Summary

RMgm-695
Malaria parasiteP. berghei
Genotype
TaggedGene model (rodent): PBANKA_1400300; 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_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; Liver stage;
Last modified: 12 August 2015, 14:22
  *RMgm-695
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-695
Principal name1531cl3
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/GameteNot tested
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.




Other mutants
RMgm-1281: An additional mutant, in which PBANKA_1400300 was c-terminally tagged with EGFP,
 


  Tagged: Mutant parasite with a tagged gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_1400300
Gene Model P. falciparum ortholog Not available
Gene productPIR protein BIR protein
Gene product: Alternative name
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
CCGCGGCGATAATTATAAATAATATGGTAGATTGAATAGCGTTATTGTTCTAATATATAT
AATATTGTATCAATGATTAAAACTGAAATATGTTAAATTTGGGAAACATGTAAATTATAT
ATTAATTCAAAGTATACAGAAAAGGTATGTAATAATTAATTTAATTAGAATAATATTTTT
ATTAGGTTATTATATATATATATAAAATCACAAATATATTGTTATTGCTAAATAATATAT
ATTCTAAAATACTTAATTTAAAAACTATGAAACGAGGGAATATTATACATTGGTTTAAAG
TATTTTTATTAAGCGCATTAATTATTCCGTTGTAATGTACAGTGATATAGTAGTAACACT
AATAATAGTAGTAATAGCAATAGATAAACGTATTAAATACGAACAAATCATTAAATTCAT
TTGATTCGAATACGTTGATATATATATTAATTATATATATTATTAAACTTATAATAATAT
TAAAATTGTATGTATAAAAGATCAAATGAAATATTATAATACTTATTTAAGTAAGCTTTA
ATGCGATAATATTATAATTAACACTGCCAAGCAAAATAATATTAATAATTCTATAGTTTT
CCATCATAGAACTAAATATAGTTTATTATAAAATATAAAAGCAATCTATATATTTAGAAA
ATAATTCTAAGGCATTTTTAATGTATGCATGAATTGAAAGCTTTAATGGTAGAAAACATA
AAACATAATTAAACTATTTAGAATAAGTAAATCTTATATGGCTATATCCTTGTCTTAAAA
AACATACTTCCCTTATCTCTCCTCTCAATGTGCAATATACCAATCTAATACACATAGTTT
TCTATTATACTTTAAAATTTGCATCAATAATTATTTATGTGTTATATATTTAATATATAC
TAAGTATTATTTTAAAAAAAATATGCATTTAAAGACATATTTATGCTTATAAATAACATA
ATAAATACGGTTTTAGTGCTAATTGCCGTTTTAAACCGTGGGAAATATGTGCAAAATATA
TTATAAAATCACCCAATAGTGTATATTTTTAAATTGAAGTATATATGAATAAATATAAAG
TTATTTATTTATTAAAATGGACTATGAATTTATTCATATTAAATAAAAATAATGTTAAAA
TTATGATATGCAATTAAAAAAGGGAGCAATGCAACTTATTTTGTAAATGTTTTAATTTTA
GAAAAAACTATATTATATATTATAAGAAACAAGTATATACATATTTAATATATTTAAAAA
AATTACTATTTATATACTTTTAAGGATTATAGTAATAATAGGATTTTTTATTTTTTAGAT
TTATACATTATTTAAGTTTCAGGACATTGTTTATGTTCTATATTAAAGCATATGTATAAG
ATATATTTTAAATTCATTATAAAAGTATATTAATTTTTATAATAATGGTCATATGAGTGT
TATTATGGATAAAAATTTATGCAAAATTGTATTATATATACATATGATAATAAATGTTTA
AACAAACCCTCAAAATATGATAATTTATCTAACTACCATTAAAGTATATATTGTTCCATA
