatcagg gccaccatgg tgtcgatggc
     1741 gtgagaatcg tcattctcca gcagctttgg caggatgttg gagccatccc tgaacacgat
     1801 gcccttctcc tccagcaggg cgatcagctc gttggcagga tacaggtccc ggtatctgcc
     1861 ggtgaatctg tgattctcga tcactggcac gattctcttg ccggcgatga aaggggtgcc
     1921 cttggcgtca aactgtgtct cgttcttctc gaacacgata tcccatgcag gcataaagcc
     1981 gggcaggccc ctctggaagg acagatttct gttcatctta aagtgcagga tgaagtcgcc
     2041 ggttttcacg tcgtagtgca gaaagtcgaa gccctccagg aagtgcttgc ggctctcgtg
     2101 attcttgatg gttttccaca cgaaggggtc cacgaagccg gtcaggggat cgatcttaga
     2161 tgtatatggg gcaggcacgt aaaacaggaa gccagactgg gtgcccatct tggcaaagga
     2221 ggtgaactgg tctgtcagct ggtatgggtt cagcacgcct cccactttct ctgctggata
     2281 gtccttcagc accaggcaat tcagcttatc gatcagcatc ttctcgaact gctggtacac
     2341 ggccttctcg gcgatgccgg tcctcttgct cttaaagccg aaattcaggt tctccagcac
     2401 caccacggcc tggtagtgga tcatcaggtc cacgatctcg tggatgacct ggctcagata
     2461 gccctgcttc agatccttga ttgtgcccac cacagaccag gcctgccttg ctgccaccct
     2521 ctccttctcc ctgttgtcca gcttcttctg gtaatcaaac tgctggatgg tgttcaggct
     2581 ccgctgctcc aggatcttgc cggtggagtc gatcactgtg atatagatca ggtttctctc
     2641 gccccgatcg atgccgatga taggtgtctc ggggtgctcc ttcaggtagg cattcaccct
     2701 ctggttgaac ttagatgggg aattggcggc ctgatagttc agtgtgatag gcacgtggaa
     2761 aaagaacttg tcgctggtaa agcgcctatc cttgatgatc tcgtgagaca cctccttggt
     2821 gatcacgttg ggcagcaggg ccctggcctc atcagacagg tcgtgggaca gtctgtgatt
     2881 cacatagtcg tacagctcct ggtacagggt gtcggggatt ggggttttct gatccttcag
     2941 cttcttgttc agcatcttct ctcccagccg gtgtgccatc ctcttcatcc tggacttagg
     3001 gcggtagaac agctcggcct ggccattcag cttgatgctt gtcttggcca ggttctctgg
     3061 agaaaacagg ccggtccaat acagtgtgtg cagattaggc ttgccgtggt ggcccttggc
     3121 aaagtccttg ttatagatct ggaacaggta cagcttgcct gtctccacgg catccatgat
     3181 ctccttctcg gcgattctct ggaagctgat gtggtacagc aggggattca gctcggcata
     3241 gtactcgccc aggtccttat actgagagga tggccgcagg ctagacagat cgatagaggt
     3301 tgtcttggta tacttggaca gaaaatccct tgtgaagtcg atccacttgc acagggcctc
     3361 tctgtagccc ttctggtcgc cggttttctt ggcgtaggct gtctgaaact tctttggctc
     3421 cttctcagga ttgttcaggt cgtagatctc ctttgtgatc tccagaggct cgatgaaatt
     3481 gttggacagc aggatggggg ttgtgtgggt ctgaaagtgg gctgtcacgg ccttcagctg
     3541 ggtgctgcac tttgggatca tcttggcggc atcagggaag tagtcatagt acatcttatc
     3601 aaagccctcg ctggttttct ctgtgggctc gaagctcagg gccttatacc tgcccttctg
     3661 ctttggcatg atgcccagat agtacaggcc gttcttcaca aacaggatgg cgccattgtt
     3721 cttctcctta ttcacgtccc agccagaggc cagtgtaggc atctgaaagt tcagcttgaa
     3781 cttctccacg gagtagggct tcttggtggc ataatttctg gccttgttgt agaagctcag
     3841 agaaggctcc atctccagct tgatgccggt cagccgggca gagaactcgg ggtccacctc
     3901 gttggactca tccacggcaa accagtccag caggtggtac aggcccagca ggctgtccag
     3961 ctgagacttc aggatctcct tctcctcctg cttcttcagg gttgtaggca gtggctgatc
     4021 cagggcggcg tgtgcgtggg acaggatctc gctggttttc tgcttgaagg cctcgctcag
     4081 ctccttgcct gcggcagaga tgatctcctg caggttgata tcctcgtgct tcaggctgcg
     4141 ctgcaccttc tccttggcag acttggtgat cttgcctgtc agctcggaga ttctccgctc
     4201 atacagggca ttcctcagtg tatcccagtg gtcgcacagg gcgctgctga ttgtctccag
     4261 cttcttgtgg ctgatgaaga tgtgtgtcag gtcgatgctg ttcagctcgt taaacagggc
     4321 ctcggctgtc tccagcacgt tctcgtttct cagcagtgtc ttgtacttgc agaaggactg
     4381 gatcacttcc tcgtcgctct taaactcctc caggatgaaa gacagggtgt tcctatcgga
     4441 caggatctgc ttaaacaggg ggatgaatct gtgtggcagg gaggcgatga tgtgggctgt
     4501 ctcatcattc ttctggatgg ccagattcag cacctcgttc aggcccttga tcttctcggt
     4561 gcctgcctcc cgagagattc ctcccagcag ctggttatac aggtcgatct gggtctgtgt
     4621 cagcagctgg ttataaaaag ggaaggaaaa cacctcctcg atggaggtgc tcacgaagat
     4681 gccgatggcc ttcttcacgt tctcaaagtg ctcccgcagg ctgggcacgg cggtgatcag
     4741 gcgtgtgaag atgtgacaat tctccttaaa cttggggaag ttgtcctgca cgatgcggtg
     4801 tgggatggct gtgctgatat cctcggcgct gaacacgttc ttcctgttct cataaaagcc
     4861 ggagaagtag gttgtaaact tgtcgaagct ccgcagcagg gcgttctcgt gctcggttgt
     4921 ggtcacggtg cccagctgct tcagcacctt gccattaaac agctcggcct tgaacaggcc
     4981 cttgtagatc tcggcgtgtc tcttattgat ggcatcggtc aggttgtctg tccggccgat
     5041 gaagtagtcg tggatggcat tgcgatatgt ggcctgctcc tcgatcaggg cgttccttgt
     5101 ctcctcggtt ttctcctttc tataggagtc gatggcggcg ctcaggttct cccaatccag
     5161 ctgcaccagc tgcaggcact ggtcggcata ggtcttgtag atccgatcga tgatgggctt
     5221 cagctccttg tagtgatcat tgcgggcctt gtcctcctcg atgaagccct gctcctggat
     5281 gtgcttcagg gtcttgccct gtgggatcag ctcaaaccgc agtgtcttgc tcacctgata
     5341 caggttggta aagccctcga actgtgtcat ctcgagttta ttcgaaatgt gggaagaaaa
     5401 aaaatataat aaaaaaggaa aatttctttt taaagtaaaa tatttatgta aatatttata
     5461 tattttatta tatatatatt attatatata tatatatata t
//
