Coding

Part:BBa_K1321009

Designed by: Chris N Micklem   Group: iGEM14_Imperial   (2014-10-08)

Nickel Binding Protein (NiBP) Freiburg format (RFC[25])

**Important note**: Please be aware we discovered that this part is out of frame, leading to an incorrect translation and read-through of the stop-codon. This also means any subsequent fusion proteins to the C-terminal are out of frame. This is because there is a deletion of a base compared to the equivalent coding sequence from Heliobacter pylori (GenBank CP000012.1 bp1391851-1392033, reverse complement strand). This existed already in BBa_K1151001 from which we cloned our part. To correct the frame-shift, a 'C' needs to be added between bases 49 and 50.


This part is BBa_K1151001 in Frieburg (RFC[25]) format to allow for easy use in fusion proteins. Note that the start and stop codon, plus 6 bp either side of the sequence, are included the RFC25 prefix and suffix which is not shown. It is a histidine-rich metal binding protein which binds up to 5 Ni2+ ions (please see the original BBa_K1151001 biobrick page from team Università del Salento Lecce 2013 for more detailed information).

For expression purposes this part is available with T7 promoter (BBa_K1321163) or LacI promoter (BBa_K1321162). We used this part to create a [http://2014.igem.org/Team:Imperial/Functionalisation library of fusion proteins with cellulose-binding domains].


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
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None