RAS Clone Collections- RAS and RAS Pathway Constructs
(Kit #
1000000070, 1000000089
)
Depositing Lab: Dominic Esposito
The Esposito Lab has provided 2 kits for both Ras wild type and mutant constructs.
Kit #1000000070
The RAS Pathway 2.0 Clone Collection is a set of vectors for use with the Gateway® Cloning Platform (Life Technologies) to permit construction of RAS pathway gene expression vectors for in vitro and in vivo research. While these clones can be used for standard Gateway cloning, their utility is greatly enhanced with the use of the FNLCR Combinatorial Cloning Platform (CCP) which allows simplified construction of vectors with different elements (see Wall VE et al, 2014 for more information).
Kit #1000000089
This collection contains 61 wild type and mutant RAS genes as Gateway entry clones. It is comprised of wild type HRAS, NRAS, KRAS4a and KRAS4b gene plasmids as well as different mutant versions of each gene.
These kits will be sent as bacterial glycerol stocks in a 96-well plate format.
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | ||
|---|---|---|---|---|---|---|
| Kit | 1000000070 | RAS Pathway 2.0 Clone Collection- Wild type genes (360 constructs) | 1 | $1320 | Add to Cart | |
| Kit | 1000000089 | RAS Mutant Clone Collection- Wild type and mutant variations (61 constructs) | 1 | $440 | Add to Cart | |
Description
Kit #1000000070
This collection contains 180 genes in the “RAS pathway version 2.0”, a crowdsourced pathway map of the RAS signaling pathway generated on the RASCentral website of the Frederick National Laboratory RAS Initiative. In each case, the primary splice variant was chosen to represent the most commonly found transcript by RNAseq data across all TCGA tumor samples. Many of these transcript variants are not available in cDNA or ORF libraries, or are available only in mutated forms. The sequences in this collection encode wild type proteins, and match the RefSeq data for the particular splice variants with the exception of known SNPs or cases in which the RefSeq data is not consistent with genome sequence data (see pathway map below).
Some uses of this system are:
- Construction of protein expression constructs with various fusion tags
- Generation of expression constructs with different promoters for in vivo expression
- Production of clones with fluorescent tags for localization experiments
- Generation of constructs for making mutant cell lines or transgenic animals
- Production of vectors for shRNA or miRNA delivery
Kit #1000000089
This collection contains 61 wild type and mutant RAS genes as Gateway entry clones. It is comprised of wild type HRAS, NRAS, KRAS4a and KRAS4b gene plasmids as well as different mutant versions of each gene.
Depositor Information
Kit #1000000070
RAS Pathway Clone Collection 2.0 Overview 709.2 KBAnnotated Plasmid Maps 1.1 MB
Gateway Recombinational Cloning
For more information on the Gateway system, please see:
DNA cloning using in vitro site-specific recombination. Hartley JL, Temple GF, Brasch MA. Genome Res.2000. Nov;10(11):1788-95. PubMed PMID 11076863
For more information on the FNLCR Combinatorial Cloning Platform (CCP):
Combinatorial assembly of clone libraries using site-specific recombination. Wall VE, Garvey LA, Mehalko JL, Procter LV, Esposito D. Methods Mol Biol. 2014;1116:193-208. doi: 10.1007/978-1-62703-764-8_14. PubMed PMID 24395366
How to Cite these Kits
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which they were created, and include Addgene in the Materials and Methods of your future publications.
For your Materials and Methods section:
“The RAS Clone Collection was a gift from Dominic Esposito (Addgene kit # ).”
RAS Pathway 2.0 Clone Collection - #1000000070
- Resistance Color Key
Each circle corresponds to a specific antibiotic resistance in the kit plate map wells.
- Inventory
Searchable and sortable table of all plasmids in kit. The Well column links to its kit plate map on the right, as well as lists the plasmid well location in its plate. The Plasmid column links to a plasmid's individual web page.
- Kit Plate Map
96-well plate map for plasmid layout. Hovering over a well reveals the plasmid name, while clicking on a well opens the plasmid page.
Resistance Color Key
| Spectinomycin | |
| Kanamycin |
Inventory
Kit Plate Map - #1000000070
RAS mutant clone collection - #1000000089
- Resistance Color Key
Each circle corresponds to a specific antibiotic resistance in the kit plate map wells.
