pTSA-38
(Plasmid #17328)

• Depositing Lab: Takeshi Sano
• Publication: Sano et al Proc Natl Acad Sci U S A. 1995 Apr 11. 92(8):3180-4.

Backbone

• Vector backbone: pET-3a
• Backbone manufacturer: Novagen
• Backbone size w/o insert (bp): 4600
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: streptavidin
• Alt name: Stv-38
• Species: Streptomyces avidinii
• Insert Size (bp): 400
• Mutation: Truncated to include only amino acids 16-133 of mature streptavidin. Trp-120 is replaced by Phe.
• GenBank ID: X03591

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NdeI (not destroyed)
• 3′ cloning site: NdeI (not destroyed)
• 5′ sequencing primer: T7

Terms and Licenses

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

Depositor Comments

pTSA-38 is a derivative of pTSA-13 with a W120F mutation.

In brief, mpSA-29 was generated by inserting
a 480-bp fragment of pTSA-13 carrying the entire coding
sequence into M13mpl8. Oligonucleotide-directed in vitro
mutagenesis was performed on mpSA-29 by using the
30-nt oligonucleotide, 5'-ACCAGCGTGGACTTGAAG-
GCGTTGGCCTCG-3', to convert the TGG codon for Trp-
120 to TTC for Phe (mpSA-36). Then, a 405-bp fragment of
mpSA-36 carrying the entire coding sequence was cloned into
pET-3a. The resulting expression vector, pTSA-38, encodes a streptavidin W120F mutant.