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General Purpose Fission Yeast Vectors

plasmid name base plasmid yeast markers yeast origin other features and comments refs/sources
pAL19 pUC LEU2 ars1 blue/white T. Carr (at ATCC)
paR3 arg3+ ars1 Waddell
pBG1 pUC his3+ ars1 blue/white Burke
(at ATCC)
(sequence)
pDBlet bluescript ura4+ 2x ars3002 dual ars for increased stability Brun
pDB248X pBR LEU2 2 micron Beach
pEA500 pSP2 his7+ ars1 Apolinaro
(at ATCC)
pFL20 pBR322 URA3 ars1 stb Losson
pIRT2
pIRT2U pUC LEU2
ura4+ ars1 Hindley
pIRT2-CAN1 pUC LEU2 CAN1 ars1 CAN1 for counter- selection in can1-1 strains Ekwall
pJK148
pJK210 bluescript leu1+
ura4+ — blue/white integrating Keeney
(at ATCC)
pON163 ura4+ ars1 positive selection for inserts with kanamycin Weilguny
pNPT/ADE1-3 pART adh-neoR ade1+ ars1 can be selected for with G418 M. Moser
pSP1
pSP2 LEU2
URA3 ars1 complements leu1 strains
complements ura4 strains Cottarel (a)
(at ATCC)
pSP3
pSP4 HIS3
LYS2 ars1 complements his5 strains
complements lys1 strains Cottarel (b)
pUR18
pUR19 pUC ura4+ ars1 blue/white Barbet
(at ATCC)
pZA57 pEA500 his7+ ade1+ ars1 pZA58 has ade1+ in opposite orientation M. Moser
pWH5 LEU2 2µ First generation vector used in early cloning expts. Now replaced by plasmids such as pAL19, etc Sequence

 

Fission Yeast Expression Vectors

name base plasmid yeast markers yeast origin promoter references and sources
For more information about promoters, go to the working with pombe page
pART1 pIRT/pUC LEU2 ars adh promoter McLeod
pCHY21 URA3 ars1 fbp promoter Hoffman
pEVP11 LEU2 2 µ adh promoter Russell
REP1 ,REP3,
REP4 pUC LEU2
ura4+ ars1 nmt1 full strength, Full repression: 15uM thiamine (5ug/ml). Full induction: no thiamine. Takes about 18 hours. Partial induction: 0.05uM thiamine (0.016ug/ml). For more information about titrating nmt, and the different levels of expression, visit the working with pombe page. Maundrell (a), (b)
Sequences:
REP1
REP3
REP41,
REP42 REP1,2 LEU2
ura4+ ars1 nmt med strength (nmt*) Basi
REP41sequence
REP81 ,
REP82 REP1,2 LEU2
ura4+ ars1 nmt low strength (nmt**) Basi
REP81sequence
RIP derived from above: integrating vectors lacking ars1. RIP-s vectors contain sup3-5 on PstI fragment. Maundrell
REP3X
REP4XREP41X
REP81XREP42X
REP82XRIP3X/s
RIP4X/s The REP-X derivatives are derived from the REP/RIP series above. The original REP/RIP vectors contain an ATG at the 5′ end of the polylinker; the X-series removes it and adds a XhoI site. Note that you must supply an ATG on your clone if using the X-series.
Important: the polylinker in the 40-X and 80-X series is not quite the same as the polylinker in the 3X and 4X series. See the map. Forsburg (a)
(at ATCC)
pYZ1N
pYZ41N
pYZ81N Derived from the REP series, with a lacZ alpha peptide in the polylinker to allow blue/white selection for inserts. Zhao (b)
pSLF101
pSLF102 pUC LEU2
ura4+ ars1 CaMV with tet operator, repressed by adh-tet repressor. In the pSLF series, the repressor is provided by integrated pSLF104.
In pMLtetON, the repressor is co-expressed on the same plasmid. Use of the analogue anhydrotetracycline at 6uM leads to induction within 3 hrs (max 12hrs). Martin Luetzelberger and Erler et al. (2006; Yeast 23: 813–823)
Martin tells us expression is in the REP41-81 range. Forsburg (a)
(at ATCC)
pSLF104 pUC sup3-5 —
pMLtetON leu1+; integrate by linearizing with Tth111I or BsiWI Dr Martin Luetzelberger ; sequencehere
pSM1/2 LEU2 2 micron SV40 promoter Russell
pTLM2/pAL7 (pAU5) pSV40-neoR none hCMV promoter; titrating G418 allows increase in copy number Tohda
p2UG URA3 2 micron GRE elements regulated by adh-glucocorticoid receptor (pART) Picard
pART1/N795 LEU2 ars1
pSK174
and derivatives ura4+ ars1 nmt1-lacO hybrid promoter, co-expresses lacI. Inducible by IPTG in minus thiamine conditions Kjaerulff
There are two commercially available pombe expression systems, from Asahi Glass Co. and Stratagene. This page provides information about them. Obligatory Disclaimer: We are not endorsing any product.

