Advanced Gateway Destination Vectors
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- Amie Knight
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1 Description: The Susan Lindquist laboratory of the Howard Hughes Medical Institute and the Whitehead Institute for Biomedical Research has deposited a set of Gateway destination vectors for highthroughput genetic analysis in S. cerevisiae. The plasmids are divided into three plates. Detailed information can be found at: Reference: A suite of Gateway cloning vectors for high-throughput genetic analysis in Saccharomyces cerevisiae. Simon Alberti, Aaron D. Gitler, and Susan Lindquist. Yeast, Handling and Storage: Store glycerol stocks at -80 o C and minimize freeze-thaw cycles. To access a plasmid, keep the plate on dry ice to prevent thawing. Using a sterile pipette tip, puncture the seal above an individual well and spread a portion of the glycerol stock onto an agar plate. To patch the hole, use sterile tape or a portion of a fresh aluminum seal. Note: These plasmid constructs are being distributed to non-profit institutions for the purpose of basic research. Please contact Susan Lindquist to determine proper attribution on any publications arising from the use of these plasmids. Please contact Addgene at [email protected] with any questions. [email protected]
2 Plate 3 GAL1 GPD No tag ECFP EGFP EYFP Cerulean Integrating CEN 2 µ attr1 attr2 padvancedgateway Destination Vector No tag ECFP EGFP EYFP DsRed Cerulean HA TAP HIS3 TRP1 LEU2 URA3 Plate 1 Plate 2 Alberti et al., Yeast 2007 The set of padvancedgateway Destination Vectors allows for a choice of: 1. Promoter: constitutive (GPD) or inducible (GAL1) 2. N-terminal protein tag: ECFP, EGFP, EYFP, Cerulean 3. C-terminal protein tag: ECFP, EGFP, EYFP, DsRed, Cerulean, HA, TAP 4. Integrating or extra-chromosomal: Integrating: pag303, 304, 305, 306 Low-copy number (CEN): pag413, 414, 415, 416 High-copy number (2µ): pag423, 424, 425, Selectable marker: HIS3: pag303, 413, 423 TRP1: pag304, 414, 424 LEU2: pag305, 415, 425 URA3: pag306, 416, 426 * Please see individual plasmid pages for detailed maps and full sequences.
