Overview of Streptomyces vectors

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Plasmids

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