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|10x UAS element and basal promoter for Gal4 response||DSC
 
|10x UAS element and basal promoter for Gal4 response||DSC
 
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!228||p5E-MCS
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!228||[[p5E-MCS]]
 
|multi-cloning site from pBluescript||BM
 
|multi-cloning site from pBluescript||BM
 
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!381||p5E-Fse-Asc
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!381||[[p5E-Fse-Asc]]
 
|restriction sites for 8-cutters FseI and AscI||EF
 
|restriction sites for 8-cutters FseI and AscI||EF
  

Revision as of 23:46, 12 February 2007

Here is a list of the constructs in the Tol2kit. We have included crudely annotated sequences and maps for several constructs, 2/7/07. Better-formatted information for all sequences is planned Real Soon Now.

Constructs in the Tol2kit v1 (Feb 2007)

# name insert made by
5' entry clones, attL4-R1 (kan resistant)
299 p5E-bactin2 5.3 kb beta-actin promoter (ubiquitous) KMK
380 p5E-h2afx 1 kb histone2A-X promoter (quasi-ubiquitous) KMK
382 p5E-CMV/SP6 1 kb CMV/SP6 cassette from pCS2+ KMK
222 p5E-hsp70 1.5 kb hsp70 promoter for heat-shock induction BM
327 p5E-UASE1B 10x UAS element and basal promoter for Gal4 response DSC
228 p5E-MCS multi-cloning site from pBluescript BM
381 p5E-Fse-Asc restriction sites for 8-cutters FseI and AscI EF
middle entry clones, attL1-L2 (kan resistant)
383 pME-EGFP EGFP KMK
384 pME-EGFPCAAX membrane-localized (prenylated) EGFP; fused to 21 aa of H-ras KMK
385 pME-nlsEGFP nuclear-localized EGFP KMK
386 pME-mCherry monomeric red fluorophore mCherry KMK
232 pME-mCherryCAAX prenylated mCherry KMK
233 pME-nlsmCherry nuclear-localized mCherry KMK
234 pME-H2AmCherry mCherry fused to zebrafish histone H2A.F/Z KMK
387 pME-Gal4VP16 Gal4 DNA binding domain fused to VP16 transactivation domain KMK
3' entry clones, attR2-L3 (kan resistant)
302 p3E-polyA SV40 late polyA signal KMK
229 p3E-MTpA 6x myc tag for C-terminal fusions, plus SV40 late polyA BM
366 p3E-EGFPpA EGFP for C-terminal fusions, plus SV40 late polyA MEH
388 p3E-mCherrypA mCherry for C-terminal fusions, plus SV40 late polyA MEH
389 p3E-IRES-EGFPpA IRES driving EGFP plus SV40 late polyA KMK
390 p3E-IRES-EGFPCAAXpA IRES driving prenylated EGFP plus SV40 late polyA KMK
391 p3E-IRES-nlsEGFPpA IRES driving nuclear EGFP plus SV40 late polyA KMK
destination vectors, attR4-R3 (amp/chlor resistant; grow in ccdB-tolerant cells)
392 pDestTol2pA* attR4-R3 gate with SV40 polyA flanked by Tol2 inverted repeats CG
393 pDestTol2CG* pDestTol2pA with cmlc2:egfp transgenesis marker EF
394 pDestTol2pA2 pDestTol2pA with ~2 kb extraneous sequence removed EF
395 pDestTol2CG2 pDestTol2CG2 with ~2 kb extraneous sequence removed EF
transposase clones
396 pCS2FA-transposase for in vitro transcription of capped Tol2 transposase mRNA KMK

Notes

  • # is the Chien lab stock number
  • made by shows who made the construct:
    • CG: Clemens Grabher, Look lab
    • BM: Ben Mangum, Chien lab
    • DSC, Doug Campbell, Chien lab
    • EF, Esther Fujimoto, Chien lab
    • KMK, Kristen Kwan, Chien lab
    • MEH, Melissa Hardy, Chien lab
  • pDestTol2pA and pDestTol2CG have been superseded by pDestTol2pA2 and pDestTol2CG2, and are no longer being distributed.