3 Plate 1 A 303GAL-ccdB 413GAL-ccdB 423GAL-ccdB 303GAL-ccdB-ECFP 413GAL-ccdB-ECFP 423GAL-ccdB-ECFP B 303GPD-ccdB 413GPD-ccdB 423GPD-ccdB 303GPD-ccdB-ECFP 413GPD-ccdB-ECFP 423GPD-ccdB-ECFP C 304GAL-ccdB 414GAL-ccdB 424GAL-ccdB 304GAL-ccdB-ECFP 414GAL-ccdB-ECFP 424GAL-ccdB-ECFP D 304GPD-ccdB 414GPD-ccdB 424GPD-ccdB 304GPD-ccdB-ECFP 414GPD-ccdB-ECFP 424GPD-ccdB-ECFP E 305GAL-ccdB 415GAL-ccdB 425GAL-ccdB 305GAL-ccdB-ECFP 415GAL-ccdB-ECFP 425GAL-ccdB-ECFP F 305GPD-ccdB 415GPD-ccdB 425GPD-ccdB 305GPD-ccdB-ECFP 415GPD-ccdB-ECFP 425GPD-ccdB-ECFP G 306GAL-ccdB 416GAL-ccdB 426GAL-ccdB 306GAL-ccdB-ECFP 416GAL-ccdB-ECFP 426GAL-ccdB-ECFP H 306GPD-ccdB 416GPD-ccdB 426GPD-ccdB 306GPD-ccdB-ECFP 416GPD-ccdB-ECFP 426GPD-ccdB-ECFP A 303GAL-ccdB-EGFP 413GAL-ccdB-EGFP 423GAL-ccdB-EGFP 303GAL-ccdB-EYFP 413GAL-ccdB-EYFP 423GAL-ccdB-EYFP B 303GPD-ccdB-EGFP 413GPD-ccdB-EGFP 423GPD-ccdB-EGFP 303GPD-ccdB-EYFP 413GPD-ccdB-EYFP 423GPD-ccdB-EYFP C 304GAL-ccdB-EGFP 414GAL-ccdB-EGFP 424GAL-ccdB-EGFP 304GAL-ccdB-EYFP 414GAL-ccdB-EYFP 424GAL-ccdB-EYFP D 304GPD-ccdB-EGFP 414GPD-ccdB-EGFP 424GPD-ccdB-EGFP 304GPD-ccdB-EYFP 414GPD-ccdB-EYFP 424GPD-ccdB-EYFP E 305GAL-ccdB-EGFP 415GAL-ccdB-EGFP 425GAL-ccdB-EGFP 305GAL-ccdB-EYFP 415GAL-ccdB-EYFP 425GAL-ccdB-EYFP F 305GPD-ccdB-EGFP 415GPD-ccdB-EGFP 425GPD-ccdB-EGFP 305GPD-ccdB-EYFP 415GPD-ccdB-EYFP 425GPD-ccdB-EYFP G 306GAL-ccdB-EGFP 416GAL-ccdB-EGFP 426GAL-ccdB-EGFP 306GAL-ccdB-EYFP 416GAL-ccdB-EYFP 426GAL-ccdB-EYFP H 306GPD-ccdB-EGFP 416GPD-ccdB-EGFP 426GPD-ccdB-EGFP 306GPD-ccdB-EYFP 416GPD-ccdB-EYFP 426GPD-ccdB-EYFP
4 Plate 2 A 303GAL-ccdB-DsRed 413GAL-ccdB-DsRed 423GAL-ccdB-DsRed 303GAL-ccdB-Cer 413GAL-ccdB-Cer 423GAL-ccdB-Cer B 303GPD-ccdB-DsRed 413GPD-ccdB-DsRed 423GPD-ccdB-DsRed 303GPD-ccdB-Cer 413GPD-ccdB-Cer 423GPD-ccdB-Cer C 304GAL-ccdB-DsRed 414GAL-ccdB-DsRed 424GAL-ccdB-DsRed 304GAL-ccdB-Cer 414GAL-ccdB-Cer 424GAL-ccdB-Cer D 304GPD-ccdB-DsRed 414GPD-ccdB-DsRed 424GPD-ccdB-DsRed 304GPD-ccdB-Cer 414GPD-ccdB-Cer 424GPD-ccdB-Cer E 305GAL-ccdB-DsRed 415GAL-ccdB-DsRed 425GAL-ccdB-DsRed 305GAL-ccdB-Cer 415GAL-ccdB-Cer 425GAL-ccdB-Cer F 305GPD-ccdB-DsRed 415GPD-ccdB-DsRed 425GPD-ccdB-DsRed 305GPD-ccdB-Cer 415GPD-ccdB-Cer 425GPD-ccdB-Cer G 306GAL-ccdB-DsRed 416GAL-ccdB-DsRed 426GAL-ccdB-DsRed 306GAL-ccdB-Cer 416GAL-ccdB-Cer 426GAL-ccdB-Cer H 306GPD-ccdB-DsRed 416GPD-ccdB-DsRed 426GPD-ccdB-DsRed 306GPD-ccdB-Cer 416GPD-ccdB-Cer 426GPD-ccdB-Cer A 303GAL-ccdB-HA * 413GAL-ccdB-HA 423GAL-ccdB-HA 303GAL-ccdB-TAP 413GAL-ccdB-TAP 423GAL-ccdB-TAP B 303GPD-ccdB-HA * 413GPD-ccdB-HA 423GPD-ccdB-HA 303GPD-ccdB-TAP 413GPD-ccdB-TAP 