Forsburg Lab pombe pages:
Index of fission yeast vectors

General information about plasmids

S. pombe plasmids consist of a bacterial origin of replication and selectable marker (usually an antibiotic resistance gene), a yeast selectable marker (usually a metabolic marker) and an autonomous replication sequence (ars) which is responsible for high frequency of transformation. There are no single copy centromere plasmids, as there are in budding yeast, because the fission yeast centromere is too big to be easily encompassed on a shuttle vector. Other features may include various promoters and fusion or tagging sequences.

About this list
The accuracy of this list is not guaranteed, and it is not exhaustive, although we try to keep it up to date. More information about vectors, markers, and expression levels is available on our pombe methods page. (For a detailed directory to our site, start with our home page).

We've divided pombe plasmids into four categories:

• General purpose vectors
• Expression vectors
• Tagging and fusion vectors (GFP, GST, HA....)
• Special purpose vectors

Plasmid sequences
Some of following plasmids have sequences on this site (also linked from the table). These were sent to us by others; we are not responsible for their accuracy. Other sequences are linked from NCBI or elsewhere.

Sequences on this site		Sequences elsewhere
pBG1 pFS119 pFS118 pFS118 pFS118 pFS118 pWH5 REP1 REP3 REP41 REP81 pART1	pFA6a-13Myc-kanMX6 pFA6a-3HA-kanMX6 pFA6a-GFP(S65T)-kanMX6 pFA6a-GST-kanMX6 pFA6a-kanMX6 pFA6a-kanMX6-P3nmt1 pFA6a-kanMX6-P3nmt1-3HA pFA6a-kanMX6-P3nmt1-GFP pFA6a-kanMX6-P3nmt1-GST	pJK210 pJK148 pDBLet

Choosing the right promoter
Our pombe technology page describes relative promoter activity for several cloned promoters. Note that the nmt promoter in particular can be modulated simply by varying the amount of thiamine in the media. Full induction: no thiamine. Full repression: 15uM thiamine (5ug/ml). Partial induction: 0.05uM thiamine (0.016ug/ml). It's important to note that the fully repressed nmt promoter allows a low level of expression; this may be sufficient to complement, if your protein is reasonably stable. Thus, "promoter shut-off" experiments in pombe typically only work with rather unstable proteins.

GFP fusion library
The Hiraoka lab published a paper on Large-scale screening of intracellular protein localization in living fission yeast cells by the use of a GFP-fusion genomic DNA library (Genes to Cells (2000) 5, 169-190). All the information is on their excellent web site.

Also, see this S. pombe post genome database with results from a systematic analysis of protein expression and localization.

Feedback
If you have suggestions or additions or you find errors, please email us. We will try to keep this index up-to-date but we need your help to do it. If you have made plasmids that you are willing to share, send us the specifications and the reference and we'll add them to the list. Our thanks to those who have contributed already.

Getting plasmids
There are a number of places that now provide pombe strains and plasmids to the community. There is a list on our pombeweb page.

If you want particular plasmids, check out the resources above, or contact the lab that built the vector in question (or an author of the paper). Our lab does not send out plasmids built by other groups, so please only ask us for our own plasmids.

© S. L. Forsburg .