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http://www.addgene.org/46886

Vector Backbone Description: Backbone Size:8353; Vector Backbone:pMSP3535; Vector Types:Bacterial Expression, Nisin-inducible expression; Gram-positive Bacterial Shuttle Vector; Bacterial Resistance:Erythromycin
Defining Citation: PMID:10964628
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/25819

Genetic Insert: qQint
Vector Backbone Description: Backbone Size:6559; Vector Backbone:pAT19; Vector Types:bacterial gene inactivation; Bacterial Resistance:Erythromycin
Defining Citation: PMID:19775243
Comments: Between Addgene sequence and author sequence, there is a 12 nucleotide mismatched region. It is in the intron sequence, which needs to be replaced in order to use the plasmid and should not be of concern.

Proper citation: RRID:Addgene_25819 Copy   

•   Source: Addgene
  

http://www.addgene.org/177208

Species: Branchiostoma lanceolatum
Genetic Insert: mNeonGreen
Vector Backbone Description: Vector Backbone:pmr101A; Vector Types:Bacterial Expression; Bacterial Resistance:Erythromycin
Comments: Braatsch S, Helmark S, Kranz H, Koebmann B, Jensen PR. Escherichia coli strains with promoter libraries constructed by Red/ET recombination pave the way for transcriptional fine-tuning. Biotechniques. 2008 Sep;45(3):335-7. doi: 10.2144/000112907. PMID: 18778259.

Proper citation: RRID:Addgene_177208 Copy   

•   Source: Addgene
  

http://www.addgene.org/177206

Species: Discosoma
Genetic Insert: mCherry
Vector Backbone Description: Vector Backbone:pmr101A; Vector Types:Bacterial Expression; Bacterial Resistance:Erythromycin
Comments: Braatsch S, Helmark S, Kranz H, Koebmann B, Jensen PR. Escherichia coli strains with promoter libraries constructed by Red/ET recombination pave the way for transcriptional fine-tuning. Biotechniques. 2008 Sep;45(3):335-7. doi: 10.2144/000112907. PMID: 18778259.

Proper citation: RRID:Addgene_177206 Copy   

•   Source: Addgene
  

http://www.addgene.org/153516

Species: inactivated dCas9 from Streptococcus pneumoniae
Genetic Insert: dCas9Str
Vector Backbone Description: Backbone Marker:Gary Dunny; Backbone Size:8514; Vector Backbone:pMSP3545; Vector Types:Bacterial Expression; Bacterial Resistance:Erythromycin
Defining Citation: PMID:33082254

Proper citation: RRID:Addgene_153516 Copy   

•   Source: Addgene
  

http://www.addgene.org/141095

Species: S. pyogenes
Genetic Insert: Cas9 and tracrRNA
Vector Backbone Description: Vector Backbone:colE1, pAM beta; Vector Types:Bacterial Expression; Bacterial Resistance:Erythromycin
Defining Citation: PMID:30156038

Proper citation: RRID:Addgene_141095 Copy   

•   Source: Addgene
  

http://www.addgene.org/135656

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
Defining Citation: PMID:31826971
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_135656 Copy   

•   Source: Addgene
  

http://www.addgene.org/135659

Species: P1 phage
Genetic Insert: Cre recombinase
Vector Backbone Description: Backbone Size:6500; Vector Backbone:pAT19; Vector Types:Bacterial Expression, Cre/Lox; Bacterial Resistance:Erythromycin
Defining Citation: PMID:31826971
Comments: Cre protein under the control of the gram + PpagA promoter. Leibig M, Krismer B, Kolb M, Friede A, Götz F, Bertram R. 2008. Marker removal in staphylococci via Cre recombinase and different lox sites. Appl Environ Microbiol 74:1316– 1323.

Proper citation: RRID:Addgene_135659 Copy   

•   Source: Addgene
  

http://www.addgene.org/135662

Species: Synthetic
Genetic Insert: Lox511/66 and LoxFas/71
Vector Backbone Description: Vector Backbone:pAT19; Vector Types:Bacterial Expression, Clostridial vector, pAMβ1 origin; Bacterial Resistance:Erythromycin
Defining Citation: PMID:31826971
Comments: Cargo can be inserted between the lox sites using the SpeI and XhoI sites.

