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Here is a list of the constructs in the Tol2kit.
+
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.
  
−
Sequences and maps coming Real Soon Now.
+
[[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.
  
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'''Constructs in the Tol2kit v1 (Jan 2007)'''
+
'''Constructs in the Tol2kit v1.2 (Nov 2007)'''
−
 
 
−
"#" is the Chien lab stock number; "made by" indicates who made the construct: CG, Clemens Grabher, Look lab; BM, Ben Mangum; DSC, Doug Campbell; EF, Esther Fujimoto, KMK, Kristen Kwan; MEH, Melissa Hardy (all Chien lab).
 
  
 
{| border="1"
 
{| border="1"
  
 
|+  
 
|+  
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! # !! name !! insert !! made by
+
! # !! name !! insert !! size (bp) !! made by !! ver !! MTA?
−
 
 
 
|-
 
|-
−
! colspan="4" style="background:#ffdead;" |5' entry clones, attL4-R1 (kan resistant)
+
! colspan="7" style="background:#ffdead;" |5' entry clones, attL4-R1 (kan resistant)
 
|-
 
|-
−
!299||p5E-''bactin2''
+
!299||[[p5E-bactin2|p5E-''bactin2'']]
−
|5.3 kb beta-actin promoter (ubiquitous)||KMK
+
|5.3 kb beta-actin promoter (ubiquitous)||7950||KMK||1.0||
 
|-
 
|-
−
!380||p5E-''h2afx''
+
!380||[[p5E-h2afx|p5E-''h2afx'']]
−
|1 kb histone2A-X promoter (quasi-ubiquitous)||KMK
+
|1 kb histone2A-X promoter (quasi-ubiquitous)||3604||KMK||1.0||
 
|-
 
|-
−
!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||1.0||
 
|-
 
|-
−
!222||p5E-''hsp70''
+
!222||[[p5E-hsp70l|p5E-''hsp70l'']]
−
|1.5 kb hsp70 promoter for heat-shock induction||BM
+
|1.5 kb hsp70l promoter for heat-shock induction||4163||BM||1.0||
 
|-
 
|-
−
!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||1.0||
 
|-
 
|-
−
!228||p5E-MCS
+
!228||[[p5E-MCS]]
−
|multi-cloning site from pCS2+||BM
+
|multiple-cloning site from pBluescript||2810||BM||1.0||
 
|-
 
|-
−
!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||1.0||
  
 
|-
 
|-
−
! colspan="4" style="background:#ffdead;" |middle entry clones, attL1-L2 (kan resistant)
+
! colspan="7" style="background:#ffdead;" |middle entry clones, attL1-L2 (kan resistant)
 +
|-
 +
!383||[[pME-EGFP]]
 +
|EGFP||3327||KMK||1.0||
 
|-
 
|-
−
!383||pME-EGFP
+
!384||[[pME-EGFPCAAX]]
−
|EGFP||KMK
+
|membrane-localized (prenylated) EGFP; fused to 21 aa of H-ras||3345||KMK||1.0||
 
|-
 
|-
−
!384||pME-EGFPCAAX
+
!385||[[pME-nlsEGFP]]
−
|membrane-localized (prenylated_ EGFP; fused to 21 aa of H-ras||KMK
+
|nuclear-localized EGFP||3342||KMK||1.0||
 
|-
 
|-
−
!385||pME-nlsEGFP
+
!386||[[pME-mCherry]]
−
|nuclear-localized EGFP||KMK
+
|monomeric red fluorophore mCherry||3261||KMK||1.0||
 
|-
 
|-
−
!386||pME-mCherry
+
!232||[[pME-mCherryCAAX H80D]]**
−
|monomeric red fluorophore mCherry||KMK
+
|prenylated mCherry (deleterious H80D mutation; superseded by 450)||3321||KMK||1.0||
 
|-
 
|-
−
!232||pME-mCherryCAAX
+
!233||[[pME-nlsmCherry]]
−
|prenylated mCherry||KMK
+
|nuclear-localized mCherry||3288||KMK||1.0||
 
|-
 
|-
−
!233||pME-nlsmCherry
+
!234||[[pME-H2AmCherry]]
−
|nuclear-localized mCherry||KMK
+
|mCherry fused to zebrafish histone H2A.F/Z||3651||KMK||1.0||
 
|-
 
|-
−
!234||pME-H2AmCherry
+
!387||[[pME-Gal4VP16]]
−
|mCherry fused to zebrafish histone H2A.F/Z||KMK
+
|Gal4 DNA binding domain fused to VP16 transactivation domain||3204||EF||1.0||
 
|-
 
|-
−
!387||pME-Gal4VP16
+
!237||[[pME-MCS]]
−
|Gal4 DNA binding domain fused to VP16 transactivation domain||KMK
+
|multiple-cloning site from pBluescript||2765||BM||1.1||
−
 
