From Tol2Kit
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Here is a list of the constructs in the Tol2kit.
 
Here is a list of the constructs in the Tol2kit.
We have included crudely annotated sequences and maps for several constructs, 2/7/07.
+
We have included crudely annotated sequences and maps for several constructs, 2/16/07. Sizes in bp have been added for all constructs, 2/20/07.
 
Better-formatted information for all sequences is planned Real Soon Now.
 
Better-formatted information for all sequences is planned Real Soon Now.
  
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|+  
 
|+  
! # !! name !! insert !! made by
+
! # !! name !! insert !! size (bp) !! made by
  
 
|-
 
|-
! colspan="4" style="background:#ffdead;" |5' entry clones, attL4-R1 (kan resistant)
+
! colspan="5" style="background:#ffdead;" |5' entry clones, attL4-R1 (kan resistant)
 
|-
 
|-
 
!299||p5E-''bactin2''
 
!299||p5E-''bactin2''
|5.3 kb beta-actin promoter (ubiquitous)||KMK
+
|5.3 kb beta-actin promoter (ubiquitous)||7950||KMK
 
|-
 
|-
 
!380||p5E-''h2afx''
 
!380||p5E-''h2afx''
|1 kb histone2A-X promoter (quasi-ubiquitous)||KMK
+
|1 kb histone2A-X promoter (quasi-ubiquitous)||3604||KMK
 
|-
 
|-
 
!382||p5E-CMV/SP6
 
!382||p5E-CMV/SP6
|1 kb CMV/SP6 cassette from pCS2+||KMK
+
|1 kb CMV/SP6 cassette from pCS2+||3704||KMK
 
|-
 
|-
 
!222||p5E-''hsp70''
 
!222||p5E-''hsp70''
|1.5 kb hsp70 promoter for heat-shock induction||BM
+
|1.5 kb hsp70 promoter for heat-shock induction||4163||BM
 
|-
 
|-
!327||p5E-UASE1B
+
!327||p5E-UAS
|10x UAS element and basal promoter for Gal4 response||DSC
+
|10x UAS element and basal promoter for Gal4 response||3127||DSC
 
|-
 
|-
 
!228||[[p5E-MCS]]
 
!228||[[p5E-MCS]]
|multi-cloning site from pBluescript||BM
+
|multi-cloning site from pBluescript||2810||BM
 
|-
 
|-
 
!381||[[p5E-Fse-Asc]]
 
!381||[[p5E-Fse-Asc]]
|restriction sites for 8-cutters FseI and AscI||EF
+
|restriction sites for 8-cutters FseI and AscI||2663||EF
  
 
|-
 
|-
! colspan="4" style="background:#ffdead;" |middle entry clones, attL1-L2 (kan resistant)
+
! colspan="5" style="background:#ffdead;" |middle entry clones, attL1-L2 (kan resistant)
 
|-
 
|-
 
!383||pME-EGFP
 
!383||pME-EGFP
|EGFP||KMK
+
|EGFP||3327||KMK
 
|-
 
|-
 
!384||pME-EGFPCAAX
 
!384||pME-EGFPCAAX
|membrane-localized (prenylated) EGFP; fused to 21 aa of H-ras||KMK
+
|membrane-localized (prenylated) EGFP; fused to 21 aa of H-ras||3345||KMK
 
|-
 
|-
 
!385||pME-nlsEGFP
 
!385||pME-nlsEGFP
|nuclear-localized EGFP||KMK
+
|nuclear-localized EGFP||3342||KMK
 
|-
 
|-
 
!386||pME-mCherry
 
!386||pME-mCherry
|monomeric red fluorophore mCherry||KMK
+
|monomeric red fluorophore mCherry||3261||KMK
 
|-
 
|-
 
!232||pME-mCherryCAAX
 
!232||pME-mCherryCAAX
|prenylated mCherry||KMK
+
|prenylated mCherry||3321||KMK
 
|-
 
|-
 
!233||pME-nlsmCherry
 
!233||pME-nlsmCherry
|nuclear-localized mCherry||KMK
+
|nuclear-localized mCherry||3288||KMK
 
