Part:BBa_K3347010
Dehalogenase type 2, variant 2
This part was generated modification of BBa_K3347003, DeHa2.1.gene. This sequence was codon optimized for E. coli using snapgene for purposes of creating a large quantity of protein using E. coli as an expression chassis. In addition to the untagged variant shown here, we designed two additional, 6xHis tagged variants (BBa_K3347012 & BBa_K3347014) for use in affinity purification for future proteomic studies.
Naming notation for Dehalogenase genes and expression blocks
Plasmid notation:
Represented by pXX_DeHa
Where XX can represent:
C: pSB1C3
A: pSB1A3
K: pSB1K3
KB: pSRKBB
Dehalogenase gene & Expression Block Notation
Represented by DeHaX.Y.Z
Where,
X is the gene type (1 or 2 ; referencing the two dehalogenase genes in D. acidovorans)
Y is the variant (native variant = 1, E. coli codon optimized = 2, etc.)
Z shows the tags/modifications on the protein (0 = no modifications, HisC = 6xHis tag on carboxyl terminal, HisN = 6xHis tag on amine terminal, gene, etc.)
Note: If Z is replaced by the 'gene' modifier, the Dehalogenase gene sequence is being referenced. In all other cases, the fully assembled expression block is referenced. Example: DeHa1.1.gene references BBa_K3347000 while DeHa1.1.0 references BBa_K3347001.
Example notations
pKB_DeHa2.2.HisN: Dehalogenase type II, variant 2 (E. coli optimized) with a N terminal 6xHis tag all inside the pSRKBB vector
pC_DeHa1.1.0: Dehalogenase type I, variant 1 with no further tags or modifications all inside the pSB1C3 vector
This part is intended for high expression of dehalogenase in the chassis, and thus is utilized in a T7-based expression vector inside BL21-DE3 (pLysS) E. coli (BBa_K3347011).
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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