RMgmDB - Rodent Malaria genetically modified Parasites

Summary

RMgm-694
Malaria parasiteP. berghei
Genotype
TaggedGene model (rodent): PBANKA_1246161; Gene model (P.falciparum): Not available; Gene product: fam-a protein (Pb-fam-1.3)
Name tag: mCherry
Transgene
 Transgene not Plasmodium: A fusion of GFP (gfp-mu3) and Luciferase Firefly (LucIAV)
Promoter: Gene model: PBANKA_1133300; Gene model (P.falciparum): PF3D7_1357100; Gene product: elongation factor 1-alpha (eef1a)
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: 27 July 2015, 18:24
  *RMgm-694
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 676m1cl1 (RMgm-29)
Other information parent line676m1cl1 (RMgm-29) is a reference ANKA mutant line which expresses GFP-luciferase under control of a constitutive promoter. This reference line does not contain a drug-selectable marker (PubMed: PMID: 16242190).
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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-694
Principal name1702
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_1246161
Gene Model P. falciparum ortholog Not available
Gene productfam-a protein
Gene product: Alternative namePb-fam-1.3
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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/construct(Linear) plasmid 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
CCGCGGTGGCGGCCGCTCTAGAACTAGTAACATCAATCCGACCCAGATATTAAAAAAATG
CATTATATAGCTCGTAAATGCTATGAGGTATCAATTAACGAACGATATTAAAGTTATAAA
TCAAATTAAAATAAAAAAATTATTATAATTTATATACATATATTTCTCTTTGTTTCCATT
AGTATAATGAAATAATAAACATGTCATGGGACCCCGATTATGCAAATTTTTTAAATACTG
GCTCTTTTAAAAGTATACAAAACATAAATAATTAGTCAAATTAATATATTATTGTTACTA
TTTTTAGAAATTTCATGATTTATTGTTGACATTGTTATATGATACTATATATTGTTTCCC
ATATCTTCAAATTCATAATATTTTAATTTACATAATGCAAAATTATTATTATTTTTAAAA
GTTGTCCGTGTGTACAATCCAAATTTAGTAATGGTACAACAACGTTACGAAGATAGTTCT
ATGAACTGTCAGAAATATTTTTATGCTTTAGCTGCATATGTTGAAGTAAGGAAAATTTAT
TTTTCGTCCCCCCCCAAATCATATTTTTCATATAATTCACAATATTTTATGCAATATATA
TTATTAATTTTGTAGATATCAAAAGACAAAACTATAATTGTCAGGGTTTCAGCAGATATA
AATGATCACAACCCTTCCAATGAAAAATATAAAAACGAAATAGTAAAAAGTGCAAGTTTA
TTCAAAACTGAGATTGATTCTGAAGATGATATTAGAAAAGGAGAATTAAAAAAAATGTTT
GTTAACATAGCTGGATACCTCATTGAAAATAAAGGCGATTGTGTTAATATTACCTACGTT
GAATCTGTAATCAATATCCACATTTTAATAACATAATAATTTATTTCAACATTTGTTAAA