|-
 
|-
 
!234||pME-H2AmCherry
 
!234||pME-H2AmCherry
|mCherry fused to zebrafish histone H2A.F/Z||KMK
+
|mCherry fused to zebrafish histone H2A.F/Z||3651||KMK
 
|-
 
|-
 
!387||pME-Gal4VP16
 
!387||pME-Gal4VP16
|Gal4 DNA binding domain fused to VP16 transactivation domain||KMK
+
|Gal4 DNA binding domain fused to VP16 transactivation domain||3204||EF
  
 
|-
 
|-
! colspan="4" style="background:#ffdead;" |3' entry clones, attR2-L3 (kan resistant)
+
! colspan="5" style="background:#ffdead;" |3' entry clones, attR2-L3 (kan resistant)
 
|-
 
|-
 
!302||p3E-polyA
 
!302||p3E-polyA
|SV40 late polyA signal||KMK
+
|SV40 late polyA signal||2838||KMK
 
|-
 
|-
 
!229||p3E-MTpA
 
!229||p3E-MTpA
|6x myc tag for C-terminal fusions, plus SV40 late polyA||BM
+
|6x myc tag for C-terminal fusions, plus SV40 late polyA||3151||BM
 
|-
 
|-
 
!366||p3E-EGFPpA
 
!366||p3E-EGFPpA
|EGFP for C-terminal fusions, plus SV40 late polyA||MEH
+
|EGFP for C-terminal fusions, plus SV40 late polyA||3634||MEH
 
|-
 
|-
 
!388||p3E-mCherrypA
 
!388||p3E-mCherrypA
|mCherry for C-terminal fusions, plus SV40 late polyA||MEH
+
|mCherry for C-terminal fusions, plus SV40 late polyA||3586||MEH
 
|-
 
|-
 
!389||[[p3E-IRES-EGFPpA]]
 
!389||[[p3E-IRES-EGFPpA]]
|IRES driving EGFP plus SV40 late polyA||KMK
+
|IRES driving EGFP plus SV40 late polyA||4219||KMK
 
|-
 
|-
 
!390||[[p3E-IRES-EGFPCAAXpA]]
 
!390||[[p3E-IRES-EGFPCAAXpA]]
|IRES driving prenylated EGFP plus SV40 late polyA||KMK
+
|IRES driving prenylated EGFP plus SV40 late polyA||4250||KMK
 
|-
 
|-
 
!391||[[p3E-IRES-nlsEGFPpA]]
 
!391||[[p3E-IRES-nlsEGFPpA]]
|IRES driving nuclear EGFP plus SV40 late polyA||KMK
+
|IRES driving nuclear EGFP plus SV40 late polyA||4248||KMK
  
 
|-
 
|-
! colspan="4" style="background:#ffdead;" |destination vectors, attR4-R3 (amp/chlor resistant; grow in ccdB-tolerant cells)
+
! colspan="5" style="background:#ffdead;" |destination vectors, attR4-R3 (amp/chlor resistant; grow in ccdB-tolerant cells)
 
|-
 
|-
 
!392||pDestTol2pA*
 
!392||pDestTol2pA*
|attR4-R3 gate with SV40 polyA flanked by Tol2 inverted repeats||CG
+
|attR4-R3 gate with SV40 polyA flanked by Tol2 inverted repeats||7897||CG
 
|-
 
|-
 
!393||pDestTol2CG*
 
!393||pDestTol2CG*
|pDestTol2pA with cmlc2:egfp transgenesis marker||EF
+
|pDestTol2pA with cmlc2:egfp transgenesis marker||9702||EF
 
|-
 
|-
 
!394||[[pDestTol2pA2]]
 
!394||[[pDestTol2pA2]]
|pDestTol2pA with ~2 kb extraneous sequence removed||EF
+
|pDestTol2pA with ~2 kb extraneous sequence removed||5877||EF
 
|-
 
|-
 
!395||[[pDestTol2CG2]]
 
