TgTn(pb-Gt(ROSA)26Sor/CAG-EYFP*,-tdTomato*,-KDR*,-mEYFP*,-Kdr*,-mKate2*)#Ben
Transgene Detail

Summary

• Symbol: TgTn(pb-Gt(ROSA)26Sor/CAG-EYFP*,-tdTomato*,-KDR*,-mEYFP*,-Kdr*,-mKate2*)#Ben
• Name: transgenic transposon concatemer, Rui Benedito
• MGI ID: MGI:6761550
• Synonyms: iMb-Vegfr2-Mosaic, TgBAC(Rosa)26Sor^{(iMb-Vegfr2-Mosaic)}
• Transgene: TgTn(pb-Gt(ROSA)26Sor/CAG-EYFP*,-tdTomato*,-KDR*,-mEYFP*,-Kdr*,-mKate2*)#Ben Location: unknown
• Alliance: TgTn(pb-Gt(ROSA)26Sor/CAG-EYFP*,-tdTomato*,-KDR*,-mEYFP*,-Kdr*,-mKate2*)#Ben page

Transgene origin

• Germline Transmission: Earliest citation of germline transmission: J:252386
• Parent Cell Line: G4 (ES Cell)
• Strain of Origin: (129S6/SvEvTac x C57BL/6NCrl)F1

Transgene description

• Transgene Type: Transgenic (Conditional ready, Epitope tag, Inserted expressed sequence, Reporter, Transposon concatemer)
• Mutation: Transposon insertion
• TgTn(pb-Gt(ROSA)26Sor/CAG-EYFP*,-tdTomato*,-KDR*,-mEYFP*,-Kdr*,-mKate2*)#Ben expresses 2 genes
  Transgene expresses:
  

  Organism

  Expressed Gene

  Homolog in Mouse

  Note

  mouse

  Kdr (MGI:96683)

  sequence coding for mutated mouse peptide with p.Y1173F substitution is downstream in-frame with T2A sequence which is downstream in-frame with mEYFP gene

  human

  KDR (3791)

  Kdr	(MGI:96683)

  sequence coding for human peptide where extra-cellular domain was deleted is downstream in-frame with T2A sequence which is downstream in-frame with tdTomato gene

• Mutation details: This allele has three membrane-targeted fluorescent protein sequences - each uniquely flanked with pairs of incompatible lox sites (in the same direction) inserted between exons 1 and 2 of the Gt(ROSA)26Sor locus on BAC RP23-401D9. PiggyBAC transposon inverted repeats were added to the BAC to facilitate single-copy and complete insertion into the genome with the PiggyBAC system. The targeting vector is designed with (from 5' to 3') two copies of chicken beta-globin HS4 insulator element (2xINS; increases downstream gene expression and minimizes regulatory interference), a CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter (from pCAGGS), a loxN site, a lox2272 site, a loxP site, the non-cytotoxic/non-fluorescent Phialidium yellow fluorescent protein variant PhiYFP with nuclear localization signal (N-PhiMut), bovine GH polyA signal sequence, a second loxN site, the MbTomato gene (non-oligomerizing tandem-dimer DsRed fluorescent protein variant tdTomato with N-terminal membrane-targeting tag), in-frame Thosea asigna virus 2A cleavage site (T2A; mediates ribosomal skipping) followed by in-frame mutated human KDR cDNA for the active form (where sequence coding for the extra-cellular domain was deleted), bovine GH polyA signal sequence, a second lox2272 site, the MbYFP gene (monomeric enhanced yellow fluorescent protein [mYFP^A206K] with N-terminal membrane-targeting tag), in-frame T2A sequence followed by in-frame mutated mouse Kdr cDNA (with a p.Y1173F mutation that renders the residue non-phosphorylatable and abolishes the peptide's downstream signalling upon binding Vegf), bovine GH polyA signal sequence, a second loxP site, the MbKate2 gene (monomeric far-red fluorescent protein mKate2 with N-terminal membrane-targeting tag), bovine GH polyA signal sequence, and a second 2xINS sequence. (J:252386)

Find Mice (IMSR)

• Mouse strains and cell lines available from the International Mouse Strain Resource (IMSR)
• Carrying this Mutation: Mouse Strains: 0 strains available Cell Lines: 0 lines available

References

• Original: J:252386 Pontes-Quero S, et al., Dual ifgMosaic: A Versatile Method for Multispectral and Combinatorial Mosaic Gene-Function Analysis. Cell. 2017 Aug 10;170(4):800-814.e18
• All: 2 reference(s)