- Inventory
Searchable and sortable table of all plasmids in kit. The Well column lists the plasmid well location in its plate. The Plasmid column links to a plasmid's individual web page.
- Kit Plate Map
96-well plate map for plasmid layout. Hovering over a well reveals the plasmid name, while clicking on a well opens the plasmid page.
Resistance Color Key
| Spectinomycin |
Inventory
| Well | Plasmid | Resistance |
|---|---|---|
| A / 1 | Hs.KRAS4b |
|
| A / 2 | Hs.KRAS4b G12C |
|
| A / 3 | Hs.KRAS4b G12D |
|
| A / 4 | Hs.KRAS4b G12V |
|
| A / 5 | Hs.KRAS4b G13D |
|
| A / 6 | Hs.KRAS4b Q61L |
|
| A / 7 | Hs.KRAS4b Q61R |
|
| A / 8 | Hs.KRAS4b S181A |
|
| A / 9 | Hs.KRAS4b S181D |
|
| A / 10 | Hs.KRAS4b S181E |
|
| A / 11 | Hs.KRAS4b C185S |
|
| A / 12 | Hs.KRAS4b Q61H |
|
| B / 1 | Hs.KRAS4b M188L |
|
| B / 2 | Hs.KRAS4b G12A |
|
| B / 3 | Hs.KRAS4b G12R |
|
| B / 4 | Hs.KRAS4b G12S |
|
| B / 5 | Hs.KRAS4b G13C |
|
| B / 6 | Hs.KRAS4b K117N |
|
| B / 7 | Hs.KRAS4b A146V |
|
| B / 8 | Hs.KRAS4b R68S |
|
| B / 9 | Hs.KRAS4b G12F |
|
| B / 10 | Hs.KRAS4b A146T |
|
| B / 11 | Hs.KRAS4b Y64A |
|
| B / 12 | Hs.KRAS4b K104A |
|
| C / 1 | Hs.KRAS4b K104Q |
|
| C / 2 | Hs.KRAS4b K147L |
|
| C / 3 | Hs.KRAS4b K147Q |
|
| C / 4 | Hs.KRAS4b S17N |
|
| C / 5 | Hs.KRAS4b T35S |
|
| C / 6 | Hs.KRAS4b E37G |
|
| C / 7 | Hs.KRAS4b Y40C |
|
| C / 8 | Hs.KRAS4b M72C |
|
| C / 9 | Hs.KRAS4b Y40F |
|
| C / 10 | Hs.KRAS4b R73E |
|
| C / 11 | Hs.KRAS4b R73K |
|
| C / 12 | Hs.KRAS4b Q61K |
|
| D / 1 | Hs.KRAS4b G12D/C185S |
|
| D / 2 | Hs.KRAS4a |
|
| D / 3 | Hs.KRAS4a G12C |
|
| D / 4 | Hs.KRAS4a G12D |
|
| D / 5 | Hs.KRAS4a G12V |
|
| D / 6 | Hs.KRAS4a G13D |
|
| D / 7 | Hs.KRAS4a Q61L |
|
| D / 8 | Hs.KRAS4a Q61R |
|
| D / 9 | Hs.NRAS |
|
| D / 10 | Hs.NRAS G12V |
|
| D / 11 | Hs.NRAS G12C |
|
| D / 12 | Hs.NRAS G12D |
|
| E / 1 | Hs.NRAS G13D |
|
| E / 2 | Hs.NRAS Q61L |
|
| E / 3 | Hs.NRAS Q61R |
|
| E / 4 | Hs.NRAS Q61K |
|
| E / 5 | Hs.HRAS |
|
| E / 6 | Hs.HRAS G12C |
|
| E / 7 | Hs.HRAS G12D |
|
| E / 8 | Hs.HRAS G12V |
|
| E / 9 | Hs.HRAS G13D |
|
| E / 10 | Hs.HRAS Q61L |
|
| E / 11 | Hs.HRAS Q61K |
|
| E / 12 | Hs.HRAS Q61R |
|
| F / 1 | Hs.HRAS C186S |
|