Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Jordan Berg
Date created: 2014-10-05 11:00:00
Date modified: 2015-05-08 01:10:02
Alpha Amylase with TolB Signaling Sequence and PstI Site Removed
| Types | DnaRegion |
| Roles | sequence_feature DNA |
| Sequences | BBa_K1356000_sequence (Version 1) |
Description
This is the alpha amylase taken from part BBa_K1195001. Attached to it is a TolB signaling sequence meant to all the gene product to be expressed extracellularly in N. multiformis in the break down of biofilm in wastewater treatment plants. Additionally, the PstI site originally found in the BBa_K1195001 part was removed using site-directed mutagenesis. The restriction site was changed from "CTGCAG" to "CTCCAG". This gene is located in the standard iGem pSB1C3 plasmid backbone.Amylase is an enzyme naturally synthesized by bacteria, such as E. coli, fungi, and even in humans in saliva and the pancreas. This enzyme catalyzes the hydrolysis of starches into sugars and breaks down the components of bacterial extracellular polymeric substance (EPS), which contains extracellular DNA, polysaccharides, and proteins.
Notes
The TolB signaling sequence was synthesized using RNA primers overlap-extension PCR owing it the signaling sequence's large size. The mutation was done through mutagenic PCR.Source
The original amylase we modified was from part BBa_K1195001 in the iGem parts registry.| Sequence Annotation | Location | Component / Role(s) |
| DsbA Signaling Sequence Alpha Amylase Mutated PstI Restriction Site for Removal | 1,101 102,1589 977,977 | sequence_feature feature/misc feature/protein CDS sequence_feature feature/misc |