RMgmDB - Rodent Malaria genetically modified Parasites

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Summary

RMgm-698
Malaria parasiteP. berghei
Genotype
TaggedGene model (rodent): PBANKA_0216000; Gene model (P.falciparum): Not available; Gene product: PIR protein BIR protein
Name tag: mCherry
Transgene
 Transgene not Plasmodium: A fusion of GFP (gfp-mu3) and Luciferase Firefly (LucIAV)
Promoter: Gene model: PBANKA_0915000; Gene model (P.falciparum): PF3D7_1133400; Gene product: apical membrane antigen 1 (AMA-1)
3'UTR: Gene model: PBANKA_0719300; Gene product: bifunctional dihydrofolate reductase-thymidylate synthase, putative (dhfr/ts)
Replacement locus: Gene model: PBANKA_0306000; Gene product: 6-cysteine protein (230p)
Phenotype Asexual bloodstage;
Last modified: 2 December 2012, 15:29
  *RMgm-698
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Gene tagging, Introduction of a transgene
Reference (PubMed-PMID number) Reference 1 (PMID number) : 23197789
MR4 number
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Parent parasite used to introduce the genetic modification
Rodent Malaria ParasiteP. berghei
Parent strain/lineP. berghei ANKA
Name parent line/clone P. berghei ANKA 1037m1f1mocl1 (RMgm-32)
Other information parent lineP. berghei ANKA 1037m1f1mocl1 (1037cl1; RMgm-32) is a reference ANKA mutant line which expresses GFP-luciferase under control of a schizont-specific promoter. This reference line does not contain a drug-selectable marker (PubMed: PMID: 20019192).
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The mutant parasite was generated by
Name PI/ResearcherE.M. Pasini, C.J. Janse, B.M.D. Franke-Fayard
Name Group/DepartmentDepartment of Parasitology
Name InstituteBiomedical Primate Research Centre
CityRijswijk
CountryThe Netherlands
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Name of the mutant parasite
RMgm numberRMgm-698
Principal name1892
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationNo
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Phenotype
Asexual blood stageThe tagged-protein is exported into the cytoplasm of the host erythrocyte and shows a diffuse or patchy localization in the cytoplasm
Gametocyte/GameteNot tested
Fertilization and ookineteNot tested
OocystNot tested
SporozoiteNot tested
Liver stageNot tested
Additional remarks phenotype

Mutant/mutation

Protein (function)

Phenotype

Additional information
The mutant parasite expresses an mCherry-tagged protein. The protein has been selected for tagging in a screen for putative exported proteins of P. berghei.

The genotype of the parasites has not been analysed in detail.
Southern analysis of PFG-separated chromosomes (to show integration into the chromosome on which the target gene is located) has been performed (see below). This analysis provided evidence for tagging of the correct gene. The construct used aims at integration of the tagging construct by single-cross-over integration resulting in the presence of a tagged copy of the endogenous gene.