 

Tagging / Fusion Vectors

plasmid name base plasmid yeast markers promoter, other features references and source
Forsburg lab tagging series (compatible polylinkers, modular design)
pSLF172
pSLF272
pSLF372 REP4X
REP42X
REP82X ura4+ nmt/nmt*/nmt** expression vectors allowing triple HA epitope tag to be fused to C-terminus. Include stop codon, no ATG. Forsburg (b)
(at ATCC)
pSLF173
pSLF273
pSLF373 REP4X
REP42X
REP82X ura4+ nmt/nmt*/nmt** expression vectors allowing triple HA epitope tag to be fused to N-terminus. Contain ATG codon, no stop.
pDS472
pDS473 REP4X ura4+ full strength nmt expression vectors allowing GST tag to be fused to C-terminus (472) or N terminus (473). Compatible with pSLF172/173 series.
pSGP72
pSGP73 REP3X LEU2 LEU2 versions of 3xHA tagging vectors pSLF172 and pSLF173 with full strength nmt promoter S.G. Pasion
(Forsburg lab)
pSGP572
pSGP573 REP4X ura4+ full strength nmt expression vectors allowing GFP to be fused to C-terminus (572) or N terminus (573). Compatible with pSLF172/173 series. Pasion
pSLF972
pJAH1172 REP4X
REP3X ura4+
LEU2 full strength nmt expression vectors allowing 3xV5 tag (from paramyxovirus SV5 p/k protein) fusion, C terminal only right now. Compatible with pSLF172/173 series. Antibody to V5 is commercially available from Invitrogen (R960) or Serotec (MCA1360) (Forsburg lab)
pSLF1072
pSLF1073 REP4X ura4+ full strength nmt expression vectors allowing 8XHis HA double tag fusion at C-terminus (1072) or N-terminus (1073). Compatible with pSLF172/173 series. (Forsburg lab)
pNCH1472 REP4X ura4+ full strength nmt expression vector with 12x myc tag fusion at C-terminus. Compatible with pSLF172/173 series. (Forsburg lab)
Hagan/Carr tagging series (compatible polylinkers)
REP41MH-N
REP42MH-N REP41
REP42 LEU2
ura4+ N-terminal tag with 6xHis 2xMyc in mid strength (nmt*) nmt vectors with ura4+ or LEU2 markers Craven
REP41HA-N
REP42HA-N REP41
REP42 LEU2
ura4+ N-terminal tag with 3xHA in mid strength (nmt*) nmt vectors with ura4+ or LEU2 markers
REP41Pk-N
REP42Pk-N
REP81PK-N REP41
REP42
REP81 LEU2
ura4+
LEU2 N-terminal tag with 3xPk tag (from virus SV5 P-protein) in mid strength (nmt*) nmt vectors with ura4+ or LEU2 markers. Antibody to V5 is commercially available from Invitrogen (R960) or Serotec (MCA1360)
REP41GFP/EGFP-N
REP42GFP/EGFP-N REP41
REP42 LEU2
ura4+ N-terminal tag with GFP or enhanced GFP in mid strength (nmt*) nmt vectors with ura4+ or LEU2 markers
REP41Pk-C
REP42Pk-C
REP81Pk-C REP41
REP42
REP81 LEU2
ura4+
LEU2 C-terminal tag with 3xPk tag (from virus SV5 P-protein) in mid strength (nmt*) nmt vectors with ura4+ or LEU2 markers. Antibody to V5 is commercially available from Invitrogen (R960) or Serotec (MCA1360)
REP41GFP/EGFP-C
REP42GFP/EGFP-C REP41
REP42 LEU2
ura4+ C-terminal tag with GFP or enhanced GFP in mid strength (nmt*) nmt vectors with ura4+ or LEU2 markers
Bähler PCR tagging series
KS-ura4 Bluescript KS- ura4+ used as a PCR template, followed by transformation for gene deletion Bähler et alSequences:
kanMX6
3HA-kanMX6
13Myc-kanMX6
GST-kanMX6
GFP(S65T)-kanMX6
P3nmt1
P3nmt1-3HA
P3nmt1-GFP
P3nmt1-GST
pFA6a-kanMX6 pFA (Wach et al., 1994, Yeast 10:1793; Wach, 1996, Yeast 12: 259) kanMX6 used as a PCR template, followed by transformation for gene deletion
pFA6a-3HA-kanMX6
pFA6a-13Myc-kanMX6
pFA6a-GST-kanMX6
pFA6a-GFP(S65T)-kanMX6 pFA (Wach et al., 1994, Yeast 10:1793; Wach, 1996, Yeast 12: 259) kanMX6 used as PCR template followed by yeast transformation for C-terminal tagging of proteins by 3HA, 13Myc, GST, or GFP, respectively, at their normal chromosomal locations
pFA6a-kanMX6-P3nmt1, pFA (Wach et al., 1994, Yeast 10:1793; Wach, 1996, Yeast 12: 259) kanMX6 used as PCR template followed by yeast transformation for expression of proteins under the nmt1 promoter (3 different strengths) or N-terminal tagging of proteins by 3HA, GST, or GFP, respectively, at their normal chromosomal locations.
Promoter: nmt1 (full strength: P3nmt1; medium strength: P41nmt1; low strength: P81nmt1)
Others in series: pFA6a-kanMX6-P41nmt1
pFA6a-kanMX6-P81nmt1
pFA6a-kanMX6-P3nmt1-3HA
pFA6a-kanMX6-P41nmt1-3HA
pFA6a-kanMX6-P81nmt1-3HA
pFA6a-kanMX6-P3nmt1-GST
pFA6a-kanMX6-P41nmt1-GST
pFA6a-kanMX6-P81nmt1-GST
pFA6a-kanMX6-P3nmt1-GFP
pFA6a-kanMX6-P41nmt1-GFP
pFA6a-kanMX6-P81nmt1-GFP
Bahler lab urg1+ promoter-tagging series
pFA6a-kanMX6-Purg1 pFA (Wach et al., 1994, Yeast 10:1793; Wach, 1996, Yeast 12: 259) kanMX6 PCR template for expression under urg1+ promoter and/or N-terminal tagging.
Others in the series: pFA6a-natMX6-Purg1
pFA6a-kanMX6-Purg1-3HA
pFA6a-kanMX6-Purg1-GST
pFA6a-kanMX6-Purg1-GFP Watt et al
Gould lab TAP-tagging series
Derived from the Bähler series. See sequences, maps and links on the Gould lab TAP-tag page
Also see Juraj Gregan’s Tap Tagging any ORF in pombe page
Sawin lab red-tagging series
pKS390 pFA (Wach et al., 1994, Yeast 10:1793; Wach, 1996, Yeast 12: 259) kanMX6, natMX6 used as PCR template followed by yeast transformation for expression of proteins under the nmt1 promoter (3 different strengths) or N-terminal tagging of proteins at their normal chromosomal locations.
Promoter: nmt1 (full strength: P3nmt1; medium strength: P41nmt1; low strength: P81nmt1)
Other plasmids in the series
pKS391 pFA6a-mCherry-natMX6
pKS392 pFA6a-tdTomato-kanMX6
pKS393 pFA6a-tdTomato-natMX6
pKS394 pFA6a-kanMX6-P3nmt1-mCherry
pKS395 pFA6a-kanMX6-P41nmt1-mCherry
pKS396 pFA6a-kanMX6-P81nmt1-mCherry
pKS397 pFA6a-kanMX6-P3nmt1-tdTomato
pKS398 pFA6a-kanMX6-P41nmt1-tdTomato
pKS399 pFA6a-kanMX6-P81nmt1-tdTomato Snaith
MacNeill BIFC vectors
pFA6a-VN173-kanMX6
pFA6a-VN173-natMX6
pFA6a-VC155-kanMX6
pFA6a-VC155-natMX6 pFA (Wach et al., 1994, Yeast 10:1793; Wach, 1996, Yeast 12: 259) kanMX6, natMX6 used as PCR template followed by yeast transformation
These plasmids allow expression of proteins fused to two halves of YFP, allowing for bimolecular fluoresence complementation in vivo. One putative partner needs to be fused to VN173, and one to VC155. Akman
Other tagging vectors
pARTCM pART LEU2 ars1 adh, with N-terminal cMYC tag. Cha
ng
pALL
pALU pART LEU2
ura4+ ars1 adh, with N-terminal HA1 tag. Chang
pYZ3N-GFP Derived from the REP series, with a lacZ alpha peptide in the polylinker to allow blue/white selection for inserts and GFP fusion.

