Generator

Part:BBa_K1758102:Design

Designed by: Team Bielefeld-CeBiTec 2015   Group: iGEM15_Bielefeld-CeBiTec   (2015-08-18)

Translation enhancing 5-UTR + sfGFP under control of T7 promoter


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 82


Design Notes

[http://www.ncbi.nlm.nih.gov/pubmed/23654270 Lentini et al. 2013] showed that the scar which is created by standard biobrick assembly is disadvantageous for in vitro translation when occuring between RBS and start codon.

Source

This part was created by adding 5'-UTR to BBa_I746909 via Gibson primers.

Amplification with Gibson-primer:

  • Amplification of PT7-5'-UTR-sfGFP-double terminator:

Forward primer [http://2015.igem.org/Team:Bielefeld-CeBiTec/Primers#UTR_fwd UTR_fwd]

5'-GTTTAACTTTAAAAAAAAAAAAGAAGGAGATATACATATGCGTAAAGGCGAAGAGCTGTTCAC-3'

Reverse primer [http://2015.igem.org/Team:Bielefeld-CeBiTec/Primers#UTR_fwd UTR_rev]

5'-CCTTCTTTTTTTTTTTTAAAGTTAAACAAAATTATTTCTCCCTATAGTGAGTCGTATTACTCTAGAAG-3'


Ligation via [http://2015.igem.org/Team:Bielefeld-CeBiTec/Protocols Gibson-Assembly].

References