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http://www.addgene.org/191355

Species: Synthetic
Genetic Insert: PGusA2-TetO2/1-Nanoluc, miniPthl-tetR cassette
Vector Backbone Description: Backbone Marker:Tolonen Lab; Backbone Size:5471; Vector Backbone:pQmod2E-GG (Addgene 191345); Vector Types:Bacterial Expression; Bacterial Resistance:Erythromycin
References:
Comments:

Proper citation: RRID:Addgene_191355 Copy   

•   Source: Addgene

http://www.addgene.org/191352

Species: Other
Genetic Insert: Plac-RFP (IPTG-inducible red fluorescent protein)
Vector Backbone Description: Backbone Marker:PMID 19775243; Backbone Size:6972; Vector Backbone:pQexp; Vector Types:Bacterial Expression; Bacterial Resistance:Erythromycin
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Comments:

Proper citation: RRID:Addgene_191352 Copy   

•   Source: Addgene

http://www.addgene.org/218505

Species: Other
Genetic Insert: lacI
Vector Backbone Description: Vector Backbone:p7INT; Vector Types:Bacterial Expression; Bacterial Resistance:Erythromycin
References:
Comments: This plasmid enables the constitutive expression of LacI in S. pyogenes. It can be used for inducible gene expression in combination with the Plac promoter variants described in the associated publication. Resource information: The Pveg promoter originates from the Bacillus subtilis PY79 genome. The lacI repressor is derived from pPEPY-PF6_lacI (Plasmid #85589) and originally codon-optimised for Streptococcus pneumoniae. However, codon usage is quite similar between S. pyogenes and S. pneumoniae. p7INT was a gift from Prof. Michael Federle (University of Illinois Chicago, USA). The p7INT plasmid integrates into the 3' end of the tmRNA locus in several S. pyogenes strains. It was originally described in: McShan, W.M., McLaughlin, R.E., Nordstrand, A. et al. Vectors containing streptococcal bacteriophage integrases for site-specific gene insertion. Methods Cell Sci 20, 51–57 (1998). https://doi.org/10.1023/A:1009773309163 Please download the file linked below to obtain a well annotated map of the plasmid.

Proper citation: RRID:Addgene_218505 Copy   

•   Source: Addgene

http://www.addgene.org/220306

Species:
Genetic Insert:
Vector Backbone Description: Backbone Size:4457; Vector Backbone:pERASE; Vector Types:Synthetic Biology; Bacterial Resistance:Erythromycin
References:
Comments: This plasmid allows to obtain scarless gene deletions in S. pyogenes and potentially other Gram-positive bacteria. The multiple cloning site (EcoRI, XbaI, SpeI, PstI) contains a Plac_mrfp cassette, allowing for red-white screening of clones in E. coli (DH5a). When the Plac_mrfp cassette is replaced with the insert of interest, colonies will appear white on LB agar plates. See the associated publication for more information. Resource Information: The pUC origin for replication in E. coli was taken from pUC19 (Plasmid #50005). The erythromycin resistance cassette was taken from p7INT. The MCS containing the mrfp cassette was taken from pSpy0C4. The coding sequence for the counterselection marker pheS* was taken from the native pheS coding sequence found in S. pyogenes SF370. This sequence was adapted to avoid recombination events of the plasmid at the phis locus due to high DNA sequence homology. Two mutations were introduced into the adapted pheS sequence to facilitate incorporation of the toxic phenylalanine analogue PCPA (4-chlor-DL-phenylalanine) into proteins, leading to bacterial cell death. p7INT was a gift from Prof. Michael Federle (University of Illinois Chicago, USA). The p7INT plasmid integrates into the 3' end of the tmRNA locus in several S. pyogenes strains. It was originally described in: McShan, W.M., McLaughlin, R.E., Nordstrand, A. et al. Vectors containing streptococcal bacteriophage integrases for site-specific gene insertion. Methods Cell Sci 20, 51–57 (1998). https://doi.org/10.1023/A:1009773309163 Please download the detailed plasmid map using the file linked below.

