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pTSA-38
(Plasmid #17328)

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Full plasmid sequence is not available for this item.

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 17328 Standard format: Plasmid sent in bacteria as agar stab 1 $85
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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
    • (Common Sequencing Primers)

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry
Trademarks:
  • Zeocin® is an InvivoGen trademark.

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.

How to cite this plasmid ( Back to top)

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.

  • For your Materials & Methods section:

    pTSA-38 was a gift from Takeshi Sano (Addgene plasmid # 17328 ; http://n2t.net/addgene:17328 ; RRID:Addgene_17328)

  • For your References section:

    Intersubunit contacts made by tryptophan 120 with biotin are essential for both strong biotin binding and biotin-induced tighter subunit association of streptavidin. Sano T, Cantor CR. Proc Natl Acad Sci U S A. 1995 Apr 11. 92(8):3180-4. 10.1073/pnas.92.8.3180 PubMed 7724536
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