Overview of Streptomyces vectors
From ActinoBase
Commonly Used Streptomyces Vectors
This page describes the characteristics of some of the most commonly-used plasmids developed for use in Streptomyces. They have been organized by general function (expression vectors, reporters, etc.) [Huge thanks to Mervyn Bibb for assembling the original "Streptomyces Genetic Toolkit," which has been invaluable for preparing this wiki.] See Plasmids for an alphabetical list.
This is still a work in progress, and we hope to add more plasmids soon! Please contact us if you notice any plasmids we've missed. Individual pages with more information on each plasmid will be added soon.
The following abbreviations are used in all of the tables below.
Antibiotics:
- Amp, ampicillin
- Apr, apramycin
- Cml, chloramphenicol
- Hyg, hygromycin
- Kan, kanamycin
- Spc, spectinomycin
- Str, streptomycin
- Tsr, thiostrepton
- Vio, viomycin
Strains:
- E, E. coli
- S, Streptomyces
Expression vectors
| Plasmid | Description | Resistance | Replicative status and copy number in Streptomyces | Comment |
|---|---|---|---|---|
| pIJ86 | Expression vector (ermE*p) | AprR (ES) | Conjugative, multi-copy | pIJ101 origin, constitutive expression vector |
| pIJ6021 | Expression vector (tipAp) | KanR (S), TsrR (S) | Non-conjugative, multi-copy | Tsr-inducible (tipAp) expression vector |
| pIJ6902 | Expression vector (tipAp) | AprR (ES), TsrR (S) | Conjugative, integrative (phiC31) | Tsr-inducible (tipAp) expression vector |
| pIJ8671 | Expression vector (tipAp) | AprR (ES), TsrR (S) | Conjugative, requires insert for homologous integration | Allows Tsr-inducible expression of chromosomal genes after homologous recombination |
| pIJ10257 | Expression vector (ermE*p) | HygR (ES) | Conjugative, integrative (phiBT1) | Constitutive (ermE*p) expression vector |
| pIJ10727 | Expression vector (aphp) | AprR (ES) | Conjugative, multi-copy | pIJ101 origin, derived from pIJ86, constitutive (aphp) expression vector |
| pIJ12477 | Expression vector (ermE*p) | AprR (ES), KanR (ES) | Conjugative, multi-copy | pIJ101 origin, derived from pIJ86 by addition of neo from SuperCos1, constitutive (ermE*p) expression vector |
| pIJ12551 | Expression vector (ermE*p) | AprR (ES) | Conjugative, integrative (phiC31) | Constitutive (ermE*p) expression vector |
| pIJ12552 | Expression vector (ermE*p) | AprR (ES) | Conjugative, integrative (phiC31) | ermE*p in opposite orientation to pIJ12551, constitutive expression vector |
| pIJ10790 (pSS90) | Expression vector (kasO*p) | HygR (ES) | Conjugative, integrative (phiBT1) | pIJ10257 derivative with kasO*p instead of ermE*p, constitutive expression vector |
| pIJ12901 | Expression vector (tipAp) | KanR (ES), TsrR (S) | Conjugative, multi-copy, unstable | pSG5 origin, temperature-sensitive, Tsr-inducible (tipAp) expression vector |
| pIJ13042 | Expression vector (ermE*p) | AprR (ES), TsrR (S) | Conjugative, multi-copy | pIJ101 origin, derived from pIJ86 by addition of tsr, constitutive (ermE*p) expression vector |
| pIJ12739 | Expression vector (tipAp) | AprR (ES), TsrR (S) | Conjugative, multi-copy, unstable | pSG5 origin, temperature-sensitive, Tsr-inducible (tipAp) Meganuclease gene |
| pIJ12742 | Expression vector (ermE*p) | AprR (ES), TsrR (S) | Conjugative, multi-copy, unstable | pSG5 origin, temperature-sensitive, ermE*p constitutive Meganuclease gene |
Mutational plasmids
| Plasmid | Description | Resistance | Replicative status and copy number in Streptomyces | Comment |
|---|---|---|---|---|
| pIJ12738 | Insertional vector with SceI site | AprR (ES) | Conjugative, requires insert for homologous integration | Vector for integration by homologous recombination before Meganuclease cleavage |
| pIJ12739 | Meganuclease delivery plasmid | AprR (ES), TsrR (S) | Conjugative, multi-copy, unstable | pSG5 origin, temperature-sensitive, Tsr-inducible (tipAp) Meganuclease gene |
| pIJ12742 | Meganuclease delivery plasmid | AprR (ES), TsrR (S) | Conjugative, multi-copy, unstable | pSG5 origin, temperature-sensitive, ermE*p constitutive Meganuclease gene |
| pIJ12900 | Deletion/replacement vector | KanR (ES), TsrR (S) | Non-conjugative, multi-copy, unstable | pSG5 origin, temperature-sensitive, useful for deletion after introduction of homologous flanking regions and AprR/oriT |
| pIJ12901 | Deletion/replacement vector | KanR (ES), TsrR (S) | Conjugative, multi-copy, unstable | pIJ12900 with oriT, useful for generation of unmarked deletions |