 

Special Purpose Vectors

plasmid name base plasmid yeast markers yeast origin other features references and source
pCRR1 pBR LEU2 ars1 E. coli markers supF and tetR Zhao
To determine rate of mutagenesis: passage plasmid through pombe and transform into E. coli strain KS40/pKY241 to quantitate
pCF83
pCF85 pIRT2U
pIRT2 ura4+
LEU2 ars1 Promoter reporter construct: S.cerevisaie CYC1 TATA cloned upstream of lacZ C. Fankhauser
REP3X-lacZ
REP41X-lacZ
REP81X lacZ REP3X
REP41X
REP81X LEU2 ars1 nmt-lacZ reporter fusions Forsburg (a)
pDM291 pSP72 hisG-ura4+-hisG disruption cassette, allowing recovery and re-use of the ura4 marker in disrupted strains McNabb
pFS119
pFS118 ? ade6+
ura4+ ars1 contains adh-thymidine kinase, allowing counterselection with FuDR (see Kiely et al for the method) Sivakumar et al
pJAH29
pJAH31 pJK148
pEA2 leu1+
his7+ integration vectors System for labeling cells with BudR. One plasmid expresses adh-tk, and one expresses the hENT nucleoside transporter. These plasmids are also available already integrated into the genome of Forsburg lab strain FY2317 Hodson et al
pFS177
pFS181
pFS255 pART1
pJK148
pFA6a-kanMX LEU2 ars1
leu1
kanR episomal adh-hENT1
integrating adh-hENT1
adh1-tk integrating System for labeling cells with BudR. One plasmid expresses adh-tk, and one expresses the hENT nucleoside transporter. Sivakumar et al
available fromaddgene