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Here is a list of the constructs in the Tol2kit. Click on any construct name (second column) to see more information. 12/18/07: Clone pages now included annotated sequence in Genbank format.
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Here is a list of the constructs in the Tol2kit. Click on any construct name (second column) to see more information. 12/18/07: Clone pages now include annotated sequence in Genbank format.
  
 
[[inserts and ends|Here]] are insert sequences for entry clones, and 5' and 3' ends for destination clones, along with a description of how to assemble predicted sequences for expression clones.
 
[[inserts and ends|Here]] are insert sequences for entry clones, and 5' and 3' ends for destination clones, along with a description of how to assemble predicted sequences for expression clones.

Revision as of 04:18, 19 December 2007

Here is a list of the constructs in the Tol2kit. Click on any construct name (second column) to see more information. 12/18/07: Clone pages now include annotated sequence in Genbank format.

Here are insert sequences for entry clones, and 5' and 3' ends for destination clones, along with a description of how to assemble predicted sequences for expression clones.

Constructs in the Tol2kit v1.2 (Nov 2007)

# name insert size (bp) made by ver MTA?
5' entry clones, attL4-R1 (kan resistant)
299 p5E-bactin2 5.3 kb beta-actin promoter (ubiquitous) 7950 KMK 1.0
380 p5E-h2afx 1 kb histone2A-X promoter (quasi-ubiquitous) 3604 KMK 1.0
382 p5E-CMV/SP6 1 kb CMV/SP6 cassette from pCS2+ 3704 KMK 1.0
222 p5E-hsp70 1.5 kb hsp70 promoter for heat-shock induction 4163 BM 1.0
327 p5E-UAS 10x UAS element and basal promoter for Gal4 response 3127 DSC 1.0
228 p5E-MCS multiple-cloning site from pBluescript 2810 BM 1.0
381 p5E-Fse-Asc restriction sites for 8-cutters FseI and AscI 2663 EF 1.0
middle entry clones, attL1-L2 (kan resistant)
383 pME-EGFP EGFP 3327 KMK 1.0
384 pME-EGFPCAAX membrane-localized (prenylated) EGFP; fused to 21 aa of H-ras 3345 KMK 1.0
385 pME-nlsEGFP nuclear-localized EGFP 3342 KMK 1.0
386 pME-mCherry monomeric red fluorophore mCherry 3261 KMK 1.0
232 pME-mCherryCAAX H80D** prenylated mCherry (deleterious H80D mutation; superseded by 450) 3321 KMK 1.0
233 pME-nlsmCherry nuclear-localized mCherry 3288 KMK 1.0
234 pME-H2AmCherry mCherry fused to zebrafish histone H2A.F/Z 3651 KMK 1.0
387 pME-Gal4VP16 Gal4 DNA binding domain fused to VP16 transactivation domain 3204 EF 1.0
237 pME-MCS multiple-cloning site from pBluescript 2765 BM 1.1
450 pME-mCherryCAAX prenylated mCherry 3321 KMK/SYC 1.2
455 pME-EGFP no stop EGFP, no stop (to make N-terminal fusions) 3324 KMK 1.2
456 pME-mCherry no stop mCherry, no stop (to make N-terminal fusions) 3258 KMK 1.2
3' entry clones, attR2-L3 (kan resistant)
302 p3E-polyA SV40 late polyA signal 2838 KMK 1.0
229 p3E-MTpA 6x myc tag for C-terminal fusions, plus SV40 late polyA 3151 BM 1.0
366 p3E-EGFPpA EGFP for C-terminal fusions, plus SV40 late polyA 3634 MEH 1.0
388 p3E-mCherrypA mCherry for C-terminal fusions, plus SV40 late polyA 3586 MEH 1.0
389 p3E-IRES-EGFPpA IRES driving EGFP plus SV40 late polyA 4219 KMK 1.0
390 p3E-IRES-EGFPCAAXpA IRES driving prenylated EGFP plus SV40 late polyA 4250 KMK 1.0
391 p3E-IRES-nlsEGFPpA IRES driving nuclear EGFP plus SV40 late polyA 4248 KMK 1.0
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 7903 CG 0.9 KK
393 pDestTol2CG* pDestTol2pA with cmlc2:egfp transgenesis marker 9708 EF 0.9 KK
394 pDestTol2pA2 pDestTol2pA with ~2 kb extraneous sequence removed 5883 EF 1.0 KK
395 pDestTol2CG2 pDestTol2CG2 with ~2 kb extraneous sequence removed 7796 EF 1.0 KK
donor vectors (kan/chlor resistant; grow in ccdB-tolerant cells)
218 pDONR221 middle donor vector; attP1-P2 flanking chlor/ccdB cassette Invitrogen 1.1
219 pDONRP4-P1R middle donor vector; attP4-P1R flanking chlor/ccdB cassette Invitrogen 1.1
220 pDONRP2R-P3 middle donor vector; attP2R-P3 flanking chlor/ccdB cassette Invitrogen 1.1
transposase clones
396 pCS2FA-transposase for in vitro transcription of capped Tol2 transposase mRNA 6034 KMK 1.0 KK

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
    • SYC, Seok-yong Choi, Chitnis lab
  • ver shows the version of the Tol2kit in which this construct first appeared.
  • *:pDestTol2pA and pDestTol2CG have been superseded by pDestTol2pA2 and pDestTol2CG2. They are not formally part of the v1.0 release, and are no longer being distributed.
  • **:clone 232 has been superseded by clone 450, which lacks the H80D mutation in mCherryCAAX.
  • MTA? indicates whether an MTA is associated with each clone. "KK", Koichi Kawakami.