Summary

RMgm-690
Malaria parasiteP. berghei
Genotype
TaggedGene model (rodent): PBANKA_0836800; Gene model (P.falciparum): Not available; Gene product: erythrocyte membrane associated protein 1 fam-a protein (EMAP1: erythrocyte membrane associated protein 1)
Name tag: mCherry
Phenotype Asexual bloodstage; Gametocyte/Gamete; Liver stage;
Last modified: 12 August 2015, 13:33
  *RMgm-690
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Gene tagging
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 cl15cy1
Other information parent lineA reference wild type clone from the ANKA strain of P. berghei (PubMed: PMID: 17406255).
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-690
Principal name1477cl3
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 location at the surface membrane of infected erythrocytes
Gametocyte/GameteGametocytes: the tagged-protein is exported into the cytoplasm of the host erythrocyte and shows a location at the surface membrane of infected erythrocytes
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

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) and/or diagnostic PCR (for correct integration of the construct) have 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 location at the surface membrane of infected erythrocytes.

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_0836800 was c-terminally tagged with EGFP, has been made in P. berghei ANKA 1037m1f1mocl1 (1941; RMgm-1283). 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-489: mutant lacking expression of PBANKA_0836800 and expressing GFP-luciferase under control of a schizont-specific promoter
RMgm-691: P. berghei K173 mutant expressing an mCherry-tagged version of PBANKA_0836800
RMgm-692: mutant lacking expression of PBANKA_010060 (SMAC) and expressing GFP-luciferase under control of a schizont-specific promoter (RMgm-662). This mutant is also expressing an mCherry-tagged version of PBANKA_0836800
RMgm-1283: An additional mutant, in which PBANKA_0836800 was c-terminally tagged with EGFP, has been made in P. berghei ANKA 1037m1f1mocl1.


  Tagged: Mutant parasite with a tagged gene
Details of the target gene
Gene Model of Rodent Parasite PBANKA_0836800
Gene Model P. falciparum ortholog Not available
Gene producterythrocyte membrane associated protein 1 fam-a protein
Gene product: Alternative nameEMAP1: erythrocyte membrane associated protein 1
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
CCGCGGTGGCGGCCGCTCTAGAACTAGTCACAAGTGTGCACTGATCCTGAAGAAATCAAA
AAAGTAACGGAAATTATGGACGAAGCTGTATATCGCTTAAAATACCATTCTAATTTTAAT
AATGGTTATAATCATTATTATGTTGGCAATGAAGAGTTATCTTTATGTTTTACGGAAGAC
AACGGAATAAAATTTGAAAGATTCCATCTTGTGATCCCCAATCCAGATAATGTGCGAAAT
AAGAAACATATTTTTTTATGTTATAAAATTAAGTTCAAATAAAAAAATATAATTATTATA
TATATCCATATATTATTTTTTGTTGCCATTAGTATGAAGAAATAGTAAAAATGTTTTACA
CATTTAGAGGCATTCCCTATATGGATAACTTCGATTTTACGGGTATATGAAAAATAATCA
ATTAAGCATTTTAATATATTATTATTATTATTATTATTATTATTATTACTCTCATGTTAT
ATTCATTGTTTTATTTTTTAATTTTATATGCTACCATTTTTCATCCGTCATATGTCATTT
TTTGTAATATATTAACTTACATGATGAAAAATTATAATTTTTTTTAGAAAAATTGGTTCA
