RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-360
Malaria parasiteP. berghei
Genotype
DisruptedGene model (rodent): PBANKA_1301300; Gene model (P.falciparum): PF3D7_1474900; Gene product: trailer hitch homolog, putative (CITH; CAR-I/Trailer Hitch Homolog)
Transgene
Transgene not Plasmodium: RFP
Promoter: Gene model: PBANKA_1319500; Gene model (P.falciparum): PF3D7_1455800; Gene product: LCCL domain-containing protein (LAP4; LCCL/lectin adhesive-like protein 4; CCp2)
3'UTR: Gene model: PBANKA_1359600; Gene product: transmission blocking target antigen precursor 6-cysteine protein (P48/45)
Replacement locus: Gene model: PBANKA_0306000; Gene product: 6-cysteine protein (230p)
Transgene
Transgene not Plasmodium: GFP (gfp-mut3)
Promoter: Gene model: PBANKA_0416100; Gene model (P.falciparum): PF3D7_0905300; Gene product: dynein heavy chain, putative
3'UTR: Gene model: PBANKA_1010600; Gene product: calmodulin, putative (cam)
Replacement locus: Gene model: PBANKA_0306000; Gene product: 6-cysteine protein (230p)
Phenotype Gametocyte/Gamete; Fertilization and ookinete;
Last modified: 21 February 2010, 13:37
  *RMgm-360
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Gene disruption, Introduction of a transgene, Introduction of a transgene
Reference (PubMed-PMID number) Reference 1 (PMID number) : 20169188
MR4 number
Parent parasite used to introduce the genetic modification
Rodent Malaria ParasiteP. berghei
Parent strain/lineP. berghei ANKA
Name parent line/clone P. berghei ANKA 820cl1m1cl1 (RMgm-164)
Other information parent lineP. berghei ANKA 820cl1m1cl1 (RMgm-164) is a reference ANKA mutant line which expresses GFP under control of a male and RFP under control of a female gametocyte specific promoter (PubMed: PMID: 19438517).
The mutant parasite was generated by
Name PI/ResearcherG.R. Mair; C.J. Janse; A.P. Waters
Name Group/DepartmentLeiden Malaria Research Group
Name InstituteLeiden University Medical Center
CityLeiden
CountryThe Netherlands
Name of the mutant parasite
RMgm numberRMgm-360
Principal name934cl1
Alternative nameΔpbcith
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Phenotype
Asexual blood stageNot different from wild type
Gametocyte/GameteNormal production of gametocytes and gametes. Northern analysis of mRNA of gametocytes showed a nearly complete loss of transcripts (mRNA) of genes that are translationally repressed (for example genes encoding the ookinete surface proteins P25, P28).
Microarray analysis of mRNA of gametocytes showed a significant reduction (> 2x) of transcripts of a large number of genes.
Fertilization and ookineteNormal production of gametocytes and gametes. Normal fertilization. A complete block of development of fertilized female gametes (zygotes) into mature ookinetes.
OocystNot tested
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

Mutant/mutation
The mutant lacks expression of CITH (CAR-I/Trailer Hitch Homolog) and expresses GFP under control of a male and RFP under control of a female gametocyte specific promoter.

Protein (function)
CITH shows homology with proteins that co-locolaize with germ cell and P granules in other organisms such as worm CAR-I and fly Trailer Hitch but also Xenopus Rap55. The Plasmodium protein contains both the conserved LSM14 domain and the extended FDF motif, known to compete with the enhancer of mRNA decapping EDC3 for binding to DDX6 helicases.

Phenotype
The phenotype analyses indicate an essential role of CITH in the development of fertilized zygotes. Fertilisation and meiosis (meiotic DNA replication) is comparable to these processes in wild type parasites but the development of zygotes into ookinetes is completely blocked.
Through RNA-immunoprecipitation, and global expression profiling of CITH-KO mutants evidence is presented that CITH is a crucial repressor of maternally supplied mRNAs and plays an important role in translational repression of mRNAs in gametocytes.

Additional information
In Plasmodium CITH was identified in immunoprecipitation experiments with the protein DOZI (protein development of zygote inhibited; ATP-dependent RNA helicase, putative). DOZI is an RNA helicase that is highly up-regulated in female gametocytes and shows high sequence homology to the DDX6 family of RNA helicases.  It contains domains involved in RNA-binding and RNA-unwinding activity.  In other eukaryotes the DDX6 family of DEAD-box RNA helicases is tightly linked both to storage of mRNAs encoding proteins associated with progression through meiosis into translationally silent mRNPs and with the transport of mRNA to degradation centers in the cell (P-bodies). These helicases are found in organisms as diverse as yeast (e.g., Dhh1p) and humans (e.g., RCK/p54).
RNA immunoprecipitation experiments indicate that CITH together with DOZI resides in a stable, translationally quiescent P-body-like structure (containing translationally repressed mRNA's).

The cith gene has been disrupted in a reporter line (RMgm-164) expressing GFP under control of a male and RFP under control of a female gametocyte specific promoter. This reporter line mutant can used to determine fertilisation of female gametes and meiosis in zygotes by FACS analysis. Female gametes/zygotes express RFP allowing for FACS selection of the RFP-positive females to be analysed. Before FACS analysis live cells are stained with the DNA specific dye Hoechst 32258 to measure DNA content and determine the ploïdy of female cells by FACS analysis (Ponzi, M. et al., 2009, Cell. Microbiol 11, 1272-88).

Other mutants
RMgm-358: A mutant that expresses a c-terminally GFP-tagged version of CITH
RMgm-359: A mutant lacking expression of CITH which expresses GFP under the constitutive eef1a promoter


  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_1301300
Gene Model P. falciparum ortholog PF3D7_1474900
Gene producttrailer hitch homolog, putative
Gene product: Alternative nameCITH; CAR-I/Trailer Hitch Homolog
Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system
Type of plasmid/construct usedPlasmid double cross-over
PlasmoGEM (Sanger) construct/vector usedNo
Modified PlasmoGEM construct/vector usedNo
Plasmid/construct map
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Plasmid/construct sequence
