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Plasmid

Part:BBa_K1930007:Design

Designed by: Barbora Waclawikova   Group: iGEM16_Groningen   (2016-10-16)
Revision as of 11:06, 17 October 2016 by Barawaclawik (Talk | contribs) (→‎References)


Ciprofloxacin resistance cassette in integration plasmid (BBa_K823023)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 536
    Illegal BamHI site found at 7181
    Illegal XhoI site found at 4689
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 2243
    Illegal NgoMIV site found at 4653
    Illegal NgoMIV site found at 5792
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 5616


Design Notes

This part is an integration plasmid (BBa_K823023) carrying ciprofloxacin resistance cassette (BBa_K1930004). BBa_K823023 is an available BioBrick from iGEM Munich 2012. It is an integration plasmid for Bacillus subtilis, which can be used for cloning in E. coli as well. Ciprofloxacin resistance cassette is containing a PAtpI constitutive promoter, ciprofloxacin resistance gene qnrS1 and double terminator.

The promoter PAtpI (BBa_K1930005) is a constitutive promoter which has its origin in Bacillus subtilis. It is responsible for the expression of atpA gene (ATP synthesis) during the first 30 minutes of the germination of B. subtilis. atpA gene is part of an operon (atpI-atpB-atpE-atpF-atpH-atpA-atpG-atpD-atpC), therefore the promoter region in front of the first protein coding gene (atpI) in this operon was chosen.

Ciprofloxacin resistance gene qnrS1 is found naturally in E. coli, and other Gram-negative strains. qnr genes code for pentapeptide repeat proteins. These proteins reduce susceptibility to quinolones by protecting the complex of DNA and DNA gyrase enzyme from the inhibitory effect of quinolones.

Double terminator (BBa_B0015) is the most commonly used terminator. For more information see here: https://parts.igem.org/Part:BBa_B0015.

For more information about designing this part see: [http://2016.igem.org/Team:Groningen/Experiments Experiments Plasmid construction]

Source

Ciprofloxacin resistance cassette: Ordered as gBlock. IDT (Integrated DNA technologies).

BBa_K823023: Created by iGEM Munich 2012. https://parts.igem.org/Part:BBa_K823023.


References

<p> Quinolone resistance

Gene ID