Ordering

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Backbone

• Vector backbone
  pRS404
  (Search Vector Database)
• Backbone manufacturer
  Stratagene
• Backbone sizew/o insert(bp)4274
• Total vector size (bp)5320
• Modifications to backbone
  CYC1 transcriptional terminator present downstream of inserts (between XhoI and PvuII sites).
• Vector type
  Yeast Expression, Synthetic Biology ; Expression regulator/reporter
• Selectable markers
  TRP1

Growth in Bacteria

• Bacterial Resistance(s)
  Ampicillin
• Growth Temperature
  37°C
• Growth Strain(s)
  XL10 Gold
• Copy number
  High Copy

Gene/Insert1

• Gene/Insert name
  PTETREG promoter
• Alt name
  Yeast ADH1 transcriptional term. and two copies of the tetO2 operator site upstream of the PCYC1 TATA box and minimal promoter
• Alt name
  rtTA-MF inducible promoter
• Species
  Synthetic
• Insert Size (bp)
  504
• PromoterPTETREG

Cloning Informationfor Gene/Insert 1

• Cloning methodRestriction Enzyme
• 5′ cloning siteAflII(not destroyed)
• 3′ cloning siteBamHI(not destroyed)
• 5′ sequencing primerF1ori-R (AGGGAAGAAAGCGAAAGGAG)
• 3′ sequencing primerTet-R (GGCGAGTTTACGGGTTGTTA)
(Common Sequencing Primers)

Gene/Insert2

• Gene/Insert name
  rtetR-M2::FFF
• Alt name
  modified rtTA transactivator (rtTA-MF)
• Alt name
  rtetR-M2 transactivator variant augmented with the short FFF activation domain
• Alt name
  Three VP16-derived "F-type" minimal acidic activation domains
• Species
  Synthetic; E. Coli
• Insert Size (bp)
  750
• Mutation
  Compared to rtTA, rtTA-M2 has S12G, E19G, A56P, D148E, and H179R

Cloning Informationfor Gene/Insert 2

• Cloning methodRestriction Enzyme
• 5′ cloning siteBamHI(not destroyed)
• 3′ cloning siteXhoI(not destroyed)
• 5′ sequencing primerTetR-F1 (TTGATCACCAAGGTGCAGAG)
(Common Sequencing Primers)

Resource Information

• Supplemental Documents
  • pDN-T2dMFot.PDW
• A portion of this plasmid was derived from a plasmid made by
  PTETREG promoter from plasmid pBB247 from Attila Becskei and Luis Serrano, EMBL Heidelberg, Germany.Modified Tet transactivator rtetR-M2 from plasmid pCM190-M2 from Wolfgang Hiller, Friedrich-Alexander-Universität Erlangen-Nürnberg, Germany.VP16-derived "F-type" minimal acidic activation domain described by Hermann Bujard, ZMBH, Heidelberg, Germany.
• Terms and Licenses
  • UBMTA
• Industry Terms
  • Not Available to Industry