Part:BBa_K3076200

Coding sequence of Vanabin2 gene from Ascidia sydneiensis samea

Description

This part contains the coding sequence of the vanabin2 gene which encodes a vanadium-binding protein (VB). The gene was identified from a vanadium-rich sea squirt and it has a function of helping the organism to accumulate metal ion for physiological purpose. The reason for the accumulation is still unsolved.

Although it’s a “vanadium” binding protein, literature reported that copper (II) ions inhibits the binding of vanadium by competing for the binding sites. [1] It showed that VB actually has a higher affinity for copper. Therefore, the VB seems to be an ideal protein for our project to enhance the metal accumulating ability of E. coli.

Usage and Biology

We synthesized the vanabin2 CDS fragment by IDT gBlock and planned to assemble the fragment into pET28a expression vector by Hifi assembly. However, due to time constraints, the work is still in progress.

Source of the part

This part contains the coding sequence of the vanabin2 gene. It was identified from the literature review [1] and extracted from NCBI (ACCESSION: AB088204) starting from start codon to stop codon. Then the sequence was codon-optimized for E. coli by the ThermoFisher GeneArt platform.

Design notes

Overlapping sequence of pET28a multiple cloning site was added to flank the CDS for cloning by Hifi assembly.

Reference

[1] Ueki, T., Sakamoto, Y., Yamaguchi, N., & Michibata, H. (2003). Bioaccumulation of Copper Ions by Escherichia coli Expressing Vanabin Genes from the Vanadium-Rich Ascidian Ascidia sydneiensis samea. Applied and Environmental Microbiology, 69(11), 6442–6446. doi: 10.1128/aem.69.11.6442-6446.2003