Composite

Part:BBa_K1119011:Design

Designed by: Wai In Hui   Group: iGEM13_Hong_Kong_HKUST   (2013-10-24)


Human Elongation Factor-1alpha Promoter - GFP - hGH polyA tail


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 326
    Illegal AgeI site found at 83
    Illegal AgeI site found at 1067
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1470
    Illegal BsaI.rc site found at 972


Design Notes

There are NgoMIV & AgeI sites around GFP reporter (BBa_K648013), which make the part incompatible with RFC25 standard as shown by Part Registry, but users can perform in-frame protein fusion using the mentioned sites.

Source

The EF-1alpha promoter (BBa_K1119010) sequence was cloned out from plasmid iDUET101a (http://www.addgene.org/17629/) using PCR with primers that includes prefix and suffix in RFC10 standard.

GFP reporter in RFC25 format from partsregistry(BBa_K648013)

hGH polyA terminator in RFC10 format from partsregistry (BBa_K404108)

References

Qin, Jane Yuxia, Li Zhang, et al. "Systematic Comparison of Constitutive Promoters and the Doxycycline-Inducible Promoter." PLoS ONE. 5.5 (2010) <http://www.plosone.org/article/info:doi/10.1371/journal.pone.0010611>.

Zhou, B. Y., Ye, Z., Chen, G., Gao, Z. P., Zhang, Y. A., & Cheng, L. (2007). Inducible and reversible transgene expression in human stem cells after efficient and stable gene transfer. Stem Cells, 25(3), 779-789. doi:10.1634/stemcells.2006-0128 <http://onlinelibrary.wiley.com/doi/10.1634/stemcells.2006-0128/abstract>