Part:BBa_K299025

RBS measurement device pT7 J61100 GFP

This part is licensed under
Creative BioCommons

We planned to use this device to determine BBa_J61100 strength relative to BBa_B0034 using cell-free system.

Authors

Clonning:

• Ania Puławska
• Milena Bażlekowa

The lab work was supervised by:

• Michał Lower
• Ania Olchowik

Construct design

The device consists of BBa_I719005 T7 promoter followed by BBa_J61100 Anderson's RBS and BBa_I13401 reporter module. GFPmut3b was used as reporter protein. Device was created by cloning BBa_I13401 behind ExpressIt! vector containing appropriate RBS.

Note about parts cotaining Anderson's RBS

In the physical DNA after J61100 in composite parts there is Aactag sequence instead of the biobrick scar Tactag. J61100 was originally designed in a way that its last nucleotide "A" is also first nucleotide of the suffix (wchich should be T)- it modifies the suffix. The sequence of J61101 in the registry does not include its last nucleotide. If last nucleotide were included the registry sequence would show the additional nucleotide, now it shows T instead of A.

The seguence of the J612100 does not match its physical DNA

To go around this problem in out bricks we used scarless assembly option and manually included the scars:

BBa_K299800 DNA standard biobrick scar (if next part starts with ATG)

BBa_K299801 DNA standard biobrick scar (if next sequence does NOT start with ATG)

BBa_K299802 DNA Aactag - like a biobrick scar but first T changed to A

BBa_K299803 DNA Gactag - like a biobrick scar but first T changed to G

BBa_K299804 DNA Cactag - like a biobrick scar but first T changed to C.

Sequence and Features

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 BsaI.rc site found at 693