RMgmDB - Rodent Malaria genetically modified Parasites

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Summary

RMgm-781

Malaria parasite	P. berghei
Genotype
Tagged	Gene model (rodent): PBANKA_1425900; Gene model (P.falciparum): PF3D7_0811600; Gene product: conserved Plasmodium protein, unknown function Name tag: mCherry
Phenotype	Asexual bloodstage;

Last modified: 11 August 2014, 19:07

*RMgm-781

Successful modification	The parasite was generated by the genetic modification
The mutant contains the following genetic modification(s)	Gene tagging
Reference (PubMed-PMID number)	Not published (yet)
MR4 number
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Parent parasite used to introduce the genetic modification
Rodent Malaria Parasite	P. berghei
Parent strain/line	P. berghei ANKA
Name parent line/clone	P. berghei ANKA cl15cy1
Other information parent line	A reference wild type clone from the ANKA strain of P. berghei (PubMed: PMID: 17406255)
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The mutant parasite was generated by
Name PI/Researcher	J.A. Eken; O. Klop; B.M.D. Franke-Fayard; C.J. Janse
Name Group/Department	Leiden Malaria Research Group
Name Institute	Leiden University Medical Center (LUMC)
City	Leiden
Country	The Netherlands
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Name of the mutant parasite
RMgm number	RMgm-781
Principal name	2191
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modification	No
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Phenotype
Asexual blood stage	Expression of the mCherry-tagged protein in blood stages
Gametocyte/Gamete	Not tested
Fertilization and ookinete	Not tested
Oocyst	Not tested
Sporozoite	Not tested
Liver stage	Not tested
Additional remarks phenotype	Mutant/mutation The mutant expresses a C-terminal mCherry-tagged version of PBANKA_142590. The mutant is not cloned and genotype analysis indicates the presence of episomes (see Figure below). We included this mutant in the database since it shows a clear expression pattern in merozoites of schizonts (see Figure below). Protein (function) Phenotype The protein is expressed in blood stages (in merozoites of schizonts). No evidence for transport into the host RBC (see Figure below). The phenotype and expression have not been analysed in detail. Additional information Figure: mCherry::PBANKA_PBANKA_1425900 expression in merozoites of schizonts. Nuclear staining with Hoechst (blue); erthrocyte membrane staining with anti-TER antibodies (green). Southern analysis of separated chromosomes (right) confirms integration of the construct into chromosome 14 (hybridisation with the 3'UTR Pbdhfr/ts gene recognizing the integrated construct in chr. 14 and the endogenous dhfr/ts gene on chr. 7); however, the strong additional hybridisation indicates also the presence of episomes/plasmids Other mutants

Tagged: Mutant parasite with a tagged gene

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Details of the target gene

Gene Model of Rodent Parasite PBANKA_1425900

Gene Model P. falciparum ortholog PF3D7_0811600

Gene product conserved Plasmodium protein, unknown function

Gene product: Alternative name

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Details of the genetic modification