AAAATATTGTTTTAAACAAATGTGTAGATATTTATAATTTGCAAAATATTTAAACATTTT
ATATATTCATCCATTTATGTTTTTTTTTCTTAGATTAATAGACATTGTACCATTGGATCC
GGTGGAGGTGGAAGATCCATGGTGAGCAAGGGCGAGGAGGATAACATGGCCATCATCAAG
GAGTTCATGCGCTTCAAGGTGCACATGGAGGGCTCCGTGAACGGCCACGAGTTCGAGATC
GAGGGCGAGGGCGAGGGCCGCCCCTACGAGGGCACCCAGACCGCCAAGCTGAAGGTGACC
AAGGGTGGCCCCCTGCCCTTCGCCTGGGACATCCTGTCCCCTCAGTTCATGTACGGCTCC
AAGGCCTACGTGAAGCACCCCGCCGACATCCCCGACTACTTGAAGCTGTCCTTCCCCGAG
GGCTTCAAGTGGGAGCGCGTGATGAACTTCGAGGACGGCGGCGTGGTGACCGTGACCCAG
GACTCCTCCCTGCAGGACGGCGAGTTCATCTACAAGGTGAAGCTGCGCGGCACCAACTTC
CCCTCCGACGGCCCCGTAATGCAGAAGAAGACCATGGGCTGGGAGGCCTCCTCCGAGCGG
ATGTACCCCGAGGACGGCGCCCTGAAGGGCGAGATCAAGCAGAGGCTGAAGCTGAAGGAC
GGCGGCCACTACGACGCTGAGGTCAAGACCACCTACAAGGCCAAGAAGCCCGTGCAGCTG
CCCGGCGCCTACAACGTCAACATCAAGTTGGACATCACCTCCCACAACGAGGACTACACC
ATCGTGGAACAGTACGAACGCGCCGAGGGCCGCCACTCCACCGGCGGCATGGACGAGCTG
TACAAGTAATCTAGAATTATTAATATATATGAATATATATACATCGTTGTATGCGTAATA
TACGTGTAACTGTTTTTTTTAATTTTTATATTAACTTTTCCTTATACATTTTAATGCCAT
TAACTAATAATATTTTACTTGAAAAAATTCTAAATTTTTATAACTCTAATATAAAATCTG
TGAATAATAAAAAATTAGCATAAAGTACGAATTAAACATCGAATATTTGTTTTAAAGGGG
CACACATTATGTGCATATAATTTTATTCTTTGAATTTAATTTTTTGCTTATAATTAGATT
AAATTAATATAATTTTTTTATTGAAAATAAAAACTTATTTTAAAAAATTAACAAATGGAA
GTTAATGAAAAATACGAAAAAAGTAAAGGGTTAATTCTTATATGGTCGGTACCGAGCTCG
AATTCGATATCGAAATTGAAGGAAAAAACATCATTTGTGTTTATATGTTTTTTTTTAATT
TTTCAACTTTTTTTATTTAACTATGCTAATCATATTATAAACATTTTAAAAATCAAGTCC
AACTATTTATGAATCATTGAAGAGACAACATAATTATAACAGTAAAAATAATAATAATAA
TAATAATTTTGTCTTATTGTTTCAAACATATTATGTTCCATAAATATCTAAATTTTTAAT
ATGGTCATAAATTCATAGGATTTAATTTATATAAAATATGGCTATTCATACTAGCCATTT
TATGTGTGTAAAGTTTTTTTTTTCTGAACAAAAAATAAAATGTTCATAATTTTAGCTATT
TACATGCATGTGCATGCACAAAAAAAAATATGCACACAACATACACATTTTTACAGTTAT
AAATACAATCAATTGGTATAAATATATAAGTAAGAAAAACGGGATCAAAGCTAGCTTCTG
TATTTCCGCTAGACAGCCATCTCCATCTGGATTCGTCCGTGCGGGACGTAGCCCACGACC
TCAAAATCCTCGGCGGTGAAATCGTCGATTTCCTTGATGCGTTCCTTGTTGAGGATGTTC
ACAATGGGGAACGGTCTCGGTTCTCTCCGCAGCTGCTCTTTTAAAGCCTCGACATGGTTC
GTGTAGACATGCGTGTTCCCCATGAAGTGAATGAACTCCTTAGGTTTTAGGTTGCAGACG
TGTGCAACCATGAGCGTCAAAAGCGAATAGGAAGCGATGTTGAAGGGGACGCCGAGGCCG
ACATCGCACGACCGCTGATACATGATGCACGACAGCTCCTTCTGGTCGTTCACGTAGAAC
TGGCACAACAAGTGACAAGGCGGCAGCGCCATTTCGTCCAGCGCTGCAGGATTCCAGGCA
GTCATGAGCATGCGACGATCTGTTGGATTCGTTCTCAGCATCTGGATCACATTCTTCAGC
