Difference between revisions of "Part:BBa J435114"

 
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One-Step Cloning and Chromosomal Integration of DNA
 
One-Step Cloning and Chromosomal Integration of DNA
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For propagation, please grow this plasmid in the original strain (E811) In that strain, the strong promoter pE will be repressed, thus avoiding possible toxicity and accumulation of unwanted mutations in the plasmid. Use 50ug/ml ampicillin (100ug/ml may inhibit growth)
  
 
https://freegenes.github.io/genes/BBF10K_000016.html
 
https://freegenes.github.io/genes/BBF10K_000016.html

Latest revision as of 16:25, 23 May 2023


pE-FLP (AmpR Low-copy)

Can be used to remove the integration module (including the antibiotic resistance cassette) from pOSIP integrants.

One-Step Cloning and Chromosomal Integration of DNA

For propagation, please grow this plasmid in the original strain (E811) In that strain, the strong promoter pE will be repressed, thus avoiding possible toxicity and accumulation of unwanted mutations in the plasmid. Use 50ug/ml ampicillin (100ug/ml may inhibit growth)

https://freegenes.github.io/genes/BBF10K_000016.html


One-step cloning and chromosomal integration of DNA. St-Pierre F, Cui L, Priest DG, Endy D, Dodd IB, Shearwin KE. ACS Synth Biol. 2013 Sep 20;2(9):537-41. doi: 10.1021/sb400021j. Epub 2013 May 20. 10.1021/sb400021j PubMed 24050148

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 837
    Illegal SpeI site found at 1155
    Illegal SpeI site found at 2573
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 837
    Illegal SpeI site found at 1155
    Illegal SpeI site found at 2573
    Illegal NotI site found at 2892
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 837
    Illegal BglII site found at 25
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 837
    Illegal SpeI site found at 1155
    Illegal SpeI site found at 2573
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 837
    Illegal SpeI site found at 1155
    Illegal SpeI site found at 2573
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 3538