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Genetic Insert: See comments
Vector Backbone Description: Vector Backbone:N/A; Vector Types:Synthetic Biology; Bacterial Resistance:None
Defining Citation: PMID:21189298
Comments: MG1655 + lacIq intergrated at intS + Δ hfq + Δ micC
Proper citation: RRID:Addgene_60372 Copy
Genetic Insert: pTet--Cas9 cassette integrated at 186 primary attB site
Vector Backbone Description: Vector Backbone:na; Vector Types:; Bacterial Resistance:None
Defining Citation: PMID:27060147
Proper citation: RRID:Addgene_78552 Copy
Genetic Insert: none
Vector Backbone Description: Vector Backbone:none; Vector Types:; Bacterial Resistance:None
Defining Citation: PMID:26251500
Comments: The genotype of the strain relative to MG1655 is complex, since there are many single nucleotide variations and several 10-20kb differences (Brown & Jun (2015) Genome Announcements, Lyons et. al. (2011) PLoS ONE). There are a particular abundance of differences in the rDNA operons, which in several cases align more closely to E. coli strains DH10B, MDS42, and W.
Particularly relevant genotypes are: λ+ gal+ eut+ pyrE+ ilvG+ rpoS33Am glnV(SupAm) evgA::IS1 Δrfb
Proper citation: RRID:Addgene_67755 Copy
Vector Backbone Description: Vector Backbone:none; Vector Types:Synthetic Biology; Bacterial Resistance:None
Defining Citation: PMID:27084942
Proper citation: RRID:Addgene_69778 Copy
Vector Backbone Description: Vector Backbone:none; Vector Types:Synthetic Biology; Bacterial Resistance:None
Defining Citation: PMID:27084942
Proper citation: RRID:Addgene_69775 Copy
Vector Backbone Description: Vector Backbone:none; Vector Types:Synthetic Biology; Bacterial Resistance:None
Defining Citation: PMID:27084942
Proper citation: RRID:Addgene_69774 Copy
Vector Backbone Description: Vector Backbone:none; Vector Types:Synthetic Biology; Bacterial Resistance:None
Defining Citation: PMID:27084942
Proper citation: RRID:Addgene_69782 Copy
Vector Backbone Description: Vector Backbone:none; Vector Types:Synthetic Biology; Bacterial Resistance:None
Defining Citation: PMID:27084942
Proper citation: RRID:Addgene_69781 Copy
Genetic Insert: KI (37)-Oplac2 strain
Vector Backbone Description: Vector Backbone:n/a; Vector Types:; Bacterial Resistance:None
Defining Citation: PMID:22605776
Comments: To test whether the mRNA secondary structure of our codon redesigned sequences affected expression, we substituted the first 37 nucleotides of Oplac2 with those of KIlac, producing and KI(37)-Oplac2.
Proper citation: RRID:Addgene_52704 Copy
Genetic Insert: degtag strain
Vector Backbone Description: Vector Backbone:n/a; Vector Types:; Bacterial Resistance:None
Defining Citation: PMID:22605776
Comments: To generate the degtag strain, we introduced the ssrA tag to the C-terminus of LacZ, thus targeting the protein for degradation by the ClpXP and ClpAP proteases.
Proper citation: RRID:Addgene_52705 Copy
Genetic Insert: KI (37)-Oplac1 strain
Vector Backbone Description: Vector Backbone:n/a; Vector Types:; Bacterial Resistance:None
Defining Citation: PMID:22605776
Comments: To test whether the mRNA secondary structure of our codon redesigned sequences affected expression, we substituted the first 37 nucleotides of OpLac1 with those of KIlac.
Proper citation: RRID:Addgene_52703 Copy
Genetic Insert: OpLac2-Δ6 strain
Vector Backbone Description: Vector Backbone:na; Vector Types:; Bacterial Resistance:None
Defining Citation: PMID:22605776
Comments: Oplac2Δ6 was erroneously synthesized missing the first 6 nucleotides of Oplac2. These deletions correspond to the first 2 N-terminal amino acid residues (Methionine and Threonine), and instead begin at the Methionine at position 3.
Proper citation: RRID:Addgene_52700 Copy
Genetic Insert: Oplac1 (37)-KI strain
Vector Backbone Description: Vector Backbone:na; Vector Types:; Bacterial Resistance:None
Defining Citation: PMID:22605776
Comments: To test whether the mRNA secondary structure of our codon redesigned sequences affected expression, we substituted the first 37 nucleotides of KILac with those of OpLac1.
Proper citation: RRID:Addgene_52701 Copy
Genetic Insert: S56L strain
Vector Backbone Description: Vector Backbone:n/a; Vector Types:; Bacterial Resistance:None
Defining Citation: PMID:22605776
Comments: Impaired LacY function
Proper citation: RRID:Addgene_52709 Copy
Genetic Insert: ΔZ strain
Vector Backbone Description: Vector Backbone:n/a; Vector Types:; Bacterial Resistance:None
Defining Citation: PMID:22605776
Comments: lacZ deletion
Proper citation: RRID:Addgene_52706 Copy
Genetic Insert: G794D strain
Vector Backbone Description: Vector Backbone:n/a; Vector Types:; Bacterial Resistance:None
Defining Citation: PMID:22605776
Proper citation: RRID:Addgene_52710 Copy
Genetic Insert: KIlac strain
Vector Backbone Description: Vector Backbone:n/a; Vector Types:; Bacterial Resistance:None
Defining Citation: PMID:22605776
Comments: KIlac refers to the knock-in of the wild-type operon into the attTn7 locus; this strain was used as the reference strain for all analyses. We constructed all knock-in strains using the approach of McKenzie and Craig. The knock-in contained the entire lac operon starting 75bp upstream of lacI and ending 100bp downstream of lacA. The ΔZYA strain was transformed with pGRG37. The plasmid carried both the lac operon and the transposon genes tnsABCD, which allow for a site-specific insertion at attTn7.
Proper citation: RRID:Addgene_52696 Copy
Genetic Insert: OpLac1 strain
Vector Backbone Description: Vector Backbone:na; Vector Types:; Bacterial Resistance:None
Defining Citation: PMID:22605776
Comments: The sequence Oplac1 was synthesized using preferred codons at every position (GENEART), thus optimizing the codon adaptation index.
Proper citation: RRID:Addgene_52697 Copy
Genetic Insert: OpLac2 strain
Vector Backbone Description: Vector Backbone:na; Vector Types:; Bacterial Resistance:None
Defining Citation: PMID:22605776
Comments: Oplac2 was designed by a proprietary optimization strategy (developed by DNA 2.0) that incorporated codons for a subset of amino acids read by tRNAs that are the most highly charged during amino acid starvation.
Proper citation: RRID:Addgene_52699 Copy
Genetic Insert: MG1655 Z1 malE
Vector Backbone Description: Vector Backbone:n/a; Vector Types:; Bacterial Resistance:None
Defining Citation: PMID:26900850
Comments: F-, lambda-, rph-1, laciq, PN25-tetR, SpR, malE-
Proper citation: RRID:Addgene_65915 Copy
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