pST5832
(Plasmid #36256)

• Depositing Labs: Carolyn Bertozzi, Jessica Seeliger
• Publication: Seeliger et al PLoS One. 2012;7(1):e29266. Epub 2012 Jan 18.

Backbone

• Vector backbone: pMV261
• Backbone manufacturer: Stover et al., 1991 Nature 351:456
• Total vector size (bp): 4357
• Modifications to backbone: The plasmid is derived from pMV261 (http://www.lablife.org/p?a=vdb_view&id=g2.7281169b992e40c865698d3455e3633c24b04b31). It contains a modified promoter and MCS (see reference).
• Vector type: Bacterial Expression, Synthetic Biology ; Mycobacteria replicating vector

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: lacZ
• Alt name: beta-D-galactosidase
• Alt name: bgal
• Species: E. coli
• Insert Size (bp): 3075

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: XbaI (not destroyed)
• 3′ cloning site: HindIII (not destroyed)
• 5′ sequencing primer: M13_for20
• 3′ sequencing primer: rrnB-T1-term-Rev

Resource Information

• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

The plasmid is high copy in E. coli, but low copy in mycobacteria.

Cloning was done via assembly PCR, sot he XbaI-HindIII fragment comprises the promoter-riboswitch. The insert is 3361bp including the promoter. There are mismatches between Addgene's quality control sequence and the depositor sequence, but they are not known to affect plasmid function.