Part:BBa_K4414042

P_RPBSA-GFP-P2A-PuroR

Usage and Biology

P_RPBSA is a promoter. GFP is an extensive use of fluorescent bioimaging in the fields of biochemistry, biotechnology, cell and developmental biology. In comparison with other fluorescent tags, GFP-like proteins have the advantage of forming internal chromophore without requiring accessory cofactors, enzymes or substrates other than molecular oxygen, making possible chromophore formation in live organisms, tissues and cells.[1][2]. Puro, produced by Streptomyces alboniger, a peptide based nucleoside antibiotic that inhibits peptide based transfer in prokaryotic cells and eukaryotic cells. PuroR is pac resistance gene that can resist the lethal effect of puromycin. In our experiments,we use it to select transfected cells carrying pac resistance gene. GFP and Puro(ORF) becomes an ORF through the 2A polypeptide chain ligation, and the mRNA is translated into 1 fusion protein. Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 513
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 513
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 513
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 513
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 606
  Illegal BsaI.rc site found at 541
  Illegal BsaI.rc site found at 1895
  

Reference

[1].Cubitt, A. B., Heim, R., Adams, S. R., Boyd, A. E., Gross, L. A., & Tsien, R. Y. (1995). Understanding, improving and using green fluorescent proteins. Trends in Biochemical Sciences, 20(11), 448–455. https://doi.org/10.1016/s0968-0004(00)89099-4. [2].Stepanenko, O. V., Verkhusha, V. V., Kuznetsova, I. M., Uversky, V. N., & Turoverov, K. K. (2008). Fluorescent Proteins as Biomarkers and Biosensors: Throwing Color Lights on Molecular and Cellular Processes. Current Protein & Peptide Science, 9(4), 338–369.