Proper citation: RRID:Addgene_220306 Copy   

•   Source: Addgene

http://www.addgene.org/209986

Species: Other
Genetic Insert: resistance cassette (ermBL)
Vector Backbone Description: Vector Backbone:pLP5; Vector Types:Bacterial Expression; Bacterial Resistance:Erythromycin
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Comments:

Proper citation: RRID:Addgene_209986 Copy   

•   Source: Addgene

http://www.addgene.org/209979

Species: Other
Genetic Insert: resistance cassette (ermBL)
Vector Backbone Description: Vector Backbone:pLP2; Vector Types:Bacterial Expression; Bacterial Resistance:Erythromycin
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Comments:

Proper citation: RRID:Addgene_209979 Copy   

•   Source: Addgene

http://www.addgene.org/223200

Species:
Genetic Insert: N/A
Vector Backbone Description: Vector Backbone:pJC005.em; Vector Types:CRISPR; Bacterial Resistance:Erythromycin
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Comments:

Proper citation: RRID:Addgene_223200 Copy   

•   Source: Addgene

http://www.addgene.org/223201

Species:
Genetic Insert: N/A
Vector Backbone Description: Vector Backbone:pJC005.em; Vector Types:CRISPR; Bacterial Resistance:Erythromycin
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Comments:

Proper citation: RRID:Addgene_223201 Copy   

•   Source: Addgene

http://www.addgene.org/46886

Species:
Genetic Insert:
Vector Backbone Description: Backbone Size:8353; Vector Backbone:pMSP3535; Vector Types:Bacterial Expression, Other, Nisin-inducible expression; Gram-positive Bacterial Shuttle Vector; Bacterial Resistance:Erythromycin
References:
Comments: This plasmid utilizes the NIsin Controlled Expression (NICE) system. NICE® is a registered trademark of NIZO food research BV, P.O.Box 20, 6710 BA Ede, The Netherlands. pMSP3535 contains the Nisin-inducible PnisA promoter, and the pAMB1 replicon for expression in gram-positive bacteria. For protein expression in E. faecalis, induce with 10-25 ng/mL nisin (Sigma, N-5764). See associated article for instructions on how to make nisin solution. Note: Nucleotide sequence encoding the signal sequence and one-third of the mature NisA (nisin) peptide remains associated with the PnisA promoter. The depositing laboratory recommends adding of a Stop codon to the 5' end of inserted sequences to avoid unintentional gene fusions. Addgene's quality control sequencing finds discrepancies with the depositor's provided sequencew which includes a mutation in RepE- F229L. This however is thought to not affect plasmid function.

Proper citation: RRID:Addgene_46886 Copy   

•   Source: Addgene

http://www.addgene.org/229148

Species: Other
Genetic Insert: mCherry, DinJ/YafQ
Vector Backbone Description: Backbone Marker:Addgene ; Vector Backbone:pLp_3050sNuc; Vector Types:Bacterial Expression; Bacterial Resistance:Erythromycin
References:
Comments: Addgene observed a ~1.4k base pair insertion downstream of the T7 terminator. This discrepancy does not affect plasmid function.

Proper citation: RRID:Addgene_229148 Copy   

•   Source: Addgene

http://www.addgene.org/229145

Species: Other
Genetic Insert: mCherry
Vector Backbone Description: Backbone Marker:Addgene ; Backbone Size:2800; Vector Backbone:pLp_3050sNuc; Vector Types:Bacterial Expression; Bacterial Resistance:Erythromycin
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Comments:

Proper citation: RRID:Addgene_229145 Copy   

•   Source: Addgene

http://www.addgene.org/98841

Species:
Genetic Insert: tracr/Cas9
Vector Backbone Description: Backbone Size:6419; Vector Backbone:pTRKL2; Vector Types:; Bacterial Resistance:Erythromycin
References:
Comments: Note from the paper of O'Sullivan, and Klaenhammer: We found that BHI (Difco Laboratories, Detroit, MI, USA) was an excellent selection medium for ErR in E. coli. Unlike LB medium, BHI plates containing 150µg Er/ml afforded a clean selection, with no background colonies appearing even upon prolonged incubaction of one week or more. O'Sullivan, D.J., T.R. Klaenhammer. 1993. High- and low-copy-number Lactococcus shuttle cloning vectors with features for clone screening. Gene 137:227-231.