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GCGGATAACAATTTCACACAGGAAACAGCTATGACCATGATTACGCCAAGCTCGAAATTA
ACCCTCACTAAAGGGAACAAAAGCTGGTACCTTGACATATCAATTTGTGTACATAAAAAA
AACAAATAATAAAGTATATTTTAGACCCTTCATAATTATATATATTATGATAAAAATATA
ATTATTTCCTTTTGAAAGATATTATAACTGGAAAAATATATTACTAACCCATAAAACAAT
TTATTATTTATTCCATTTTGTTTTGTTTACGTTTATTTTATATATATTATGTATATTTAT
CCACGTATATAATTATTTAATTATGTATTATTATATGCATTAGCTATGTATGCAAAATAT
ATATATTATTATTTAATTTTGGTATAAATTAGCTATTTTTCATTTAAGTTTTGATTGAAA
TATTATACTACATTTTTTTCATTCATTATTTTACTATTTAAACATTTCCCATCGCCTGCA
ATATATATATGTATGATATTATACTATATACTTTTAATTTATGCTTAATGCTATACTTTA
TGCTAAATTATATCATATCCATATAACATTTTTATTTGCTTAGAGTTTTTAACAATGCAT
GGCTTTTTTTTTTTATGCTTATTCTTTCCATAAAAAGAGAGTAAATTCCTTTTTAAAATG
TTATGAAGCATTCATTCAAATATCGATAATATTATTTTATTTTTTAGCTTTTACATTTTG
TGTTTAAATATATATACTACGCATGCATATGTATATATAATAATATGCGTGATGTGTATA
GTAAAACGAGCACAGTGAAATACACATATTAAAGCATATTATCATAATAATGTGCATAGG
CATAATTTTATATATTTAGAGAAATAGTGCAATGCATATTAAAATACACATTTGTAAGCA
TATTATATTTGGTATAATTTTCATTAACATATATATATTATCGTTTCCAGCTTACATAAT
ATATAGGGTTATTAGCAATTTAGGATATAAACTTATATATATTTTTTTCGTTTAAAACAA
ATTGATAAGGGGTAAAAGCTTGCATGCCTGCAGGTCAACAATAAATAATAAATAAATATT
GTGGAAATAAAATAACATATAATTATTTTTAATACATTGATTTCCCTTTTATTTTTTTAA
ATTTCATTGATATAAAAATATATAATAATAACATATATGATTTCAAATTAATCTTTTCAA
AAATGGTGTTTATTTTTGTATGTTTGTGTATGAATTAATCACATAACACATCTATTAAAT
TGAGTTGGTAATATAGACACAAATAAATATATATATTTTTATAGCTTAAAAGTGTGTTAT
GAATATTTTAAGCATATTTTCTTTTTCTTTGGATTGTGTAAAATGAACTCATATAATGCG
TTTTTTTGTTTTTGTTATTTTGTCATTTTGTTATTTTGCTATTTTATGGATTAATTTTTG
TTTATAAAATGGGAAATAATTTTAACATATTTAAATAAATGGAGAAAAAATATAAAATAA
TTATAAAAAAAAGTTAATACACATTTTTTCCTGTTATAGACCTTATATTTATTTATCCAT
ATATATATATATATATATATATATATATATATACATACCAAGTGAATTAAGAGGAAAGCT
AATTTATTATTCAGAATAATATATGAACTATATATAATTTTTATTATTTTGGTGTATATT
AATCTGTCTATATGCATACATGCAATAATTTATCGACTTATATATCAAATAACATAAAAT
AGAAGTGTTTTAAATTATGGATATATGCTCAATATTCATTTTTTTTAATAAGTTAGCTAT
ATTTAAATTATACATTTTATATATGGTCTCTTTTTTTTTTAAATATTATTTAAGTGATCA
TGAAAATATAAATAATTTTTTTTTATTTAATATCCTTTTGCTTGCATGTGGTAAATGGAA
ATTTGGATGTGTTTTGAARGTTCGGATATAGTTGTATGGACATATAATATATTTTGTGAA
AAATTGGTTTTATGTTTATACTTATGCCAATACTTTTTGAGTAAAACAAAGCAAGTGCTT
ATAAATAATTAAAGCCAATTTTATAATATATATTTTTTTATTTAATTTGAATTTAGTAGT
ATAATTTTTTATGGTAAGTGCTCAAAGAGAGTTGCTTATAAAGTATGGTTTGTTTCTTTT
TCGCCATTTTGAATTACACATTAAAAATATATAGATACATATATTATAATATGAAATCAT
TAATAATTTAGGGAAATTCTACAAATTTAAAAACGAATAAAATAATTGTTTTTCATCATG
CCATAACACAATATTGATATATACATGTACAAACATTTTTTTTATTTGGAAAATATAAAT
TATATAAAAAAAAATGTATAGTATACAAAATGAGCATATTCACACGGGGTGGACGTTCAT
TTTTTCATTTTTCCCCTGTTTTTTATGAGTATATGATAAAATTTTATGAACATTTACACA
AAATGAAAATGGATATATAGGAAAAATGGAGCGGTATTTCATTTATCTTTGATTGTCATT
TGGATATTATATTACCYTGGGTAGGCAATTAAAAATGTTAAATAACAATTTAAGGAAATT
ATATTTTATATATTAAAATTAACACTGTATTATATGATTCGCTTATAAAAGCCACTCTTT
CCCCATGCAAAGCTGTTTAATATCAATTTTAACAAATTACACACATGTTAATATATTTAT
ATATATAATTTATATATTTATAATTTATATATTTATATTTTTATTATTTATATATTTATT
ATTTATTGTGTGTGTCAATTCGGGTAGGATATACCTCTTTTTTATTGTTTAAAGCGATTT
GTATTCTAAAATATAAAGRATTTGAAAAAGAGAAAGATAGAATATGATCCCATCATATAT
AGCCCTATAATTTTTATTTAGCAGCGAATTAATTTTTCTATTAAGTTTATGTGTAATTAA
AATAACGGAATATATATAATACAATAAAAAAGTGCATAAATTAAAATTTTTTCAATTAAA
TTTTTTTTTTTAAGGGGTTATATAATATTAAATATATAAAATACGATTATATATTTTTGC
TACAATTTTTTATATTAAGATATAAATAGTAAATAAATGGTATTATATGGCATGTAATAT
ATAAATTTTTTCCAATTTTTATTTTATATACACTTTTCCTTTTTTTGTCATAAAACTTAA
ACAATTTACACATTCATTTTAAAAATTGACTATTTGTTTCAACATTTTTTGAGTTTCCGT
TTTATAATAGTATTTTCATTTGTATATTGCTTATATATATAAATACACACCTAAATGTTA
CAAAGGATCAATGCATAAACCGGTGTGTCTGGTCGTCGCGATGACCCCCAAGAGGGGCAT
CGGCATCAACAACGGCCTCCCGTGGCCCCACTTGACCACAGATTTCAAACACTTTTCTCG
TGTGACAAAAACGACGCCCGAAGAAGCCAGTCGCCTGAACGGGTGGCTTCCCAGGAAATT
TGCAAAGACGGGCGACTCTGGACTTCCCTCTCCATCAGTCGGCAAGAGATTCAACGCCGT
TGTCATGGGACGGAAAACCTGGGAAAGCATGCCTCGAAAGTTTAGACCCCTCGTGGACAG
ATTGAACATCGTCGTTTCCTCTTCCCTCAAAGAAGAAGACATTGCGGCGGAGAAGCCTCA
AGCTGAAGGCCAGCAGCGCGTCCGAGTCTGTGCTTCACTCCCAGCAGCTCTCAGCCTTCT
GGAGGAAGAGTACAAGGATTCTGTCGACCAGATTTTTGTCGTGGGAGGAGCGGGACTGTA
CGAGGCAGCGCTGTCTCTGGGCGTTGCCTCTCACCTGTACATCACGCGTGTAGCCCGCGA
GTTTCCGTGCGACGTTTTCTTCCCTGCGTTCCCCGGAGATGACATTCTTTCAAACAAATC
AACTGCTGCGCAGGCTGCAGCTCCTGCCGAGTCTGTGTTCGTTCCCTTTTGTCCGGAGCT
CGGAAGAGAGAAGGACAATGAAGCGACGTATCGACCCATCTTCATTTCCAAGACCTTCTC
AGACAACGGGGTTCCCTACGACTTTGTGGTTCTCGAGAAGAGAAGGAAGACTGACGACGC
AGCCACTGCGGAACCGAGCAACGCAATGAGCTCCTTGACGTCCACGAGGGAGACAACTCC
CGTGCACGGGTTGCAGGCTCCTTCTTCGGCCGCAGCCATTGCCCCGGTGTTGGCGTGGAT
GGACGAAGAAGACCGGAAAAAACGCGAGCAAAAGGAACTGATTCGGGCCGTTCCGCATGT
TCACTTTAGAGGCCATGAAGAGTTCCAGTACCTTGATCTCATTGCCGACATTATTAACAA
TGGAAGGACAATGGATGACCGAACGGGCGTTGGTGTCATCTCCAAATTCGGCTGCACTAT
GCGCTACTCGCTGGATCAGGCCTTTCCACTTCTCACCACAAAGCGTGTGTTCTGGAAAGG
GGTCCTCGAAGAGTTGCTGTGGTTCATTCGCGGCGACACGAACGCAAACCATCTTTCTGA
GAAGGGCGTGAAGATCTGGGACAAGAATGTGACACGCGAGTTCCTCGATTCGCGCAATCT
CCCCCACCGAGAGGTCGGAGACATCGGCCCGGGCTACGGCTTCCAGTGGAGACACTTCGG
CGCGGCATACAAAGACATGCACACAGACTACACAGGGCAGGGCGTCGACCAGCTGAAGAA
TGTGATCCAGATGCTGAGAACGAATCCAACAGATCGTCGCATGCTCATGACTGCCTGGAA
TCCTGCAGCGCTGGACGAAATGGCGCTGCCGCCTTGTCACTTGTTGTGCCAGTTCTACGT
GAACGACCAGAAGGAGCTGTCGTGCATCATGTATCAGCGGTCGTGCGATGTCGGCCTCGG
CGTCCCCTTCAACATCGCTTCCTATTCGCTTTTGACGCTCATGGTTGCACACGTCTGCAA
CCTAAAACCTAAGGAGTTCATTCACTTCATGGGGAACACGCATGTCTACACGAACCATGT
CGAGGCTTTAAAAGAGCAGCTGCGGAGAGAACCGAGACCGTTCCCCATTGTGAACATCCT
CAACAAGGAACGCATCAAGGAAATCGACGATTTCACCGCCGAGGATTTTGAGGTCGTGGG
CTACGTCCCGCACGGACGAATCCAGATGGAGATGGCTGTCTAGCGGAAATACAGAAGCTA
GCTTTGATCCCGTTTTTCTTACTTATATATTTATACCAATTGATTGTATTTATAACTGTA
AAAATGTGTATGTTGTGTGCATATTTTTTTTTGTGCATGCACATGCATGTAAATAGCTAA
AATTATGAACATTTTATTTTTTGTTCAGAAAAAAAAAACTTTACACACATAAAATGGCTA
GTATGAATAGCCATATTTTATATAAATTAAATCCTATGAATTTATGACCATATTAAAAAT
TTAGATATTTATGGAACATAATATGTTTGAAACAATAAGACAAAATTATTATTATTATTA
TTATTTTTACTGTTATAATTATGTTGTCTCTTCAATGATTCATAAATAGTTGGACTTGAT
TTTTAAAATGTTTATAATATGATTAGCATAGTTAAATAAAAAAAGTTGAAAAATTAAAAA
AAAACATATAAACACAAATGATGTTTTTTCCTTCAATTTCGATATCGAATTCCAAAGATG
TATTATCTGCAAATAGTAACATACAGGTAGGAGGATATGATAAAAAGTCTAGTTTCTTTG
ATAATATTAGTTGTGAAACATTAGACAAACAACAAGGAAAAGATGAAAAGGTGGACAGAG
AAAAATTAAGGATGCTAGATGTTGATACTTTCGGTATAGCAGCGGCACATTATAGGACTA
ATATGCATAATAGAAATAATAACAGAAATAAAGGAAGAAATTCTAAGAATAATAAAATGA
TGGGCAACTTTAATTATAATTATTATAACAGAAACCAAAATCCATTTAACAGATACCCAG
CCTATTAATATATATATTATCTACTAATTAATTTTAATTTTATTCGTTTATATTAAGTAA
ACACTTTCCTTTTAAGAGTCATAACAAATATATATGTACCCAAAAAAAACAGTATAATTC
AATTTTTAAATATTTACAAATGGAATTACACCTATATACAAATACTGCGGCCGCCACCGC
GGTGGAGCTCCAATTCGCCCTATAGTGAGTCGTATTACGCGCGCTCACTGGCCGTCGTTT
