rEcoli XpS
(Bacterial strain #192872)

• Purpose: MG1655-C321 -- All 321 UAG codons changed to UAA
• Depositing Lab: Jesse Rinehart
• Publication: Gassaway et al Nat Methods. 2022 Oct 24. pii: 10.1038/s41592-022-01638-5. doi: 10.1038/s41592-022-01638-5.

Backbone

• Vector backbone: N/A

Growth in Bacteria

• Bacterial Resistance(s): Bleocin (Zeocin), 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): MG1655-C321
• Growth instructions: This strain can be grown in standard LB but must be supplemented with d-biotin and Serine when grown in minimal media.
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: MG1655-C321 mutS+, λ-, Δ(ybhB-bioAB)::zeoR, ΔprfA, ΔserB
• Species: E. coli

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

MG1655-C321 -- All 321 UAG codons changed to UAA.
mutS+ -- Restores normal spontaneous mutation frequency.
(λ-) – Deleted Lambda Phage operon to allow growth at 37C.
Δ(ybhB-bioAB)::zeoR – Deletion of biotin biosynthetic operon using Zeocin cassette; strain is d-biotin auxotroph.
ΔprfA – Deletion of polypeptide release factor 1 (RF1) to prevent translation termination at UAG codons.
ΔserB – Deletion of phosphoserine phosphatase to provide intracellular source of phosphoserine; strain is Serine auxotroph.
The genome sequence of a similar strain (C321.deltaA) was deposited at NCBI (accession # CP006698.1)