Phenotype analyses of the blood stages by fluorescence microscopy show that the tagged-protein is exported into the cytoplasm of the host erythrocyte and shows a diffuse or patchy localization in the cytoplasm.



Other mutants

  Tagged: Mutant parasite with a tagged gene
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Details of the target gene
Gene Model of Rodent Parasite PBANKA_0216000
Gene Model P. falciparum ortholog Not available
Gene productPIR protein BIR protein
Gene product: Alternative name
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Details of the genetic modification
Name of the tagmCherry
Details of taggingC-terminal
Additional remarks: tagging
Commercial source of tag-antibodiesClontech
Type of plasmid/constructPlasmid single 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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CTAAATTGTAAGCGTTAATATTTTGTTAAAATTCGCGTTAAATTTTTGTTAAATCAGCTC
ATTTTTTAACCAATAGGCCGAAATCGGCAAAATCCCTTATAAATCAAAAGAATAGACCGA
GATAGGGTTGAGTGTTGTTCCAGTTTGGAACAAGAGTCCACTATTAAAGAACGTGGACTC
CAACGTCAAAGGGCGAAAAACCGTCTATCAGGGCGATGGCCCACTACGTGAACCATCACC
CTAATCAAGTTTTTTGGGGTCGAGGTGCCGTAAAGCACTAAATCGGAACCCTAAAGGGAG
CCCCCGATTTAGAGCTTGACGGGGAAAGCCGGCGAACGTGGCGAGAAAGGAAGGGAAGAA
AGCGAAAGGAGCGGGCGCTAGGGCGCTGGCAAGTGTAGCGGTCACGCTGCGCGTAACCAC
CACACCCGCCGCGCTTAATGCGCCGCTACAGGGCGCGTCCCATTCGCCATTCAGGCTGCG
CAACTGTTGGGAAGGGCGATCGGTGCGGGCCTCTTCGCTATTACGCCAGCTGGCGAAAGG
GGGATGTGCTGCAAGGCGATTAAGTTGGGTAACGCCAGGGTTTTCCCAGTCACGACGTTG
TAAAACGACGGCCAGTGAGCGCGCGTAATACGACTCACTATAGGGCGAATTGGAGCTCCA
CCGCGGCATATTTTATTACAACACATTCCTTATGTGCAAAATAGCATTATGACAAATTGT
TTACAGATTAAAAATAAAATTCAGCACAATGACTCTCAAAGTGGTATACACATTTTTTAA
TAAAATGAATTTTATTTTATTGTTTTGCATTGTTTATAAATTAACTTAAATTTCAATAAA
ATTTCATTAATATATTTTTTATTAATTTTTATAATATTTTCGCAGTGTAAAGCATTTAAG
GACATTGAGGAATATTTGCCTGATAATTTTTCTTTCGAGAAGAATCATACCGATACTGAA
ATATACAATGCTTATTGTCCTATTAGCAAGGAAACAGGGAAAGGAAAATGTGGAACAATT
GGTCAAGTAGTTAGCGCTGTTACTACATTATTAATTAATAATTTAATCACCGGAGATGAA
TATATAGAATCTGAAAATGAAAATAACGAATATATCACGTATATTATTCTATGGTTAAGT
GATAAAATGAAATTAATTAAAACATGGGAATACGGAAGCGTATCAGATTTTGATACCACG
TTTATAAAAGATGGTGAATATTACAAAGAGCATATTGATCAAATCAAAAAAAAAGAAAAC
ATAATGAATGTTAAAATTGATGAAATGCATGAACTTTATGAGTTACTTAATGATCTGTGT
AATGTAATTACTAAATATACCAACGATTCTTCAAATTGCTCCGATTGTTCAAATTTTACT
AGTAATTGGGAAGAAAAATCCAATAAACTTGTTAATAAAAAAATTAAGGTTTTTGAAGAT
GAGTATTATTGTGATGTGTTGTTGACTTTAAAAAATGCTTATGAGAAATTTAAAAGAGAT
AATAATATTCAAAATAAACTTCCAAAAATTAAAGAGATAGAAGAAATAAATAACTGTAAG
AAATTATGTAAAGAAGCAACAAAATCATGGAGAATTAAAAGTGTGGATTTCCAACAAATT
AAGGAAAAAAAAAATATTGAGAAAAGCAAAGATAACTCGAGATATATAGATGTTGCTAAA
AATAAATTTGAAACGTATAGTTCATTTTTTAGTATTATATTTACTAATATTGGCAATAAC
TTATATGAAAAGGTGTTTCCAGCGCTAACAAATGTTTACGGTAAATTTATAAATTTTACT