TGGTCGACGCCCTGCCCTGTGTAGTCTGTGTGCATGTCTTTGTATGCCGCGCCGAAGTGT
CTCCACTGGAAGCCGTAGCCCGGGCCGATGTCTCCGACCTCTCGGTGGGGGAGATTGCGC
GAATCGAGGAACTCGCGTGTCACATTCTTGTCCCAGATCTTCACGCCCTTCTCAGAAAGA
TGGTTTGCGTTCGTGTCGCCGCGAATGAACCACAGCAACTCTTCGAGGACCCCTTTCCAG
AACACACGCTTTGTGGTGAGAAGTGGAAAGGCCTGATCCAGCGAGTAGCGCATAGTGCAG
CCGAATTTGGAGATGACACCAACGCCCGTTCGGTCATCCATTGTCCTTCCATTGTTAATA
ATGTCGGCAATGAGATCAAGGTACTGGAACTCTTCATGGCCTCTAAAGTGAACATGCGGA
ACGGCCCGAATCAGTTCCTTTTGCTCGCGTTTTTTCCGGTCTTCTTCGTCCATCCACGCC
AACACCGGGGCAATGGCTGCGGCCGAAGAAGGAGCCTGCAACCCGTGCACGGGAGTTGTC
TCCCTCGTGGACGTCAAGGAGCTCATTGCGTTGCTCGGTTCCGCAGTGGCTGCGTCGTCA
GTCTTCCTTCTCTTCTCGAGAACCACAAAGTCGTAGGGAACCCCGTTGTCTGAGAAGGTC
TTGGAAATGAAGATGGGTCGATACGTCGCTTCATTGTCCTTCTCTCTTCCGAGCTCCGGA
CAAAAGGGAACGAACACAGACTCGGCAGGAGCTGCAGCCTGCGCAGCAGTTGATTTGTTT
GAAAGAATGTCATCTCCGGGGAACGCAGGGAAGAAAACGTCGCACGGAAACTCGCGGGCT
ACACGCGTGATGTACAGGTGAGAGGCAACGCCCAGAGACAGCGCTGCCTCGTACAGTCCC
GCTCCTCCCACGACAAAAATCTGGTCGACAGAATCCTTGTACTCTTCCTCCAGAAGGCTG
AGAGCTGCTGGGAGTGAAGCACAGACTCGGACGCGCTGCTGGCCTTCAGCTTGAGGCTTC
TCCGCCGCAATGTCTTCTTCTTTGAGGGAAGAGGAAACGACGATGTTCAATCTGTCCACG
AGGGGTCTAAACTTTCGAGGCATGCTTTCCCAGGTTTTCCGTCCCATGACAACGGCGTTG
AATCTCTTGCCGACTGATGGAGAGGGAAGTCCAGAGTCGCCCGTCTTTGCAAATTTCCTG
GGAAGCCACCCGTTCAGGCGACTGGCTTCTTCGGGCGTCGTTTTTGTCACACGAGAAAAG
TGTTTGAAATCTGTGGTCAAGTGGGGCCACGGGAGGCCGTTGTTGATGCCGATGCCCCTC
TTGGGGGTCATCGCGACGACCAGACACACCGGTTTATGCATTGATCCTTTGTAACATTTA
GGTGTGTATTTATATATATAAGCAATATACAAATGAAAATACTATTATAAAACGGAAACT
CAAAAAATGTTGAAACAAATAGTCAATTTTTAAAATGAATGTGTAAATTGTTTAAGTTTT
ATGACAAAAAAAGGAAAAGTGTATATAAAATAAAAATTGGAAAAAATTTATATATTACAT
GCCATATAATACCATTTATTTACTATTTATATCTTAATATAAAAAATTGTAGCAAAAATA
TATAATCGTATTTTATATATTTAATATTATATAACCCCTTAAAAAAAAAAATTTAATTGA
AAAAATTTTAATTTATGCACTTTTTTATTGTATTATATATATTCCGTTATTTTAATTACA
CATAAACTTAATAGAAAAATTAATTCGCTGCTAAATAAAAATTATAGGGCTATATATGAT
GGGATCATATTCTATCTTTCTCTTTTTCAAATYCTTTATATTTTAGAATACAAATCGCTT
TAAACAATAAAAAAGAGGTATATCCTACCCGAATTGACACACACAATAAATAATAAATAT
ATAAATAATAAAAATATAAATATATAAATTATAAATATATAAATTATATATATAAATATA
TTAACATGTGTGTAATTTGTTAAAATTGATATTAAACAGCTTTGCATGGGGAAAGAGTGG
CTTTTATAAGCGAATCATATAATACAGTGTTAATTTTAATATATAAAATATAATTTCCTT
AAATTGTTATTTAACATTTTTAATTGCCTACCCARGGTAATATAATATCCAAATGACAAT
CAAAGATAAATGAAATACCGCTCCATTTTTCCTATATATCCATTTTCATTTTGTGTAAAT
GTTCATAAAATTTTATCATATACTCATAAAAAACAGGGGAAAAATGAAAAAATGAACGTC