TACAACGTCGTGACTGGGAAAACCCTGGCGTTACCCAACTTAATCGCCTTGCAGCACATC
CCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCCTTCCCAACAGT
TGCGCAGCCTGAATGGCGAATGGGACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTG
TGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCG
CTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGG
GGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATT
AGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGT
TGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTA
TCTCGGTCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAA
ATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGCTTACAATTT
AGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACA
TTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAA
AAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATT
TTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCA
GTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAG
TTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGC
GGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCA
GAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGT
AAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCT
GACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGT
AACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGA
CACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACT
TACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACC
ACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGA
GCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGT
AGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGA
GATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACT
TTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGA
TAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGT
AGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCA
AACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCT
TTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTA
GCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCT
AATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTC
AAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACA
GCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGA
AAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGG
AACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGT
CGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAG
CCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTT
TGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTT
TGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGA
GGAAGCGGAAGAGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTA
ATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAA
TGTGAGTTAGCTCACTCATTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTAT
GTTGTGTGGAATTGTGA
Restriction sites to linearize plasmid
Partial or complete disruption of the geneComplete
Additional remarks partial/complete disruption
Selectable marker used to select the mutant parasitetgdhfr
Promoter of the selectable markerpbdhfr
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modification
Additional remarks selection procedure
Primer information: Primers used for amplification of the target sequences  Click to view information
Primer information: Primers used for amplification of the target sequences  Click to hide information
Sequence Primer 1aaaGGTACCTTGACATATCAATTTGTG
Additional information primer 12773 (Asp718); 5' target Fw
Sequence Primer 2aaaAAGCTTTTACCCCTTATCAATTTG
Additional information primer 22774 (HindIII); 5' target Rv
Sequence Primer 3aaaGAATTCCAAAGATGTATTATCTGC
Additional information primer 32775 (EcoRI); 3' target Fw
Sequence Primer 4aaaGCGGCCGCAGTATTTGTATATAGGTG
Additional information primer 42776 (NotI); 3' target Rv
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6

  Transgene: Mutant parasite expressing a transgene
Type and details of transgene
Is the transgene Plasmodium derived Transgene: not Plasmodium
Transgene nameRFP
Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system
Type of plasmid/constructPlasmid double cross-over
PlasmoGEM (Sanger) construct/vector usedNo
Modified PlasmoGEM construct/vector usedNo
Plasmid/construct map
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Plasmid/construct sequence
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AATTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCGTTACCCAACTT
AATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACC
GATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGCGCCTGATGCGGTATTTT
CTCCTTACGCATCTGTGCGGTATTTCACACCGCATATGGTGCACTCTCAGTACAATCTGC
TCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGA
CGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGC
ATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAAGGGCCTCGTGATA
CGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACT
TTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATG
TATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT
ATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCT
GTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCA
CGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCC
GAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCC
CGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTG
GTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTA
TGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATC
GGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTT
GATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATG
CCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCT
TCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGC
TCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCT
CGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTAC
ACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCC
TCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGAT
TTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATG
ACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATC
AAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAA
CCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAG
GTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTA
GGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTA
CCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAG
TTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTG
GAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCATTGAGAAAGCGCCACG
CTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAG
CGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGC
CACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA
AACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATG
TTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCT
GATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAA
GAGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGG
CACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAG
CTCACTCATTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGA
ATTGTGAGCGGATAACAATTTCACACAGGAAACAGCTATGACCATGATTACGCCAAGCTT
CCGCGGGTATATGGTAAAGAACCTACTAACACAATAAAATATTTAAATAATGTATTTCCT
ATAAATAAATTTACAGATTTATTTTTTAATACAAAAGATATAGATATACCAGAAATAAAT
GATCAGTTTAAAGGTTTTAAATTCTTTATGACATCATTTATAAATCATGGATCATATCCA
CTAACAATAGAATGTGGTGTAACAAATGGTGGAACTAGTTATAAAAGAGCAATTATTTTA
TTGCATGTTCGAACTGATTTAAAAGATAGACCAGTTTCATTTTGTGATTTTCGAAAAGGA
GAATTATATAATTATTTGAATGCTTATACTGAAGGGGATGTATGCATAATAATTTCCAAA
TCAAATACAAGTTTTGGTTTTAGATGCCCAGTAAATACAAAAAAAATGCCAAAAAATTGT
TTTACGCAAGTATATGAAAAAGGGTATCTAAATGACGCCAATAAAATTAATACTAAAAAT
GTTATTAACTATTCATTTGAAAATCCAGAATATGCGCTAGCTGGTTYTAATTATACATTA
ACAAAATCGTATCAATTTGAATGTCATTGTGTAGATAAAGAAACAGAACAAATTGTAAAA
ACGGTTTTAGTCAAATATGTAAATGAAGATGAAATATATGATTATAATGATTTTCCAATG
GTGAATCACAAACCTATTATTGCACATCCAAATAAAACACATCAAGCTTGCATGCCTGCA
GGAATTCGATGGCCGCTCTAGCTTTGATCCCGTTTTTCTTACTTATATATTTATACCAAT
TGATTGTATTTATAACTGTAAAAATGTGTATGTTGTGTGCATATTTTTTTTTGTGCATGC
ACATGCATGTAAATAGCTAAAATTATGAACATTTTATTTTTTGTTCAGAAAAAAAAAACT
TTACACACATAAAATGGCTAGTATGAATAGCCATATTTTATATAAATTAAATCCTATGAA
TTTATGACCATATTAAAAATTTAGATATTTATGGAACATAATATGTTTGAAACAATAAGA
CAAAATTATTATTATTATTATTATTTTTACTGTTATAATTATGTTGTCTCTTCAATGATT
CATAAATAGTTGGACTTGATTTTTAAAATGTTTATAATATGATTAGCATAGTTAAATAAA
AAAAGTTGAAAAATTAAAAAAAAACATATAAACACAAATGATGTTTTTTCCTTCAATTTC
GATTGATAATTATGCAGCCCAGCTTAATTCTTTTCGAGCTCTTTATGCTTAAGTTTACAA
TTTAATATTCATACTTTAAGTATTTTTTGTAGTATCCTAGATATTGTGCTTTAAATGCTC
ACCCCTCAAAGCACCAGTAATATTTTCATCCACTGAAATACCATTAAATTTTCAAAAAAA
TACTATGCATATAATGTTATACATATAAACATAAAACGCCATGTAAATCAAAAAATATAT
AAAAATATGTATAAAAATAAATATGCACTAAATATAAGCTAATTATGCATAAAAATTAAA
GTGCCCTTTATTAACTAGAACTAGTCGTAATTATTTATATTTCTATGTTATAAAAAAATC
CTCATATAATAATATAATTAATATATGTAATGTTTTTTTTATTTTATAATTTTAATATAA
AATAATATGTAAATTAATTCAAAAAATAAATATAATTGTTGTGAAACAAAAAACGTAATT
TTTTCATTTGCCTTCAAAATTTAAATTTATTTTAATATTTCCTAAAATATATATACTTTG
TGTATAAATATATAAAAATATATATTTGCTTATAAATAAATAAAAAATTTTATAAAACAT
AGGGGGATCCATGGTTGGTTCGCTAAACTGCATCGTCGCTGTGTCCCAGAACATGGGCAT
CGGCAAGAACGGGGACCTGCCCTGGCCACCGCTCAGGAACGAATTTAGATATTTCCAGAG
AATGACCACAACCTCTTCAGTAGAAGGTAAACAGAATCTGGTGATTATGGGTAAGAAGAC
CTGGTTCTCCATTCCTGAGAAGAATCGACCTTTAAAGGGTAGAATTAATTTAGTTCTCAG
CAGAGAACTCAAGGAACCTCCACAAGGAGCTCATTTTCTTTCCAGAAGTCTAGATGATGC
CTTAAAACTTACTGAACAACCAGAATTAGCAAATAAAGTAGACATGGTCTGGATAGTTGG
TGGCAGTTCTGTTTATAAGGAAGCCATGAATCACCCAGGCCATCTTAAACTATTTGTGAC
AAGGATCATGCAAGACTTTGAAAGTGACACGTTTTTTCCAGAAATTGATTTGGAGAAATA
TAAACTTCTGCCAGAATACCCAGGTGTTCTCTCTGATGTCCAGGAGGAGAAAGGCATTAA
GTACAAATTTGAAGTATATGAGAAGAATGATGCTAGCGGAGGAGGTGGATCTGGTGGAGG
TGGAAGTGCTAGCGTGACAGGGGGAATGGCAAGCAAGTGGGATCAGAAGGGTATGGACAT
TGCCTATGAGGAGGCGGCCTTAGGTTACAAAGAGGGTGGTGTTCCTATTGGCGGATGTCT
TATCAATAACAAAGACGGAAGTGTTCTCGGTCGTGGTCACAACATGAGATTTCAAAAGGG
ATCCGCCACACTACATGGTGAGATCTCCACTTTGGAAAACTGTGGGAGATTAGAGGGCAA
AGTGTACAAAGATACCACTTTGTATACGACGCTGTCTCCATGCGACATGTGTACAGGTGC
CATCATCATGTATGGTATTCCACGCTGTGTTGTCGGTGAGAACGTTAATTTCAAAAGTAA
GGGCGAGAAATATTTACAAACTAGAGGTCACGAGGTTGTTGTTGTTGACGATGAGAGGTG
TAAAAAGATCATGAAACAATTTATCGATGAAAGACCTCAGGATTGGTTTGAAGATATTGG
TGAGGCTTCGGAACCATTTAAGAACGTCTACTTGCTACCTCAAACAAACCAATTGCTGGG
TTTGTACACCATCATCAGAAATAAGAATACAACTAGACCTGATTTCATTTTCTACTCCGA
TAGAATCATCAGATTGTTGGTTGAAGAAGGTTTGAACCATCTACCTGTGCAAAAGCAAAT
TGTGGAAACTGACACCAACGAAAACTTCGAAGGTGTCTCATTCATGGGTAAAATCTGTGG
TGTTTCCATTGTCAGAGCTGGTGAATCGATGGAGCAAGGATTAAGAGACTGTTGTAGGTC
TGTGCGTATCGGTAAAATTTTAATTCAAAGGGACGAGGAGACTGCTTTACCAAAGTTATT
CTACGAAAAATTACCAGAGGATATATCTGAAAGGTATGTCTTCCTATTAGACCCAATGCT
GGCCACCGGTGGTAGTGCTATCATGGCTACAGAAGTCTTGATTAAGAGAGGTGTTAAGCC
AGAGAGAATTTACTTCTTAAACCTAATCTGTAGTAAGGAAGGGATTGAAAAATACCATGC
CGCCTTCCCAGAGGTCAGAATTGTTACTGGTGCCCTCGACAGAGGTCTAGATGAAAACAA
GTATCTAGTTCCAGGGTTGGGTGACTTTGGTGACAGATACTACTGTGTTTAACTCGATCC
CGTTTTTCTTACTTATATATTTATACCAATTGATTGTATTTATAACTGTAAAAATGTGTA
TGTTGTGTGCATATTTTTTTTTGTGCATGCACATGCATGTAAATAGCTAAAATTATGAAC
ATTTTATTTTTTGTTCAGAAAAAAAAAACTTTACACACATAAAATGGCTAGTATGAATAG
CCATATTTTATATAAATTAAATCCTATGAATTTATGACCATATTAAAAATTTAGATATTT
ATGGAACATAATATGTTTGAAACAATAAGACAAAATTATTATTATTATTATTATTTTTAC
TGTTATAATTATGTTGTCTCTTCAATGATTCATAAATAGTTGGACTTGATTTTTAAAATG
TTTATAATATGATTAGCATAGTTAAATAAAAAAAGTTGAAAAATTAAAAAAAAACATATA
AACACAAATGATGTTTTTTCCTTCAATTTCGGGTACCGACCATATAAGAATTAACCCTTT
ACTTTTTTCGTATTTTTCATTAACTTCCATTTGTTAATTTTTTAAAATAAGTTTTTATTT
TCAATAAAAAAACTATATTAATTTAATCTAATTATAAGCAAAAAATTAAATTCAAAGAAT
AAAATTATATGCACATAATGTGTGCCCCTTTAAAACAAATATTCGATGTTTAATTCGTAC
TTTATGCTAATTTTTTATTATTCACAGATTTTATATTAGAGTTATAAAAATTTAGAATTT
TTTCAAGTAAAATATTATTAGTTAATGGCATTAAAATGTATAAGGAAAAGTTAATATAAA
AATTAAAAAAAACAGTTACACGTATATTACGCATACAACGATGTATATATATTCATATAT
ATTAATAATTCTAGAGTCGCGGCCGCTTTACTTGTACAGCTCGTCCATGCCGAGAGTGAT
CCCGGCGGCGGTCACGAACTCCAGCAGGACCATGTGATCGCGCTTCTCGTTGGGGTCTTT
GCTCAGGGCGGACTGGGTGCTCAGGTAGTGGTTGTCGGGCAGCAGCACGGGGCCGTCGCC
GATGGGGGTGTTCTGCTGGTAGTGGTCGGCGAGCTGCACGCTGCCGTCCTCGATGTTGTG
GCGGATCTTGAAGTTCACCTTGATGCCGTTCTTCTGCTTGTCGGCCATGATATAGACGTT
GTGGCTGTTGTAGTTGTACTCCAGCTTGTGCCCCAGGATGTTGCCGTCCTCCTTGAAGTC
GATGCCCTTCAGCTCGATGCGGTTCACCAGGGTGTCGCCCTCGAACTTCACCTCGGCGCG
GGTCTTGTAGTTGCCGTCGTCCTTGAAGAAGATGGTGCGCTCCTGGACGTAGCCTTCGGG
CATGGCGGACTTGAAGAAGTCGTGCTGCTTCATGTGGTCGGGGTAGCGGCTGAAGCACTG
CACGCCGTAGGTCAGGGTGGTCACGAGGGTGGGCCAGGGCACGGGCAGCTTGCCGGTGGT
GCAGATGAACTTCAGGGTCAGCTTGCCGTAGGTGGCATCGCCCTCGCCCTCGCCGGACAC
GCTGAACTTGTGGCCGTTTACGTCGCCGTCCAGCTCGACCAGGATGGGCACCACCCCGGT
GAACAGCTCCTCGCCCTTGCTCACCATGGTGGCGACCGGTGGATCCTTTTTATCATTTGG
ATAATTAATTCTTATATTTATTCTGAATTAAATCAAACTTCCTATTCATATGTACATGCA
TCTATATGTATGAAATACATATAATACTACTATATACCACCAAATTACATATCCAAATAA
CTTGTGCCCTTAACACTGCATATATATGTTTGTCCATATATTTGCAAATATCTATATATA
CTATAAACCTTATGCACAAACCCCTTTTTCGCATATAAAAATGAATATATATATATATAT
ATATATATACTGTCATATGAAAATATATATCCATTTATTTATATTTATATTTATATTATC
TTGCATAAACATTTTATAATATATTTTCTTTAATATAACTTTCCCATATACGTACACACA
TATATAAATATATAGTATGTGTACATATCTACTTATTTTATTGCATATTATATCCCCTTT
TTTTCTCCAATTTTTTTTTTTTATACATGTAAAAGTCGAAAATTCCCATTTATAATAAAC
