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.(Cubitt et al,. 1995;Stepanenko et al, 2008). 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
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 513
- 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 513
- 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 513
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 513
- 1000INCOMPATIBLE 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. https://doi.org/10.2174/138920308785132668
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