CACCCCGTGTGAATATGCTCATTTTGTATACTATACATTTTTTTTTATATAATTTATATT
TTCCAAATAAAAAAAATGTTTGTACATGTATATATCAATATTGTGTTATGGCATGATGAA
AAACAATTATTTTATTCGTTTTTAAATTTGTAGAATTTCCCTAAATTATTAATGATTTCA
TATTATAATATATGTATCTATATATTTTTAATGTGTAATTCAAAATGGCGAAAAAGAAAC
AAACCATACTTTATAAGCAACTCTCTTTGAGCACTTACCATAAAAAATTATACTACTAAA
TTCAAATTAAATAAAAAAATATATATTATAAAATTGGCTTTAATTATTTATAAGCACTTG
CTTTGTTTTACTCAAAAAGTATTGGCATAAGTATAAACATAAAACCAATTTTTCACAAAA
TATATTATATGTCCATACAACTATATCCGAACYTTCAAAACACATCCAAATTTCCATTTA
CCACATGCAAGCAAAAGGATATTAAATAAAAAAAAATTATTTATATTTTCATGATCACTT
AAATAATATTTAAAAAAAAAAGAGACCATATATAAAATGTATAATTTAAATATAGCTAAC
TTATTAAAAAAAATGAATATTGAGCATATATCCATAATTTAAAACACTTCTATTTTATGT
TATTTGATATATAAGTCGATAAATTATTGCATGTATGCATATAGACAGATTAATATACAC
CAAAATAATAAAAATTATATATAGTTCATATATTATTCTGAATAATAAATTAGCTTTCCT
CTTAATTCACTTGGTATGTATATATATATATATATATATATATATATATATATGGATAAA
TAAATATAAGGTCTATAACAGGAAAAAATGTGTATTAACTTTTTTTTATAATTATTTTAT
ATTTTTTCTCCATTTATTTAAATATGTTAAAATTATTTCCCATTTTATAAACAAAAATTA
ATCCATAAAATAGCAAAATAACAAAATGACAAAATAACAAAAACAAAAAAACGCATTATA
TGAGTTCATTTTACACAATCCAAAGAAAAAGAAAATATGCTTAAAATATTCATAACACAC
TTTTAAGCTATAAAAATATATATATTTATTTGTGTCTATATTACCAACTCAATTTAATAG
ATGTGTTATGTGATTAATTCATACACAAACATACAAAAATAAACACCATTTTTGAAAAGA
TTAATTTGAAATCATATATGTTATTATTATATATTTTTATATCAATGAAATTTAAAAAAA
TAAAAGGGAAATCAATGTATTAAAAATAATTATATGTTATTTTATTTCCACAATATTTAT
TTATTATTTATTGTTGACCTGCAGGCATGCAAGCTTATCGATACCGTCGACCTCGAGGGG
GGGCCCGGTACCCAGCTTTTGTTCCCTTTAGTGAGGGTTAATTGCGCGCTTGGCGTAATC
ATGGTCATAGCTGTTTCCTGTGTGAAATTGTTATCCGCTCACAATTCCACACAACATACG
AGCCGGAAGCATAAAGTGTAAAGCCTGGGGTGCCTAATGAGTGAGCTAACTCACATTAAT
TGCGTTGCGCTCACTGCCCGCTTTCCAGTCGGGAAACCTGTCGTGCCAGCTGCATTAATG
AATCGGCCAACGCGCGGGGAGAGGCGGTTTGCGTATTGGGCGCTCTTCCGCTTCCTCGCT
CACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGC
GGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGG
CCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCG
CCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGG
ACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGAC
CCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCA
TAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGT
GCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTC
CAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAG
AGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACAC
TAGAAGGACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGT
TGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAA
GCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGG
GTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAA
AAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTAT
ATATGAGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCACCTATCTCAGC
GATCTGTCTATTTCGTTCATCCATAGTTGCCTGACTCCCCGTCGTGTAGATAACTACGAT
ACGGGAGGGCTTACCATCTGGCCCCAGTGCTGCAATGATACCGCGAGACCCACGCTCACC
GGCTCCAGATTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCC
TGCAACTTTATCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGTAAGTAG
TTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCATTGCTACAGGCATCGTGGTGTCACG
CTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAAGGCGAGTTACATG
ATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAG
TAAGTTGGCCGCAGTGTTATCACTCATGGTTATGGCAGCACTGCATAATTCTCTTACTGT
CATGCCATCCGTAAGATGCTTTTCTGTGACTGGTGAGTACTCAACCAAGTCATTCTGAGA
ATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAATACGGGATAATACCGCGCC
ACATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTTCTTCGGGGCGAAAACTCTC
AAGGATCTTACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTGCACCCAACTGATC
TTCAGCATCTTTTACTTTCACCAGCGTTTCTGGGTGAGCAAAAACAGGAAGGCAAAATGC
CGCAAAAAAGGGAATAAGGGCGACACGGAAATGTTGAATACTCATACTCTTCCTTTTTCA
ATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAATGTAT
TTAGAAAAATAAACAAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCAC
Restriction sites to linearize plasmid NdeI
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 1AACTAGTAACATCAATCCGACCCAG
Additional information primer 1L5800 (SpeI); PB403064.00.0 Forward
Sequence Primer 2AGGATCCAATGGTACAATGTCTATTAATCTAAG
Additional information primer 2L5801 (BamHI); PB403064.00.0 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/construct(Linear) plasmid 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
GCCCAGCTTAATTCTTTTCGAGCTCTTTATGCTTAAGTTTACAATTTAATATTCATACTT
TAAGTATTTTTTGTAGTATCCTAGATATTGTGCTTTAAATGCTCACCCCTCAAAGCACCA
GTAATATTTTCATCCACTGAAATACCATTAAATTTTCAAAAAAATACTATGCATATAATG
TTATACATATAAACATAAAACGCCATGTAAATCAAAAAATATATAAAAATATGTATAAAA
ATAAATATGCACTAAATATAAGCTAATTATGCATAAAAATTAAAGTGCCCTTTATTAACT
AGTCGTAATTATTTATATTTCTATGTTATAAAAAAATCCTCATATAATAATATAATTAAT