TTAGGAGAAAATATATATTATTAACTAGTAGTGGTGAATAAGCTCTTTGAAGTCTCAAAA
CAATAAAATTTAAATGTGCATAAATAAGATTGTGAATACGCAAAAATGCAAATAAATACG
CATATTATCATTCATTTCTTAATTATTTAGAAACGGTTCGAACTGCTTCTAATCGTGTCT
TAAGACAAACCCACATCTTATCTCAAAGTTTAGCATTTCGAAGCTTATTATTGTTTCTAA
AACATCTCAACATTTTACTTACAAACTTTGAATGAAAAAAAACAATTTTATAATTATATA
CTAACCATAAAAAAATAAATACCCAAAACGAATAAAATATATTATGTACCTATTTTAAAT
GCGTTTATACCCAAATTTTCGAACCACAATTTTTATATGAACATGTTAATTTTATTTCAC
GTCCCCATTTTATATATATAAAATTTTTTTCCTATTTCTTATTACCTTTAATATTATTTA
TACTAGATACAGAAATAATTTAAAAGGTACTATTTAAAAATGTGTTACCATTTTATTTAA
CTCACAAGCCCCTTTATTATATTAAATTTAAAAGCTTATTTTATGAGGGTTTATATAAAA
ATTCATTCGGTTTTAAATGTAAATATATATATGCACATAATTTTGTATAAATAAAACATA
TATATACATTATAATTATTTATTTGACAGTTTTGAAGGATTTATGAAAAGGAAAAATTTA
TCATGTGTATATATGTTTTTATATTTTATTGACAAGAAATTATAAAGCTCGGAAAATTAA
TAGAAAAGTAGGTGCAAGCATGCACATATAAATACATTAAACACCAAAATACTTTTCCAA
TGACTTGCCTACATGTCTTTGTACATATAATACATACATAGTATAATAAAATATGAAGCT
TTTAAATTATAATAAATTTTTATCTTCATCGGTATAAGGTTCAGATGCAGGGGCGTTATC
TTCCTCATTAGTATAAGAACCTTTTGTTTCATCATCATCATTATTATCTTCGGTACTTAA
TCTGTTTAAATATGATATCTTTATAAATGAAGCAATACCACATTTTTACACAAATATATT
ACATTTTTTATGTATTGCATATGTGAAATGTATGAGTGATATAATATGGTAGACATGAAT
AGGCTAAAACATAGATAAAACCGAACTAAACAAATAAATATAGGAAAATACCCCAATACG
AATATTTGTAACATTAAAAATTAAAAAGACACACTTTAATTAAATAAAATAGAGACGTGA
ATAAAAATGTATTTATAATAAAAGTACGTAAAAAGAAAGCGTTACCAAAACATAAATGAT
TTTTTTCTAACTAAATAAAATTGCATATGTGAAGGTATAATATTTTCCTACATAAAATGG
CGTTTATTGGTTTTTTAATATGTTTTACTTCATTTTGATTTATTACACATATATAAATTT
TTTACAAAATTACGCCATTAATATAGACATTTTTAAGAATGTTGTATTTATTGATCATAT
TCGAACTAAATTCCGTTGTAAATTCACTCAGGTGCATGCAAAAGATGCGGTATTTTTTTG
TACTATTAAAAATTTATATATACAATAATTGTTCAAAATACATAATTTTTTTTAAGCATT
TTACAAACACAAATAATTTTTTCAAAATAAAATAAATTGTTACGTATCTCGAAAATAGTG
TATTTAATATTATACAAATGTAAGGGCAATACATATATATATATTTATTTATTTAAAAGA
CAATTTATAAATGCATATTATTCATAAATTTTTATAATATTTCTCGATAATTTTGTTCGC
AATTTGATGGTGACCCCTTATACATTTGAATTTGCTTTAAAAATATTAAAAAGGAAAATT
TTTTTTTGTTATATAGAGTCATCATGCTTGATTATTTTATTTCTGAATTTAATTTATCCT
CATGTATTTATTATAACAATACGACGTGAAGTTAAAACACCAGTTGCATTGATCAATTTA
AAATTTTGATAAAAAGTATTTAAAAATAAAAGGTTTATAAATAGGGGCTTATTCATTTAT
ATTTCTTTTTATTATATTTTCTTTATATTTTTCAACAAATATTTATAATTCGTAATTTTT
TCGTCAATTTAATATATTTGCCTTTAAGTTTTTCGTAAATTACACTTCAGTAATAATAAT
AAAGCTACATTTTATAGTGCGCATTGTTAAGCATATAATCGATAGGACTTTTAATATATA
TTTTGTTTTTACACAAAATAAGAAGAAGGTCCCTACATTTTTTTCGTTAAAAAGGAAATC
TCAATAATATTAAATTGTGTGTAATTTTTTTTGATGTATACAAATATGACGTGTATATAC
GCAAAGGTGCTTGTAAAACAAATGGTATTAAGCTGTGTGGTCACATTTGTAGTTACATAG
CAATTATATATATATCATTTCTAATTTTAATTCAATTTAATTTCGGTATTAACTATGCAT
ATAAACATATTGTTATTATTATCTTGCCTATCATAATAAATTTTGTGTCTATATAGAAAT
GTATTATCTATTTTTTAGCGAAGTAGCATAAGTATTATTTATATTCAAAGAAATAAATAA
AGGTATTAAAATTAATACGAAAATAAAAATTAAAGGATGATGGCGATTATTAGTTCTTTT
TAATTCCATCATCTATCCTTATATGTAGTATATTGGGGTTAGAGATAGTGGGTTATGAAT
ACCAAAAATATAGAAAATTGCAAAGGAGTTGATTTATTCTCTGTTGTATAAGAATGTACA
CAGTTAAAGTTAGTATATATGTGTATATTTTTCAGTCTATATAAATATACTTTTGTTTTC
TCATTGGTATGTTCAATTTTTTATATTATTTTACGTGTCGATGGATACAATTTAGTATTT
TTTTTAATTTTGTATTTTTGAATTATATTAATCTAGATTATTCATATATTTTTTCATATG
CAGATAGAAAAAGGTTTCATACGTTAGTTTATCCTACAAATTGAATATTAACACATACAT
ATATTTATATTTTTTATTAATAGAAGGATCCAAAATGAGTAGATCTTCTAAGAACGTCAT
CAAGGAATTCATGAGATTCAAGGTTAAAATGGAAGGTACTGTTAACGGCCACGAATTCGA
AATCGAAGGTGAAGGTGAGGGTAGACCATATGAAGGTCACAACACAGTCAAGTTGAAGGT
TACTAAGGGTGGTCCACTGCCATTCGCTTGGGACATCTTGTCTCCACAATTCCAATACGG
TTCTAAGGTCTACGTCAAGCACCCAGCTGACATTCCAGACTACAAGAAGTTGTCCTTCCC
AGAAGGTTTCAAGTGGGAAAGGATCATGAACTTCGAAGACGGTGGCGTTGTTACTGTTAC
TCAAGACTCCTCCTTGCAAGACGGTTGTTTCATCTACAAGGTCAAGCTCATTGGTGTCAA
CTTCCCATCTGACGGTCCAGTCATGCAAAAGAAGACTATGGGTTGGGAAGCTTCTACCGA
ACGTTTGTACCCAAGAGACGGTGTCTTGAAGGGTGAAATCCACAAGGCCTTGAAGTTGAA
GGACGGTGGTCACTACTTGGTCGAATTCAAGTCTATCTACAAGGCCAAGAAGCAAGTCCA
ATTGCCAGGCTATTACTACGTTGACTCTAAGTTGGACATCATCTCTCACAACGAAGACTA
CACTATCGTCGAACAATACGAACGTACTGAAGGTAGACACCACTTGTTCTTGTAAACTAG
TTCTAGAAGTAGTGTGTAGCGTATTCTTTTATTTTACAGAAAATGTATATTATCATTTTT
CGCATTTCATCGTTTAAACAAGTACATGAACGTGTGCATGCAATTATTTCTGTGTTTGGA
TTTGTATTCAAATCATCATGAACTTTTTTTGAATATTCGTTACAAGCCATAATAGAATAT
ACACCATCTACAATATAATTTTTTTTTCACTATTTTATATTTCATCCTATTGTGTATGTA
TATTCCTTTAATAAAAAATCCATATGCTTAACATTCACATATATTAATAATTTTAATTAA
TTTTTTTTTTTGCACAGATAAAACTTTTCTTTTTAAAAAAAAGGTATATACACTCTTTTT
TACCCCACATTTAACTGATTTTCACTCACCATTACATATAGCTATATATTGCATATTACT
ATTATGTAAGGCAAATGTTTATTATTATTGCTGTTATTTTTTATTTATATATTTTTTTCC
ATATATCAATTTAAGGAGAAATAAAATAATTTGCTATTTGAAAAAATATAAAACAAATAT
CTAGTCGTGCAATACTGAATTCGAATACTTAAAACAAAAAATAAATTAATTAAATACCCT
TTTAATATTGTAGTTAAAATCGATGTGTTTAGAGAATATTTTTTATTGCTATTTATCTTC
TTTTTTTTATTTTTCTTCCATGTTTTCTCATTCAGCTTTCTATGTATTTTAGTAATATTC
TCCACACACAAACTATAAATTTTCTTATGAAAAATCACAAAATTTAGCAAATAGCGCTTA
GGCATGTACTCGTAAATAAGCATGTGCATAGTATAATACACTTCTCATCAATTTTGAGAG
GATTTTATTTAATTTCGGCACGCCCCTCTCAACTTGTGTTTATCACATATTCAATACTCC
ATAAAAAATGATAGCATCATTTCGATATGCGGGTACCGAGCTCGAATTCTCTTGAGCCCG
TTAATGAAATAGATACAATTCATTCATGTTATATACATCTAGAACATAATCTGAATATGG
TTCAAGTTAAATGTCCAAAAATTATAAAAAGTGATGATATTTTTGATGGTAATACCATAA
TAGACACCAAGGTAACATCACGAAGTAGTCAACAAAATAATTTTTATTTAGAAAATACAG
ATGTTGAACCAGAAGAAATAGAGAAATATAAAAATATAGAATACATACCAGAAAACGATG
AAGTAATGCATCTAGACAAAAAAGAAAAGCTAGATGATATATTACCAGGTGTTATCATAT
TAGATAAAAATAAAATGTTCAAAGAAAAAGGACATTTCACTTTTGTTACTCCATTAATTG
TAGAAAAGGTATTAATATTAAAAATATATTGTGATAATACTAAAACAATAATTAATAATA
TGAAAGGGAAAAAAGGTATTACAGTAATAAGGATTTCTCAAAATACAACAAAAAATAAAT
TTTATGGATGTGACTTTTCAGGTAATTCTAAAAAAACATTTTACTATTCCAATGTTTATG
ATTTAGAAAAAAAAAATGAGTTTTGTGAAATAGAATTAAAAGAAAATATAGTAGTTAGCT
TAAATTGTCCAACTGGTAAAATTAATCCAAAAAATTGTTTTAGAAATGTATATATAAAAA
GTAATATGAATGAACAAACAACCGAAAATATAGAAAATATATTTAACGAAATAAAAGTTA
TAGATGCAGATTATTTTATAAATAATTCATCAACCTTTTTGATGATTTCCAAAATTACAA
AAAAAGAGTTTGATTTTTATTGTACATGTGAAGATTATAAAACCAAAAATATAGGAACAA
TATATATTAAAAATTATGAATATCTAGATTCAAAACCTAAATATAAAAATAAACAAATTT
CCTATATAGATGTAGTTCCATACCCGCGGGGAAAGGGCG
Restriction sites to linearize plasmid KspI (SacII)
Selectable marker used to select the mutant parasitehdhfr/yfcu
Promoter of the selectable markereef1a
Selection (positive) procedurepyrimethamine
Selection (negative) procedure5-fluorocytosine (5-FC)
Additional remarks genetic modification
Additional remarks selection procedure
Other details transgene
Promoter
Gene Model of Parasite PBANKA_1319500
Gene Model P. falciparum ortholog PF3D7_1455800
Gene productLCCL domain-containing protein
Gene product: Alternative nameLAP4; LCCL/lectin adhesive-like protein 4; CCp2
