Summary

RMgm-700
Malaria parasiteP. berghei
Genotype
TaggedGene model (rodent): PBANKA_0722600; Gene model (P.falciparum): Not available; Gene product: fam-b protein (Pb-fam-3.2)
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; Gametocyte/Gamete; Liver stage;
Last modified: 12 August 2015, 13:57
  *RMgm-700
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
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).
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
Name of the mutant parasite
RMgm numberRMgm-700
Principal name1731cl4
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
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/GameteGametocytes: The tagged-protein is exported into the cytoplasm of the host erythrocyte and shows a diffuse or patchy localization in the cytoplasm
Fertilization and ookineteNot tested
OocystNot different from wild type
SporozoiteNot different from wild type
Liver stageThe tagged protein is expressed in late liver stages (schizonts) where it is located in the parasitophorous vacuole.
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.

The protein is not detected in oocysts and in sporozoites.

The tagged protein is expressed in late liver stages (schizonts) where it is located in the parasitophorous vacuole.

An additional mutant, in which PBANKA_0722600 was c-terminally tagged with EGFP, has been made in P. berghei ANKA 1037m1f1mocl1 (1942; RMgm-1282). P. 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).



Other mutants
RMgm-1283: An additional mutant, in which PBANKA_0836800 was c-terminally tagged with EGFP,


  Tagged: Mutant parasite with a tagged gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_0722600
Gene Model P. falciparum ortholog Not available
Gene productfam-b protein
Gene product: Alternative namePb-fam-3.2
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
CCGCGGGTACTATAACACGTGTTTAATGAATATGAAAAAATAAAGCGCTTGCTATTAAGT
ATATTTCATTTTCTGGTTTCATATAATAATAAATTAATAATTTGTTACTAATTTTTTATT
TTACATGTATTTTTCACTGTAATTTATACGAGGTATACTTGATAATATGAATATTTTATA
TTATTTTAATATAAATATAGTATATTTGATGGTGTATATTCTGAGATATATTTTTAAATA
AAATTATTATATACCAATTTATGGAGAGGCCATATATTTTAAAAAACGAAGAAAAATAAA
ATGTAGAATATTATTAAATAATATTTAAAATTGGAGAAATATTATAATAAAATATAATAA
ATTAGAAACGATTTAAATGTAAATATCAATGGTGCTTTGCTAATGTTTTCATTATGTGTT
TGTATTTATTCATATAAAAAAATATATCCAATATTTTATTATTTTAATAGATATTTATTA
ATGTGTTCATATGTGAACAATAATAGGAAAATATAAGTTTATAAAATATTAAAAACGTAC
CACTAAAATCACCAACGTATTATAAATTATATTAAAAGCCTATTTGTATACCAATTTTCC
AAAAAATATATTTTATGGAAAATTATTATATACATATATTTTTTTATAATTAAAATATTA