+
|-
 +
!450||[[pME-mCherryCAAX]]**
 +
|prenylated mCherry (some preps contam w/ H80D, superseded by #550)||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||
 +
|-
 +
!550||[[pME-mCherryCAAX]]
 +
|prenylated mCherry||3321||KMK/SYC||1.2||
 
|-
 
|-
−
! colspan="4" style="background:#ffdead;" |3' entry clones, attR2-L3 (kan resistant)
+
! colspan="7" 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||1.0||
 
|-
 
|-
−
!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||1.0||
 
|-
 
|-
−
!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||1.0||
 
|-
 
|-
−
!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||1.0||
 
|-
 
|-
−
!389||p3E-IRES-EGFPpA
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!389||[[p3E-IRES-EGFPpA]]
−
|IRES driving EGFP plus SV40 late polyA||KMK
+
|IRES driving EGFP plus SV40 late polyA||4219||KMK||1.0||
 
|-
 
|-
−
!390||p3E-IRES-EGFPCAAXpA
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!390||[[p3E-IRES-EGFPCAAXpA]]
−
|IRES driving prenylated EGFP plus SV40 late polyA||KMK
+
|IRES driving prenylated EGFP plus SV40 late polyA||4250||KMK||1.0||
 
|-
 
|-
−
!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||1.0||
  
 
|-
 
|-
−
! colspan="4" style="background:#ffdead;" |destination vectors, attR4-R3 (amp/chlor resistant; grow in ccdB-tolerant cells)
+
! colspan="7" style="background:#ffdead;" |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
 +
 
 
|-
 
|-
−
!392||pDestTol2pA
+
! colspan="7" style="background:#ffdead;" |donor vectors (kan/chlor resistant; grow in ccdB-tolerant cells)
−
|attR4-R3 gate with SV40 polyA flanked by Tol2 inverted repeats||CG
 
 
|-
 
|-
−
!393||pDestTol2CG
+
!218||pDONR221
−
|pDestTol2pA with cmlc2:egfp transgenesis marker||EF
+
|middle donor vector; attP1-P2 flanking chlor/ccdB cassette||||Invitrogen||1.1||
 
|-
 
|-
−
!394||pDestTol2pA2
+
!219||pDONRP4-P1R
−
|pDestTol2pA with ~2 kb extraneous sequence removed||EF
+
|5' donor vector; attP4-P1R flanking chlor/ccdB cassette||||Invitrogen||1.1||
 
|-
 
|-
−
!395||pDestTol2CG2
+
!220||pDONRP2R-P3
−
|pDestTol2CG2 with ~2 kb extraneous sequence removed||EF
+
|3' donor vector; attP2R-P3 flanking chlor/ccdB cassette||||Invitrogen||1.1||
  
 
|-
 
|-
−
! colspan="4" style="background:#ffdead;" |transposase clones
+
! colspan="7" 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||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 was superseded by clone 450, which lacks the H80D mutation in mCherryCAAX. Subsequently, some preps of clone 450 turned out to still be clone 232, so we cleaned it up again, and clone 450 has been superseded by 550.
 +
* '''MTA?''' indicates whether an MTA is associated with each clone. "KK", Koichi Kawakami.

Latest revision as of 11:50, 12 October 2009

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-hsp70l 1.5 kb hsp70l 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 (some preps contam w/ H80D, superseded by #550) 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
550 pME-mCherryCAAX prenylated mCherry 3321 KMK/SYC 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 5' donor vector; attP4-P1R flanking chlor/ccdB cassette Invitrogen 1.1
220 pDONRP2R-P3 3' 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 was superseded by clone 450, which lacks the H80D mutation in mCherryCAAX. Subsequently, some preps of clone 450 turned out to still be clone 232, so we cleaned it up again, and clone 450 has been superseded by 550.
  • MTA? indicates whether an MTA is associated with each clone. "KK", Koichi Kawakami.
Retrieved from "http://tol2kit.genetics.utah.edu/index.php?title=List_of_entry_and_destination_vectors&oldid=466"