Primer information details of the primers used for amplification of the promoter sequence  Click to view information
Primer information details of the primers used for amplification of the promoter sequence  Click to hide information
Sequence Primer 1
Additional information primer 1
Sequence Primer 2
Additional information primer 2
3'-UTR
Gene Model of Parasite PBANKA_1359600
Gene producttransmission blocking target antigen precursor 6-cysteine protein
Gene product: Alternative nameP48/45
Primer information details of the primers used for amplification the 3'-UTR sequences  Click to view information
Primer information details of the primers used for amplification the 3'-UTR sequences  Click to hide information
Sequence Primer 13’-cggGGTACCCGCATATCGAAATGATGCTATC
Additional information primer 13’utr of the p48/45 gene (PB001525.02.0; 3’48/45; Asp718/XbaI; 1882
Sequence Primer 23’- gcTCTAGAAGTAGTGTGTAGCGTATTC
Additional information primer 23’utr of the p48/45 gene (PB001525.02.0; 3’48/45; Asp718/XbaI;1881
Insertion/Replacement locus
Replacement / InsertionReplacement locus
Gene Model of Parasite PBANKA_0306000
Gene product6-cysteine protein
Gene product: Alternative name230p
Primer information details of the primers used for amplification of the target sequences  Click to view information
Primer information details of the primers used for amplification of the target sequences  Click to hide information
Sequence Primer 15'-cccaagcttccgcgggtatatggtaaagaacctactaacac
Additional information primer 1primer L1345: 0.7kb 5'region of PB000214.00.0 (230p gene)
Sequence Primer 25'-cccaagcttgatgtgttttatttggatgtgc
Additional information primer 2primer L1346: 0.7kb 5'region of PB000214.00.0 (230p gene)
Sequence Primer 35'-ccggaattctcttgagcccgttaatg
Additional information primer 3primer L1347: 1kb 3'region of PB000214.00.0 (230p gene)
Sequence Primer 45'-tccccgcgggtatggaactacatctatatagg
Additional information primer 4primer L1348: 1kb 3'region of PB000214.00.0 (230p gene)

  Transgene: Mutant parasite expressing a transgene
Type and details of transgene
Is the transgene Plasmodium derived Transgene: not Plasmodium
Transgene nameGFP (gfp-mut3)
Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system
Type of plasmid/constructPlasmid double cross-over
PlasmoGEM (Sanger) construct/vector usedNo
Modified PlasmoGEM construct/vector usedNo
Plasmid/construct map
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Plasmid/construct sequence
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AATTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCGTTACCCAACTT
AATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACC
GATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGCGCCTGATGCGGTATTTT
CTCCTTACGCATCTGTGCGGTATTTCACACCGCATATGGTGCACTCTCAGTACAATCTGC
TCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGA
CGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGC
ATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAAGGGCCTCGTGATA
CGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACT
TTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATG
TATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT
ATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCT
GTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCA
CGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCC
GAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCC
CGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTG
GTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTA
TGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATC
GGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTT
GATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATG
CCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCT
TCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGC
TCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCT
CGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTAC
ACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCC
TCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGAT
TTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATG
ACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATC
AAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAA
CCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAG
GTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTA
GGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTA
CCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAG
TTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTG
GAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCATTGAGAAAGCGCCACG
CTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAG
CGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGC
CACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA
AACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATG
TTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCT
GATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAA
GAGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGG
CACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAG
CTCACTCATTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGA
ATTGTGAGCGGATAACAATTTCACACAGGAAACAGCTATGACCATGATTACGCCAAGCTT
CCGCGGGTATATGGTAAAGAACCTACTAACACAATAAAATATTTAAATAATGTATTTCCT
ATAAATAAATTTACAGATTTATTTTTTAATACAAAAGATATAGATATACCAGAAATAAAT
GATCAGTTTAAAGGTTTTAAATTCTTTATGACATCATTTATAAATCATGGATCATATCCA
CTAACAATAGAATGTGGTGTAACAAATGGTGGAACTAGTTATAAAAGAGCAATTATTTTA
TTGCATGTTCGAACTGATTTAAAAGATAGACCAGTTTCATTTTGTGATTTTCGAAAAGGA
GAATTATATAATTATTTGAATGCTTATACTGAAGGGGATGTATGCATAATAATTTCCAAA
TCAAATACAAGTTTTGGTTTTAGATGCCCAGTAAATACAAAAAAAATGCCAAAAAATTGT
TTTACGCAAGTATATGAAAAAGGGTATCTAAATGACGCCAATAAAATTAATACTAAAAAT
GTTATTAACTATTCATTTGAAAATCCAGAATATGCGCTAGCTGGTTYTAATTATACATTA
ACAAAATCGTATCAATTTGAATGTCATTGTGTAGATAAAGAAACAGAACAAATTGTAAAA
ACGGTTTTAGTCAAATATGTAAATGAAGATGAAATATATGATTATAATGATTTTCCAATG
GTGAATCACAAACCTATTATTGCACATCCAAATAAAACACATCAAGCTTGCATGCCTGCA
GGAATTCGATGGCCGCTCTAGCTTTGATCCCGTTTTTCTTACTTATATATTTATACCAAT
TGATTGTATTTATAACTGTAAAAATGTGTATGTTGTGTGCATATTTTTTTTTGTGCATGC
ACATGCATGTAAATAGCTAAAATTATGAACATTTTATTTTTTGTTCAGAAAAAAAAAACT
TTACACACATAAAATGGCTAGTATGAATAGCCATATTTTATATAAATTAAATCCTATGAA
TTTATGACCATATTAAAAATTTAGATATTTATGGAACATAATATGTTTGAAACAATAAGA
CAAAATTATTATTATTATTATTATTTTTACTGTTATAATTATGTTGTCTCTTCAATGATT
CATAAATAGTTGGACTTGATTTTTAAAATGTTTATAATATGATTAGCATAGTTAAATAAA
AAAAGTTGAAAAATTAAAAAAAAACATATAAACACAAATGATGTTTTTTCCTTCAATTTC