Reporter vectors
| Plasmid | Description | Resistance | Replicative status and copy number in Streptomyces | Comment |
|---|---|---|---|---|
| pIJ486/7 | KanR reporter plasmid | TsrR (S) | Non-conjugative, multi-copy | Kan reporter plasmids |
| pIJ8630 | EGFP reporter plasmid | AprR (ES) | Conjugative, integrative (phiC31) | Cloning sites: BglII, KpnI, XbaI, NdeI |
| pIJ8660 | EGFP reporter plasmid | AprR (ES) | Conjugative, integrative (phiC31) | Cloning sites: BglII, EcoRV, BamHI, XbaI, NsiI, KpnI, NdeI |
| pIJ10500 | Triple flag-tag plasmid | HygR (ES) | Conjugative, integrative (phiBT1) | Flag (3x) cassette in EcoRV site of pMS82 |
| pIJ10740 | Positive control for pGUS assays | AprR (ES) | Conjugative, integrative (phiC31) | pGUS with ermE*p |
| pIJ10741 | Positive control for pIJ10742 assays | HygR (ES), SpcR (E) | Conjugative, integrative (phiBT1) | pIJ10742 with ermE*p |
| pIJ10742 | Beta-glucuronidase reporter plasmid | HygR (ES), SpcR (E) | Conjugative, integrative (phiBT1) | pMS82 with gusA (beta-glucuronidase assays) |
Cloning vectors
| Plasmid | Description | Resistance | Replicative status and copy number in Streptomyces | Comment |
|---|---|---|---|---|
| pIJ10706 | Cloning vector with MCS | HygR (ES) | Conjugative, integrative (phiC31) | pSET152 derivative with AprR replaced with HygR |
| pIJ10750 (pSS10) | Cloning vector with MCS | HygR (ES) | Conjugative, integrative (phiBT1) | pMS82 derivative with additional cloning sites (AprR gene promoter reads into MCS) |
| pIJ10770 (pSS170) | Cloning vector with MCS | HygR (ES) | Conjugative, integrative (phiBT1) | pIJ10750 derivative with AprR gene promoter removed |
| pIJ12330 | Cloning vector with MCS | HygR (ES), AprR (ES) | Conjugative, integrative (phiC31) | pIJ10706 derivative with additional resistance marker - useful for establishing conjugation/selection conditions |
| pIJ12331 | Cloning vector with MCS | HygR (ES), SpcR/StrR (S) | Conjugative, integrative (phiC31) | pIJ10706 derivative with additional resistance marker - useful for establishing conjugation/selection conditions |
| pIJ12332 | Cloning vector with MCS | HygR (ES), VioR (S) | Conjugative, integrative (phiC31) | pIJ10706 derivative with additional resistance marker - useful for establishing conjugation/selection conditions |
| pIJ12333 | Cloning vector with MCS | HygR (ES), TsrR (S) | Conjugative, integrative (phiC31) | pIJ10706 derivative with additional resistance marker - useful for establishing conjugation/selection conditions |
| pSET152 | Cloning vector | AprR (ES) | Conjugative, integrative (phiC31) |
Cosmid vector
| Plasmid | Description | Resistance | Replicative status and copy number in Streptomyces | Comment |
|---|---|---|---|---|
| pIJ10702 | Conjugative, integrative cosmid vector | AprR (ES), Amp (E ) | Conjugative, integrative (phiC31) | Derived from SuperCos1 |
Vectors used for PCR targeting
| Plasmid | Description | Resistance | Replicative status and copy number in Streptomyces | Comment |
|---|---|---|---|---|
| pIJ773 | PCR-targeting - AprR, oriT cassette | AprR (ES), AmpR (E ) | Non-conjugative, multi-copy in E. coli | With FRT sites |
| pIJ773_del_oriT | PCR-targeting - AprR cassette | AprR (ES), AmpR (E ) | Non-conjugative, multi-copy in E. coli | With FRT sites |
| pIJ778 | PCR-targeting - SpcR/StrR, oriT cassette | SpcR/StrR (S), AmpR/SpcR (E ) | Non-conjugative, multi-copy in E. coli | With FRT sites |
| pIJ799 | PCR-targeting - AprR, oriT cassette | AprR (ES), AmpR (E ) | Non-conjugative, multi-copy in E. coli | No FRT sites, flanked by bla sequences (to replace bla by AprR-oriT to construct conjugative SuperCos (including flipped) derivatives |
| pIJ10700 | PCR-targeting - HygR, oriT cassette | HygR (ES), AmpR (E ) | Non-conjugative, multi-copy in E. coli | With FRT sites |
| pIJ10701 | PCR-targeting - HygR, oriT cassette | HygR (ES), AmpR (E ) | Non-conjugative, multi-copy in E. coli | No FRT sites, flanked by bla sequences (to replace bla by HygR-oriT to construct conjugative SuperCos (including flipped) derivatives |
Other vectors
| Plasmid | Description | Resistance | Replicative status and copy number in Streptomyces | Comment |
|---|---|---|---|---|
| pKC1132 | Suicide vector with oriT | AprR (ES) | Conjugative, non-integrative | Blue/white screening in E. coli for insertion of deletion/disruption cassette |