CGAATATTATTCAAATTTATCATTGATGTATAATAGTTATGAAAGCCGAACCACACCATT
CTGTGGATATAGTTTTGTTTTATCTGCAGATATTCAAGTAAGGAAAACTTCTTTTCATTT
ACCCCTCTATATATCATATTTTTCATATTTACACAATTTATTCAAATTCAAATACATTTG
TATTAATTTTGTAGGTATCAGAAGACATAACAATGTTTGTTCAAACATCAGTAGATATAG
ATGACCACAACGAGTTCGATACCGAAAAATATAAAAACACTATTGTAGAAAGTGCAAATT
CATTAAATATTGATGTTTATGCGAACGAATATGACCAAATGGAAAACAAAACAAAATTGT
TTGTTAATTTATCTGGAATTATCGTTAAAAAAAAAAGCGACAGTGTTCGTGTTACATACG
TCAACTCTGTAAGCAATATAAAAATTATAATAACATAATAATTTTATTTTATCATTATCA
AAGATATATACCTTTTCTATAATTTGCCATATTTTAATATTTCTATTCATCTACAAACAA
ATTGACCTATTTTATATTTCTATTCATTCTTTTCTTTTTCCGTAGATTCATAATGGTGAC
TCCTTTCCAAAAGATTTTTGTGACAAACAATTTAGAGTAAAAAATATGCTATATTTGATC
GATTTATTAAAAAATAGTTTTAAAGGATCCGGTGGAGGTGGAAGATCCATGGTGAGCAAG
GGCGAGGAGGATAACATGGCCATCATCAAGGAGTTCATGCGCTTCAAGGTGCACATGGAG
GGCTCCGTGAACGGCCACGAGTTCGAGATCGAGGGCGAGGGCGAGGGCCGCCCCTACGAG
GGCACCCAGACCGCCAAGCTGAAGGTGACCAAGGGTGGCCCCCTGCCCTTCGCCTGGGAC
ATCCTGTCCCCTCAGTTCATGTACGGCTCCAAGGCCTACGTGAAGCACCCCGCCGACATC
CCCGACTACTTGAAGCTGTCCTTCCCCGAGGGCTTCAAGTGGGAGCGCGTGATGAACTTC
GAGGACGGCGGCGTGGTGACCGTGACCCAGGACTCCTCCCTGCAGGACGGCGAGTTCATC
TACAAGGTGAAGCTGCGCGGCACCAACTTCCCCTCCGACGGCCCCGTAATGCAGAAGAAG
ACCATGGGCTGGGAGGCCTCCTCCGAGCGGATGTACCCCGAGGACGGCGCCCTGAAGGGC
GAGATCAAGCAGAGGCTGAAGCTGAAGGACGGCGGCCACTACGACGCTGAGGTCAAGACC
ACCTACAAGGCCAAGAAGCCCGTGCAGCTGCCCGGCGCCTACAACGTCAACATCAAGTTG
GACATCACCTCCCACAACGAGGACTACACCATCGTGGAACAGTACGAACGCGCCGAGGGC
CGCCACTCCACCGGCGGCATGGACGAGCTGTACAAGTAATCTAGAATTATTAATATATAT
GAATATATATACATCGTTGTATGCGTAATATACGTGTAACTGTTTTTTTTAATTTTTATA
TTAACTTTTCCTTATACATTTTAATGCCATTAACTAATAATATTTTACTTGAAAAAATTC
TAAATTTTTATAACTCTAATATAAAATCTGTGAATAATAAAAAATTAGCATAAAGTACGA
ATTAAACATCGAATATTTGTTTTAAAGGGGCACACATTATGTGCATATAATTTTATTCTT
TGAATTTAATTTTTTGCTTATAATTAGATTAAATTAATATAATTTTTTTATTGAAAATAA
AAACTTATTTTAAAAAATTAACAAATGGAAGTTAATGAAAAATACGAAAAAAGTAAAGGG
TTAATTCTTATATGGTCGGTACCGAGCTCGAATTCGATATCGAAATTGAAGGAAAAAACA
TCATTTGTGTTTATATGTTTTTTTTTAATTTTTCAACTTTTTTTATTTAACTATGCTAAT
CATATTATAAACATTTTAAAAATCAAGTCCAACTATTTATGAATCATTGAAGAGACAACA
TAATTATAACAGTAAAAATAATAATAATAATAATAATTTTGTCTTATTGTTTCAAACATA
TTATGTTCCATAAATATCTAAATTTTTAATATGGTCATAAATTCATAGGATTTAATTTAT
ATAAAATATGGCTATTCATACTAGCCATTTTATGTGTGTAAAGTTTTTTTTTTCTGAACA
AAAAATAAAATGTTCATAATTTTAGCTATTTACATGCATGTGCATGCACAAAAAAAAATA
TGCACACAACATACACATTTTTACAGTTATAAATACAATCAATTGGTATAAATATATAAG
TAAGAAAAACGGGATCAAAGCTAGCTTCTGTATTTCCGCTAGACAGCCATCTCCATCTGG
ATTCGTCCGTGCGGGACGTAGCCCACGACCTCAAAATCCTCGGCGGTGAAATCGTCGATT
TCCTTGATGCGTTCCTTGTTGAGGATGTTCACAATGGGGAACGGTCTCGGTTCTCTCCGC
AGCTGCTCTTTTAAAGCCTCGACATGGTTCGTGTAGACATGCGTGTTCCCCATGAAGTGA
ATGAACTCCTTAGGTTTTAGGTTGCAGACGTGTGCAACCATGAGCGTCAAAAGCGAATAG
GAAGCGATGTTGAAGGGGACGCCGAGGCCGACATCGCACGACCGCTGATACATGATGCAC
GACAGCTCCTTCTGGTCGTTCACGTAGAACTGGCACAACAAGTGACAAGGCGGCAGCGCC
ATTTCGTCCAGCGCTGCAGGATTCCAGGCAGTCATGAGCATGCGACGATCTGTTGGATTC
GTTCTCAGCATCTGGATCACATTCTTCAGCTGGTCGACGCCCTGCCCTGTGTAGTCTGTG
TGCATGTCTTTGTATGCCGCGCCGAAGTGTCTCCACTGGAAGCCGTAGCCCGGGCCGATG
TCTCCGACCTCTCGGTGGGGGAGATTGCGCGAATCGAGGAACTCGCGTGTCACATTCTTG
TCCCAGATCTTCACGCCCTTCTCAGAAAGATGGTTTGCGTTCGTGTCGCCGCGAATGAAC
CACAGCAACTCTTCGAGGACCCCTTTCCAGAACACACGCTTTGTGGTGAGAAGTGGAAAG
