From Tol2Kit
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| − | 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 | + | 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. | [[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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|1 kb CMV/SP6 cassette from pCS2+||3704||KMK||1.0|| | |1 kb CMV/SP6 cassette from pCS2+||3704||KMK||1.0|| | ||
|- | |- | ||
| − | !222||[[p5E- | + | !222||[[p5E-hsp70l|p5E-''hsp70l'']] |
| − | |1.5 kb | + | |1.5 kb hsp70l promoter for heat-shock induction||4163||BM||1.0|| |
|- | |- | ||
!327||[[p5E-UAS]] | !327||[[p5E-UAS]] | ||
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|multiple-cloning site from pBluescript||2765||BM||1.1|| | |multiple-cloning site from pBluescript||2765||BM||1.1|| | ||
|- | |- | ||
| − | !450||[[pME-mCherryCAAX]] | + | !450||[[pME-mCherryCAAX]]** |
| − | |prenylated mCherry||3321||KMK/SYC||1.2|| | + | |prenylated mCherry (some preps contam w/ H80D, superseded by #550)||3321||KMK/SYC||1.2|| |
|- | |- | ||
!455||[[pME-EGFP no stop]] | !455||[[pME-EGFP no stop]] | ||
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!456||[[pME-mCherry no stop]] | !456||[[pME-mCherry no stop]] | ||
|mCherry, no stop (to make N-terminal fusions)||3258||KMK||1.2|| | |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) | ! colspan="7" style="background:#ffdead;" |3' entry clones, attR2-L3 (kan resistant) | ||
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!219||pDONRP4-P1R | !219||pDONRP4-P1R | ||
| − | | | + | |5' donor vector; attP4-P1R flanking chlor/ccdB cassette||||Invitrogen||1.1|| |
|- | |- | ||
!220||pDONRP2R-P3 | !220||pDONRP2R-P3 | ||
| − | | | + | |3' donor vector; attP2R-P3 flanking chlor/ccdB cassette||||Invitrogen||1.1|| |
|- | |- | ||
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* '''ver''' shows the version of the Tol2kit in which this construct first appeared. | * '''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. | * *: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 | + | * **: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. | * '''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.