Name of the tag mCherry

Details of tagging C-terminal

Additional remarks: tagging

Commercial source of tag-antibodies

Type of plasmid/construct (Linear) plasmid single cross-over

PlasmoGEM (Sanger) construct/vector used No

Modified PlasmoGEM construct/vector used No

Plasmid/construct map

Plasmid/construct sequence

CTAAATTGTAAGCGTTAATATTTTGTTAAAATTCGCGTTAAATTTTTGTTAAATCAGCTC

ATTTTTTAACCAATAGGCCGAAATCGGCAAAATCCCTTATAAATCAAAAGAATAGACCGA

GATAGGGTTGAGTGTTGTTCCAGTTTGGAACAAGAGTCCACTATTAAAGAACGTGGACTC

CAACGTCAAAGGGCGAAAAACCGTCTATCAGGGCGATGGCCCACTACGTGAACCATCACC

CTAATCAAGTTTTTTGGGGTCGAGGTGCCGTAAAGCACTAAATCGGAACCCTAAAGGGAG

CCCCCGATTTAGAGCTTGACGGGGAAAGCCGGCGAACGTGGCGAGAAAGGAAGGGAAGAA

AGCGAAAGGAGCGGGCGCTAGGGCGCTGGCAAGTGTAGCGGTCACGCTGCGCGTAACCAC

CACACCCGCCGCGCTTAATGCGCCGCTACAGGGCGCGTCCCATTCGCCATTCAGGCTGCG

CAACTGTTGGGAAGGGCGATCGGTGCGGGCCTCTTCGCTATTACGCCAGCTGGCGAAAGG

GGGATGTGCTGCAAGGCGATTAAGTTGGGTAACGCCAGGGTTTTCCCAGTCACGACGTTG

TAAAACGACGGCCAGTGAGCGCGCGTAATACGACTCACTATAGGGCGAATTGGAGCTCCA

CCGCGGTGGCGGCCGCTCTAGAACTAGACATCTTCATTTGACAACTATTATGGAACTTAT

TTATTAGCTAAAAGCGATATAAAATTAAAATATGTCTATGAATATATGTTAAAATATGAT

AAAAAATATAAAAATGCAAGATATATAGAATTTATAATGAAAAATGAAATGCATGGAGAT

ATACATGACCAATTAAATAATATAGAAAACAGTTTATCATGTTTATTTAGCTTAAATGAT

AAAATTAAAATTTCTTATATAATCGACTATTGTAATTATGATAAAAAATCATTTTTCCTT

TTTAACAAATCGTATAAATCTAAAAATATTTTTTCAGTACCATCACAAGATGCATGTACA

ACTCCAAAATATAGTTATCTCAAATTATGCCAAAATGCATCTCTCCTTAGGAAATTTTTT

ACGAACAAAATTGATACTGAAATTGGTCATATACATAAAAAGGAAATAAAAAGAATATCA

ACAATGAAAAGAAGTCTTGAGAGTAATATTGATATTATGAATATTTTTTCCATATCAAAT

GACTCATTAGTTTCGATAATTCATGATAAAAATGAAGACATAACTACGTTTGATGTGAAT

GCGTGCTTTATTGCATCAGCCAAACCCACACTTGGAAATATATTTAACGTGAATAGTCGA

ATGGATCCAGAAACTATGCGCACTAATATCAATAACTCAATATTTTGTACTCCCGTAACT

GTTCCGGTTATATTAAACAGACCAATAATGAAAAGTATAAATGATATATACATAAGAGCA

ATTATTGATATTATAAAAAATATCGAATTTAGAGAGCTTATGTCGATACCTGAAAAATCA

TATCCTTATCATTCCTTTATATATTTTTTTGATAAATTTGCTTATGTTTATAAAAAAAAA

AAATGGTATAAAAATATAAATGAAGTAAATATGTTTATCCCACCCCAAACAATTAAATGG

AATATGTTTTATTATTTATTAAGACATGATATATCTACAGTATACAATAATGAAGAATTT

TTATATGATTTCTTTTACGGAAAAAAATCTACTTCTGTCAAACCACTAGTTAAAAATATA

TTGAAACAATTCATGAGTCATATTACCCTCTTCTTCTATCTTTATAAAGTGGATGAATCT

ACTGGTAATAGATCCGGTGGAGGTGGAAGATCCATGGTGAGCAAGGGCGAGGAGGATAAC

ATGGCCATCATCAAGGAGTTCATGCGCTTCAAGGTGCACATGGAGGGCTCCGTGAACGGC

CACGAGTTCGAGATCGAGGGCGAGGGCGAGGGCCGCCCCTACGAGGGCACCCAGACCGCC

AAGCTGAAGGTGACCAAGGGTGGCCCCCTGCCCTTCGCCTGGGACATCCTGTCCCCTCAG

TTCATGTACGGCTCCAAGGCCTACGTGAAGCACCCCGCCGACATCCCCGACTACTTGAAG

CTGTCCTTCCCCGAGGGCTTCAAGTGGGAGCGCGTGATGAACTTCGAGGACGGCGGCGTG

GTGACCGTGACCCAGGACTCCTCCCTGCAGGACGGCGAGTTCATCTACAAGGTGAAGCTG

CGCGGCACCAACTTCCCCTCCGACGGCCCCGTAATGCAGAAGAAGACCATGGGCTGGGAG

GCCTCCTCCGAGCGGATGTACCCCGAGGACGGCGCCCTGAAGGGCGAGATCAAGCAGAGG

CTGAAGCTGAAGGACGGCGGCCACTACGACGCTGAGGTCAAGACCACCTACAAGGCCAAG

