Vector table
Search for general purpose, expression vectors, tagging constructs, or speciality plasmids. Send additions!The Vector Table
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 |
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|---|---|---|---|---|---|
| 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 REP41 sequence |
| REP81 , REP82 | REP1,2 | LEU2 ura4+ | ars1 | nmt low strength (nmt**) | Basi
REP81 sequence |
| 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 ; sequence here | |||
| 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 | |||
| 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 |
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|---|---|---|---|---|---|
| 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 al
Sequences: 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. | Chang |
| 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. | Zhao (b) | |||
SPECIAL PURPOSE VECTORS |
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|---|---|---|---|---|---|
| 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 from addgene |