GATAACACGATTAGTTATGTGAATGAGCAACTTAAAAAAGCAATAGAAGCTTATACATTC
AATAACGATGTACCTGAGGAAACAGACCCAGGAGGTGACCAATTATCATCTCAAGAAACT
CCATCTGAAGCTTCATCACCATCAAGTTCTACGAAACAAACTAGAACATCAGAATTGCCT
CAGAAATCATCTGAAAAAAAAAATTATGATCAAAAGAATGAAGAAGGTTCTAAAAAAACA
ATGCCAAGTTTAGTGATTAAACATGAAAATTCAGTAACCGAAGTAAAATTAAATGGAGCA
ACAGGAATAGATGACAATCCACTCAACTTATACAAGAAAATTGGAATTTCAATTATAATG
CTTTTAATACCCATTGCTTTAGCTATTATGTACAAGGTAAATAAAAATAAAACTGTGAAG
TATACAATTTTTAAAATATTTCTTCACTATAAATATTATATATTTTCCATAATTTTATGT
TAGTATTTTCCATTTGGATGGAGAAAGGAATTGAAGAAAAAAAAAAACATGAAAAAGGTT
ATAAATATGTTTGGTGTAAATGAAGCGACAAAAAGAGTTATAAACCCAACTAATCGAAAA
AACAAAGTGCAAATAATTATAAATTTATCTAAAAAAAAACAGGACAAAAAGTTTACAAAT
TCATCTACTCAAAAAAAACAGGACAAAAAGTTTACATATTCATCTACTCAAAAAAAGCAG
AATAAACAGTTTATAAATTCCATTTATTGGGAAAAATATCCATTATTAAATATATATAAA
CTTATGGAGGCCGATGCTGTACCATTTATTATTTTGTATTTGTTATTTATTTTTTATGTT
TATAAAAGAAAAGACGATTCTTTAGAAGGATCCGGTGGAGGTGGAAGATCCATGGTGAGC
AAGGGCGAGGAGGATAACATGGCCATCATCAAGGAGTTCATGCGCTTCAAGGTGCACATG
GAGGGCTCCGTGAACGGCCACGAGTTCGAGATCGAGGGCGAGGGCGAGGGCCGCCCCTAC
GAGGGCACCCAGACCGCCAAGCTGAAGGTGACCAAGGGTGGCCCCCTGCCCTTCGCCTGG
GACATCCTGTCCCCTCAGTTCATGTACGGCTCCAAGGCCTACGTGAAGCACCCCGCCGAC
ATCCCCGACTACTTGAAGCTGTCCTTCCCCGAGGGCTTCAAGTGGGAGCGCGTGATGAAC
TTCGAGGACGGCGGCGTGGTGACCGTGACCCAGGACTCCTCCCTGCAGGACGGCGAGTTC
ATCTACAAGGTGAAGCTGCGCGGCACCAACTTCCCCTCCGACGGCCCCGTAATGCAGAAG
AAGACCATGGGCTGGGAGGCCTCCTCCGAGCGGATGTACCCCGAGGACGGCGCCCTGAAG
GGCGAGATCAAGCAGAGGCTGAAGCTGAAGGACGGCGGCCACTACGACGCTGAGGTCAAG
ACCACCTACAAGGCCAAGAAGCCCGTGCAGCTGCCCGGCGCCTACAACGTCAACATCAAG
TTGGACATCACCTCCCACAACGAGGACTACACCATCGTGGAACAGTACGAACGCGCCGAG
GGCCGCCACTCCACCGGCGGCATGGACGAGCTGTACAAGTAATCTAGAATTATTAATATA
TATGAATATATATACATCGTTGTATGCGTAATATACGTGTAACTGTTTTTTTTAATTTTT
ATATTAACTTTTCCTTATACATTTTAATGCCATTAACTAATAATATTTTACTTGAAAAAA
TTCTAAATTTTTATAACTCTAATATAAAATCTGTGAATAATAAAAAATTAGCATAAAGTA
CGAATTAAACATCGAATATTTGTTTTAAAGGGGCACACATTATGTGCATATAATTTTATT
CTTTGAATTTAATTTTTTGCTTATAATTAGATTAAATTAATATAATTTTTTTATTGAAAA
TAAAAACTTATTTTAAAAAATTAACAAATGGAAGTTAATGAAAAATACGAAAAAAGTAAA
GGGTTAATTCTTATATGGTCGGTACCGAGCTCGAATTCGATATCGAAATTGAAGGAAAAA
ACATCATTTGTGTTTATATGTTTTTTTTTAATTTTTCAACTTTTTTTATTTAACTATGCT
AATCATATTATAAACATTTTAAAAATCAAGTCCAACTATTTATGAATCATTGAAGAGACA
ACATAATTATAACAGTAAAAATAATAATAATAATAATAATTTTGTCTTATTGTTTCAAAC
ATATTATGTTCCATAAATATCTAAATTTTTAATATGGTCATAAATTCATAGGATTTAATT
TATATAAAATATGGCTATTCATACTAGCCATTTTATGTGTGTAAAGTTTTTTTTTTCTGA
ACAAAAAATAAAATGTTCATAATTTTAGCTATTTACATGCATGTGCATGCACAAAAAAAA
ATATGCACACAACATACACATTTTTACAGTTATAAATACAATCAATTGGTATAAATATAT
AAGTAAGAAAAACGGGATCAAAGCTAGCTTCTGTATTTCCGCTAGACAGCCATCTCCATC
TGGATTCGTCCGTGCGGGACGTAGCCCACGACCTCAAAATCCTCGGCGGTGAAATCGTCG
ATTTCCTTGATGCGTTCCTTGTTGAGGATGTTCACAATGGGGAACGGTCTCGGTTCTCTC
CGCAGCTGCTCTTTTAAAGCCTCGACATGGTTCGTGTAGACATGCGTGTTCCCCATGAAG
TGAATGAACTCCTTAGGTTTTAGGTTGCAGACGTGTGCAACCATGAGCGTCAAAAGCGAA
TAGGAAGCGATGTTGAAGGGGACGCCGAGGCCGACATCGCACGACCGCTGATACATGATG
