Size of standard vector

After consideration of inclusion of incP and incN sites into the basic standard vector, I’ve come to the conclusion that the standard plasmid is too large at it’s current time. To remove unnecessary parts, I am going to synthesize the new ends at the end of the incP incN oriT synthesis and RE clone over the KG1 construct.oriT_N-P-1 Map

Conjugation

http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0148989

  • Could this method be taken advantage of or somehow manipulated for more organisms?

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC262305/

  • Looks like I need incN for these

I should rethink the standardization to perhaps include these sites, as they would be useful. Would the simplicity of not having to add another plasmid be worth the 300~ extra base pairs? Hum. Also, since I am redoing the backbone, I ought to redo the transcriptional termination sites. Lots of work

Planned tests

Planned tests to do-

  • Can synthesized DNA fragments be used efficiently in GoldenGate reactions?
  • Can BAR or hygromycin be used efficiency for soil based plant selection?
    • For development of plant genetic transformation method without tissue culture

Planned cloning-

  • Clone RFP and F1-ori into pKGs2 for GoldenGate reaction to create full basic set of vectors
    • Amp, Kan, Spec series with and without F1 ori
      • Cam is only used for basic parts
      • All cloning can be done with GoldenGates
  • Finish cloning and sequencing of pKGs1
  • Sequence M13sc