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Species: Synthetic
Genetic Insert: N Terminal fragment of T7 RNAP split betwen 179-180
Vector Backbone Description: Backbone Size:5500; Vector Backbone:pBAD33; Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:23479654
Proper citation: RRID:Addgene_60720 Copy
Species: Synthetic
Genetic Insert: Residues 1-179 of T7 RNAP
Vector Backbone Description: Backbone Size:5500; Vector Backbone:pBAD33; Vector Types:Bacterial Expression, Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:23479654
Proper citation: RRID:Addgene_60721 Copy
Species: Synthetic
Genetic Insert: Residues 1-179 of split T7 RNAP
Vector Backbone Description: Backbone Size:5500; Vector Backbone:pBAD33; Vector Types:Bacterial Expression, Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:23479654
Proper citation: RRID:Addgene_60728 Copy
Species: Synthetic
Genetic Insert: Residues 1-179 of split T7 RNAP
Vector Backbone Description: Backbone Size:5500; Vector Backbone:pBAD33; Vector Types:Bacterial Expression, Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:23479654
Proper citation: RRID:Addgene_60727 Copy
Species: Synthetic
Genetic Insert: Residues 1-179 of split T7 RNAP
Vector Backbone Description: Backbone Size:5500; Vector Backbone:pBAD33; Vector Types:Bacterial Expression, Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:23479654
Proper citation: RRID:Addgene_60730 Copy
Species: Streptococcus pyogenes
Genetic Insert: Cas9, tracrRNA, crRNA
Vector Backbone Description: Vector Backbone:Custom pR1162-CmR; Vector Types:Bacterial Expression, CRISPR; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:25240928
Proper citation: RRID:Addgene_61271 Copy
Species: E. coli
Genetic Insert: SCRIBE_kanR(ON)
Vector Backbone Description: Backbone Marker:Expressys; Vector Backbone:pZE32; Vector Types:Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:25395541
Proper citation: RRID:Addgene_61450 Copy
Genetic Insert: none
Vector Backbone Description: Vector Backbone:none; Vector Types:; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:21925267
Comments: Genotype = cyo ilvE avtA aspC hisG argH metA lysA
Precursor strain = ML31
Selective amino acid labeling (and/or requirement) = (Ala#), Ile, Leu*, Tyr#, Val, His, Arg, Met, Lys
ML40(K1) contains an extra pACYC plasmid carrying the cat cassette plus extra tRNA genes (argU, ileY, leuW) from Agilent Technologies Inc.
*In the presence of 0.4-1 mM Tyr, tyrB is repressed and Leu is required for growth in minimal medium. This strategy can only be applicable for a short-term cultivation but not suitable for a long-term cultivation for heterologous expression of foreign genes [Methods 55, 370-378 (2011)].
#In the presence of 0.4-1 mM Tyr, tyrB is repressed and Tyr is required for growth in minimal medium. Under these conditions, this strain can also be used for selective labeling of the input Tyr and/or Ala label(s), although minor diffusion of the input label(s) can occur. Note that this strategy can only be applicable for a short-term cultivation but not suitable for a long-term cultivation for heterologous expression of foreign geness [Methods 55, 370-378 (2011)].
Each target gene was deleted from the chromosome of C43 (DE3) E. coli strain using λ-Red recombination system.
Supplemental documents contain a list of PCR primers used for verification of each knocked-out gene as well as an image showing PCR results for this strain.
Proper citation: RRID:Addgene_61920 Copy
Vector Backbone Description: Backbone Marker:Bacillus Genetic Stock Center; Backbone Size:4217; Vector Backbone:pNW33N; Vector Types:Bacterial Expression, Synthetic Biology; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:22578246
Proper citation: RRID:Addgene_61805 Copy
Vector Backbone Description: Backbone Marker:Structural Genomics Consortium; Backbone Size:8362; Vector Backbone:pACYC-LIC+; Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol
Proper citation: RRID:Addgene_62329 Copy
Genetic Insert: Cas9 nuclease
Vector Backbone Description: Vector Backbone:pdCas9-bacteria; Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:26463009
Comments: Please note that this plasmid runs as a dimer (>13kb). While this may reduce DNA yield, the plasmid still functions as expected.
Proper citation: RRID:Addgene_62655 Copy
Species: E. coli MG1655
Genetic Insert: L31-sfGFP
Vector Backbone Description: Vector Backbone:p15A oripACYC; Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:36533433
Proper citation: RRID:Addgene_185030 Copy
Species: E. coli MG1655
Genetic Insert: L31-sfGFP
Vector Backbone Description: Vector Backbone:p15A oripACYC; Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:36533433
Proper citation: RRID:Addgene_185037 Copy
Species: E. coli MG1655
Genetic Insert: L31-sfGFP
Vector Backbone Description: Vector Backbone:p15A oripACYC; Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:36533433
Proper citation: RRID:Addgene_185038 Copy
Species: E. coli MG1655
Genetic Insert: L31-sfGFP
Vector Backbone Description: Vector Backbone:p15A oripACYC; Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:36533433
Proper citation: RRID:Addgene_185036 Copy
Species: E. coli MG1655
Genetic Insert: L31-sfGFP
Vector Backbone Description: Vector Backbone:p15A oripACYC; Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:36533433
Proper citation: RRID:Addgene_185033 Copy
Species: E. coli MG1655
Genetic Insert: L31-sfGFP
Vector Backbone Description: Vector Backbone:p15A oripACYC; Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:36533433
Proper citation: RRID:Addgene_185034 Copy
Species: E. coli MG1655
Genetic Insert: L31-sfGFP
Vector Backbone Description: Vector Backbone:p15A oripACYC; Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:36533433
Proper citation: RRID:Addgene_185031 Copy
Species: E. coli MG1655
Genetic Insert: L31-sfGFP
Vector Backbone Description: Vector Backbone:p15A oripACYC; Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:36533433
Proper citation: RRID:Addgene_185032 Copy
Species: E. coli MG1655
Genetic Insert: L31-sfGFP
Vector Backbone Description: Vector Backbone:p15A oripACYC; Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:36533433
Proper citation: RRID:Addgene_185039 Copy
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