AAGCCCGTGCAGCTGCCCGGCGCCTACAACGTCAACATCAAGTTGGACATCACCTCCCAC

AACGAGGACTACACCATCGTGGAACAGTACGAACGCGCCGAGGGCCGCCACTCCACCGGC

GGCATGGACGAGCTGTACAAGTAATCTAGAATTATTAATATATATGAATATATATACATC

GTTGTATGCGTAATATACGTGTAACTGTTTTTTTTAATTTTTATATTAACTTTTCCTTAT

ACATTTTAATGCCATTAACTAATAATATTTTACTTGAAAAAATTCTAAATTTTTATAACT

CTAATATAAAATCTGTGAATAATAAAAAATTAGCATAAAGTACGAATTAAACATCGAATA

TTTGTTTTAAAGGGGCACACATTATGTGCATATAATTTTATTCTTTGAATTTAATTTTTT

GCTTATAATTAGATTAAATTAATATAATTTTTTTATTGAAAATAAAAACTTATTTTAAAA

AATTAACAAATGGAAGTTAATGAAAAATACGAAAAAAGTAAAGGGTTAATTCTTATATGG

TCGGTACCGAGCTCGAATTCGATATCGAAATTGAAGGAAAAAACATCATTTGTGTTTATA

TGTTTTTTTTTAATTTTTCAACTTTTTTTATTTAACTATGCTAATCATATTATAAACATT

TTAAAAATCAAGTCCAACTATTTATGAATCATTGAAGAGACAACATAATTATAACAGTAA

AAATAATAATAATAATAATAATTTTGTCTTATTGTTTCAAACATATTATGTTCCATAAAT

ATCTAAATTTTTAATATGGTCATAAATTCATAGGATTTAATTTATATAAAATATGGCTAT

TCATACTAGCCATTTTATGTGTGTAAAGTTTTTTTTTTCTGAACAAAAAATAAAATGTTC

ATAATTTTAGCTATTTACATGCATGTGCATGCACAAAAAAAAATATGCACACAACATACA

CATTTTTACAGTTATAAATACAATCAATTGGTATAAATATATAAGTAAGAAAAACGGGAT

CAAAGCTAGCTTCTGTATTTCCGCTAGACAGCCATCTCCATCTGGATTCGTCCGTGCGGG

ACGTAGCCCACGACCTCAAAATCCTCGGCGGTGAAATCGTCGATTTCCTTGATGCGTTCC

TTGTTGAGGATGTTCACAATGGGGAACGGTCTCGGTTCTCTCCGCAGCTGCTCTTTTAAA

GCCTCGACATGGTTCGTGTAGACATGCGTGTTCCCCATGAAGTGAATGAACTCCTTAGGT

TTTAGGTTGCAGACGTGTGCAACCATGAGCGTCAAAAGCGAATAGGAAGCGATGTTGAAG

GGGACGCCGAGGCCGACATCGCACGACCGCTGATACATGATGCACGACAGCTCCTTCTGG

TCGTTCACGTAGAACTGGCACAACAAGTGACAAGGCGGCAGCGCCATTTCGTCCAGCGCT

GCAGGATTCCAGGCAGTCATGAGCATGCGACGATCTGTTGGATTCGTTCTCAGCATCTGG

ATCACATTCTTCAGCTGGTCGACGCCCTGCCCTGTGTAGTCTGTGTGCATGTCTTTGTAT

GCCGCGCCGAAGTGTCTCCACTGGAAGCCGTAGCCCGGGCCGATGTCTCCGACCTCTCGG

TGGGGGAGATTGCGCGAATCGAGGAACTCGCGTGTCACATTCTTGTCCCAGATCTTCACG

CCCTTCTCAGAAAGATGGTTTGCGTTCGTGTCGCCGCGAATGAACCACAGCAACTCTTCG

AGGACCCCTTTCCAGAACACACGCTTTGTGGTGAGAAGTGGAAAGGCCTGATCCAGCGAG

TAGCGCATAGTGCAGCCGAATTTGGAGATGACACCAACGCCCGTTCGGTCATCCATTGTC

CTTCCATTGTTAATAATGTCGGCAATGAGATCAAGGTACTGGAACTCTTCATGGCCTCTA

AAGTGAACATGCGGAACGGCCCGAATCAGTTCCTTTTGCTCGCGTTTTTTCCGGTCTTCT

TCGTCCATCCACGCCAACACCGGGGCAATGGCTGCGGCCGAAGAAGGAGCCTGCAACCCG

TGCACGGGAGTTGTCTCCCTCGTGGACGTCAAGGAGCTCATTGCGTTGCTCGGTTCCGCA

GTGGCTGCGTCGTCAGTCTTCCTTCTCTTCTCGAGAACCACAAAGTCGTAGGGAACCCCG

TTGTCTGAGAAGGTCTTGGAAATGAAGATGGGTCGATACGTCGCTTCATTGTCCTTCTCT

CTTCCGAGCTCCGGACAAAAGGGAACGAACACAGACTCGGCAGGAGCTGCAGCCTGCGCA

GCAGTTGATTTGTTTGAAAGAATGTCATCTCCGGGGAACGCAGGGAAGAAAACGTCGCAC

GGAAACTCGCGGGCTACACGCGTGATGTACAGGTGAGAGGCAACGCCCAGAGACAGCGCT

GCCTCGTACAGTCCCGCTCCTCCCACGACAAAAATCTGGTCGACAGAATCCTTGTACTCT

TCCTCCAGAAGGCTGAGAGCTGCTGGGAGTGAAGCACAGACTCGGACGCGCTGCTGGCCT

TCAGCTTGAGGCTTCTCCGCCGCAATGTCTTCTTCTTTGAGGGAAGAGGAAACGACGATG

TTCAATCTGTCCACGAGGGGTCTAAACTTTCGAGGCATGCTTTCCCAGGTTTTCCGTCCC

ATGACAACGGCGTTGAATCTCTTGCCGACTGATGGAGAGGGAAGTCCAGAGTCGCCCGTC

