Regulatory

Part:BBa_K1300002:Design

Designed by: Katty Wu, Ellen Jorgensen   Group: iGEM14_StuyGem_NYC   (2014-06-16)


Constitutive T7 promoter + Lock (crRNA)


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
    COMPATIBLE WITH RFC[1000]


Design Notes

We first had designed the construct to have the pTet promoter. However, we realized that the pTet promoter has a lot of secondary structure. The lock (cis sequence) forms a hairpin (secondary structure) with the RBS. These two secondary structures gave the DNA synthesis companies a lot of trouble. So as a backup plan, we designed this construct. The T7 promoter is a simple, well-known constitutive promoter which does not have any secondary structure. Thus, we were able to design this construct to be synthesized as single-stranded oligos that would later be annealed.

T7 promoter + six bp of DNA (to separate the promoter from the RBS) + cis sequence

Source

This sequence was designed based on existing biobrick parts. 

T7 Promoter sequence [1] & Cis Sequence [2]

References