pPMQAK1
(Plasmid #26052)

• Depositing Lab: Peter Lindblad
• Publication: Huang et al Nucleic Acids Res. 2010 May . 38(8):2577-93.

Backbone

• Vector backbone: RSF1010 derivative pAWG1.1 + pSB1AK3-BBa_B0015
• Backbone size w/o insert (bp): 8372
• Vector type: Bacterial Expression
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin and Kanamycin, 100 & 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DB3.1
• Growth instructions: DB3.1
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: BioBrick-compatible cyanobacterial shuttle vector

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: see comments section (not destroyed)
• 3′ cloning site: see comments section (not destroyed)
• 5′ sequencing primer: Amp-R

Resource Information

• Addgene Notes:
  • ccdB sequence

Terms and Licenses

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

Depositor Comments

RSF1010 replicon from pAWG1.1 was PCR amplified with pAWG02-MunI-f and pAWG02-KpnI-r. A 2434bp fragment containing BioBrick cloning site and cassettes encoding resistance for ampicillin and kanamycin were PCR-amplified from pSB1AK3-BBa_B0015 with pSB03-KpnI-f and pSB03-MunI-r primers (see publication for primer sequences). These PCR products were cloned together to form the plasmid pPMQAK1-BBa_B0015. The BioBrick part BBa_B0015 was exchanged with part BBa_P1010 (encoding ccdB) using the EcoRI and PstI sites of the BioBrick cloning site.

pPMQAK1 multiple cloning site: AatII-EcoRI-XbaI-ccdB gene-BamHI-SpeI-PstI.

The ccdB gene is removed by cloning a BioBrick part into the cloning site.

For information on using BioBrick vectors see: Shetty RP, Endy D, Knight TF Jr. Engineering BioBrick vectors from BioBrick parts. J. Biol. Eng. (2008) 2:5