GATTGATAATTATGCAGCCCAGCTTAATTCTTTTCGAGCTCTTTATGCTTAAGTTTACAA
TTTAATATTCATACTTTAAGTATTTTTTGTAGTATCCTAGATATTGTGCTTTAAATGCTC
ACCCCTCAAAGCACCAGTAATATTTTCATCCACTGAAATACCATTAAATTTTCAAAAAAA
TACTATGCATATAATGTTATACATATAAACATAAAACGCCATGTAAATCAAAAAATATAT
AAAAATATGTATAAAAATAAATATGCACTAAATATAAGCTAATTATGCATAAAAATTAAA
GTGCCCTTTATTAACTAGAACTAGTCGTAATTATTTATATTTCTATGTTATAAAAAAATC
CTCATATAATAATATAATTAATATATGTAATGTTTTTTTTATTTTATAATTTTAATATAA
AATAATATGTAAATTAATTCAAAAAATAAATATAATTGTTGTGAAACAAAAAACGTAATT
TTTTCATTTGCCTTCAAAATTTAAATTTATTTTAATATTTCCTAAAATATATATACTTTG
TGTATAAATATATAAAAATATATATTTGCTTATAAATAAATAAAAAATTTTATAAAACAT
AGGGGGATCCATGGTTGGTTCGCTAAACTGCATCGTCGCTGTGTCCCAGAACATGGGCAT
CGGCAAGAACGGGGACCTGCCCTGGCCACCGCTCAGGAACGAATTTAGATATTTCCAGAG
AATGACCACAACCTCTTCAGTAGAAGGTAAACAGAATCTGGTGATTATGGGTAAGAAGAC
CTGGTTCTCCATTCCTGAGAAGAATCGACCTTTAAAGGGTAGAATTAATTTAGTTCTCAG
CAGAGAACTCAAGGAACCTCCACAAGGAGCTCATTTTCTTTCCAGAAGTCTAGATGATGC
CTTAAAACTTACTGAACAACCAGAATTAGCAAATAAAGTAGACATGGTCTGGATAGTTGG
TGGCAGTTCTGTTTATAAGGAAGCCATGAATCACCCAGGCCATCTTAAACTATTTGTGAC
AAGGATCATGCAAGACTTTGAAAGTGACACGTTTTTTCCAGAAATTGATTTGGAGAAATA
TAAACTTCTGCCAGAATACCCAGGTGTTCTCTCTGATGTCCAGGAGGAGAAAGGCATTAA
GTACAAATTTGAAGTATATGAGAAGAATGATGCTAGCGGAGGAGGTGGATCTGGTGGAGG
TGGAAGTGCTAGCGTGACAGGGGGAATGGCAAGCAAGTGGGATCAGAAGGGTATGGACAT
TGCCTATGAGGAGGCGGCCTTAGGTTACAAAGAGGGTGGTGTTCCTATTGGCGGATGTCT
TATCAATAACAAAGACGGAAGTGTTCTCGGTCGTGGTCACAACATGAGATTTCAAAAGGG
ATCCGCCACACTACATGGTGAGATCTCCACTTTGGAAAACTGTGGGAGATTAGAGGGCAA
AGTGTACAAAGATACCACTTTGTATACGACGCTGTCTCCATGCGACATGTGTACAGGTGC
CATCATCATGTATGGTATTCCACGCTGTGTTGTCGGTGAGAACGTTAATTTCAAAAGTAA
GGGCGAGAAATATTTACAAACTAGAGGTCACGAGGTTGTTGTTGTTGACGATGAGAGGTG
TAAAAAGATCATGAAACAATTTATCGATGAAAGACCTCAGGATTGGTTTGAAGATATTGG
TGAGGCTTCGGAACCATTTAAGAACGTCTACTTGCTACCTCAAACAAACCAATTGCTGGG
TTTGTACACCATCATCAGAAATAAGAATACAACTAGACCTGATTTCATTTTCTACTCCGA
TAGAATCATCAGATTGTTGGTTGAAGAAGGTTTGAACCATCTACCTGTGCAAAAGCAAAT
TGTGGAAACTGACACCAACGAAAACTTCGAAGGTGTCTCATTCATGGGTAAAATCTGTGG
TGTTTCCATTGTCAGAGCTGGTGAATCGATGGAGCAAGGATTAAGAGACTGTTGTAGGTC
TGTGCGTATCGGTAAAATTTTAATTCAAAGGGACGAGGAGACTGCTTTACCAAAGTTATT
CTACGAAAAATTACCAGAGGATATATCTGAAAGGTATGTCTTCCTATTAGACCCAATGCT
GGCCACCGGTGGTAGTGCTATCATGGCTACAGAAGTCTTGATTAAGAGAGGTGTTAAGCC
AGAGAGAATTTACTTCTTAAACCTAATCTGTAGTAAGGAAGGGATTGAAAAATACCATGC
CGCCTTCCCAGAGGTCAGAATTGTTACTGGTGCCCTCGACAGAGGTCTAGATGAAAACAA
GTATCTAGTTCCAGGGTTGGGTGACTTTGGTGACAGATACTACTGTGTTTAACTCGATCC
CGTTTTTCTTACTTATATATTTATACCAATTGATTGTATTTATAACTGTAAAAATGTGTA
TGTTGTGTGCATATTTTTTTTTGTGCATGCACATGCATGTAAATAGCTAAAATTATGAAC
ATTTTATTTTTTGTTCAGAAAAAAAAAACTTTACACACATAAAATGGCTAGTATGAATAG
CCATATTTTATATAAATTAAATCCTATGAATTTATGACCATATTAAAAATTTAGATATTT
ATGGAACATAATATGTTTGAAACAATAAGACAAAATTATTATTATTATTATTATTTTTAC
TGTTATAATTATGTTGTCTCTTCAATGATTCATAAATAGTTGGACTTGATTTTTAAAATG
TTTATAATATGATTAGCATAGTTAAATAAAAAAAGTTGAAAAATTAAAAAAAAACATATA
AACACAAATGATGTTTTTTCCTTCAATTTCGGGTACCGACCATATAAGAATTAACCCTTT
ACTTTTTTCGTATTTTTCATTAACTTCCATTTGTTAATTTTTTAAAATAAGTTTTTATTT
TCAATAAAAAAACTATATTAATTTAATCTAATTATAAGCAAAAAATTAAATTCAAAGAAT
AAAATTATATGCACATAATGTGTGCCCCTTTAAAACAAATATTCGATGTTTAATTCGTAC
TTTATGCTAATTTTTTATTATTCACAGATTTTATATTAGAGTTATAAAAATTTAGAATTT
TTTCAAGTAAAATATTATTAGTTAATGGCATTAAAATGTATAAGGAAAAGTTAATATAAA
AATTAAAAAAAACAGTTACACGTATATTACGCATACAACGATGTATATATATTCATATAT
ATTAATAATTCTAGAGTCGCGGCCGCTTTACTTGTACAGCTCGTCCATGCCGAGAGTGAT
CCCGGCGGCGGTCACGAACTCCAGCAGGACCATGTGATCGCGCTTCTCGTTGGGGTCTTT
GCTCAGGGCGGACTGGGTGCTCAGGTAGTGGTTGTCGGGCAGCAGCACGGGGCCGTCGCC
GATGGGGGTGTTCTGCTGGTAGTGGTCGGCGAGCTGCACGCTGCCGTCCTCGATGTTGTG
GCGGATCTTGAAGTTCACCTTGATGCCGTTCTTCTGCTTGTCGGCCATGATATAGACGTT
GTGGCTGTTGTAGTTGTACTCCAGCTTGTGCCCCAGGATGTTGCCGTCCTCCTTGAAGTC
GATGCCCTTCAGCTCGATGCGGTTCACCAGGGTGTCGCCCTCGAACTTCACCTCGGCGCG
GGTCTTGTAGTTGCCGTCGTCCTTGAAGAAGATGGTGCGCTCCTGGACGTAGCCTTCGGG
CATGGCGGACTTGAAGAAGTCGTGCTGCTTCATGTGGTCGGGGTAGCGGCTGAAGCACTG
CACGCCGTAGGTCAGGGTGGTCACGAGGGTGGGCCAGGGCACGGGCAGCTTGCCGGTGGT
GCAGATGAACTTCAGGGTCAGCTTGCCGTAGGTGGCATCGCCCTCGCCCTCGCCGGACAC
GCTGAACTTGTGGCCGTTTACGTCGCCGTCCAGCTCGACCAGGATGGGCACCACCCCGGT
GAACAGCTCCTCGCCCTTGCTCACCATGGTGGCGACCGGTGGATCCTTTTTATCATTTGG
ATAATTAATTCTTATATTTATTCTGAATTAAATCAAACTTCCTATTCATATGTACATGCA
TCTATATGTATGAAATACATATAATACTACTATATACCACCAAATTACATATCCAAATAA
CTTGTGCCCTTAACACTGCATATATATGTTTGTCCATATATTTGCAAATATCTATATATA
CTATAAACCTTATGCACAAACCCCTTTTTCGCATATAAAAATGAATATATATATATATAT
ATATATATACTGTCATATGAAAATATATATCCATTTATTTATATTTATATTTATATTATC
TTGCATAAACATTTTATAATATATTTTCTTTAATATAACTTTCCCATATACGTACACACA
TATATAAATATATAGTATGTGTACATATCTACTTATTTTATTGCATATTATATCCCCTTT
TTTTCTCCAATTTTTTTTTTTTATACATGTAAAAGTCGAAAATTCCCATTTATAATAAAC
TTAGGAGAAAATATATATTATTAACTAGTAGTGGTGAATAAGCTCTTTGAAGTCTCAAAA
CAATAAAATTTAAATGTGCATAAATAAGATTGTGAATACGCAAAAATGCAAATAAATACG
CATATTATCATTCATTTCTTAATTATTTAGAAACGGTTCGAACTGCTTCTAATCGTGTCT
TAAGACAAACCCACATCTTATCTCAAAGTTTAGCATTTCGAAGCTTATTATTGTTTCTAA
AACATCTCAACATTTTACTTACAAACTTTGAATGAAAAAAAACAATTTTATAATTATATA
CTAACCATAAAAAAATAAATACCCAAAACGAATAAAATATATTATGTACCTATTTTAAAT
GCGTTTATACCCAAATTTTCGAACCACAATTTTTATATGAACATGTTAATTTTATTTCAC
GTCCCCATTTTATATATATAAAATTTTTTTCCTATTTCTTATTACCTTTAATATTATTTA
TACTAGATACAGAAATAATTTAAAAGGTACTATTTAAAAATGTGTTACCATTTTATTTAA
CTCACAAGCCCCTTTATTATATTAAATTTAAAAGCTTATTTTATGAGGGTTTATATAAAA
ATTCATTCGGTTTTAAATGTAAATATATATATGCACATAATTTTGTATAAATAAAACATA
TATATACATTATAATTATTTATTTGACAGTTTTGAAGGATTTATGAAAAGGAAAAATTTA
TCATGTGTATATATGTTTTTATATTTTATTGACAAGAAATTATAAAGCTCGGAAAATTAA
TAGAAAAGTAGGTGCAAGCATGCACATATAAATACATTAAACACCAAAATACTTTTCCAA
TGACTTGCCTACATGTCTTTGTACATATAATACATACATAGTATAATAAAATATGAAGCT
TTTAAATTATAATAAATTTTTATCTTCATCGGTATAAGGTTCAGATGCAGGGGCGTTATC
TTCCTCATTAGTATAAGAACCTTTTGTTTCATCATCATCATTATTATCTTCGGTACTTAA
TCTGTTTAAATATGATATCTTTATAAATGAAGCAATACCACATTTTTACACAAATATATT
ACATTTTTTATGTATTGCATATGTGAAATGTATGAGTGATATAATATGGTAGACATGAAT
AGGCTAAAACATAGATAAAACCGAACTAAACAAATAAATATAGGAAAATACCCCAATACG
AATATTTGTAACATTAAAAATTAAAAAGACACACTTTAATTAAATAAAATAGAGACGTGA
ATAAAAATGTATTTATAATAAAAGTACGTAAAAAGAAAGCGTTACCAAAACATAAATGAT
TTTTTTCTAACTAAATAAAATTGCATATGTGAAGGTATAATATTTTCCTACATAAAATGG
CGTTTATTGGTTTTTTAATATGTTTTACTTCATTTTGATTTATTACACATATATAAATTT
TTTACAAAATTACGCCATTAATATAGACATTTTTAAGAATGTTGTATTTATTGATCATAT
TCGAACTAAATTCCGTTGTAAATTCACTCAGGTGCATGCAAAAGATGCGGTATTTTTTTG
TACTATTAAAAATTTATATATACAATAATTGTTCAAAATACATAATTTTTTTTAAGCATT
TTACAAACACAAATAATTTTTTCAAAATAAAATAAATTGTTACGTATCTCGAAAATAGTG
TATTTAATATTATACAAATGTAAGGGCAATACATATATATATATTTATTTATTTAAAAGA