AAAACGGTGTATTTTTTAGAGAAAAGTTAATATTTCTTTGAAAAAAAACATAATATATAT
ATCCATAAGCAATTTTAATGAAATAACGAAATTTATTATAACAATTACATTAATTAATTT
TACAAAAAGTGTAAAAAGTTAATAAATGAAATAAATTCGTTTTTTTTATAGATATATTCA
TTTATTCATAAATTTTAGAACAAAAAACTAGTAAATATTACTATTTTGTGAATAAATATT
AAGTTACCGACATTACCTCAAACTTCATATATTAGTTCTTTTATTTAATAAAGGTAAAAA
TGAGAGTCAGTGTTTTAAAATATGTTATTTTTTCAATTGTTATTTGTTCTTTTGAATATG
CCAAAAATGTAAGTTACACTTTATTTTCTTTTATTATTAATAATATTTCATTATAGTTTT
TGTATTTATCGCTTTACATTAACCTTATATTATTGTCTCATGTATTGACAAGGTACTTAA
GTTAAAGAAGCAATTAAAATTAATTACATATATGTTAATAAATATTTCATTTCTTTGTAG
GAATTATACTTTGTAAACGATAAAGGGATATACCTTGAAGGGAATGTAATAAACTTTAGA
AATAATAGGATATTAGAAGAAGCGGACAATCAAATTGATTTAAATGAATTTTATCAATCA
ACTTTGAGTCTTGCAAATCAATTTAATGGTTGTAATGATGGTAAAAAGGAAATAGAACAC
CTTCAAAATATTATAAATTCACATATAAAGAAGCATAAAGAAAATAATACATTACCCAAT
TTAAATAATTTAGATAAGAAAACTAAAAAGCTAATTGATGACCTTCGAACAGAATTAGAA
GAAGTAAAAAGAGAGATTGATAATAAAAGGAATGGTGAATTAGCAATACATTCGATAGAT
GATAAAATAATAATAAAAAAAGATGAAAATGGTTCTGTATCAGAACATGAAGACTTTAAA
CAATTGAGAAATTCTGAAAATATTTTGGAAATTGATGAAGATATTTTTGATAATGAATAT
TGTGAAATTAAATCAAGTGAAAATTATAGGAAATTCCAAATTAATAAAAATATTAAAAAA
AATGAAAAATTAATTAAGAAGTTGATGATTTTGTCTGTTACAATTTTGTCTGTGGCAATA
GCATCAGGAACAGTGAGCTTACTATTGCTACTTGCACCGTTGGGGGGGTTAATATTTAGG
AACTGGCGGAGAATCAGGAAATATAGATTTAGATTAGGATCCGGTGGAGGTGGAAGATCC
ATGGTGAGCAAGGGCGAGGAGGATAACATGGCCATCATCAAGGAGTTCATGCGCTTCAAG
GTGCACATGGAGGGCTCCGTGAACGGCCACGAGTTCGAGATCGAGGGCGAGGGCGAGGGC
CGCCCCTACGAGGGCACCCAGACCGCCAAGCTGAAGGTGACCAAGGGTGGCCCCCTGCCC
TTCGCCTGGGACATCCTGTCCCCTCAGTTCATGTACGGCTCCAAGGCCTACGTGAAGCAC
CCCGCCGACATCCCCGACTACTTGAAGCTGTCCTTCCCCGAGGGCTTCAAGTGGGAGCGC
GTGATGAACTTCGAGGACGGCGGCGTGGTGACCGTGACCCAGGACTCCTCCCTGCAGGAC
GGCGAGTTCATCTACAAGGTGAAGCTGCGCGGCACCAACTTCCCCTCCGACGGCCCCGTA
ATGCAGAAGAAGACCATGGGCTGGGAGGCCTCCTCCGAGCGGATGTACCCCGAGGACGGC
GCCCTGAAGGGCGAGATCAAGCAGAGGCTGAAGCTGAAGGACGGCGGCCACTACGACGCT
GAGGTCAAGACCACCTACAAGGCCAAGAAGCCCGTGCAGCTGCCCGGCGCCTACAACGTC
AACATCAAGTTGGACATCACCTCCCACAACGAGGACTACACCATCGTGGAACAGTACGAA
CGCGCCGAGGGCCGCCACTCCACCGGCGGCATGGACGAGCTGTACAAGTAATCTAGAATT
ATTAATATATATGAATATATATACATCGTTGTATGCGTAATATACGTGTAACTGTTTTTT
TTAATTTTTATATTAACTTTTCCTTATACATTTTAATGCCATTAACTAATAATATTTTAC
TTGAAAAAATTCTAAATTTTTATAACTCTAATATAAAATCTGTGAATAATAAAAAATTAG
CATAAAGTACGAATTAAACATCGAATATTTGTTTTAAAGGGGCACACATTATGTGCATAT
AATTTTATTCTTTGAATTTAATTTTTTGCTTATAATTAGATTAAATTAATATAATTTTTT
TATTGAAAATAAAAACTTATTTTAAAAAATTAACAAATGGAAGTTAATGAAAAATACGAA
AAAAGTAAAGGGTTAATTCTTATATGGTCGGTACCGAGCTCGAATTCGATATCGAAATTG
AAGGAAAAAACATCATTTGTGTTTATATGTTTTTTTTTAATTTTTCAACTTTTTTTATTT
AACTATGCTAATCATATTATAAACATTTTAAAAATCAAGTCCAACTATTTATGAATCATT
