pSL-MS2-24X
(Plasmid #27120)

• Depositing Lab: Robert Singer
• Publication: Bertrand et al Mol Cell. 1998 Oct . 2(4):437-45.

Backbone

• Vector backbone: pSL1180
• Backbone manufacturer: Pharmacia
• Backbone size w/o insert (bp): 3422
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): Stbl2
• Growth instructions: Maintain in Stbl2 cells. Grow at 30C for 2 days.
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: MS2 binding sites (24)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: BglII (not destroyed)
• 5′ sequencing primer: M13_Rev

Terms and Licenses

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

Depositor Comments

Loop sequence: acatgaggatcacccatgt

This plasmid has been replaced by a more stable version of the 24X MS2 repeats that is less prone to recombination. Please see pCR4-24XMS2SL-stable (Addgene plasmid# 31865).

Note that this plasmid requires special handling during transformation and amplification because the bacteria can excise the MS2 binding site repeats. To reduce the chance of recombination, maintain the plasmid in Stbl2 cells and grow the bacteria at 30C (for at least two days). To prepare plasmid DNA, perform mini preps from 10 colonies, run double digests with BamHI and BglII for each colony to screen for colonies that contain 24 repeats. A correct double digest should show bands at ~3.4kb and 1.3kb. Select a positive colony containing 24 repeats for a larger plasmid prep and verify the plasmid again with a double digest.