TTTGCAAATTTCCTGGGAAGCCACCCGTTCAGGCGACTGGCTTCTTCGGGCGTCGTTTTT

GTCACACGAGAAAAGTGTTTGAAATCTGTGGTCAAGTGGGGCCACGGGAGGCCGTTGTTG

ATGCCGATGCCCCTCTTGGGGGTCATCGCGACGACCAGACACACCGGTTTATGCATTGAT

CCTTTGTAACATTTAGGTGTGTATTTATATATATAAGCAATATACAAATGAAAATACTAT

TATAAAACGGAAACTCAAAAAATGTTGAAACAAATAGTCAATTTTTAAAATGAATGTGTA

AATTGTTTAAGTTTTATGACAAAAAAAGGAAAAGTGTATATAAAATAAAAATTGGAAAAA

ATTTATATATTACATGCCATATAATACCATTTATTTACTATTTATATCTTAATATAAAAA

ATTGTAGCAAAAATATATAATCGTATTTTATATATTTAATATTATATAACCCCTTAAAAA

AAAAAATTTAATTGAAAAAATTTTAATTTATGCACTTTTTTATTGTATTATATATATTCC

GTTATTTTAATTACACATAAACTTAATAGAAAAATTAATTCGCTGCTAAATAAAAATTAT

AGGGCTATATATGATGGGATCATATTCTATCTTTCTCTTTTTCAAATYCTTTATATTTTA

GAATACAAATCGCTTTAAACAATAAAAAAGAGGTATATCCTACCCGAATTGACACACACA

ATAAATAATAAATATATAAATAATAAAAATATAAATATATAAATTATAAATATATAAATT

ATATATATAAATATATTAACATGTGTGTAATTTGTTAAAATTGATATTAAACAGCTTTGC

ATGGGGAAAGAGTGGCTTTTATAAGCGAATCATATAATACAGTGTTAATTTTAATATATA

AAATATAATTTCCTTAAATTGTTATTTAACATTTTTAATTGCCTACCCARGGTAATATAA

TATCCAAATGACAATCAAAGATAAATGAAATACCGCTCCATTTTTCCTATATATCCATTT

TCATTTTGTGTAAATGTTCATAAAATTTTATCATATACTCATAAAAAACAGGGGAAAAAT

GAAAAAATGAACGTCCACCCCGTGTGAATATGCTCATTTTGTATACTATACATTTTTTTT

TATATAATTTATATTTTCCAAATAAAAAAAATGTTTGTACATGTATATATCAATATTGTG

TTATGGCATGATGAAAAACAATTATTTTATTCGTTTTTAAATTTGTAGAATTTCCCTAAA

TTATTAATGATTTCATATTATAATATATGTATCTATATATTTTTAATGTGTAATTCAAAA

TGGCGAAAAAGAAACAAACCATACTTTATAAGCAACTCTCTTTGAGCACTTACCATAAAA

AATTATACTACTAAATTCAAATTAAATAAAAAAATATATATTATAAAATTGGCTTTAATT

ATTTATAAGCACTTGCTTTGTTTTACTCAAAAAGTATTGGCATAAGTATAAACATAAAAC

CAATTTTTCACAAAATATATTATATGTCCATACAACTATATCCGAACYTTCAAAACACAT

CCAAATTTCCATTTACCACATGCAAGCAAAAGGATATTAAATAAAAAAAAATTATTTATA

TTTTCATGATCACTTAAATAATATTTAAAAAAAAAAGAGACCATATATAAAATGTATAAT

TTAAATATAGCTAACTTATTAAAAAAAATGAATATTGAGCATATATCCATAATTTAAAAC

ACTTCTATTTTATGTTATTTGATATATAAGTCGATAAATTATTGCATGTATGCATATAGA

CAGATTAATATACACCAAAATAATAAAAATTATATATAGTTCATATATTATTCTGAATAA

TAAATTAGCTTTCCTCTTAATTCACTTGGTATGTATATATATATATATATATATATATAT

ATATATATGGATAAATAAATATAAGGTCTATAACAGGAAAAAATGTGTATTAACTTTTTT

TTATAATTATTTTATATTTTTTCTCCATTTATTTAAATATGTTAAAATTATTTCCCATTT

TATAAACAAAAATTAATCCATAAAATAGCAAAATAACAAAATGACAAAATAACAAAAACA

AAAAAACGCATTATATGAGTTCATTTTACACAATCCAAAGAAAAAGAAAATATGCTTAAA

ATATTCATAACACACTTTTAAGCTATAAAAATATATATATTTATTTGTGTCTATATTACC

AACTCAATTTAATAGATGTGTTATGTGATTAATTCATACACAAACATACAAAAATAAACA

CCATTTTTGAAAAGATTAATTTGAAATCATATATGTTATTATTATATATTTTTATATCAA

TGAAATTTAAAAAAATAAAAGGGAAATCAATGTATTAAAAATAATTATATGTTATTTTAT

TTCCACAATATTTATTTATTATTTATTGTTGACCTGCAGGCATGCAAGCTTATCGATACC

GTCGACCTCGAGGGGGGGCCCGGTACCCAGCTTTTGTTCCCTTTAGTGAGGGTTAATTGC

GCGCTTGGCGTAATCATGGTCATAGCTGTTTCCTGTGTGAAATTGTTATCCGCTCACAAT

TCCACACAACATACGAGCCGGAAGCATAAAGTGTAAAGCCTGGGGTGCCTAATGAGTGAG

CTAACTCACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGGGAAACCTGTCGTG

CCAGCTGCATTAATGAATCGGCCAACGCGCGGGGAGAGGCGGTTTGCGTATTGGGCGCTC

TTCCGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATC

AGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAA

CATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTT

TTTCCATAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTG

GCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCG

CTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAG

CGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTC

CAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAA

CTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGG

TAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCC

TAACTACGGCTACACTAGAAGGACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTAC

CTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGG

TTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTT

GATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGT

CATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAA

ATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGA

GGCACCTATCTCAGCGATCTGTCTATTTCGTTCATCCATAGTTGCCTGACTCCCCGTCGT

GTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCCCAGTGCTGCAATGATACCGCG

AGACCCACGCTCACCGGCTCCAGATTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCGA

GCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGA

AGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCATTGCTACAGG

CATCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATC

AAGGCGAGTTACATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCC

GATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGTTATGGCAGCACTGCA

TAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTGGTGAGTACTCAAC

CAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAATACG

GGATAATACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTTCTTC

GGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCG

TGCACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCGTTTCTGGGTGAGCAAAAAC

AGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATGTTGAATACTCAT

ACTCTTCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATA

CATATTTGAATGTATTTAGAAAAATAAACAAATAGGGGTTCCGCGCACATTTCCCCGAAA

AGTGCCAC

Restriction sites to linearize plasmid BamHI

Selectable marker used to select the mutant parasite tgdhfr

Promoter of the selectable marker pbdhfr

Selection (positive) procedure pyrimethamine

Selection (negative) procedure No

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 1	atctagaCATCTTCATTTGACAACTATTATGG
Additional information primer 1	7123 (XbaI); PBANKA_142590 forward
Sequence Primer 2	cagatctATTACCAGTAGATTCATCCAC
Additional information primer 2	6888 (BglII); PBANKA_142590 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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