TTATCTTTAAATTTATATTAAGAATAATAAAAATAATTTTATTTACAGAAATTTGCTGTA
TATAGGGTTAAATAATTGTATTACCTATTTTAGTATATATATAACATGATATCCCCTTAT
CCTAGAAACTATAAATTATATTCATCATAATGGATGATGCTCTAGTATGTACATTTTTTT
ATGTTATATTTTCTATAATCTTCATTAATTTTTATTTTAATATCATTTTCTTAATACATA
TTTTTATCATTTTTTTCTAAATGGTTTCTTAGTGTCAAAACTTTGATGTTTTGAGGATGC
ATTTACCCGCTAAATTAGGCGGAACCGAAAAATGTGAACTTAAAAATATTAGTGATTTCC
AAAATTACTGCCCTAATAAAAACTGCAATACTGATCTCGAAAAAATTACGGTTGGATTTT
TATGGTTACTTGGAAAATATTATTCTATATCCCAAAATAGAAGTTATAATGAAAATAATA
CTAATCCATTTTTTCTATATATGATTTCATGGTTTAGTTACAAATTAAGTCAAATCACAG
GGCACGATTCCACCACAATAAACGATTTTTATAATAAAAATATAATAAATAATGATAAAT
ATGAAAAATTTATAAGTGATTCCAGTAAATTTACAAAACTTAATGAAGAATTAAATACAA
AAAGTGATTTTTTGAATATTAATATTGAAGATCTGTCTAAATTTTATGATGCATTTAAAC
TATTATGTAGTATGCATGATAAAGTTGCAAGGAATCAAACAGGCGACATACTGTTAAATA
ATGCTACTAGTTTTGTTGAGAAATATACAGAGCTAAACAATAGATATAATAATGGTATCC
CACATACTAAAATATTGTCTGTTTTATTAACTGGTTATAATAATTTAAAAAATGATCATA
CATATTTTCCATCCATTCTAACAATAGAACAAAAACATCATGTACAAAGTTCTGGAGGTA
ATTATGGACAAAGTTCTGAACTAATTTCTGAAATTACATCAAGTTCGTCGATAGGAAACA
AATTATTTACAGTTTTATCGATATTTGGTGCAATAGCATTTTTTTTAGGAATTTCTTATA
AGGTAAATAATAAGGAATTTAAAAATAATTTTCATTATATATATAGAAATGTTAACAAAA
AAATCATACGTTTCTTAACATTTTATATTAGTATTCGTTATTTGGATTTCGGAAACGAGT
TCAAAAACAACATTTAAGAGGAACGTTAAAAAATATAAAGAAGAAAATGAATTATCGGGG
ATCCGGTGGAGGTGGAAGATCCATGGTGAGCAAGGGCGAGGAGGATAACATGGCCATCAT
CAAGGAGTTCATGCGCTTCAAGGTGCACATGGAGGGCTCCGTGAACGGCCACGAGTTCGA
GATCGAGGGCGAGGGCGAGGGCCGCCCCTACGAGGGCACCCAGACCGCCAAGCTGAAGGT
GACCAAGGGTGGCCCCCTGCCCTTCGCCTGGGACATCCTGTCCCCTCAGTTCATGTACGG
CTCCAAGGCCTACGTGAAGCACCCCGCCGACATCCCCGACTACTTGAAGCTGTCCTTCCC
CGAGGGCTTCAAGTGGGAGCGCGTGATGAACTTCGAGGACGGCGGCGTGGTGACCGTGAC
CCAGGACTCCTCCCTGCAGGACGGCGAGTTCATCTACAAGGTGAAGCTGCGCGGCACCAA
CTTCCCCTCCGACGGCCCCGTAATGCAGAAGAAGACCATGGGCTGGGAGGCCTCCTCCGA
GCGGATGTACCCCGAGGACGGCGCCCTGAAGGGCGAGATCAAGCAGAGGCTGAAGCTGAA
GGACGGCGGCCACTACGACGCTGAGGTCAAGACCACCTACAAGGCCAAGAAGCCCGTGCA
GCTGCCCGGCGCCTACAACGTCAACATCAAGTTGGACATCACCTCCCACAACGAGGACTA
CACCATCGTGGAACAGTACGAACGCGCCGAGGGCCGCCACTCCACCGGCGGCATGGACGA
GCTGTACAAGTAATCTAGAATTATTAATATATATGAATATATATACATCGTTGTATGCGT
AATATACGTGTAACTGTTTTTTTTAATTTTTATATTAACTTTTCCTTATACATTTTAATG
CCATTAACTAATAATATTTTACTTGAAAAAATTCTAAATTTTTATAACTCTAATATAAAA
TCTGTGAATAATAAAAAATTAGCATAAAGTACGAATTAAACATCGAATATTTGTTTTAAA
GGGGCACACATTATGTGCATATAATTTTATTCTTTGAATTTAATTTTTTGCTTATAATTA
GATTAAATTAATATAATTTTTTTATTGAAAATAAAAACTTATTTTAAAAAATTAACAAAT
GGAAGTTAATGAAAAATACGAAAAAAGTAAAGGGTTAATTCTTATATGGTCGGTACCGAG
CTCGAATTCGATATCGAAATTGAAGGAAAAAACATCATTTGTGTTTATATGTTTTTTTTT