CAATTTATAAATGCATATTATTCATAAATTTTTATAATATTTCTCGATAATTTTGTTCGC
AATTTGATGGTGACCCCTTATACATTTGAATTTGCTTTAAAAATATTAAAAAGGAAAATT
TTTTTTTGTTATATAGAGTCATCATGCTTGATTATTTTATTTCTGAATTTAATTTATCCT
CATGTATTTATTATAACAATACGACGTGAAGTTAAAACACCAGTTGCATTGATCAATTTA
AAATTTTGATAAAAAGTATTTAAAAATAAAAGGTTTATAAATAGGGGCTTATTCATTTAT
ATTTCTTTTTATTATATTTTCTTTATATTTTTCAACAAATATTTATAATTCGTAATTTTT
TCGTCAATTTAATATATTTGCCTTTAAGTTTTTCGTAAATTACACTTCAGTAATAATAAT
AAAGCTACATTTTATAGTGCGCATTGTTAAGCATATAATCGATAGGACTTTTAATATATA
TTTTGTTTTTACACAAAATAAGAAGAAGGTCCCTACATTTTTTTCGTTAAAAAGGAAATC
TCAATAATATTAAATTGTGTGTAATTTTTTTTGATGTATACAAATATGACGTGTATATAC
GCAAAGGTGCTTGTAAAACAAATGGTATTAAGCTGTGTGGTCACATTTGTAGTTACATAG
CAATTATATATATATCATTTCTAATTTTAATTCAATTTAATTTCGGTATTAACTATGCAT
ATAAACATATTGTTATTATTATCTTGCCTATCATAATAAATTTTGTGTCTATATAGAAAT
GTATTATCTATTTTTTAGCGAAGTAGCATAAGTATTATTTATATTCAAAGAAATAAATAA
AGGTATTAAAATTAATACGAAAATAAAAATTAAAGGATGATGGCGATTATTAGTTCTTTT
TAATTCCATCATCTATCCTTATATGTAGTATATTGGGGTTAGAGATAGTGGGTTATGAAT
ACCAAAAATATAGAAAATTGCAAAGGAGTTGATTTATTCTCTGTTGTATAAGAATGTACA
CAGTTAAAGTTAGTATATATGTGTATATTTTTCAGTCTATATAAATATACTTTTGTTTTC
TCATTGGTATGTTCAATTTTTTATATTATTTTACGTGTCGATGGATACAATTTAGTATTT
TTTTTAATTTTGTATTTTTGAATTATATTAATCTAGATTATTCATATATTTTTTCATATG
CAGATAGAAAAAGGTTTCATACGTTAGTTTATCCTACAAATTGAATATTAACACATACAT
ATATTTATATTTTTTATTAATAGAAGGATCCAAAATGAGTAGATCTTCTAAGAACGTCAT
CAAGGAATTCATGAGATTCAAGGTTAAAATGGAAGGTACTGTTAACGGCCACGAATTCGA
AATCGAAGGTGAAGGTGAGGGTAGACCATATGAAGGTCACAACACAGTCAAGTTGAAGGT
TACTAAGGGTGGTCCACTGCCATTCGCTTGGGACATCTTGTCTCCACAATTCCAATACGG
TTCTAAGGTCTACGTCAAGCACCCAGCTGACATTCCAGACTACAAGAAGTTGTCCTTCCC
AGAAGGTTTCAAGTGGGAAAGGATCATGAACTTCGAAGACGGTGGCGTTGTTACTGTTAC
TCAAGACTCCTCCTTGCAAGACGGTTGTTTCATCTACAAGGTCAAGCTCATTGGTGTCAA
CTTCCCATCTGACGGTCCAGTCATGCAAAAGAAGACTATGGGTTGGGAAGCTTCTACCGA
ACGTTTGTACCCAAGAGACGGTGTCTTGAAGGGTGAAATCCACAAGGCCTTGAAGTTGAA
GGACGGTGGTCACTACTTGGTCGAATTCAAGTCTATCTACAAGGCCAAGAAGCAAGTCCA
ATTGCCAGGCTATTACTACGTTGACTCTAAGTTGGACATCATCTCTCACAACGAAGACTA
CACTATCGTCGAACAATACGAACGTACTGAAGGTAGACACCACTTGTTCTTGTAAACTAG
TTCTAGAAGTAGTGTGTAGCGTATTCTTTTATTTTACAGAAAATGTATATTATCATTTTT
CGCATTTCATCGTTTAAACAAGTACATGAACGTGTGCATGCAATTATTTCTGTGTTTGGA
TTTGTATTCAAATCATCATGAACTTTTTTTGAATATTCGTTACAAGCCATAATAGAATAT
ACACCATCTACAATATAATTTTTTTTTCACTATTTTATATTTCATCCTATTGTGTATGTA
TATTCCTTTAATAAAAAATCCATATGCTTAACATTCACATATATTAATAATTTTAATTAA
TTTTTTTTTTTGCACAGATAAAACTTTTCTTTTTAAAAAAAAGGTATATACACTCTTTTT
TACCCCACATTTAACTGATTTTCACTCACCATTACATATAGCTATATATTGCATATTACT
ATTATGTAAGGCAAATGTTTATTATTATTGCTGTTATTTTTTATTTATATATTTTTTTCC
ATATATCAATTTAAGGAGAAATAAAATAATTTGCTATTTGAAAAAATATAAAACAAATAT
CTAGTCGTGCAATACTGAATTCGAATACTTAAAACAAAAAATAAATTAATTAAATACCCT
TTTAATATTGTAGTTAAAATCGATGTGTTTAGAGAATATTTTTTATTGCTATTTATCTTC
TTTTTTTTATTTTTCTTCCATGTTTTCTCATTCAGCTTTCTATGTATTTTAGTAATATTC
TCCACACACAAACTATAAATTTTCTTATGAAAAATCACAAAATTTAGCAAATAGCGCTTA
GGCATGTACTCGTAAATAAGCATGTGCATAGTATAATACACTTCTCATCAATTTTGAGAG
GATTTTATTTAATTTCGGCACGCCCCTCTCAACTTGTGTTTATCACATATTCAATACTCC
ATAAAAAATGATAGCATCATTTCGATATGCGGGTACCGAGCTCGAATTCTCTTGAGCCCG
TTAATGAAATAGATACAATTCATTCATGTTATATACATCTAGAACATAATCTGAATATGG
TTCAAGTTAAATGTCCAAAAATTATAAAAAGTGATGATATTTTTGATGGTAATACCATAA
TAGACACCAAGGTAACATCACGAAGTAGTCAACAAAATAATTTTTATTTAGAAAATACAG
ATGTTGAACCAGAAGAAATAGAGAAATATAAAAATATAGAATACATACCAGAAAACGATG
AAGTAATGCATCTAGACAAAAAAGAAAAGCTAGATGATATATTACCAGGTGTTATCATAT
TAGATAAAAATAAAATGTTCAAAGAAAAAGGACATTTCACTTTTGTTACTCCATTAATTG
TAGAAAAGGTATTAATATTAAAAATATATTGTGATAATACTAAAACAATAATTAATAATA
TGAAAGGGAAAAAAGGTATTACAGTAATAAGGATTTCTCAAAATACAACAAAAAATAAAT
TTTATGGATGTGACTTTTCAGGTAATTCTAAAAAAACATTTTACTATTCCAATGTTTATG
ATTTAGAAAAAAAAAATGAGTTTTGTGAAATAGAATTAAAAGAAAATATAGTAGTTAGCT
TAAATTGTCCAACTGGTAAAATTAATCCAAAAAATTGTTTTAGAAATGTATATATAAAAA
GTAATATGAATGAACAAACAACCGAAAATATAGAAAATATATTTAACGAAATAAAAGTTA
TAGATGCAGATTATTTTATAAATAATTCATCAACCTTTTTGATGATTTCCAAAATTACAA
AAAAAGAGTTTGATTTTTATTGTACATGTGAAGATTATAAAACCAAAAATATAGGAACAA
TATATATTAAAAATTATGAATATCTAGATTCAAAACCTAAATATAAAAATAAACAAATTT
CCTATATAGATGTAGTTCCATACCCGCGGGGAAAGGGCG
Restriction sites to linearize plasmid KspI (SacII)
Selectable marker used to select the mutant parasitehdhfr/yfcu
Promoter of the selectable markereef1a
Selection (positive) procedurepyrimethamine
Selection (negative) procedure5-fluorocytosine (5-FC)
Additional remarks genetic modification
Additional remarks selection procedure
Other details transgene
Promoter
Gene Model of Parasite PBANKA_0416100
Gene Model P. falciparum ortholog PF3D7_0905300
Gene productdynein heavy chain, putative
Gene product: Alternative name
Primer information details of the primers used for amplification of the promoter sequence  Click to view information
Primer information details of the primers used for amplification of the promoter sequence  Click to hide information
Sequence Primer 1
Additional information primer 1
Sequence Primer 2
Additional information primer 2
3'-UTR
Gene Model of Parasite PBANKA_1010600
Gene productcalmodulin, putative
Gene product: Alternative namecam
Primer information details of the primers used for amplification the 3'-UTR sequences  Click to view information
Primer information details of the primers used for amplification the 3'-UTR sequences  Click to hide information
Sequence Primer 1tgcTCTAGAATTATTAATATATATGAATATATATACATCGTTGTATGCG
Additional information primer 12566, Asp718
Sequence Primer 2cggGGTACCGACCATATAAGAATTAACCC
Additional information primer 22576, XbaI
Insertion/Replacement locus
Replacement / InsertionReplacement locus
Gene Model of Parasite PBANKA_0306000
Gene product6-cysteine protein
Gene product: Alternative name230p
Primer information details of the primers used for amplification of the target sequences  Click to view information
Primer information details of the primers used for amplification of the target sequences  Click to hide information
Sequence Primer 15'-cccaagcttccgcgggtatatggtaaagaacctactaacac
Additional information primer 1primer L1345: 0.7kb 5'region of PB000214.00.0 (230p gene)
Sequence Primer 25'-cccaagcttgatgtgttttatttggatgtgc
Additional information primer 2primer L1346: 0.7kb 5'region of PB000214.00.0 (230p gene)
Sequence Primer 35'-ccggaattctcttgagcccgttaatg
Additional information primer 3primer L1347: 1kb 3'region of PB000214.00.0 (230p gene)
Sequence Primer 45'-tccccgcgggtatggaactacatctatatagg
Additional information primer 4primer L1348: 1kb 3'region of PB000214.00.0 (230p gene)