Plasmids   Select Another Resource Report Type

http://www.addgene.org/196026

Species: E. coli
Genetic Insert: MlaD
Vector Backbone Description: Vector Backbone:pET21a; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:37100290
Comments: Please visit https://www.biorxiv.org/content/10.1101/2022.12.09.519820v1 for bioRxiv preprint.

Proper citation: RRID:Addgene_196026 Copy   

•   Source: Addgene

http://www.addgene.org/195692

Species: E. coli
Genetic Insert: MlaFEDCB
Vector Backbone Description: Vector Backbone:pBAD-His; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:37100290
Comments: Please visit https://www.biorxiv.org/content/10.1101/2022.12.09.519820v1 for bioRxiv preprint.

Proper citation: RRID:Addgene_195692 Copy   

•   Source: Addgene

http://www.addgene.org/195687

Species: E. coli
Genetic Insert: MlaD
Vector Backbone Description: Vector Backbone:pET21a; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:37100290
Comments: Please visit https://www.biorxiv.org/content/10.1101/2022.12.09.519820v1 for bioRxiv preprint.

Proper citation: RRID:Addgene_195687 Copy   

•   Source: Addgene

http://www.addgene.org/195684

Species: E. coli
Genetic Insert: MlaC
Vector Backbone Description: Vector Backbone:pET21a; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:37100290
Comments: Please visit https://www.biorxiv.org/content/10.1101/2022.12.09.519820v1 for bioRxiv preprint.

Proper citation: RRID:Addgene_195684 Copy   

•   Source: Addgene

http://www.addgene.org/195682

Species: E. coli
Genetic Insert: MlaC
Vector Backbone Description: Vector Backbone:pET21a; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:37100290
Comments: Please visit https://www.biorxiv.org/content/10.1101/2022.12.09.519820v1 for bioRxiv preprint.

Proper citation: RRID:Addgene_195682 Copy   

•   Source: Addgene

http://www.addgene.org/195683

Species: E. coli
Genetic Insert: MlaC
Vector Backbone Description: Vector Backbone:pET21a; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:37100290
Comments: Please visit https://www.biorxiv.org/content/10.1101/2022.12.09.519820v1 for bioRxiv preprint.

Proper citation: RRID:Addgene_195683 Copy   

•   Source: Addgene

http://www.addgene.org/174431

Species: E. coli
Genetic Insert: β-galactosidase α gene fragment
Vector Backbone Description: Backbone Size:35555; Vector Backbone:pAd5-Blue; Vector Types:Mammalian Expression, Adenoviral; Bacterial Resistance:Kanamycin
Defining Citation: PMID:28039799

Proper citation: RRID:Addgene_174431 Copy   

•   Source: Addgene

http://www.addgene.org/192981

Species: E. coli
Genetic Insert: T7 polymerase
Vector Backbone Description: Backbone Size:5739; Vector Backbone:pSRKGm; Vector Types:Bacterial Expression; Bacterial Resistance:Gentamicin
Defining Citation: PMID:36645284
Comments: The T7 polymerase gene (1) was cloned from the chromosome of BL21(DE3).

Proper citation: RRID:Addgene_192981 Copy   

•   Source: Addgene

http://www.addgene.org/78204

Species: E. coli
Genetic Insert: ssb (W88C)
Vector Backbone Description: Vector Backbone:pET22b; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:20350930
Comments: Please note- The numbering system used was for the mature natural wild-type ssb protein. The N-terminal methionine is quantitatively cleaved off in E. coli and was not numbered so all mutations will be off by 1 nucleotide (e.g., W89C = W88C). The expressed protein is a single-cysteine variant of E. coli single-stranded DNA-binding protein, EcSSB, for subsequent labeling at the cysteine. The adduct with Cy3B maleimide has been used as a biosensor for single-stranded DNA in TIRF microscopy measurements.

