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Type and details of transgene |
Is the transgene Plasmodium derived |
Transgene: not Plasmodium |
Transgene name | mCherry |
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Details of the genetic modification |
Inducable system used | No |
Additional remarks inducable system |
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Type of plasmid/construct | (Linear) plasmid double cross-over |
PlasmoGEM (Sanger) construct/vector used | No |
Modified PlasmoGEM construct/vector used | No
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Plasmid/construct map |
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Plasmid/construct sequence |

GGCCGCATGGTGAGCAAGGGCGAGGAGGATAACATGGCCATCATCAAGGAGTTCATGCGC
TTCAAGGTGCACATGGAGGGCTCCGTGAACGGCCACGAGTTCGAGATCGAGGGCGAGGGC
GAGGGCCGCCCCTACGAGGGCACCCAGACCGCCAAGCTGAAGGTGACCAAGGGTGGCCCC
CTGCCCTTCGCCTGGGACATCCTGTCCCCTCAGTTCATGTACGGCTCCAAGGCCTACGTG
AAGCACCCCGCCGACATCCCCGACTACTTGAAGCTGTCCTTCCCCGAGGGCTTCAAGTGG
GAGCGCGTGATGAACTTCGAGGACGGCGGCGTGGTGACCGTGACCCAGGACTCCTCCCTG
CAGGACGGCGAGTTCATCTACAAGGTGAAGCTGCGCGGCACCAACTTCCCCTCCGACGGC
CCCGTAATGCAGAAGAAGACCATGGGCTGGGAGGCCTCCTCCGAGCGGATGTACCCCGAG
GACGGCGCCCTGAAGGGCGAGATCAAGCAGAGGCTGAAGCTGAAGGACGGCGGCCACTAC
GACGCTGAGGTCAAGACCACCTACAAGGCCAAGAAGCCCGTGCAGCTGCCCGGCGCCTAC
AACGTCAACATCAAGTTGGACATCACCTCCCACAACGAGGACTACACCATCGTGGAACAG
TACGAACGCGCCGAGGGCCGCCACTCCACCGGCGGCATGGACGAGCTGTACAAGTAATCT
GATATCTATAATTCATTATGAGTAGTGTAATTCAGAAAGAGAACAAATTGTTTTATGATA
AATATATTTATAAAATAAAAAGTAAGAGAAAAGGAAAAATAATGTTATAGAGAAAAAAAG
GGAAAACGTTATAAAAAAATAGCATCTTAACTATTATTATTTTACTTTATGTGTGTATAA
AACTTTTACAATATATTTTTTTATTTTGAACGTATATAAAAATATTTCCTTTAATTTGTA
GTGTGCAAATAATTTATAAAAATAATTTTTTTAAATATAAATATATTGTCTCGAAAAGAA
ATGAAAATGGATTCTAGCAAATAAACAAGTCTGGCTTCGAAAAGAAATATAAGGCAAAAA
TTATATATAATATATTTGAAAATTATATTTGTGAGAGTGTGTATTATTTTTGTTTGAAAG
ATAATTTTATGGGTTTTTGAGAGTGATTTTTAGTGTTTTATTTGTTTTTTTTTAATAATT
TGTAAGTTTTGTCAATGTAGAGAGAATTATACTTCATAACGTATTATTTGAACGATTAAA
AAATATTTTTTTTAAACAGTGAAATATAAATATGAATGGAAGCAGCCGTATATGCAAACT
TTTTTACATATGTTGTATGCATAACATGCGTTTTTATTTTTTACGAAGTATACATAATTC
CCTTTAAAATATAACTTTATATAATTTGTTTTTATTGTAATGGATATAGTATTGTATAAA
TTTTATATTTATTTCCTAAAGACTTTTTCATAAAAATTGTTGGTGTTCCAGATATATTAT
TTTTAAAAGTCAATATATCAAGTTGTTATATTGTCTTAGAGTTCAAAAGTTTGTCCTAAT
TGAATTTTTTATAACTTTATAATATTTATTGTTTTACTTTGGACCATTAAGTATATATAA
GTAGTAACTTATTAAAATAATAAATTATATGTATCATAATTCATGGCTTTATAGATTCTG
CATTAACCTTAAATATGAAAAACACAAAAATGATGACAAGCATTAAAGCGAAAGGTACCG
AGCTCGAATTCTCTTGAGCCCGTTAATGAAATAGATACAATTCATTCATGTTATATACAT
CTAGAACATAATCTGAATATGGTTCAAGTTAAATGTCCAAAAATTATAAAAAGTGATGAT
ATTTTTGATGGTAATACCATAATAGACACCAAGGTAACATCACGAAGTAGTCAACAAAAT
AATTTTTATTTAGAAAATACAGATGTTGAACCAGAAGAAATAGAGAAATATAAAAATATA
GAATACATACCAGAAAACGATGAAGTAATGCATCTAGACAAAAAAGAAAAGCTAGATGAT
ATATTACCAGGTGTTATCATATTAGATAAAAATAAAATGTTCAAAGAAAAAGGACATTTC
ACTTTTGTTACTCCATTAATTGTAGAAAAGGTATTAATATTAAAAATATATTGTGATAAT
ACTAAAACAATAATTAATAATATGAAAGGGAAAAAAGGTATTACAGTAATAAGGATTTCT
CAAAATACAACAAAAAATAAATTTTATGGATGTGACTTTTCAGGTAATTCTAAAAAAACA
TTTTACTATTCCAATGTTTATGATTTAGAAAAAAAAAATGAGTTTTGTGAAATAGAATTA
AAAGAAAATATAGTAGTTAGCTTAAATTGTCCAACTGGTAAAATTAATCCAAAAAATTGT
TTTAGAAATGTATATATAAAAAGTAATATGAATGAACAAACAACCGAAAATATAGAAAAT
ATATTTAACGAAATAAAAGTTATAGATGCAGATTATTTTATAAATAATTCATCAACCTTT
TTGATGATTTCCAAAATTACAAAAAAAGAGTTTGATTTTTATTGTACATGTGAAGATTAT
AAAACCAAAAATATAGGAACAATATATATTAAAAATTATGAATATCTAGATTCAAAACCT
AAATATAAAAATAAACAAATTTCCTATATAGATGTAGTTCCATACCCGCGGGGAAAGGGC
GAATTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCGTTACCCAACT
TAATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCAC
CGATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGCGCCTGATGCGGTATTT
TCTCCTTACGCATCTGTGCGGTATTTCACACCGCATATGGTGCACTCTCAGTACAATCTG
CTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTG
ACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTG
CATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAAGGGCCTCGTGAT
ACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCAC
TTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATAT
GTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAG
TATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCC
TGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGC
ACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCC
CGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATC
CCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTT
GGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATT
ATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGAT
CGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCT
TGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGAT
GCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGC
TTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCG
CTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTC
TCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTA
CACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGC
CTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGA
TTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCAT
GACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGAT
CAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAA
ACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAA
GGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTT
AGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTT
ACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATA
GTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTT
GGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCATTGAGAAAGCGCCAC
GCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGA
GCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCG
CCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAA
AAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACAT
GTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGC
TGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGA
AGAGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTG
GCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTA
GCTCACTCATTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGG
AATTGTGAGCGGATAACAATTTCACACAGGAAACAGCTATGACCATGATTACGCCAAGCT
TCCGCGGGTATATGGTAAAGAACCTACTAACACAATAAAATATTTAAATAATGTATTTCC
TATAAATAAATTTACAGATTTATTTTTTAATACAAAAGATATAGATATACCAGAAATAAA
TGATCAGTTTAAAGGTTTTAAATTCTTTATGACATCATTTATAAATCATGGATCATATCC
ACTAACAATAGAATGTGGTGTAACAAATGGTGGAACTAGTTATAAAAGAGCAATTATTTT
ATTGCATGTTCGAACTGATTTAAAAGATAGACCAGTTTCATTTTGTGATTTTCGAAAAGG
AGAATTATATAATTATTTGAATGCTTATACTGAAGGGGATGTATGCATAATAATTTCCAA
ATCAAATACAAGTTTTGGTTTTAGATGCCCAGTAAATACAAAAAAAATGCCAAAAAATTG
TTTTACGCAAGTATATGAAAAAGGGTATCTAAATGACGCCAATAAAATTAATACTAAAAA
TGTTATTAACTATTCATTTGAAAATCCAGAATATGCGCTAGCTGGTTYTAATTATACATT
AACAAAATCGTATCAATTTGAATGTCATTGTGTAGATAAAGAAACAGAACAAATTGTAAA
AACGGTTTTAGTCAAATATGTAAATGAAGATGAAATATATGATTATAATGATTTTCCAAT
GGTGAATCACAAACCTATTATTGCACATCCAAATAAAACACATCAAGCTTGCATGCCTGC
AGGGTGATAGTGTAGATTTTTTTGTTTGACCATTGCATCATTGTCTTTTATTCATCGATT
TTATTATGTTATATATATTTTTTTAAATATCATAATATTTTTTCAATATAGCTTCTTTGA
GCAAATACTGAACAATAAGGAATGCTTCTATGCATCCGTGGATGTATATGTACGTAATAA
AATAATTTTTTGTCCATAAAAATATCTTTAACAGCTGTTATACAAAAGCATTGAATTGGC
TGATACATTTTTGTCGTAAATAAAATAAATGAATTAATGGTACAAAATTACAATAAAAAG
GAAGCGAATTTATTGCATTTTGTTTAATTTATGTCCTTTCCATTTTATTAATACAATTAT
GGTACTAGTAATTCGTATTAAACATATATTTTGCATGTACCAAATGTTTGGGAAATAAGC
ATATGGATCCCACTAAATTTAATAATATTCTGCGATTTTTCTTATATTTACTATTAAATA
TAATGGATTCATTTTTTGATGCATGCAATTTTTTCTTTTAATGTATTAATTAGTGTGACA
ATTTTGAAAAACTTTACTCATTTATTTATTTTCCTTATTATATTGGATTTATAATACCAG
TATTATTATATAAACCACATTAAATAATTTGTAATTTATTCGAGGGTATCAAAAAAAACA
TATACAAAATGCATATATCCACATATGGCTCATTATAGGGTGAATAAAAATGGGAACAGC
AGATTAATTATTATATCATGAAAGTAATGAAAAAATTTAAATTATATGGATATATACATA
TATAATATGTTAAAAAATAATAATAAATAAATAAAATGTATTATAAATCTTATAAAATAA
GTTTTGTTGTGAATAATCCACGAAATATGCCATAAATAGACACTGAACAAATTAGTGGTT
CTTAATATTATTTTGGATACATGCGGATATTATCATTGTCAGATGATTTATTTTTTGTTA
TTTTTAAATTATAAATATTCATATTTTATAGTCCTCAAAAAATAGGATGTTTTATTTTTT
TATAGCTATATTTTATGGTTGATCCTTTCCTTTTATGGTGTTTCATAAAAATTTTATTGA
GCTATATACAATCCAATAAAAAAAGGCAATTGAATTTTGAAATATATTAAACTATTTTTA
TAATAAAATAAATATAATTTTTAAATAATGTTATATTATATATAATTTTTATCTTTACAC
AGAAATTTTTTTTTAGAGTCCAATATATATAATTAGTTATATATATAGAGATATACACTA
CATACACCACCATAAATAATTATAAGGAAAACCAGTTATTTTAATTTTAACTGAAGAAAT
TAAATAAATATATAAAAAAGAAAAGAACAAAAAATAAAACGACAGCAACCTTAAAATTTT
ATTATTATTACATATTTATACATAAAAAAAATAAAATACTTAGACAAAAAATAATAAATA
