pMS33
(Plasmid #32296)

• Depositing Lab: Elisabeth Raleigh
• Publication: Sibley et al Nucleic Acids Res. 2012 Feb;40(3):e19. doi: 10.1093/nar/gkr1085. Epub 2011 Nov 28.

Backbone

• Vector backbone: pMS26
• Backbone size w/o insert (bp): 13150
• Modifications to backbone: USERBstBI fragment replaced with lacZ amplicon
• Vector type: Bacterial Expression ; bacterial gene addition

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): DH10B
• Growth instructions: Following transformation and plating on rich Ampicillin (100 ug/ml) at 30C, restreak at 42C without drug on plates containing rhamnose and Xgal to obtain strains with insertion of mTn7(phi-lacZMS33) into attTn7
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: lacZ ORF with short 5'UTR
• Species: E. coli
• Insert Size (bp): 3204
• GenBank ID: AAC73447.1 Gene ID: 945006
• Promoter: rhaBp

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: USERBstBI (destroyed during cloning)
• 3′ cloning site: USERBstBI (destroyed during cloning)
• 5′ sequencing primer: GATCTAAACTATGACAATAAAG
• 3′ sequencing primer: na

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Industry MTA

Depositor Comments

Amplification and sequencing primers for verifying insertion into attTn7:

glmS-proximal (Tn7R side)
5’-AATCTGTAACGTTCCGGGTTC-3’ OR
5'-GATGCTGGTGGCGAAGCTGT-3'

pstS-proximal (Tn7L side)
5’-CATTAATAACGAAGAGATGAC-3’ OR
5'-GATGACGGTTTGTCACATGGA-3'