Proper citation: RRID:Addgene_78204 Copy   

•   Source: Addgene

http://www.addgene.org/78203

Species: E. coli
Genetic Insert: ssb (G26C)
Vector Backbone Description: Vector Backbone:pET22b; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:18599625
Comments: Please note: The numbering system used was for the mature natural wild-type ssb protein. The N-terminal methionine is quantitatively cleaved off in E. coli and was not numbered so all mutations will be off by 1 nucleotide (e.g., G27C = G26C). The expressed protein is a single-cysteine variant of E. coli single-stranded DNA-binding protein, EcSSB, for subsequent labeling at the cysteine with one IDCC (N-[2-(iodoacetamido)ethyl]-7-diethylaminocoumarin-3-carboxamide) per subunit. This adduct is then used as a fluorescent single-stranded DNA (ssDNA) biosensor, having ~6-fold fluorescence increase on binding ssDNA with very high affinity.

Proper citation: RRID:Addgene_78203 Copy   

•   Source: Addgene

http://www.addgene.org/78200

Species: E. coli
Genetic Insert: pstS (A17C,I76G,A197C)
Vector Backbone Description: Vector Backbone:pET22b; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:26199994
Comments: Please note- The numbering system used was for the mature natural wild-type PstS protein. The plasmid produces an extra N-terminal methionine so all mutations will be off by 1nt (e.g., A197C =A198C) PstS ORF encoding the mature E. coli (A17C, I76G, A197C)PstS protein (PBPw) containing a stop codon at the end of the gene in pET22b (inserted between Nde1 and Xho1 sites in the MCS). Expression produces MEA at the N-terminus of the mature PstS: So there is an extra M at the N-terminus over the natural, mature PstS. A stop codon was also added at the end of the sequence BEFORE the plasmid-derived His tag. This gives PLY at the C-terminus at the C-terminus of the expressed protein, as in the natural PstS. The amino-acid numbering has been left as based on positions in the natural, mature protein. Usage The expressed protein is a weak-binding variant of E. coli phosphate binding protein, PstS. It contains two cysteines (A17C, A197C) and a mutation to weaken binding (I76G). The gene encodes the mature protein without the N-terminal signal peptide, but with an N-terminal methionine. Subsequently the cysteines are labeled with 2 6-iodoactemidotetramethylrhodamines per molecule. This adduct is then used as a phosphate biosensor, having up to 9-fold fluorescence intensity increase on binding inorganic phosphate, with a low dissociation constant so inorganic phosphate can be measured in the tens of micromolar range.

Proper citation: RRID:Addgene_78200 Copy   

•   Source: Addgene

http://www.addgene.org/78202

Species: E. coli
Genetic Insert: parM (K33A D63C T174A T175N D224C C287A)
Vector Backbone Description: Vector Backbone:pHis17; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:20158267
Comments: The plasmid is used to express the plasmid segregation protein, ParM, from E. col. The protein is modified for use as the scaffold of an ADP biosensor, ATR-ParM. Two cysteine mutations are for label attachment (D63C and D224C); there is a mutation to inhibit filament formation (K33A), two mutations to weaken ATP binding (T174A and T175N), a mutation to change the exposed cysteine (C287A) and a C-terminal His6 tag to aid purification. When labelled with two iodoacetamidotetramethylrhodamine (IATR) fluorophores, this biosensor has a fluorescence intensity change of up to 20-fold and can be used to measure ADP in the range of hundreds of nanomolar to ~100 μM, in the presence of up to millimolar ATP. The biosensor can also be used for measurement of other nucleoside diphosphates e.g. GDP, UDP and others.