CACGACAGCTCCTTCTGGTCGTTCACGTAGAACTGGCACAACAAGTGACAAGGCGGCAGC
GCCATTTCGTCCAGCGCTGCAGGATTCCAGGCAGTCATGAGCATGCGACGATCTGTTGGA
TTCGTTCTCAGCATCTGGATCACATTCTTCAGCTGGTCGACGCCCTGCCCTGTGTAGTCT
GTGTGCATGTCTTTGTATGCCGCGCCGAAGTGTCTCCACTGGAAGCCGTAGCCCGGGCCG
ATGTCTCCGACCTCTCGGTGGGGGAGATTGCGCGAATCGAGGAACTCGCGTGTCACATTC
TTGTCCCAGATCTTCACGCCCTTCTCAGAAAGATGGTTTGCGTTCGTGTCGCCGCGAATG
AACCACAGCAACTCTTCGAGGACCCCTTTCCAGAACACACGCTTTGTGGTGAGAAGTGGA
AAGGCCTGATCCAGCGAGTAGCGCATAGTGCAGCCGAATTTGGAGATGACACCAACGCCC
GTTCGGTCATCCATTGTCCTTCCATTGTTAATAATGTCGGCAATGAGATCAAGGTACTGG
AACTCTTCATGGCCTCTAAAGTGAACATGCGGAACGGCCCGAATCAGTTCCTTTTGCTCG
CGTTTTTTCCGGTCTTCTTCGTCCATCCACGCCAACACCGGGGCAATGGCTGCGGCCGAA
GAAGGAGCCTGCAACCCGTGCACGGGAGTTGTCTCCCTCGTGGACGTCAAGGAGCTCATT
GCGTTGCTCGGTTCCGCAGTGGCTGCGTCGTCAGTCTTCCTTCTCTTCTCGAGAACCACA
AAGTCGTAGGGAACCCCGTTGTCTGAGAAGGTCTTGGAAATGAAGATGGGTCGATACGTC
GCTTCATTGTCCTTCTCTCTTCCGAGCTCCGGACAAAAGGGAACGAACACAGACTCGGCA
GGAGCTGCAGCCTGCGCAGCAGTTGATTTGTTTGAAAGAATGTCATCTCCGGGGAACGCA
GGGAAGAAAACGTCGCACGGAAACTCGCGGGCTACACGCGTGATGTACAGGTGAGAGGCA
ACGCCCAGAGACAGCGCTGCCTCGTACAGTCCCGCTCCTCCCACGACAAAAATCTGGTCG
ACAGAATCCTTGTACTCTTCCTCCAGAAGGCTGAGAGCTGCTGGGAGTGAAGCACAGACT
CGGACGCGCTGCTGGCCTTCAGCTTGAGGCTTCTCCGCCGCAATGTCTTCTTCTTTGAGG
GAAGAGGAAACGACGATGTTCAATCTGTCCACGAGGGGTCTAAACTTTCGAGGCATGCTT
TCCCAGGTTTTCCGTCCCATGACAACGGCGTTGAATCTCTTGCCGACTGATGGAGAGGGA
AGTCCAGAGTCGCCCGTCTTTGCAAATTTCCTGGGAAGCCACCCGTTCAGGCGACTGGCT
TCTTCGGGCGTCGTTTTTGTCACACGAGAAAAGTGTTTGAAATCTGTGGTCAAGTGGGGC
CACGGGAGGCCGTTGTTGATGCCGATGCCCCTCTTGGGGGTCATCGCGACGACCAGACAC
ACCGGTTTATGCATTGATCCTTTGTAACATTTAGGTGTGTATTTATATATATAAGCAATA
TACAAATGAAAATACTATTATAAAACGGAAACTCAAAAAATGTTGAAACAAATAGTCAAT
TTTTAAAATGAATGTGTAAATTGTTTAAGTTTTATGACAAAAAAAGGAAAAGTGTATATA
AAATAAAAATTGGAAAAAATTTATATATTACATGCCATATAATACCATTTATTTACTATT
TATATCTTAATATAAAAAATTGTAGCAAAAATATATAATCGTATTTTATATATTTAATAT
TATATAACCCCTTAAAAAAAAAAATTTAATTGAAAAAATTTTAATTTATGCACTTTTTTA
TTGTATTATATATATTCCGTTATTTTAATTACACATAAACTTAATAGAAAAATTAATTCG
CTGCTAAATAAAAATTATAGGGCTATATATGATGGGATCATATTCTATCTTTCTCTTTTT
CAAATYCTTTATATTTTAGAATACAAATCGCTTTAAACAATAAAAAAGAGGTATATCCTA
CCCGAATTGACACACACAATAAATAATAAATATATAAATAATAAAAATATAAATATATAA
ATTATAAATATATAAATTATATATATAAATATATTAACATGTGTGTAATTTGTTAAAATT
GATATTAAACAGCTTTGCATGGGGAAAGAGTGGCTTTTATAAGCGAATCATATAATACAG
TGTTAATTTTAATATATAAAATATAATTTCCTTAAATTGTTATTTAACATTTTTAATTGC
CTACCCARGGTAATATAATATCCAAATGACAATCAAAGATAAATGAAATACCGCTCCATT
TTTCCTATATATCCATTTTCATTTTGTGTAAATGTTCATAAAATTTTATCATATACTCAT
AAAAAACAGGGGAAAAATGAAAAAATGAACGTCCACCCCGTGTGAATATGCTCATTTTGT
ATACTATACATTTTTTTTTATATAATTTATATTTTCCAAATAAAAAAAATGTTTGTACAT
GTATATATCAATATTGTGTTATGGCATGATGAAAAACAATTATTTTATTCGTTTTTAAAT
TTGTAGAATTTCCCTAAATTATTAATGATTTCATATTATAATATATGTATCTATATATTT
TTAATGTGTAATTCAAAATGGCGAAAAAGAAACAAACCATACTTTATAAGCAACTCTCTT
TGAGCACTTACCATAAAAAATTATACTACTAAATTCAAATTAAATAAAAAAATATATATT
ATAAAATTGGCTTTAATTATTTATAAGCACTTGCTTTGTTTTACTCAAAAAGTATTGGCA
