Chemical Genetics Laboratory
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Plasmid

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
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