Proper citation: RRID:Addgene_78202 Copy   

•   Source: Addgene

http://www.addgene.org/78201

Species: E. coli
Genetic Insert: parM (K33A I27C T174A T175N C287A)
Vector Backbone Description: Vector Backbone:pHis17; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:19801632
Comments: The plasmid is used to express the plasmid segregation protein, ParM, from E. col. The protein is modified for use as the scaffold of an ADP biosensor, MDCC-ParM. A single cysteine mutation is for label attachment (I27C); there is a mutation to inhibit filament formation (K33A), two mutations to weaken ATP binding (T174A and T175N), a mutation to change the exposed cysteine (C287A) and a C-terminal His6 tag to aid purification. When labelled with a single N-[2-(1-maleimidyl)ethyl]-7-diethylaminocoumarin-3-carboxamide (MDCC) fluorophore, this biosensor has a fluorescence intensity change of up to 4-fold and can be used to measure ADP in the range of tens of nanomolar to tens of micromolar in the presence of up to~200 μM ATP.

Proper citation: RRID:Addgene_78201 Copy   

•   Source: Addgene

http://www.addgene.org/78199

Species: E. coli
Genetic Insert: pstS (A17C,A197C)
Vector Backbone Description: Vector Backbone:pET22b; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:26199994
Comments: Please note- The numbering system used was for the mature natural wild-type PstS protein. The plasmid produces an extra N-terminal methionine so all mutations will be off by 1nt (e.g., A197C =A198C) The expressed protein is a double-cysteine variant of E. coli phosphate binding protein. The gene encodes the mature protein without the N-terminal signal peptide, but with an N-terminal methionine, for subsequent labeling at the cysteines with two 6-iodoacetamidotetramethylrhodamines per molecule. This adduct is then used as a fluorescent phosphate biosensor, having 10-15-fold fluorescence increase on binding inorganic phosphate with very high affinity.

Proper citation: RRID:Addgene_78199 Copy   

•   Source: Addgene

http://www.addgene.org/78320

Species: E. coli
Genetic Insert: MetI
Vector Backbone Description: Backbone Marker:Novagen; Backbone Size:5791; Vector Backbone:pET19b; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:22095702

Proper citation: RRID:Addgene_78320 Copy   

•   Source: Addgene

http://www.addgene.org/78316

Species: E. coli
Genetic Insert: MetI
Vector Backbone Description: Backbone Marker:Novagen; Backbone Size:5791; Vector Backbone:pET19b; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:18621668

Proper citation: RRID:Addgene_78316 Copy   

•   Source: Addgene

http://www.addgene.org/78493

Species: E. coli
Genetic Insert: GFPmut2
Vector Backbone Description: Vector Backbone:pZA31-Luc; Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:27060147

Proper citation: RRID:Addgene_78493 Copy   

•   Source: Addgene

http://www.addgene.org/218629

Species: E. coli
Genetic Insert: mitoEcSTH
Vector Backbone Description: Backbone Marker:Clontech; Backbone Size:9227; Vector Backbone:pLVX-TetOne-Puro; Vector Types:Mammalian Expression, Lentiviral, Synthetic Biology; Bacterial Resistance:Ampicillin
Defining Citation: PMID:37884806

Proper citation: RRID:Addgene_218629 Copy   

•   Source: Addgene

http://www.addgene.org/218770

Species: E. coli
Genetic Insert: EcNR1GT pUltraBR MjY dtyrS dtyrTV::tolC tyrU::gentR lambda::ZeoR
Vector Backbone Description: Vector Backbone:n/a; Vector Types:Bacterial Expression; Bacterial Resistance:Spectinomycin (50 ug/mL), Gentamycin (10 ug/mL)
Defining Citation: PMID:30078635

Proper citation: RRID:Addgene_218770 Copy   

•   Source: Addgene

http://www.addgene.org/218766

Species: E. coli
Genetic Insert: EcTyrRS-VSMA*
Vector Backbone Description: Backbone Size:4091; Vector Backbone:pEVOL-tac; Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:38913984
Comments: EcY-tRNA-TAG After the plasmids are propagated, they need to be gel purified by agarose gel because the ATM strains contain a plasmid in their genome.

Proper citation: RRID:Addgene_218766 Copy   

•   Source: Addgene