TAAGTATAAACATAAAACCAATTTTTCACAAAATATATTATATGTCCATACAACTATATC
CGAACYTTCAAAACACATCCAAATTTCCATTTACCACATGCAAGCAAAAGGATATTAAAT
AAAAAAAAATTATTTATATTTTCATGATCACTTAAATAATATTTAAAAAAAAAAGAGACC
ATATATAAAATGTATAATTTAAATATAGCTAACTTATTAAAAAAAATGAATATTGAGCAT
ATATCCATAATTTAAAACACTTCTATTTTATGTTATTTGATATATAAGTCGATAAATTAT
TGCATGTATGCATATAGACAGATTAATATACACCAAAATAATAAAAATTATATATAGTTC
ATATATTATTCTGAATAATAAATTAGCTTTCCTCTTAATTCACTTGGTATGTATATATAT
ATATATATATATATATATATATATATGGATAAATAAATATAAGGTCTATAACAGGAAAAA
ATGTGTATTAACTTTTTTTTATAATTATTTTATATTTTTTCTCCATTTATTTAAATATGT
TAAAATTATTTCCCATTTTATAAACAAAAATTAATCCATAAAATAGCAAAATAACAAAAT
GACAAAATAACAAAAACAAAAAAACGCATTATATGAGTTCATTTTACACAATCCAAAGAA
AAAGAAAATATGCTTAAAATATTCATAACACACTTTTAAGCTATAAAAATATATATATTT
ATTTGTGTCTATATTACCAACTCAATTTAATAGATGTGTTATGTGATTAATTCATACACA
AACATACAAAAATAAACACCATTTTTGAAAAGATTAATTTGAAATCATATATGTTATTAT
TATATATTTTTATATCAATGAAATTTAAAAAAATAAAAGGGAAATCAATGTATTAAAAAT
AATTATATGTTATTTTATTTCCACAATATTTATTTATTATTTATTGTTGACCTGCAGGCA
TGCAAGCTTATCGATACCGTCGACCTCGAGGGGGGGCCCGGTACCCAGCTTTTGTTCCCT
TTAGTGAGGGTTAATTGCGCGCTTGGCGTAATCATGGTCATAGCTGTTTCCTGTGTGAAA
TTGTTATCCGCTCACAATTCCACACAACATACGAGCCGGAAGCATAAAGTGTAAAGCCTG
GGGTGCCTAATGAGTGAGCTAACTCACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCA
GTCGGGAAACCTGTCGTGCCAGCTGCATTAATGAATCGGCCAACGCGCGGGGAGAGGCGG
TTTGCGTATTGGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCG
GCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGG
GGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAA
GGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCG
ACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCC
TGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGC
CTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTC
GGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCG
CTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCC
ACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGA
GTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGGACAGTATTTGGTATCTGCGC
TCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAAC
CACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGG
ATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTC
ACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAA
TTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTA
CCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTATTTCGTTCATCCATAGT
TGCCTGACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCCCAG
TGCTGCAATGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGCAATAAACCA
GCCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTC
TATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGT
TGTTGCCATTGCTACAGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAG
CTCCGGTTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGTGCAAAAAAGCGGT
TAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCAT
GGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGT
GACTGGTGAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTC
TTGCCCGGCGTCAATACGGGATAATACCGCGCCACATAGCAGAACTTTAAAAGTGCTCAT
CATTGGAAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGATCCAG
TTCGATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCGT
TTCTGGGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACG
GAAATGTTGAATACTCATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTA
TTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATAAACAAATAGGGGTTCC
GCGCACATTTCCCCGAAAAGTGCCAC
Restriction sites to linearize plasmid SpeI
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 1CCCCGCGGCATATTTTATTACAACACATTCC
Additional information primer 1L6333 (SacII); PBANKA_021600 Forward
Sequence Primer 2CGGATCCTTCTAAAGAATCGTCTTTTCTTTTATAAAC
Additional information primer 2L6191 (BamHI); PBANKA_021600 Reverse
Sequence Primer 3
Additional information primer 3
Sequence Primer 4
Additional information primer 4
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6
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  Transgene: Mutant parasite expressing a transgene
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Type and details of transgene
Is the transgene Plasmodium derived Transgene: not Plasmodium
Transgene nameA fusion of GFP (gfp-mu3) and Luciferase Firefly (LucIAV)
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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
GGAATTCGATATCGAATTCCCATGGAACCAAAATAACATATTAATGAATTTTCTTTTATT
GTGAAAATGTTCTTATTTGTTTTAAAAAATCAAAGTAAATTTAATTCCTTTAATTGTGCT
AAATATTGTGTTTGTATAACTATAATGACATTTCCTATGCTTTTTTAATATTATATATAA
TTTTTTGTGTTTTTTCAAAAAAAACGTGTATTTGTTTTTATTTGAGTGTACAATTTGCAT
AGTGAGTTTAAAGTGTTTGAAAATTATTTAATTTATAATTTTCATAAAACGTTTAATCAT
TTATCAAAATTGCGGTTGCAATATAGACAGAAAATATTAACTTAATTTTCATTGTTTTGT
AAAACATTTTCATTATTTTTTTTAACAAAATTAAAGTAAATGAGATGGTAAATTATTTTA
ATTATAACCATTTGTTTAATTTTTTTTATTTTATTACTATTATTTTTTTTCTTTTGTGCT
GCGCAATTAAAAAGAGAATTAAGGGTGCACCAATATATTTAAGCAATCTTTTGCGACACA
CAAGAAAAAATATAACATCAACATATTTAAAATAATGGAAATTTTTTAATAATTTATTCT
ATATATATTTAATATATGTAAAAAAACTAATATTGTAATATAATTAAATAAAACTTGAAA
TTGATATAATATGTATATATATTGCATGTTTAGCTTTAAATAATATTTAAGTATTGATTT
TCTTTACACACAAAAACATTTGCCTTTTTATGTGAGGATTATTGTTTCTTTATTTATATT
GCTATGAATTTATTCTTAGTATAAATTTATTTTTTTTCGGCTTTGAAAATAATTTTATAT
ATATAAAAAGAAAAAATATGGAAAATAAAATTCATAAATCATATTTGACACATTTCAATT
TTTTTTTCTTATTTCACAGCTAGCCATATAATAATATATATGTGTCTATGGGTTCAAATA
GCTTAATTTCAATGTGCGTGATTTTTTTTTATAAAAAAACTAATTTTATGTTTGGTTATG
CATGCAACTGTTATAAAATAAAAATTGTAAAATTTGTTTTTTTGTACACATATTTGTACT
TTTTTTAATGCACATGAAAAAAGAAAGGAAAAAAAAGGGGGAAAAAAAAGGAAAAAAAAA
GGGAAAAAAGGGAAAAAACGTACATCTACGCATTGTTATTTAGCAATAAAATAGAGAACA
AATCTATAAATATATAATGTGTACAACAAATTAATTTAATTTATATTTTTACTGTAATTT
CATAATAATTTTCATTTTTACATTTTAATGTTTTTTTAATTGGAATATAATTCAAAATGA
TTTTGACACATTTTACTTATAGAAGAAGGCTATATAATTATATAAGTGTGTATATATAAG
TATTGTATGGTAATTGTTTTATAAATTAAATTTTTAAAAAACATTATTCTATTTTATAAT