ATATGTAATGTTTTTTTTATTTTATAATTTTAATATAAAATAATATGTAAATTAATTCAA
AAAATAAATATAATTGTTGTGAAACAAAAAACGTAATTTTTTCATTTGCCTTCAAAATTT
AAATTTATTTTAATATTTCCTAAAATATATATACTTTGTGTATAAATATATAAAAATATA
TATTTGCTTATAAATAAATAAAAAATTTTATAAAACATAGGGGGATCTATGAGTAAAGGA
GAAGAACTTTTCACTGGAGTTGTCCCAATTCTTGTTGAATTAGATGGTGATGTTAATGGG
CACAAATTTTCTGTCAGTGGAGAGGGTGAAGGTGATGCAACATACGGAAAACTTACCCTT
AAATTTATTTGCACTACTGGAAAACTACCTGTTCCATGGCCAACACTTGTCACTACTTTC
GGTTATGGTGTTCAATGCTTTGCGAGATACCCAGATCATATGAAACAGCATGACTTTTTC
AAGAGTGCCATGCCCGAAGGTTATGTACAGGAAAGAACTATATTTTTCAAAGATGACGGG
AACTACAAGACACGTGCTGAAGTCAAGTTTGAAGGTGATACCCTTGTTAATAGAATCGAG
TTAAAAGGTATTGATTTTAAAGAAGATGGAAACATTCTTGGACACAAATTGGAATACAAC
TATAACTCACACAATGTATACATCATGGCAGACAAACAAAAGAATGGAATCAAAGTTAAC
TTCAAAATTAGACACAACATTGAAGATGGAAGCGTTCAACTAGCAGACCATTATCAACAA
AATACTCCAATTGGCGATGGCCCTGTCCTTTTACCAGACAACCATTACCTGTCCACACAA
TCTGCCCTTTCGAAAGATCCCAACGAAAAGAGAGACCACATGGTCCTTCTTGAGTTTGTA
ACAGCTGCTGGGATTACACATGGCATGGATGAACTATACAAAGGGATCCTGGCTAGCCAG
TCGACCTGCAGGCATGCAAGCTTGCGGCCGATCCAAATGGAAGACGCCAAAAACATAAAG
AAAGGCCCGGCGCCATTCTATCCGCTGGAAGATGGAACCGCTGGAGAGCAACTGCATAAG
GCTATGAAGAGATACGCCCTGGTTCCTGGAACAATTGCTTTTACAGATGCACATATCGAG
GTGAACATCACGTACGCGGAATACTTCGAAATGTCCGTTCGGTTGGCAGAAGCTATGAAA
CGATATGGGCTGAATACAAATCACAGAATCGTCGTATGCAGTGAAAACTCTCTTCAATTC
TTTATGCCGGTGTTGGGCGCGTTATTTATCGGAGTTGCAGTTGCGCCCGCGAACGACATT
TATAATGAACGTGAATTGCTCAACAGTATGAACATTTCGCAGCCTACCGTAGTGTTTGTT
TCCAAAAAGGGGTTGCAAAAAATTTTGAACGTGCAAAAAAAATTACCAATAATCCAGAAA
ATTATTATCATGGATTCTAAAACGGATTACCAGGGATTTCAGTCGATGTACACGTTCGTC
ACATCTCATCTACCTCCCGGTTTTAATGAATACGATTTTGTACCAGAGTCCTTTGATCGT
GACAAAACAATTGCACTGATAATGAATTCCTCTGGATCTACTGGGTTACCTAAGGGTGTG
GCCCTTCCGCATAGAACTGCCTGCGTCAGATTCTCGCATGCCAGAGATCCTATTTTTGGC
AATCAAATCATTCCGGATACTGCGATTTTAAGTGTTGTTCCATTCCATCACGGTTTTGGA
ATGTTTACTACACTCGGATATTTGATATGTGGATTTCGAGTCGTCTTAATGTATAGATTT
GAAGAAGAGCTGTTTTTACGATCCCTTCAGGATTACAAAATTCAAAGTGCGTTGCTAGTA
CCAACCCTATTTTCATTCTTCGCCAAAAGCACTCTGATTGACAAATACGATTTATCTAAT
TTACACGAAATTGCTTCTGGGGGCGCACCTCTTTCGAAAGAAGTCGGGGAAGCGGTTGCA
AAACGCTTCCATCTTCCAGGGATACGACAAGGATATGGGCTCACTGAGACTACATCAGCT
ATTCTGATTACACCCGAGGGGGATGATAAACCGGGCGCGGTCGGTAAAGTTGTTCCATTT
TTTGAAGCGAAGGTTGTGGATCTGGATACCGGGAAAACGCTGGGCGTTAATCAGAGAGGC
GAATTATGTGTCAGAGGACCTATGATTATGTCCGGTTATGTAAACAATCCGGAAGCGACC
