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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/79165

Species: E. coli
Genetic Insert: yhjH
Vector Backbone Description: Backbone Marker:Novagen; Backbone Size:3719; Vector Backbone:pCOLADuet; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:27382070

Proper citation: RRID:Addgene_79165 Copy   

•   Source: Addgene

http://www.addgene.org/79167

Species: E. coli
Genetic Insert: yhjH
Vector Backbone Description: Backbone Marker:Novagen; Backbone Size:5420; Vector Backbone:pETDuet; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:27382070

Proper citation: RRID:Addgene_79167 Copy   

•   Source: Addgene

http://www.addgene.org/23336

Species: E. coli
Genetic Insert: Enhanced Circularly Permuted Outer Membrane Protein X
Vector Backbone Description: Backbone Size:5400; Vector Backbone:pBAD33; Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:18480093

Proper citation: RRID:Addgene_23336 Copy   

•   Source: Addgene

http://www.addgene.org/207231

Species: E. coli
Genetic Insert: ykgMO-IGS
Vector Backbone Description: Backbone Marker:p15A ori plasmid; Backbone Size:4770; Vector Backbone:pJBL5034; Vector Types:Bacterial Expression, Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:37523538

Proper citation: RRID:Addgene_207231 Copy   

•   Source: Addgene

http://www.addgene.org/207232

Species: E. coli
Genetic Insert: ykgMO-IGS
Vector Backbone Description: Backbone Marker:p15A ori plasmid; Backbone Size:2269; Vector Backbone:pJBL5034; Vector Types:Bacterial Expression, Luciferase, Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:37523538

Proper citation: RRID:Addgene_207232 Copy   

•   Source: Addgene

http://www.addgene.org/209131

Species: E. coli
Genetic Insert: Enzyme BirA
Vector Backbone Description: Vector Backbone:pGEX-2T; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:9887208

Proper citation: RRID:Addgene_209131 Copy   

•   Source: Addgene

http://www.addgene.org/206495

Species: E. coli
Genetic Insert: E. coli tRNA Guanine Transglycosylase
Vector Backbone Description: Backbone Marker:NEB; Backbone Size:5874; Vector Backbone:(T7)-2 Vector; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:36988146

Proper citation: RRID:Addgene_206495 Copy   

•   Source: Addgene

http://www.addgene.org/193338

Species: E. coli
Genetic Insert: tTA2
Vector Backbone Description: Vector Backbone:AAV with CAG promter; Vector Types:Mammalian Expression, AAV; Bacterial Resistance:Ampicillin
Defining Citation: PMID:38918382
Comments: Please visit https://www.biorxiv.org/content/10.1101/2022.10.20.512984v1 for bioRxiv preprint.

Proper citation: RRID:Addgene_193338 Copy   

•   Source: Addgene

http://www.addgene.org/195199

Species: E. coli
Genetic Insert: TetR
Vector Backbone Description: Vector Backbone:pQE80; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:36246369

Proper citation: RRID:Addgene_195199 Copy   

•   Source: Addgene

http://www.addgene.org/201082

Species: E. coli
Genetic Insert: CysGA-Im7 F84A
Vector Backbone Description: Backbone Size:3904; Vector Backbone:pTrc-99a; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:33753520

Proper citation: RRID:Addgene_201082 Copy   

•   Source: Addgene

http://www.addgene.org/201081

Species: E. coli
Genetic Insert: CysGA-Im7
Vector Backbone Description: Backbone Size:3904; Vector Backbone:pTrc-99a; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:33753520

Proper citation: RRID:Addgene_201081 Copy   

•   Source: Addgene