pCR4-24XMS2SL-stable
(Plasmid #31865)

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

Backbone

• Vector backbone: pCR4Blunt-TOPO
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 3956

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: 24xMS2 Stem Loops - Stable
• Insert Size (bp): 1300

Cloning Information

• Cloning method: TOPO Cloning
• 5′ sequencing primer: M13F
• 3′ sequencing primer: M13R

Resource Information

• Addgene Notes:
  • Addgene diagnostic digest for 31865
• Articles Citing this Plasmid:
  • 35 References

Terms and Licenses

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

Depositor Comments

The Singer Lab no longer recommends this plasmid, as it has a higher rate of bacterial recombination.

For mammalian cells they recommend the following plasmids:
pUbC-FLAG-24xSuntagV4-oxEBFP-AID-baUTR1-24xMS2V5-Wpre (Plasmid #84561)
HaloTag-bActinCDS-bActinUTR-MS2V5 (Plasmid #102718)

For yeast cells they recommend the following plasmids:
pET246-pUC57 12xMS2V6 (Plasmid #104390)
pET259-pUC57 24xMS2V6 (Plasmid #104391)
pET251-pUC 12xMS2V6 Loxp KANr Loxp (Plasmid #104392)
pET264-pUC 24xMS2V6 Loxp KANr Loxp (Plasmid #104393)

This version of the 24xMS2 stem loops has been engineered to allow stable propagation in bacteria. Loss of repeats is still possible, but occurs significantly less often than with previous version. This plasmid is considered the replacement for pSL-MS2-24X (Addgene plasmid# 27120).

The 24xMS2 stem loop cassettes in this plasmid were generated from two non-identical stem-loops to reduce unnecessary redundancy and the chance of recombination in bacteria. The sequence for a single cassette (with the two non-identical stem-loops in uppercase) is: tacggtacttattgccaagaaaGCACGAGCATCAGCCGTGC ctccaggtcgaatcttcaaaCGACGACGATCACGCGTCGctccagtattccagggttcatc