AACGCCTTGATTGACAAGGATGGATGGCTACATTCTGGAGACATAGCTTACTGGGACGAA
GACGAACACTTCTTCATAGTTGACCGCTTGAAGTCTTTAATTAAATACAAAGGATATCAG
GTGGCCCCCGCTGAATTGGAATCGATATTGTTACAACACCCCAACATCTTCGACGCGGGC
GTGGCAGGTCTTCCCGACGATGACGCCGGTGAACTTCCCGCCGCCGTTGTTGTTTTGGAG
CACGGAAAGACGATGACGGAAAAAGAGATCGTGGATTACGTCGCCAGTCAAGTAACAACC
GCGAAAAAGTTGCGCGGAGGAGTTGTGTTTGTGGACGAAGTACCGAAAGGTCTTACCGGA
AAACTCGACGCAAGAAAAATCAGAGAGATCCTCATAAAGGCCAAGAAGGGCGGAAAGATC
GCCGTGTAATTCTAGAAGATCCCGTTTTTCTTACTTATATATTTATACCAATTGATTGTA
TTTATAACTGTAAAAATGTGTATGTTGTGTGCATATTTTTTTTTGTGCATGCACATGCAT
GTAAATAGCTAAAATTATGAACATTTTATTTTTTGTTCAGAAAAAAAAAACTTTACACAC
ATAAAATGGCTAGTATGAATAGCCATATTTTATATAAATTAAATCCTATGAATTTATGAC
CATATTAAAAATTTAGATATTTATGGAACATAATATGTTTGAAACAATAAGACAAAATTA
TTATTATTATTATTATTTTTACTGTTATAATTATGTTGTCTCTTCAATGATTCATAAATA
GTTGGACTTGATTTTTAAAATGTTTATAATATGATTAGCATAGTTAAATAAAAAAAGTTG
AAAAATTAAAAAAAAACATATAAACACAAATGATGTTTTTTCCTTCAATTTCGGGTACCG
AGCTCGAATTCTCTTGAGCCCGTTAATGAAATAGATACAATTCATTCATGTTATATACAT
CTAGAACATAATCTGAATATGGTTCAAGTTAAATGTCCAAAAATTATAAAAAGTGATGAT
ATTTTTGATGGTAATACCATAATAGACACCAAGGTAACATCACGAAGTAGTCAACAAAAT
AATTTTTATTTAGAAAATACAGATGTTGAACCAGAAGAAATAGAGAAATATAAAAATATA
GAATACATACCAGAAAACGATGAAGTAATGCATCTAGACAAAAAAGAAAAGCTAGATGAT
ATATTACCAGGTGTTATCATATTAGATAAAAATAAAATGTTCAAAGAAAAAGGACATTTC
ACTTTTGTTACTCCATTAATTGTAGAAAAGGTATTAATATTAAAAATATATTGTGATAAT
ACTAAAACAATAATTAATAATATGAAAGGGAAAAAAGGTATTACAGTAATAAGGATTTCT
CAAAATACAACAAAAAATAAATTTTATGGATGTGACTTTTCAGGTAATTCTAAAAAAACA
TTTTACTATTCCAATGTTTATGATTTAGAAAAAAAAAATGAGTTTTGTGAAATAGAATTA
AAAGAAAATATAGTAGTTAGCTTAAATTGTCCAACTGGTAAAATTAATCCAAAAAATTGT
TTTAGAAATGTATATATAAAAAGTAATATGAATGAACAAACAACCGAAAATATAGAAAAT
ATATTTAACGAAATAAAAGTTATAGATGCAGATTATTTTATAAATAATTCATCAACCTTT
TTGATGATTTCCAAAATTACAAAAAAAGAGTTTGATTTTTATTGTACATGTGAAGATTAT
AAAACCAAAAATATAGGAACAATATATATTAAAAATTATGAATATCTAGATTCAAAACCT
AAATATAAAAATAAACAAATTTCCTATATAGATGTAGTTCCATACCCGCGGGGAAAGGGC
G
Restriction sites to linearize plasmid ApaI, SacII
Selectable marker used to select the mutant parasitegfp (FACS)
Promoter of the selectable markereef1a
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_1133300
Gene Model P. falciparum ortholog PF3D7_1357100
Gene productelongation factor 1-alpha
Gene product: Alternative nameeef1a
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