m (Components used for the Tol2kit moved to Sources: combining with References page, renaming appropriately) |
|||
| Line 1: | Line 1: | ||
| − | + | Under construction 9/14/07. | |
| − | + | == Components == | |
| − | + | Here is the provenance of the components that we have used. For details of cloning, see the [[List of entry and destination vectors|individual clone pages]] or the full methods paper. | |
| − | + | ||
| − | + | === Gateway components === | |
| − | + | The entry clones and destination vectors are all based on (three-insert) multisite Gateway technology from Invitrogen, specifically pDONR P4-P1R, pDONR221 ''(attP1-attP2)'', pDONR P2R-P3, and pDest R4-R3. Our understanding from Invitrogen is that they do not usually require any MTAs for clones generated by individual labs using their vectors. (You still need to buy recombination enzyme from them though!) | |
| + | |||
| + | Sequence information all comes from the Invitrogen website. In some cases we have noticed point mutations in our constructs relative to their sequence. The most notable example is a [[Invitrogen ccdB|point mutation]] in the ccdB coding sequence within the destination vector. In at least a couple of cases, we have seen a mutation in clones from different labs, leading us to believe that it derived from the original Invitrogen clone. | ||
| + | |||
| + | === fluorescent proteins (FPs) === | ||
| + | |||
| + | ==== EGFP ==== | ||
| + | |||
| + | ==== mCherry ==== | ||
| + | |||
| + | === Subcellular localization tags === | ||
| + | |||
| + | ==== nls- ==== | ||
| + | |||
| + | ==== -CAAX ==== | ||
| + | |||
| + | ==== H2A- ==== | ||
| + | |||
| + | === encephalomyocarditis (EMCV) IRES === | ||
| + | |||
| + | === SV40 late polyA signal === | ||
| + | |||
| + | == References == | ||
Revision as of 13:47, 15 September 2007
Under construction 9/14/07.
Contents
Components
Here is the provenance of the components that we have used. For details of cloning, see the individual clone pages or the full methods paper.
Gateway components
The entry clones and destination vectors are all based on (three-insert) multisite Gateway technology from Invitrogen, specifically pDONR P4-P1R, pDONR221 (attP1-attP2), pDONR P2R-P3, and pDest R4-R3. Our understanding from Invitrogen is that they do not usually require any MTAs for clones generated by individual labs using their vectors. (You still need to buy recombination enzyme from them though!)
Sequence information all comes from the Invitrogen website. In some cases we have noticed point mutations in our constructs relative to their sequence. The most notable example is a point mutation in the ccdB coding sequence within the destination vector. In at least a couple of cases, we have seen a mutation in clones from different labs, leading us to believe that it derived from the original Invitrogen clone.