TTGTAAATTAAAAATATTAATAAAATAAAACGATATAAAAGGATCCATGAGTAAAGGAGA
AGAACTTTTCACTGGAGTTGTCCCAATTCTTGTTGAATTAGATGGTGATGTTAATGGGCA
CAAATTTTCTGTCAGTGGAGAGGGTGAAGGTGATGCAACATACGGAAAACTTACCCTTAA
ATTTATTTGCACTACTGGAAAACTACCTGTTCCATGGCCAACACTTGTCACTACTTTCGG
TTATGGTGTTCAATGCTTTGCGAGATACCCAGATCATATGAAACAGCATGACTTTTTCAA
GAGTGCCATGCCCGAAGGTTATGTACAGGAAAGAACTATATTTTTCAAAGATGACGGGAA
CTACAAGACACGTGCTGAAGTCAAGTTTGAAGGTGATACCCTTGTTAATAGAATCGAGTT
AAAAGGTATTGATTTTAAAGAAGATGGAAACATTCTTGGACACAAATTGGAATACAACTA
TAACTCACACAATGTATACATCATGGCAGACAAACAAAAGAATGGAATCAAAGTTAACTT
CAAAATTAGACACAACATTGAAGATGGAAGCGTTCAACTAGCAGACCATTATCAACAAAA
TACTCCAATTGGCGATGGCCCTGTCCTTTTACCAGACAACCATTACCTGTCCACACAATC
TGCCCTTTCGAAAGATCCCAACGAAAAGAGAGACCACATGGTCCTTCTTGAGTTTGTAAC
AGCTGCTGGGATTACACATGGCATGGATGAACTATACAAAGGGATCCTGGCTAGCCAGTC
GACCTGCAGGCATGCAAGCTTGCGGCCGATCCAAATGGAAGACGCCAAAAACATAAAGAA
AGGCCCGGCGCCATTCTATCCGCTGGAAGATGGAACCGCTGGAGAGCAACTGCATAAGGC
TATGAAGAGATACGCCCTGGTTCCTGGAACAATTGCTTTTACAGATGCACATATCGAGGT
GAACATCACGTACGCGGAATACTTCGAAATGTCCGTTCGGTTGGCAGAAGCTATGAAACG
ATATGGGCTGAATACAAATCACAGAATCGTCGTATGCAGTGAAAACTCTCTTCAATTCTT
TATGCCGGTGTTGGGCGCGTTATTTATCGGAGTTGCAGTTGCGCCCGCGAACGACATTTA
TAATGAACGTGAATTGCTCAACAGTATGAACATTTCGCAGCCTACCGTAGTGTTTGTTTC
CAAAAAGGGGTTGCAAAAAATTTTGAACGTGCAAAAAAAATTACCAATAATCCAGAAAAT
TATTATCATGGATTCTAAAACGGATTACCAGGGATTTCAGTCGATGTACACGTTCGTCAC
ATCTCATCTACCTCCCGGTTTTAATGAATACGATTTTGTACCAGAGTCCTTTGATCGTGA
CAAAACAATTGCACTGATAATGAATTCCTCTGGATCTACTGGGTTACCTAAGGGTGTGGC
CCTTCCGCATAGAACTGCCTGCGTCAGATTCTCGCATGCCAGAGATCCTATTTTTGGCAA
TCAAATCATTCCGGATACTGCGATTTTAAGTGTTGTTCCATTCCATCACGGTTTTGGAAT
GTTTACTACACTCGGATATTTGATATGTGGATTTCGAGTCGTCTTAATGTATAGATTTGA
AGAAGAGCTGTTTTTACGATCCCTTCAGGATTACAAAATTCAAAGTGCGTTGCTAGTACC
AACCCTATTTTCATTCTTCGCCAAAAGCACTCTGATTGACAAATACGATTTATCTAATTT
ACACGAAATTGCTTCTGGGGGCGCACCTCTTTCGAAAGAAGTCGGGGAAGCGGTTGCAAA
ACGCTTCCATCTTCCAGGGATACGACAAGGATATGGGCTCACTGAGACTACATCAGCTAT
TCTGATTACACCCGAGGGGGATGATAAACCGGGCGCGGTCGGTAAAGTTGTTCCATTTTT
TGAAGCGAAGGTTGTGGATCTGGATACCGGGAAAACGCTGGGCGTTAATCAGAGAGGCGA
ATTATGTGTCAGAGGACCTATGATTATGTCCGGTTATGTAAACAATCCGGAAGCGACCAA
CGCCTTGATTGACAAGGATGGATGGCTACATTCTGGAGACATAGCTTACTGGGACGAAGA
CGAACACTTCTTCATAGTTGACCGCTTGAAGTCTTTAATTAAATACAAAGGATATCAGGT
GGCCCCCGCTGAATTGGAATCGATATTGTTACAACACCCCAACATCTTCGACGCGGGCGT
GGCAGGTCTTCCCGACGATGACGCCGGTGAACTTCCCGCCGCCGTTGTTGTTTTGGAGCA
CGGAAAGACGATGACGGAAAAAGAGATCGTGGATTACGTCGCCAGTCAAGTAACAACCGC
GAAAAAGTTGCGCGGAGGAGTTGTGTTTGTGGACGAAGTACCGAAAGGTCTTACCGGAAA
ACTCGACGCAAGAAAAATCAGAGAGATCCTCATAAAGGCCAAGAAGGGCGGAAAGATCGC
CGTGTAATTCTAGAAGATCCCGTTTTTCTTACTTATATATTTATACCAATTGATTGTATT
TATAACTGTAAAAATGTGTATGTTGTGTGCATATTTTTTTTTGTGCATGCACATGCATGT
AAATAGCTAAAATTATGAACATTTTATTTTTTGTTCAGAAAAAAAAAACTTTACACACAT
AAAATGGCTAGTATGAATAGCCATATTTTATATAAATTAAATCCTATGAATTTATGACCA
TATTAAAAATTTAGATATTTATGGAACATAATATGTTTGAAACAATAAGACAAAATTATT
ATTATTATTATTATTTTTACTGTTATAATTATGTTGTCTCTTCAATGATTCATAAATAGT
