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! colspan="7" | '''3' entry clones, attR2-L3 (kan resistant)''' | ! colspan="7" | '''3' entry clones, attR2-L3 (kan resistant)''' | ||
|- | |- | ||
| ''' | | '''302''' | ||
| [[ | | [[p3E-polyA|'''p3E-polyA''']] | ||
| | | SV40 late polyA signal | ||
| | | 2838 | ||
| KMK | |||
| 1.0 | |||
| | |||
|- | |||
| '''229''' | |||
| [[p3E-MTpA|'''p3E-MTpA''']] | |||
| 6x myc tag for C-terminal fusions, plus SV40 late polyA | |||
| 3151 | |||
| BM | | BM | ||
| 1.1 | | 1.0 | ||
| | |||
|- | |||
| '''366''' | |||
| [[p3E-EGFPpA|'''p3E-EGFPpA''']] | |||
| EGFP for C-terminal fusions, plus SV40 late polyA | |||
| 3634 | |||
| MEH | |||
| 1.0 | |||
| | | | ||
|- | |- | ||
| ''' | | '''388''' | ||
| [[ | | [[p3E-mCherrypA|'''p3E-mCherrypA''']] | ||
| | | mCherry for C-terminal fusions, plus SV40 late polyA | ||
| | | 3586 | ||
| | | MEH | ||
| 1. | | 1.0 | ||
| | | | ||
|- | |- | ||
| ''' | | '''389''' | ||
| [[ | | [[p3E-IRES-EGFPpA|'''p3E-IRES-EGFPpA''']] | ||
| | | IRES driving EGFP plus SV40 late polyA | ||
| | | 4219 | ||
| | | KMK | ||
| 1. | | 1.0 | ||
| | | | ||
|- | |- | ||
| ''' | | '''390''' | ||
| [[ | | [[p3E-IRES-EGFPCAAXpA|'''p3E-IRES-EGFPCAAXpA''']] | ||
| | | IRES driving prenylated EGFP plus SV40 late polyA | ||
| | | 4250 | ||
| | | KMK | ||
| 1. | | 1.0 | ||
| | | | ||
|- | |- | ||
| ''' | | '''391''' | ||
| [[ | | [[p3E-IRES-nlsEGFPpA|'''p3E-IRES-nlsEGFPpA''']] | ||
| | | IRES driving nuclear EGFP plus SV40 late polyA | ||
| | | 4248 | ||
| | | KMK | ||
| 1. | | 1.0 | ||
| | | | ||
|- | |- | ||
| ''' | ! colspan="7" | '''destination vectors, attR4-R3 (amp/chlor resistant; grow in ccdB-tolerant cells)''' | ||
| [[ | |- | ||
| | | '''392''' | ||
| | | [[pDestTol2pA*|'''pDestTol2pA*''']] | ||
| | | attR4-R3 gate with SV40 polyA flanked by Tol2 inverted repeats | ||
| 7903 | |||
| CG | |||
| 0.9 | |||
| KK | |||
|- | |||
| '''393''' | |||
| [[pDestTol2CG*|'''pDestTol2CG*''']] | |||
| pDestTol2pA with cmlc2:egfp transgenesis marker | |||
| 9708 | |||
| EF | |||
| 0.9 | |||
| KK | |||
|- | |||
| '''394''' | |||
| [[pDestTol2pA2|'''pDestTol2pA2''']] | |||
| pDestTol2pA with ~2 kb extraneous sequence removed | |||
| 5883 | |||
| EF | |||
| 1.0 | |||
| KK | |||
|- | |||
| '''395''' | |||
| [[pDestTol2CG2|'''pDestTol2CG2''']] | |||
| pDestTol2CG2 with ~2 kb extraneous sequence removed | |||
| 7796 | |||
| EF | |||
| 1.0 | |||
| KK | |||
|- | |||
! colspan="7" | '''donor vectors (kan/chlor resistant; grow in ccdB-tolerant cells)''' | |||
|- | |||
| '''218''' | |||
| [[pDONR221|'''pDONR221''']] | |||
| middle donor vector; attP1-P2 flanking chlor/ccdB cassette | |||
| | |||
| Invitrogen | |||
| 1.1 | | 1.1 | ||
| | | | ||
|- | |- | ||
| ''' | | '''219''' | ||
| [[ | | [[pDONRP4-P1R|'''pDONRP4-P1R''']] | ||
| | | 5' donor vector; attP4-P1R flanking chlor/ccdB cassette | ||
| | | | ||
| | | Invitrogen | ||
| 1.1 | | 1.1 | ||
| | | | ||
|- | |- | ||
| ''' | | '''220''' | ||
| [[ | | [[pDONRP2R-P3|'''pDONRP2R-P3''']] | ||
| | | 3' donor vector; attP2R-P3 flanking chlor/ccdB cassette | ||
| | | | ||
| | | Invitrogen | ||
| 1.1 | | 1.1 | ||
| | | | ||
|- | |||
! colspan="7" | '''transposase clones''' | |||
|- | |||
| '''396''' | |||
| [[pCS2FA-transposase|'''pCS2FA-transposase''']] | |||
| for in vitro transcription of capped Tol2 transposase mRNA | |||
| 6034 | |||
| KMK | |||
| 1.0 | |||
| KK | |||
|- | |- | ||
|} | |} | ||
Line 254: | Line 316: | ||
:* '''CG''': Clemens Grabher, Look lab | :* '''CG''': Clemens Grabher, Look lab | ||
:* '''BM''': Ben Mangum, Chien lab | :* '''BM''': Ben Mangum, Chien lab | ||
:* '''DSC''': Doug Campbell, Chien lab | :* '''DSC''': Doug Campbell, Chien lab | ||
:* '''EF''': Esther Fujimoto, Chien lab | :* '''EF''': Esther Fujimoto, Chien lab | ||
:* '''KMK''': Kristen Kwan, Chien lab | :* '''KMK''': Kristen Kwan, Chien lab | ||
:* '''MEH''': Melissa Hardy, Chien lab | :* '''MEH''': Melissa Hardy, Chien lab | ||
:* '''SYC''': Seok-yong Choi, Chitnis lab | :* '''SYC''': Seok-yong Choi, Chitnis lab | ||
Latest revision as of 18:01, 29 May 2025
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.
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.