TTATAGATCGATATTAGCATACATATATACCTTTCAGCACATAATTATTACGTCTGC
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Restriction sites to linearize plasmid |
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Selectable marker used to select the mutant parasite | hdhfr/yfcu |
Promoter of the selectable marker | eef1a |
Selection (positive) procedure | No |
Selection (negative) procedure | 5-fluorocytosine (5-FC) |
Additional remarks genetic modification | DNA constructs for generation of the transgenic reporter line expressing mCherry is based on the standard GIMO DNA construct pL0043 (Lin, Annoura et al. 2011). This construct contains the 5’ and 3’ targeting sequences (TRs) for the 230p locus as well as a multiple-cloning site for integration of transgene-expression cassettes. This construct integrates by double cross-over homologous recombination and replaces the positive-negative selectable marker (hdhfr::yfcu; SM) cassette with the transgene-expression cassette in the 230p locus in P. berghei parasites of GIMOPbANKA. The mCherry expression cassette contains mCherry flanked by the 5’ and 3’UTR (promoter and transcription terminator) sequences of P. berghei uis4 (PBANKA_050120) which were amplified from P. berghei WT genomic DNA. The 5'UTR was amplified with primer pair 7169 (tatcctgcaggGTGATAGTGTAGATTTTTTTGTTTGAC) and 7170 (ataagaatgcggccgcAGACGTAATAATTATGTGCTGAAAGG). The 3'UTR was amplified with primer pair 7171 (cggatatcTATAATTCATTATGAGTAGTGTAATTCAG) and 7172 (ggccggtaccTTTCGCTTTAATGCTTGTCATC)
The mCherry-expression cassette was cloned in pL0043 resulting in plasmid pL1937. |
Additional remarks selection procedure | This reporter mutant expressing mCherry does not contain a drug-selectable marker.
The mutant has been generated in the reference line GIMOPbANKA (RMgm-687). The GIMO mother line is used for introduction of transgenes into the modified 230p locus through transfection with constructs that target the 230p locus. These constructs insert into the 230p locus (‘gene insertion’), thereby removing the hdhfr::yfcu selectable marker (‘marker out’) from the genome of the mother lines. Transgenic parasites that are marker-free are subsequently selected by applying negative drug selection using 5-FC. This selection procedure is performed in vivo in mice. |
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Other details transgene |
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Promoter |
Gene Model of Parasite |
PBANKA_0501200
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Gene Model P. falciparum ortholog |
PF3D7_1016900
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Gene product | early transcribed membrane protein 10.3 | protein of early gametocyte 4 |
Gene product: Alternative name | UIS4, up-regulated in infective sporozoites; ETRAMP10.3 |
Primer information details of the primers used for amplification of the promoter sequence 
Primer information details of the primers used for amplification of the promoter sequence
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_0501200
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Gene product | early transcribed membrane protein up-regulated in infective sporozoites |
Gene product: Alternative name | UIS4, up-regulated in infective sporozoites; ETRAMP10.3 |
Primer information details of the primers used for amplification the 3'-UTR sequences 
Primer information details of the primers used for amplification the 3'-UTR sequences
Sequence Primer 1 | |
Additional information primer 1 | |
Sequence Primer 2 | |
Additional information primer 2 | |
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Insertion/Replacement locus |
Replacement / Insertion | Replacement locus |
Gene Model of Parasite |
PBANKA_0306000
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Gene product | 6-cysteine protein |
Gene product: Alternative name | 230p |
Primer information details of the primers used for amplification of the target sequences
Primer information details of the primers used for amplification of the target sequences
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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