From 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.
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-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| 232 | pME-mCherryCAAX H80D** | prenylated mCherry (deleterious H80D mutation; superseded by 450) | 3321 | KMK | 1.0 | |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.