AATTTTTCAACTTTTTTTATTTAACTATGCTAATCATATTATAAACATTTTAAAAATCAA
GTCCAACTATTTATGAATCATTGAAGAGACAACATAATTATAACAGTAAAAATAATAATA
ATAATAATAATTTTGTCTTATTGTTTCAAACATATTATGTTCCATAAATATCTAAATTTT
TAATATGGTCATAAATTCATAGGATTTAATTTATATAAAATATGGCTATTCATACTAGCC
ATTTTATGTGTGTAAAGTTTTTTTTTTCTGAACAAAAAATAAAATGTTCATAATTTTAGC
TATTTACATGCATGTGCATGCACAAAAAAAAATATGCACACAACATACACATTTTTACAG
TTATAAATACAATCAATTGGTATAAATATATAAGTAAGAAAAACGGGATCAAAGCTAGCT
TCTGTATTTCCGCTAGACAGCCATCTCCATCTGGATTCGTCCGTGCGGGACGTAGCCCAC
GACCTCAAAATCCTCGGCGGTGAAATCGTCGATTTCCTTGATGCGTTCCTTGTTGAGGAT
GTTCACAATGGGGAACGGTCTCGGTTCTCTCCGCAGCTGCTCTTTTAAAGCCTCGACATG
GTTCGTGTAGACATGCGTGTTCCCCATGAAGTGAATGAACTCCTTAGGTTTTAGGTTGCA
GACGTGTGCAACCATGAGCGTCAAAAGCGAATAGGAAGCGATGTTGAAGGGGACGCCGAG
GCCGACATCGCACGACCGCTGATACATGATGCACGACAGCTCCTTCTGGTCGTTCACGTA
GAACTGGCACAACAAGTGACAAGGCGGCAGCGCCATTTCGTCCAGCGCTGCAGGATTCCA
GGCAGTCATGAGCATGCGACGATCTGTTGGATTCGTTCTCAGCATCTGGATCACATTCTT
CAGCTGGTCGACGCCCTGCCCTGTGTAGTCTGTGTGCATGTCTTTGTATGCCGCGCCGAA
GTGTCTCCACTGGAAGCCGTAGCCCGGGCCGATGTCTCCGACCTCTCGGTGGGGGAGATT
GCGCGAATCGAGGAACTCGCGTGTCACATTCTTGTCCCAGATCTTCACGCCCTTCTCAGA
AAGATGGTTTGCGTTCGTGTCGCCGCGAATGAACCACAGCAACTCTTCGAGGACCCCTTT
CCAGAACACACGCTTTGTGGTGAGAAGTGGAAAGGCCTGATCCAGCGAGTAGCGCATAGT
GCAGCCGAATTTGGAGATGACACCAACGCCCGTTCGGTCATCCATTGTCCTTCCATTGTT
AATAATGTCGGCAATGAGATCAAGGTACTGGAACTCTTCATGGCCTCTAAAGTGAACATG
CGGAACGGCCCGAATCAGTTCCTTTTGCTCGCGTTTTTTCCGGTCTTCTTCGTCCATCCA
CGCCAACACCGGGGCAATGGCTGCGGCCGAAGAAGGAGCCTGCAACCCGTGCACGGGAGT
TGTCTCCCTCGTGGACGTCAAGGAGCTCATTGCGTTGCTCGGTTCCGCAGTGGCTGCGTC
GTCAGTCTTCCTTCTCTTCTCGAGAACCACAAAGTCGTAGGGAACCCCGTTGTCTGAGAA
GGTCTTGGAAATGAAGATGGGTCGATACGTCGCTTCATTGTCCTTCTCTCTTCCGAGCTC
CGGACAAAAGGGAACGAACACAGACTCGGCAGGAGCTGCAGCCTGCGCAGCAGTTGATTT
GTTTGAAAGAATGTCATCTCCGGGGAACGCAGGGAAGAAAACGTCGCACGGAAACTCGCG
GGCTACACGCGTGATGTACAGGTGAGAGGCAACGCCCAGAGACAGCGCTGCCTCGTACAG
TCCCGCTCCTCCCACGACAAAAATCTGGTCGACAGAATCCTTGTACTCTTCCTCCAGAAG
GCTGAGAGCTGCTGGGAGTGAAGCACAGACTCGGACGCGCTGCTGGCCTTCAGCTTGAGG
CTTCTCCGCCGCAATGTCTTCTTCTTTGAGGGAAGAGGAAACGACGATGTTCAATCTGTC
CACGAGGGGTCTAAACTTTCGAGGCATGCTTTCCCAGGTTTTCCGTCCCATGACAACGGC
GTTGAATCTCTTGCCGACTGATGGAGAGGGAAGTCCAGAGTCGCCCGTCTTTGCAAATTT
CCTGGGAAGCCACCCGTTCAGGCGACTGGCTTCTTCGGGCGTCGTTTTTGTCACACGAGA
AAAGTGTTTGAAATCTGTGGTCAAGTGGGGCCACGGGAGGCCGTTGTTGATGCCGATGCC
CCTCTTGGGGGTCATCGCGACGACCAGACACACCGGTTTATGCATTGATCCTTTGTAACA
TTTAGGTGTGTATTTATATATATAAGCAATATACAAATGAAAATACTATTATAAAACGGA
AACTCAAAAAATGTTGAAACAAATAGTCAATTTTTAAAATGAATGTGTAAATTGTTTAAG