TGGACTTGATTTTTAAAATGTTTATAATATGATTAGCATAGTTAAATAAAAAAAGTTGAA
AAATTAAAAAAAAACATATAAACACAAATGATGTTTTTTCCTTCAATTTCGGGTACCGAG
CTCGAATTCTCTTGAGCCCGTTAATGAAATAGATACAATTCATTCATGTTATATACATCT
AGAACATAATCTGAATATGGTTCAAGTTAAATGTCCAAAAATTATAAAAAGTGATGATAT
TTTTGATGGTAATACCATAATAGACACCAAGGTAACATCACGAAGTAGTCAACAAAATAA
TTTTTATTTAGAAAATACAGATGTTGAACCAGAAGAAATAGAGAAATATAAAAATATAGA
ATACATACCAGAAAACGATGAAGTAATGCATCTAGACAAAAAAGAAAAGCTAGATGATAT
ATTACCAGGTGTTATCATATTAGATAAAAATAAAATGTTCAAAGAAAAAGGACATTTCAC
TTTTGTTACTCCATTAATTGTAGAAAAGGTATTAATATTAAAAATATATTGTGATAATAC
TAAAACAATAATTAATAATATGAAAGGGAAAAAAGGTATTACAGTAATAAGGATTTCTCA
AAATACAACAAAAAATAAATTTTATGGATGTGACTTTTCAGGTAATTCTAAAAAAACATT
TTACTATTCCAATGTTTATGATTTAGAAAAAAAAAATGAGTTTTGTGAAATAGAATTAAA
AGAAAATATAGTAGTTAGCTTAAATTGTCCAACTGGTAAAATTAATCCAAAAAATTGTTT
TAGAAATGTATATATAAAAAGTAATATGAATGAACAAACAACCGAAAATATAGAAAATAT
ATTTAACGAAATAAAAGTTATAGATGCAGATTATTTTATAAATAATTCATCAACCTTTTT
GATGATTTCCAAAATTACAAAAAAAGAGTTTGATTTTTATTGTACATGTGAAGATTATAA
AACCAAAAATATAGGAACAATATATATTAAAAATTATGAATATCTAGATTCAAAACCTAA
ATATAAAAATAAACAAATTTCCTATATAGATGTAGTTCCATACCCGCGGGGAAAGGGCG
Restriction sites to linearize plasmid KspI (SacII)
Selectable marker used to select the mutant parasitegfp (FACS)
Promoter of the selectable markerama-1
Selection (positive) procedureFACS (flowsorting)
Selection (negative) procedureNo
Additional remarks genetic modificationThe GFP-Luciferase gene (1 copy) has been inserted into the 230p locus (PB000214.00.0) by double cross-over integration
Additional remarks selection procedureThis reporter mutant expressing GFP-Luciferase does not contain a drug-selectable marker. This mutant has been selected by FACS sorting after transfection based on GFP fluorescence
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Other details transgene
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Promoter
Gene Model of Parasite PBANKA_0915000
Gene Model P. falciparum ortholog PF3D7_1133400
Gene productapical membrane antigen 1
Gene product: Alternative nameAMA-1
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
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3'-UTR
Gene Model of Parasite PBANKA_0719300
Gene productbifunctional dihydrofolate reductase-thymidylate synthase, putative
Gene product: Alternative namedhfr/ts
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 1
Additional information primer 1
Sequence Primer 2
Additional information primer 2
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 1
Additional information primer 1
Sequence Primer 2
Additional information primer 2
Sequence Primer 3
Additional information primer 3
Sequence Primer 4
Additional information primer 4
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Conceptual design of the database: Dr. C.J. Janse (Leiden Malaria Research Group, Leiden, The Netherlands).
Technical design and realization: S. Mededovic and A. van Wigcheren