423GPD-ccdB-TAP C 304GAL-ccdB-HA * 414GAL-ccdB-HA 424GAL-ccdB-HA 304GAL-ccdB-TAP 414GAL-ccdB-TAP 424GAL-ccdB-TAP D 304GPD-ccdB-HA * 414GPD-ccdB-HA 424GPD-ccdB-HA 304GPD-ccdB-TAP 414GPD-ccdB-TAP 424GPD-ccdB-TAP E 305GAL-ccdB-HA 415GAL-ccdB-HA 425GAL-ccdB-HA 305GAL-ccdB-TAP 415GAL-ccdB-TAP 425GAL-ccdB-TAP F 305GPD-ccdB-HA 415GPD-ccdB-HA 425GPD-ccdB-HA 305GPD-ccdB-TAP 415GPD-ccdB-TAP 425GPD-ccdB-TAP G 306GAL-ccdB-HA * 416GAL-ccdB-HA 426GAL-ccdB-HA 306GAL-ccdB-TAP 416GAL-ccdB-TAP 426GAL-ccdB-TAP H 306GPD-ccdB-HA * 416GPD-ccdB-HA 426GPD-ccdB-HA 306GPD-ccdB-TAP 416GPD-ccdB-TAP 426GPD-ccdB-TAP * : missing CYC1 terminator
5 Plate 3 A 303GAL-ECFP-ccdB 413GAL-ECFP-ccdB 423GAL-ECFP-ccdB 303GAL-EGFP-ccdB 413GAL-EGFP-ccdB 423GAL-EGFP-ccdB B 303GPD-ECFP-ccdB 413GPD-ECFP-ccdB 423GPD-ECFP-ccdB 303GPD-EGFP-ccdB 413GPD-EGFP-ccdB 423GPD-EGFP-ccdB C 304GAL-ECFP-ccdB 414GAL-ECFP-ccdB 424GAL-ECFP-ccdB 304GAL-EGFP-ccdB 414GAL-EGFP-ccdB 424GAL-EGFP-ccdB D 304GPD-ECFP-ccdB 414GPD-ECFP-ccdB 424GPD-ECFP-ccdB 304GPD-EGFP-ccdB 414GPD-EGFP-ccdB 424GPD-EGFP-ccdB E 305GAL-ECFP-ccdB 415GAL-ECFP-ccdB 425GAL-ECFP-ccdB 305GAL-EGFP-ccdB 415GAL-EGFP-ccdB 425GAL-EGFP-ccdB F 305GPD-ECFP-ccdB 415GPD-ECFP-ccdB 425GPD-ECFP-ccdB 305GPD-EGFP-ccdB 415GPD-EGFP-ccdB 425GPD-EGFP-ccdB G 306GAL-ECFP-ccdB 416GAL-ECFP-ccdB 426GAL-ECFP-ccdB 306GAL-EGFP-ccdB 416GAL-EGFP-ccdB 426GAL-EGFP-ccdB H 306GPD-ECFP-ccdB 416GPD-ECFP-ccdB 426GPD-ECFP-ccdB 306GPD-EGFP-ccdB 416GPD-EGFP-ccdB 426GPD-EGFP-ccdB A 303GAL-EYFP-ccdB 413GAL-EYFP-ccdB 423GAL-EYFP-ccdB 303GAL-Cer-ccdB 413GAL-Cer-ccdB 423GAL-Cer-ccdB B 303GPD-EYFP-ccdB 413GPD-EYFP-ccdB 423GPD-EYFP-ccdB 303GPD-Cer-ccdB 413GPD-Cer-ccdB 423GPD-Cer-ccdB C 304GAL-EYFP-ccdB 414GAL-EYFP-ccdB 424GAL-EYFP-ccdB 304GAL-Cer-ccdB 414GAL-Cer-ccdB 424GAL-Cer-ccdB D 304GPD-EYFP-ccdB 414GPD-EYFP-ccdB 424GPD-EYFP-ccdB 304GPD-Cer-ccdB 414GPD-Cer-ccdB 424GPD-Cer-ccdB E 305GAL-EYFP-ccdB 415GAL-EYFP-ccdB 425GAL-EYFP-ccdB 305GAL-Cer-ccdB 415GAL-Cer-ccdB 425GAL-Cer-ccdB F 305GPD-EYFP-ccdB 415GPD-EYFP-ccdB 425GPD-EYFP-ccdB 305GPD-Cer-ccdB 415GPD-Cer-ccdB 425GPD-Cer-ccdB G 306GAL-EYFP-ccdB 416GAL-EYFP-ccdB 426GAL-EYFP-ccdB 306GAL-Cer-ccdB 416GAL-Cer-ccdB 426GAL-Cer-ccdB H 306GPD-EYFP-ccdB 416GPD-EYFP-ccdB 426GPD-EYFP-ccdB 306GPD-Cer-ccdB 416GPD-Cer-ccdB 426GPD-Cer-ccdB : missing portion of the Gal1 promoter
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