GAAGAGACAACATAATTATAACAGTAAAAATAATAATAATAATAATAATTTTGTCTTATT
GTTTCAAACATATTATGTTCCATAAATATCTAAATTTTTAATATGGTCATAAATTCATAG
GATTTAATTTATATAAAATATGGCTATTCATACTAGCCATTTTATGTGTGTAAAGTTTTT
TTTTTCTGAACAAAAAATAAAATGTTCATAATTTTAGCTATTTACATGCATGTGCATGCA
CAAAAAAAAATATGCACACAACATACACATTTTTACAGTTATAAATACAATCAATTGGTA
TAAATATATAAGTAAGAAAAACGGGATCAAAGCTAGCTTCTGTATTTCCGCTAGACAGCC
ATCTCCATCTGGATTCGTCCGTGCGGGACGTAGCCCACGACCTCAAAATCCTCGGCGGTG
AAATCGTCGATTTCCTTGATGCGTTCCTTGTTGAGGATGTTCACAATGGGGAACGGTCTC
GGTTCTCTCCGCAGCTGCTCTTTTAAAGCCTCGACATGGTTCGTGTAGACATGCGTGTTC
CCCATGAAGTGAATGAACTCCTTAGGTTTTAGGTTGCAGACGTGTGCAACCATGAGCGTC
AAAAGCGAATAGGAAGCGATGTTGAAGGGGACGCCGAGGCCGACATCGCACGACCGCTGA
TACATGATGCACGACAGCTCCTTCTGGTCGTTCACGTAGAACTGGCACAACAAGTGACAA
GGCGGCAGCGCCATTTCGTCCAGCGCTGCAGGATTCCAGGCAGTCATGAGCATGCGACGA
TCTGTTGGATTCGTTCTCAGCATCTGGATCACATTCTTCAGCTGGTCGACGCCCTGCCCT
GTGTAGTCTGTGTGCATGTCTTTGTATGCCGCGCCGAAGTGTCTCCACTGGAAGCCGTAG
CCCGGGCCGATGTCTCCGACCTCTCGGTGGGGGAGATTGCGCGAATCGAGGAACTCGCGT
GTCACATTCTTGTCCCAGATCTTCACGCCCTTCTCAGAAAGATGGTTTGCGTTCGTGTCG
CCGCGAATGAACCACAGCAACTCTTCGAGGACCCCTTTCCAGAACACACGCTTTGTGGTG
AGAAGTGGAAAGGCCTGATCCAGCGAGTAGCGCATAGTGCAGCCGAATTTGGAGATGACA
CCAACGCCCGTTCGGTCATCCATTGTCCTTCCATTGTTAATAATGTCGGCAATGAGATCA
AGGTACTGGAACTCTTCATGGCCTCTAAAGTGAACATGCGGAACGGCCCGAATCAGTTCC
TTTTGCTCGCGTTTTTTCCGGTCTTCTTCGTCCATCCACGCCAACACCGGGGCAATGGCT
GCGGCCGAAGAAGGAGCCTGCAACCCGTGCACGGGAGTTGTCTCCCTCGTGGACGTCAAG
GAGCTCATTGCGTTGCTCGGTTCCGCAGTGGCTGCGTCGTCAGTCTTCCTTCTCTTCTCG
AGAACCACAAAGTCGTAGGGAACCCCGTTGTCTGAGAAGGTCTTGGAAATGAAGATGGGT
CGATACGTCGCTTCATTGTCCTTCTCTCTTCCGAGCTCCGGACAAAAGGGAACGAACACA
GACTCGGCAGGAGCTGCAGCCTGCGCAGCAGTTGATTTGTTTGAAAGAATGTCATCTCCG
GGGAACGCAGGGAAGAAAACGTCGCACGGAAACTCGCGGGCTACACGCGTGATGTACAGG
TGAGAGGCAACGCCCAGAGACAGCGCTGCCTCGTACAGTCCCGCTCCTCCCACGACAAAA
ATCTGGTCGACAGAATCCTTGTACTCTTCCTCCAGAAGGCTGAGAGCTGCTGGGAGTGAA
GCACAGACTCGGACGCGCTGCTGGCCTTCAGCTTGAGGCTTCTCCGCCGCAATGTCTTCT
TCTTTGAGGGAAGAGGAAACGACGATGTTCAATCTGTCCACGAGGGGTCTAAACTTTCGA
GGCATGCTTTCCCAGGTTTTCCGTCCCATGACAACGGCGTTGAATCTCTTGCCGACTGAT
GGAGAGGGAAGTCCAGAGTCGCCCGTCTTTGCAAATTTCCTGGGAAGCCACCCGTTCAGG
CGACTGGCTTCTTCGGGCGTCGTTTTTGTCACACGAGAAAAGTGTTTGAAATCTGTGGTC
AAGTGGGGCCACGGGAGGCCGTTGTTGATGCCGATGCCCCTCTTGGGGGTCATCGCGACG
ACCAGACACACCGGTTTATGCATTGATCCTTTGTAACATTTAGGTGTGTATTTATATATA
TAAGCAATATACAAATGAAAATACTATTATAAAACGGAAACTCAAAAAATGTTGAAACAA
ATAGTCAATTTTTAAAATGAATGTGTAAATTGTTTAAGTTTTATGACAAAAAAAGGAAAA
GTGTATATAAAATAAAAATTGGAAAAAATTTATATATTACATGCCATATAATACCATTTA
TTTACTATTTATATCTTAATATAAAAAATTGTAGCAAAAATATATAATCGTATTTTATAT
ATTTAATATTATATAACCCCTTAAAAAAAAAAATTTAATTGAAAAAATTTTAATTTATGC
