View source for List of entry and destination vectors

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.

'''Constructs in the Tol2kit v1.2 (Nov 2007)'''

{| border="1"

|+ 
! # !! name !! insert !! size (bp) !! made by !! ver !! MTA?
|-
! colspan="7" style="background:#ffdead;" |5' entry clones, attL4-R1 (kan resistant)
|-
!299||[[p5E-bactin2|p5E-''bactin2'']]
|5.3 kb beta-actin promoter (ubiquitous)||7950||KMK||1.0||
|-
!380||[[p5E-h2afx|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|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||

|-
! colspan="7" style="background:#ffdead;" |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|prenylated mCherry (deleterious H80D mutation; superseded by 450)||3321||KMK||1.0||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|prenylated mCherry (some preps contam w/ H80D, superseded by #550)||3321||KMK/SYC||1.2|||-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||
|-
! colspan="7" style="background:#ffdead;" |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||

|-
! 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

|-
! colspan="7" style="background:#ffdead;" |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||

|-
! colspan="7" style="background:#ffdead;" |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.