!395||[[pDestTol2CG2]]
|pDestTol2CG2 with ~2 kb extraneous sequence removed||EF
+
|pDestTol2CG2 with ~2 kb extraneous sequence removed||7790||EF
  
 
|-
 
|-
! colspan="4" style="background:#ffdead;" |transposase clones
+
! colspan="5" style="background:#ffdead;" |transposase clones
 
|-
 
|-
 
!396||pCS2FA-transposase
 
!396||pCS2FA-transposase
|for in vitro transcription of capped Tol2 transposase mRNA||KMK
+
|for in vitro transcription of capped Tol2 transposase mRNA||6034||KMK
 
|}
 
|}
 
'''Notes'''
 
'''Notes'''

Revision as of 20:42, 20 February 2007

Here is a list of the constructs in the Tol2kit. We have included crudely annotated sequences and maps for several constructs, 2/16/07. Sizes in bp have been added for all constructs, 2/20/07. Better-formatted information for all sequences is planned Real Soon Now.

Constructs in the Tol2kit v1 (Feb 2007)

# name insert size (bp) made by
5' entry clones, attL4-R1 (kan resistant)
299 p5E-bactin2 5.3 kb beta-actin promoter (ubiquitous) 7950 KMK
380 p5E-h2afx 1 kb histone2A-X promoter (quasi-ubiquitous) 3604 KMK
382 p5E-CMV/SP6 1 kb CMV/SP6 cassette from pCS2+ 3704 KMK
222 p5E-hsp70 1.5 kb hsp70 promoter for heat-shock induction 4163 BM
327 p5E-UAS 10x UAS element and basal promoter for Gal4 response 3127 DSC
228 p5E-MCS multi-cloning site from pBluescript 2810 BM
381 p5E-Fse-Asc restriction sites for 8-cutters FseI and AscI 2663 EF
middle entry clones, attL1-L2 (kan resistant)
383 pME-EGFP EGFP 3327 KMK
384 pME-EGFPCAAX membrane-localized (prenylated) EGFP; fused to 21 aa of H-ras 3345 KMK
385 pME-nlsEGFP nuclear-localized EGFP 3342 KMK
386 pME-mCherry monomeric red fluorophore mCherry 3261 KMK
232 pME-mCherryCAAX prenylated mCherry 3321 KMK
233 pME-nlsmCherry nuclear-localized mCherry 3288 KMK
234 pME-H2AmCherry mCherry fused to zebrafish histone H2A.F/Z 3651 KMK
387 pME-Gal4VP16 Gal4 DNA binding domain fused to VP16 transactivation domain 3204 EF
3' entry clones, attR2-L3 (kan resistant)
302 p3E-polyA SV40 late polyA signal 2838 KMK
229 p3E-MTpA 6x myc tag for C-terminal fusions, plus SV40 late polyA 3151 BM
366 p3E-EGFPpA EGFP for C-terminal fusions, plus SV40 late polyA 3634 MEH
388 p3E-mCherrypA mCherry for C-terminal fusions, plus SV40 late polyA 3586 MEH
389 p3E-IRES-EGFPpA IRES driving EGFP plus SV40 late polyA 4219 KMK
390 p3E-IRES-EGFPCAAXpA IRES driving prenylated EGFP plus SV40 late polyA 4250 KMK
391 p3E-IRES-nlsEGFPpA IRES driving nuclear EGFP plus SV40 late polyA 4248 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 7897 CG
393 pDestTol2CG* pDestTol2pA with cmlc2:egfp transgenesis marker 9702 EF
394 pDestTol2pA2 pDestTol2pA with ~2 kb extraneous sequence removed 5877 EF
395 pDestTol2CG2 pDestTol2CG2 with ~2 kb extraneous sequence removed 7790 EF
transposase clones
396 pCS2FA-transposase for in vitro transcription of capped Tol2 transposase mRNA 6034 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.