Proper citation: RRID:Addgene_135662 Copy   

•   Source: Addgene
  

http://www.addgene.org/71312

Vector Backbone Description: Backbone Size:4963; Vector Backbone:pIL253; Vector Types:Bacterial Expression, E. coli-gram positive bacteria shuttle vector; Bacterial Resistance:Erythromycin
Defining Citation: PMID:8299952

Proper citation: RRID:Addgene_71312 Copy   

•   Source: Addgene
  

http://www.addgene.org/71595

Vector Backbone Description: Backbone Size:2200; Vector Backbone:pWV01; Vector Types:integration vector for bacteria; Bacterial Resistance:Erythromycin
Defining Citation: PMID:9003306
Comments: Please note that this plasmid may require a unique bacterial strain, so make sure to confirm that you can also obtain the appropriate growth strain. Please contact us at [email protected] or contact our distributors if you have any questions. This plasmid has been found to be somewhat prone to multimerization. Multimerization often does not impact plasmid function, but may reduce transformation efficiencies. You may need to screen multiple colonies to isolate the monomeric version of this plasmid. If you still have trouble isolating the monomeric version, you might consider linearizing, gel extracting, re-ligating, and transforming the plasmid.

Proper citation: RRID:Addgene_71595 Copy   

•   Source: Addgene
  

http://www.addgene.org/71803

Species: Lactobacillus gasseri
Genetic Insert: gusA3
Vector Backbone Description: Backbone Marker:N/A; Backbone Size:4568; Vector Backbone:pTRK882; Vector Types:Bacterial Expression; Bacterial Resistance:Erythromycin
Defining Citation: PMID:21375708

Proper citation: RRID:Addgene_71803 Copy   

•   Source: Addgene
  

http://www.addgene.org/191001

Species: Discosoma sp.
Genetic Insert: red chromoprotein
Vector Backbone Description: Backbone Size:7023; Vector Backbone:pBTK402; Vector Types:Bacterial Expression, Synthetic Biology; Bacterial Resistance:Erythromycin
Defining Citation: PMID:37225918
Comments: Please visit https://www.biorxiv.org/content/10.1101/2023.02.15.528778v1 for bioRxiv preprint.

Proper citation: RRID:Addgene_191001 Copy   

•   Source: Addgene
  

http://www.addgene.org/191348

Genetic Insert: Plac-RFP (IPTG-inducible red fluorescent protein)
Vector Backbone Description: Backbone Marker:Chain Biotech; Backbone Size:3823; Vector Backbone:pMTL83241; Vector Types:Bacterial Expression; Bacterial Resistance:Erythromycin
Defining Citation: PMID:36427328

Proper citation: RRID:Addgene_191348 Copy   

•   Source: Addgene
  

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
Defining Citation: PMID:36427328

Proper citation: RRID:Addgene_191355 Copy   

•   Source: Addgene
  

http://www.addgene.org/191352

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
Defining Citation: PMID:36427328

Proper citation: RRID:Addgene_191352 Copy   

•   Source: Addgene
  

http://www.addgene.org/203682

Genetic Insert: Yellow Fluorescent Protein (YFP)
Vector Backbone Description: Vector Backbone:pNF8; Vector Types:Bacterial Expression; Bacterial Resistance:Erythromycin
Defining Citation: PMID:17014739
Comments: Optimized version of the Cyan- and Yellow fluorescent proteins were created as follows: The hsp60 promoter driven GFP+ [19] was amplified and cloned into the pUC18 plasmid. pUC-yfp++ was obtained by introducing the following mutations by site-directed mutagenesis (Stratagene): F46L, S65G, V68A, S72A, S175G, T203Y, whereas the pUC-cfp++, was obtained by introducing the K26R, Y66W, N146I, N164H, S175G, N212K mutations

Proper citation: RRID:Addgene_203682 Copy   

•   Source: Addgene
  

http://www.addgene.org/203681

Genetic Insert: Cyan Fluorescent Protein (CFP)
Vector Backbone Description: Vector Backbone:pNF8; Vector Types:Bacterial Expression; Bacterial Resistance:Erythromycin
Defining Citation: PMID:17014739
Comments: Please note that this plasmid contains an additional IS4-like element in the backbone.

Proper citation: RRID:Addgene_203681 Copy   

•   Source: Addgene
  

http://www.addgene.org/218505

Species: Escherichia coli
Genetic Insert: lacI
Vector Backbone Description: Vector Backbone:p7INT; Vector Types:Bacterial Expression; Bacterial Resistance:Erythromycin
Defining Citation: PMID:38911550
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

Vector Backbone Description: Backbone Size:4457; Vector Backbone:pERASE; Vector Types:Synthetic Biology; Bacterial Resistance:Erythromycin
Defining Citation: PMID:38911550
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