Proper citation: RRID:Addgene_98841 Copy   

•   Source: Addgene

http://www.addgene.org/133853

Species:
Genetic Insert:
Vector Backbone Description: Backbone Size:1800; Vector Backbone:pWV01; Vector Types:Bacterial Expression; Bacterial Resistance:Erythromycin
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Comments:

Proper citation: RRID:Addgene_133853 Copy   

•   Source: Addgene

http://www.addgene.org/167521

Species: Synthetic
Genetic Insert: CinRAM-KmR
Vector Backbone Description: Vector Backbone:pFCiKm7; Vector Types:Bacterial Expression, Synthetic Biology; Bacterial Resistance:Erythromycin
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Comments:

Proper citation: RRID:Addgene_167521 Copy   

•   Source: Addgene

http://www.addgene.org/182738

Species:
Genetic Insert:
Vector Backbone Description: Backbone Size:11031; Vector Backbone:pJC005.em; Vector Types:CRISPR; Bacterial Resistance:Erythromycin
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Comments:

Proper citation: RRID:Addgene_182738 Copy   

•   Source: Addgene

http://www.addgene.org/25820

Species:
Genetic Insert: pQexp
Vector Backbone Description: Backbone Size:6559; Vector Backbone:pAT19; Vector Types:Bacterial Expression; Bacterial Resistance:Erythromycin
References:
Comments: Addgene sequence has a small insertion and small deletion compared to author sequence. These differences are in regions of the plasmid backbone that were not modified and should not affect function of this plasmid.

Proper citation: RRID:Addgene_25820 Copy   

•   Source: Addgene

http://www.addgene.org/48667

Species: Other
Genetic Insert: Cas9
Vector Backbone Description: Vector Backbone:colE1-erm; Vector Types:CRISPR; Bacterial Resistance:Erythromycin
References:
Comments:

Proper citation: RRID:Addgene_48667 Copy   

•   Source: Addgene

http://www.addgene.org/122030

Species: Other
Genetic Insert: signal peptide Lp_3050
Vector Backbone Description: Backbone Size:5600; Vector Backbone:spp-based expression vector (pSIP401); Vector Types:Bacterial Expression; Bacterial Resistance:Erythromycin
References:
Comments: Plasmid for heterologous secretion of proteins in Lactobacillus plantarum and L. sakei

Proper citation: RRID:Addgene_122030 Copy   

•   Source: Addgene

http://www.addgene.org/122028

Species:
Genetic Insert: gusA
Vector Backbone Description: Backbone Size:5400; Vector Backbone:spp-based expression vector (pSIP401); Vector Types:Bacterial Expression; Bacterial Resistance:Erythromycin
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Comments:

Proper citation: RRID:Addgene_122028 Copy   

•   Source: Addgene

http://www.addgene.org/135655

Species: Synthetic
Genetic Insert: Intron containing lox site
Vector Backbone Description: Backbone Size:6500; Vector Backbone:pAT19; Vector Types:Bacterial Expression, Cre/Lox; Bacterial Resistance:Erythromycin
References:
Comments: Use in combination with pQlox66F (addgene 135657) or pQlox 66R (addgene 135658), then pQcre1 (addgene 135659). Transcription of LtrB-derived group II intron is driven by the pFer promoter. Redesign intron to target gene using TargeTron or Clostron algorithms (links below), then clone by SOE-PCR or gene synthesis ligation between NdeI and BsrGI sites. Lox66 and Lox71 must be in same orientation to delete the intervening sequence, but the directions of introns relative to one another can generate inverted repeats in the chromosome after deletion has taken place, leading to futher recombinations events. See publication for details. Intron redesign links http://www.clostron.com/clostron1.php http://www.targetrons.com/targetron_pLtrB.php Reference for intron redesign Heap JT, Pennington OJ, Cartman ST, Carter GP, Minton NP. The ClosTron: A universal gene knock-out system for the genus Clostridium. J Microbiol Methods. 2007;70(3):452-464. Perutka J, Wang W, Goerlitz D, Lambowitz AM. Use of Computer-designed Group II Introns to Disrupt Escherichia coli DExH / D-box Protein and DNA Helicase Genes. 2004:421-439.

Proper citation: RRID:Addgene_135655 Copy   

•   Source: Addgene