Difference between revisions of "Part:BBa K1321009"

Revision as of 01:37, 17 October 2014

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]

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 '''**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).
-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).
+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].
-This part is in RFC25 format and the DNA sequence shown starts after the RFC25 prefix and ends just before the RFC25 suffix. The start codon and stop codon are including in the prefix and suffix sequences.
-<br /> Additionally it is available in fusions with various cellulose binding domains as front (BBa_K1321343, BBa_K1321344, BBa_K1321345) or back inserts (BBa_K1321351, BBa_K1321368, BBa_K1321369) including versions suitable for expression using T7 (BBa_K1321352, BBa_K1321353, BBa_K1321364) or LacI (BBa_K1321362, BBa_K1321361) promoters.