GCCTGATCCAGCGAGTAGCGCATAGTGCAGCCGAATTTGGAGATGACACCAACGCCCGTT
CGGTCATCCATTGTCCTTCCATTGTTAATAATGTCGGCAATGAGATCAAGGTACTGGAAC
TCTTCATGGCCTCTAAAGTGAACATGCGGAACGGCCCGAATCAGTTCCTTTTGCTCGCGT
TTTTTCCGGTCTTCTTCGTCCATCCACGCCAACACCGGGGCAATGGCTGCGGCCGAAGAA
GGAGCCTGCAACCCGTGCACGGGAGTTGTCTCCCTCGTGGACGTCAAGGAGCTCATTGCG
TTGCTCGGTTCCGCAGTGGCTGCGTCGTCAGTCTTCCTTCTCTTCTCGAGAACCACAAAG
TCGTAGGGAACCCCGTTGTCTGAGAAGGTCTTGGAAATGAAGATGGGTCGATACGTCGCT
TCATTGTCCTTCTCTCTTCCGAGCTCCGGACAAAAGGGAACGAACACAGACTCGGCAGGA
GCTGCAGCCTGCGCAGCAGTTGATTTGTTTGAAAGAATGTCATCTCCGGGGAACGCAGGG
AAGAAAACGTCGCACGGAAACTCGCGGGCTACACGCGTGATGTACAGGTGAGAGGCAACG
CCCAGAGACAGCGCTGCCTCGTACAGTCCCGCTCCTCCCACGACAAAAATCTGGTCGACA
GAATCCTTGTACTCTTCCTCCAGAAGGCTGAGAGCTGCTGGGAGTGAAGCACAGACTCGG
ACGCGCTGCTGGCCTTCAGCTTGAGGCTTCTCCGCCGCAATGTCTTCTTCTTTGAGGGAA
GAGGAAACGACGATGTTCAATCTGTCCACGAGGGGTCTAAACTTTCGAGGCATGCTTTCC
CAGGTTTTCCGTCCCATGACAACGGCGTTGAATCTCTTGCCGACTGATGGAGAGGGAAGT
CCAGAGTCGCCCGTCTTTGCAAATTTCCTGGGAAGCCACCCGTTCAGGCGACTGGCTTCT
TCGGGCGTCGTTTTTGTCACACGAGAAAAGTGTTTGAAATCTGTGGTCAAGTGGGGCCAC
GGGAGGCCGTTGTTGATGCCGATGCCCCTCTTGGGGGTCATCGCGACGACCAGACACACC
GGTTTATGCATTGATCCTTTGTAACATTTAGGTGTGTATTTATATATATAAGCAATATAC
AAATGAAAATACTATTATAAAACGGAAACTCAAAAAATGTTGAAACAAATAGTCAATTTT
TAAAATGAATGTGTAAATTGTTTAAGTTTTATGACAAAAAAAGGAAAAGTGTATATAAAA
TAAAAATTGGAAAAAATTTATATATTACATGCCATATAATACCATTTATTTACTATTTAT
ATCTTAATATAAAAAATTGTAGCAAAAATATATAATCGTATTTTATATATTTAATATTAT
ATAACCCCTTAAAAAAAAAAATTTAATTGAAAAAATTTTAATTTATGCACTTTTTTATTG
TATTATATATATTCCGTTATTTTAATTACACATAAACTTAATAGAAAAATTAATTCGCTG
CTAAATAAAAATTATAGGGCTATATATGATGGGATCATATTCTATCTTTCTCTTTTTCAA
ATYCTTTATATTTTAGAATACAAATCGCTTTAAACAATAAAAAAGAGGTATATCCTACCC
GAATTGACACACACAATAAATAATAAATATATAAATAATAAAAATATAAATATATAAATT
ATAAATATATAAATTATATATATAAATATATTAACATGTGTGTAATTTGTTAAAATTGAT
ATTAAACAGCTTTGCATGGGGAAAGAGTGGCTTTTATAAGCGAATCATATAATACAGTGT
TAATTTTAATATATAAAATATAATTTCCTTAAATTGTTATTTAACATTTTTAATTGCCTA
CCCARGGTAATATAATATCCAAATGACAATCAAAGATAAATGAAATACCGCTCCATTTTT
CCTATATATCCATTTTCATTTTGTGTAAATGTTCATAAAATTTTATCATATACTCATAAA
AAACAGGGGAAAAATGAAAAAATGAACGTCCACCCCGTGTGAATATGCTCATTTTGTATA
CTATACATTTTTTTTTATATAATTTATATTTTCCAAATAAAAAAAATGTTTGTACATGTA
TATATCAATATTGTGTTATGGCATGATGAAAAACAATTATTTTATTCGTTTTTAAATTTG
TAGAATTTCCCTAAATTATTAATGATTTCATATTATAATATATGTATCTATATATTTTTA
ATGTGTAATTCAAAATGGCGAAAAAGAAACAAACCATACTTTATAAGCAACTCTCTTTGA
GCACTTACCATAAAAAATTATACTACTAAATTCAAATTAAATAAAAAAATATATATTATA
AAATTGGCTTTAATTATTTATAAGCACTTGCTTTGTTTTACTCAAAAAGTATTGGCATAA
GTATAAACATAAAACCAATTTTTCACAAAATATATTATATGTCCATACAACTATATCCGA
ACYTTCAAAACACATCCAAATTTCCATTTACCACATGCAAGCAAAAGGATATTAAATAAA
AAAAAATTATTTATATTTTCATGATCACTTAAATAATATTTAAAAAAAAAAGAGACCATA
TATAAAATGTATAATTTAAATATAGCTAACTTATTAAAAAAAATGAATATTGAGCATATA
TCCATAATTTAAAACACTTCTATTTTATGTTATTTGATATATAAGTCGATAAATTATTGC
ATGTATGCATATAGACAGATTAATATACACCAAAATAATAAAAATTATATATAGTTCATA
TATTATTCTGAATAATAAATTAGCTTTCCTCTTAATTCACTTGGTATGTATATATATATA
TATATATATATATATATATATATGGATAAATAAATATAAGGTCTATAACAGGAAAAAATG
TGTATTAACTTTTTTTTATAATTATTTTATATTTTTTCTCCATTTATTTAAATATGTTAA
AATTATTTCCCATTTTATAAACAAAAATTAATCCATAAAATAGCAAAATAACAAAATGAC