ACTTTTTTATTGTATTATATATATTCCGTTATTTTAATTACACATAAACTTAATAGAAAA
ATTAATTCGCTGCTAAATAAAAATTATAGGGCTATATATGATGGGATCATATTCTATCTT
TCTCTTTTTCAAATYCTTTATATTTTAGAATACAAATCGCTTTAAACAATAAAAAAGAGG
TATATCCTACCCGAATTGACACACACAATAAATAATAAATATATAAATAATAAAAATATA
AATATATAAATTATAAATATATAAATTATATATATAAATATATTAACATGTGTGTAATTT
GTTAAAATTGATATTAAACAGCTTTGCATGGGGAAAGAGTGGCTTTTATAAGCGAATCAT
ATAATACAGTGTTAATTTTAATATATAAAATATAATTTCCTTAAATTGTTATTTAACATT
TTTAATTGCCTACCCARGGTAATATAATATCCAAATGACAATCAAAGATAAATGAAATAC
CGCTCCATTTTTCCTATATATCCATTTTCATTTTGTGTAAATGTTCATAAAATTTTATCA
TATACTCATAAAAAACAGGGGAAAAATGAAAAAATGAACGTCCACCCCGTGTGAATATGC
TCATTTTGTATACTATACATTTTTTTTTATATAATTTATATTTTCCAAATAAAAAAAATG
TTTGTACATGTATATATCAATATTGTGTTATGGCATGATGAAAAACAATTATTTTATTCG
TTTTTAAATTTGTAGAATTTCCCTAAATTATTAATGATTTCATATTATAATATATGTATC
TATATATTTTTAATGTGTAATTCAAAATGGCGAAAAAGAAACAAACCATACTTTATAAGC
AACTCTCTTTGAGCACTTACCATAAAAAATTATACTACTAAATTCAAATTAAATAAAAAA
ATATATATTATAAAATTGGCTTTAATTATTTATAAGCACTTGCTTTGTTTTACTCAAAAA
GTATTGGCATAAGTATAAACATAAAACCAATTTTTCACAAAATATATTATATGTCCATAC
AACTATATCCGAACYTTCAAAACACATCCAAATTTCCATTTACCACATGCAAGCAAAAGG
ATATTAAATAAAAAAAAATTATTTATATTTTCATGATCACTTAAATAATATTTAAAAAAA
AAAGAGACCATATATAAAATGTATAATTTAAATATAGCTAACTTATTAAAAAAAATGAAT
ATTGAGCATATATCCATAATTTAAAACACTTCTATTTTATGTTATTTGATATATAAGTCG
ATAAATTATTGCATGTATGCATATAGACAGATTAATATACACCAAAATAATAAAAATTAT
ATATAGTTCATATATTATTCTGAATAATAAATTAGCTTTCCTCTTAATTCACTTGGTATG
TATATATATATATATATATATATATATATATATATGGATAAATAAATATAAGGTCTATAA
CAGGAAAAAATGTGTATTAACTTTTTTTTATAATTATTTTATATTTTTTCTCCATTTATT
TAAATATGTTAAAATTATTTCCCATTTTATAAACAAAAATTAATCCATAAAATAGCAAAA
TAACAAAATGACAAAATAACAAAAACAAAAAAACGCATTATATGAGTTCATTTTACACAA
TCCAAAGAAAAAGAAAATATGCTTAAAATATTCATAACACACTTTTAAGCTATAAAAATA
TATATATTTATTTGTGTCTATATTACCAACTCAATTTAATAGATGTGTTATGTGATTAAT
TCATACACAAACATACAAAAATAAACACCATTTTTGAAAAGATTAATTTGAAATCATATA
TGTTATTATTATATATTTTTATATCAATGAAATTTAAAAAAATAAAAGGGAAATCAATGT
ATTAAAAATAATTATATGTTATTTTATTTCCACAATATTTATTTATTATTTATTGTTGAC
CTGCAGGCATGCAAGCTTATCGATACCGTCGACCTCGAGGGGGGGCCCGGTACCCAGCTT
TTGTTCCCTTTAGTGAGGGTTAATTGCGCGCTTGGCGTAATCATGGTCATAGCTGTTTCC
TGTGTGAAATTGTTATCCGCTCACAATTCCACACAACATACGAGCCGGAAGCATAAAGTG
TAAAGCCTGGGGTGCCTAATGAGTGAGCTAACTCACATTAATTGCGTTGCGCTCACTGCC
CGCTTTCCAGTCGGGAAACCTGTCGTGCCAGCTGCATTAATGAATCGGCCAACGCGCGGG
GAGAGGCGGTTTGCGTATTGGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCGCTC
GGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCAC
AGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAA
CCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATCA
CAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGC
GTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATA
CCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTA
TCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCA
GCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGA
CTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGG
TGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGGACAGTATTTGG
TATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGG
CAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAG
AAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAA
CGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGAT
CCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTC
TGACAGTTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTATTTCGTTC
ATCCATAGTTGCCTGACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATC
TGGCCCCAGTGCTGCAATGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGC
AATAAACCAGCCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTC
CATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTT
GCGCAACGTTGTTGCCATTGCTACAGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGC
TTCATTCAGCTCCGGTTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGTGCAA
AAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTT
ATCACTCATGGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATG
CTTTTCTGTGACTGGTGAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACC
GAGTTGCTCTTGCCCGGCGTCAATACGGGATAATACCGCGCCACATAGCAGAACTTTAAA
AGTGCTCATCATTGGAAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTT
GAGATCCAGTTCGATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTT
CACCAGCGTTTCTGGGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAG
GGCGACACGGAAATGTTGAATACTCATACTCTTCCTTTTTCAATATTATTGAAGCATTTA
TCAGGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATAAACAAAT
AGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCAC
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 1CCCCGCGGGTACTATAACACGTGTTTAATGAATATG
Additional information primer 1L5715 (SacII); PBANKA_072260 Forward
Sequence Primer 2CGGATCCTAATCTAAATCTATATTTCCTGATTC
Additional information primer 2L5716 (BamHI); PBANKA_072260 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

  Transgene: Mutant parasite expressing a transgene
Type and details of transgene
Is the transgene Plasmodium derived Transgene: not Plasmodium
Transgene nameA fusion of GFP (gfp-mu3) and Luciferase Firefly (LucIAV)
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.
Other details transgene
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
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