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To download sequence files, please right-click the click button and choose "save" or "download".
All files are created using a free software "ApE".
Although you can open these files in a simple text format, we recommend to use ApE.
Some tips for the LR reaction
Please note that the LR reaction should be carried out using low concentration of entry clones and destination vectors.
Although we do not know the reason, it is very difficult to obtain correct expression plasmids when high concentration of vectors and entry clones are used in the reaction.
We usually use about 10 ng (or less) of each plasmid in the LR reaction.
The reaction volume can be reduced to at least 3 microliter for cost saving.
| pREP series (Gateway-compatible) |
|
Plasmid name
|
Backbone
|
Tag
|
Tag position
|
Promoter
|
Selection marker
|
Sequence
|
| pREP1-ccdB2 |
pREP1
|
-
|
-
|
nmt1
|
LEU2
|
|
| pREP41-ccdB2 |
pREP41
|
-
|
-
|
nmt1*
|
LEU2
|
|
| pREP81-ccdB2 |
pREP81
|
-
|
-
|
nmt1**
|
LEU2
|
|
| pHFF1-ccdB2 |
pREP1
|
His6-FLAG2
|
5'
|
nmt1
|
LEU2
|
|
| pHFF41-ccdB2 |
pREP41
|
His6-FLAG2
|
5'
|
nmt1*
|
LEU2
|
|
| pHFF81-ccdB2 |
pREP81
|
His6-FLAG2
|
5'
|
nmt1**
|
LEU2
|
|
| pHFG1-ccdB2 |
pREP1
|
His6-FLAG-GFP(S65C)
|
5'
|
nmt1
|
LEU2
|
|
| pHFG41-ccdB2 |
pREP41
|
His6-FLAG-GFP(S65C)
|
5'
|
nmt1*
|
LEU2
|
|
| pHFG81-ccdB2 |
pREP81
|
His6-FLAG-GFP(S65C)
|
5'
|
nmt1**
|
LEU2
|
|
| pFFH1-ccdB2 |
pREP1
|
FLAG2-His6
|
3'
|
nmt1
|
LEU2
|
|
| pFFH41-ccdB2 |
pREP41
|
FLAG2-His6
|
3'
|
nmt1*
|
LEU2
|
|
| pFFH81-ccdB2 |
pREP81
|
FLAG2-His6
|
3'
|
nmt1**
|
LEU2
|
|
| pGFH1-ccdB2 |
pREP1
|
GFP(S65C)-FLAG-His6
|
3'
|
nmt1
|
LEU2
|
|
| pGFH41-ccdB2 |
pREP41
|
GFP(S65C)-FLAG-His6
|
3'
|
nmt1*
|
LEU2
|
|
| pGFH81-ccdB2 |
pREP81
|
GFP(S65C)-FLAG-His6
|
3'
|
nmt1**
|
LEU2
|
|
| pHFY1-ccdB2 |
pREP1
|
His6-FLAG-YFP
|
5'
|
nmt1
|
LEU2
|
|
| pHFC1-ccdB2 |
pREP1
|
His6-FLAG-CFP
|
5'
|
nmt1
|
LEU2
|
|
| pYFH1-ccdB2 |
pREP1
|
YFP-FLAG-His6
|
3'
|
nmt1
|
LEU2
|
|
| pYFH1-ccdB2 rfA |
pREP1
|
YFP-FLAG-His6
|
3'
|
nmt1
|
LEU2
|
|
| pYFH1-ccdB2 rfC |
pREP1
|
YFP-FLAG-His6
|
3'
|
nmt1
|
LEU2
|
|
| pCFH1-ccdB2 |
pREP1
|
CFP-FLAG-His6
|
3'
|
nmt1
|
LEU2
|
|
|
|
* Medium strength nmt1 promoter, ** Low strength nmt1 promoter
|
 |
 |
|
