From Tol2Kit
Here is a list of the constructs in the Tol2kit. Click on any construct name (second column) to see more information. Here are insert sequences for entry clones, and 5' and 3' ends for destination clones (used for in silico cloning).
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| 232 | pME-mCherryCAAX H80D** | prenylated mCherry (deleterious H80D mutation; superseded by 450) | 3321 | KMK | 1.0 | |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.