TTTTATGACAAAAAAAGGAAAAGTGTATATAAAATAAAAATTGGAAAAAATTTATATATT
ACATGCCATATAATACCATTTATTTACTATTTATATCTTAATATAAAAAATTGTAGCAAA
AATATATAATCGTATTTTATATATTTAATATTATATAACCCCTTAAAAAAAAAAATTTAA
TTGAAAAAATTTTAATTTATGCACTTTTTTATTGTATTATATATATTCCGTTATTTTAAT
TACACATAAACTTAATAGAAAAATTAATTCGCTGCTAAATAAAAATTATAGGGCTATATA
TGATGGGATCATATTCTATCTTTCTCTTTTTCAAATYCTTTATATTTTAGAATACAAATC
GCTTTAAACAATAAAAAAGAGGTATATCCTACCCGAATTGACACACACAATAAATAATAA
ATATATAAATAATAAAAATATAAATATATAAATTATAAATATATAAATTATATATATAAA
TATATTAACATGTGTGTAATTTGTTAAAATTGATATTAAACAGCTTTGCATGGGGAAAGA
GTGGCTTTTATAAGCGAATCATATAATACAGTGTTAATTTTAATATATAAAATATAATTT
CCTTAAATTGTTATTTAACATTTTTAATTGCCTACCCARGGTAATATAATATCCAAATGA
CAATCAAAGATAAATGAAATACCGCTCCATTTTTCCTATATATCCATTTTCATTTTGTGT
AAATGTTCATAAAATTTTATCATATACTCATAAAAAACAGGGGAAAAATGAAAAAATGAA
CGTCCACCCCGTGTGAATATGCTCATTTTGTATACTATACATTTTTTTTTATATAATTTA
TATTTTCCAAATAAAAAAAATGTTTGTACATGTATATATCAATATTGTGTTATGGCATGA
TGAAAAACAATTATTTTATTCGTTTTTAAATTTGTAGAATTTCCCTAAATTATTAATGAT
TTCATATTATAATATATGTATCTATATATTTTTAATGTGTAATTCAAAATGGCGAAAAAG
AAACAAACCATACTTTATAAGCAACTCTCTTTGAGCACTTACCATAAAAAATTATACTAC
TAAATTCAAATTAAATAAAAAAATATATATTATAAAATTGGCTTTAATTATTTATAAGCA
CTTGCTTTGTTTTACTCAAAAAGTATTGGCATAAGTATAAACATAAAACCAATTTTTCAC
AAAATATATTATATGTCCATACAACTATATCCGAACYTTCAAAACACATCCAAATTTCCA
TTTACCACATGCAAGCAAAAGGATATTAAATAAAAAAAAATTATTTATATTTTCATGATC
ACTTAAATAATATTTAAAAAAAAAAGAGACCATATATAAAATGTATAATTTAAATATAGC
TAACTTATTAAAAAAAATGAATATTGAGCATATATCCATAATTTAAAACACTTCTATTTT
ATGTTATTTGATATATAAGTCGATAAATTATTGCATGTATGCATATAGACAGATTAATAT
ACACCAAAATAATAAAAATTATATATAGTTCATATATTATTCTGAATAATAAATTAGCTT
TCCTCTTAATTCACTTGGTATGTATATATATATATATATATATATATATATATATATGGA
TAAATAAATATAAGGTCTATAACAGGAAAAAATGTGTATTAACTTTTTTTTATAATTATT
TTATATTTTTTCTCCATTTATTTAAATATGTTAAAATTATTTCCCATTTTATAAACAAAA
ATTAATCCATAAAATAGCAAAATAACAAAATGACAAAATAACAAAAACAAAAAAACGCAT
TATATGAGTTCATTTTACACAATCCAAAGAAAAAGAAAATATGCTTAAAATATTCATAAC
ACACTTTTAAGCTATAAAAATATATATATTTATTTGTGTCTATATTACCAACTCAATTTA
ATAGATGTGTTATGTGATTAATTCATACACAAACATACAAAAATAAACACCATTTTTGAA
AAGATTAATTTGAAATCATATATGTTATTATTATATATTTTTATATCAATGAAATTTAAA
AAAATAAAAGGGAAATCAATGTATTAAAAATAATTATATGTTATTTTATTTCCACAATAT
TTATTTATTATTTATTGTTGACCTGCAGGCATGCAAGCTTATCGATACCGTCGACCTCGA
GGGGGGGCCCGGTACCCAGCTTTTGTTCCCTTTAGTGAGGGTTAATTGCGCGCTTGGCGT
AATCATGGTCATAGCTGTTTCCTGTGTGAAATTGTTATCCGCTCACAATTCCACACAACA
TACGAGCCGGAAGCATAAAGTGTAAAGCCTGGGGTGCCTAATGAGTGAGCTAACTCACAT
TAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGGGAAACCTGTCGTGCCAGCTGCATT
AATGAATCGGCCAACGCGCGGGGAGAGGCGGTTTGCGTATTGGGCGCTCTTCCGCTTCCT
CGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAA
AGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAA
AAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGC
TCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGA
CAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTC
CGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTT
CTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCT
GTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTG
AGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTA
GCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCT
ACACTAGAAGGACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAA
GAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTT
GCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTA
CGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTAT
CAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAA
GTATATATGAGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCACCTATCT
CAGCGATCTGTCTATTTCGTTCATCCATAGTTGCCTGACTCCCCGTCGTGTAGATAACTA
CGATACGGGAGGGCTTACCATCTGGCCCCAGTGCTGCAATGATACCGCGAGACCCACGCT
CACCGGCTCCAGATTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCGAGCGCAGAAGTG
GTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGTAA
GTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCATTGCTACAGGCATCGTGGTGT
CACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAAGGCGAGTTA
CATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTGTCA
GAAGTAAGTTGGCCGCAGTGTTATCACTCATGGTTATGGCAGCACTGCATAATTCTCTTA
CTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTGGTGAGTACTCAACCAAGTCATTCT
GAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAATACGGGATAATACCG
CGCCACATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTTCTTCGGGGCGAAAAC
TCTCAAGGATCTTACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTGCACCCAACT
GATCTTCAGCATCTTTTACTTTCACCAGCGTTTCTGGGTGAGCAAAAACAGGAAGGCAAA
ATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATGTTGAATACTCATACTCTTCCTTT
TTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAAT
GTATTTAGAAAAATAAACAAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCAC
Restriction sites to linearize plasmid PmeI
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 1CCCCGCGGCGATAATTATAAATAATATGGTAGATTG
Additional information primer 1L4818 (SacII); PBANKA_140030 Forward
Sequence Primer 2CGGGATCCCCGATAATTCATTTTCTTCTTT
Additional information primer 2L4819 (BamHI); PBANKA_140030 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