AAAATAACAAAAACAAAAAAACGCATTATATGAGTTCATTTTACACAATCCAAAGAAAAA
GAAAATATGCTTAAAATATTCATAACACACTTTTAAGCTATAAAAATATATATATTTATT
TGTGTCTATATTACCAACTCAATTTAATAGATGTGTTATGTGATTAATTCATACACAAAC
ATACAAAAATAAACACCATTTTTGAAAAGATTAATTTGAAATCATATATGTTATTATTAT
ATATTTTTATATCAATGAAATTTAAAAAAATAAAAGGGAAATCAATGTATTAAAAATAAT
TATATGTTATTTTATTTCCACAATATTTATTTATTATTTATTGTTGACCTGCAGGCATGC
AAGCTTATCGATACCGTCGACCTCGAGGGGGGGCCCGGTACCCAGCTTTTGTTCCCTTTA
GTGAGGGTTAATTGCGCGCTTGGCGTAATCATGGTCATAGCTGTTTCCTGTGTGAAATTG
TTATCCGCTCACAATTCCACACAACATACGAGCCGGAAGCATAAAGTGTAAAGCCTGGGG
TGCCTAATGAGTGAGCTAACTCACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTC
GGGAAACCTGTCGTGCCAGCTGCATTAATGAATCGGCCAACGCGCGGGGAGAGGCGGTTT
GCGTATTGGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCT
GCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGA
TAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGC
CGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACG
CTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGG
AAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTT
TCTCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGT
GTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTG
CGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACT
GGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTT
CTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGGACAGTATTTGGTATCTGCGCTCT
GCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCAC
CGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATC
TCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACG
TTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTA
AAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTACCA
ATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTATTTCGTTCATCCATAGTTGC
CTGACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCCCAGTGC
TGCAATGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGCAATAAACCAGCC
AGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTAT
TAATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGT
TGCCATTGCTACAGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTC
CGGTTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGTGCAAAAAAGCGGTTAG
CTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGT
TATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGAC
TGGTGAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTG
CCCGGCGTCAATACGGGATAATACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCAT
TGGAAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGTTC
GATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCGTTTC
TGGGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAA
ATGTTGAATACTCATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTG
TCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATAAACAAATAGGGGTTCCGCG
CACATTTCCCCGAAAAGTGCCAC
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 1AACTAGTCACAAGTGTGCACTGATCC
Additional information primer 1L4960 (SpeI); PBANKA_083680 Forward
Sequence Primer 2AGGATCCTTTAAAACTATTTTTTAATAAATCGATCAAATATAGC
Additional information primer 2L4601 (BamHI); PBANKA_083680 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