BBa_K2013015

BBa_K2013015 Version 1

Component

Source:
http://parts.igem.org/Part:BBa_K2013015
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Demin Xu
Date created: 2016-10-11 11:00:00
Date modified: 2016-10-15 08:14:45

protocatechuate decarboxylase



Types
DnaRegion

Roles
Translational_Unit

engineered_region

Sequences BBa_K2013015_sequence (Version 1)

Description

This part contains the sequence of coding protocatechuate decarboxylase which catalyzes protocatechuate into catechol.



Experimental Validation


This part is validated through Four ways: amplification, PCR, Enzyme cutting and Sequence.


Amplification


Enzyme:KOD


Primer-F:5′-GAATTCGCGGCCGCTTCTAGAGTACTAGAGTCACACATGAGGGTACTAGATG-3′


Primer-R:5′- CGCTACTAGTATTATTAGCTTTGCAGGTAAACCAC-3′


Results




PCR


Enzyme:Taq


Primer-F:5′-CCACCTGACGTCTAAGAAAC-3′


Primer-R:5′-GTATTACCGCCTTTGAGTGA-3′


Results



Double digestion


After the assembly ,the plasmid was transferred into the Competent E. coli top10. After culturing overnight in LB,we minipreped the plasmid for double digestion .The first cutting procedure was performed with EcoRI and NcoI restriction endonuclease. The second cutting procedure was performed with PstI and NcoI restriction endonuclease.The plasmid was cutted in a 25μL system at 37 ℃ for 1 hours. The Electrophoresis was performed on a 1% Agarose glu


Results

Notes

This is a Translational units part that encodes protocatechuate decarboxylase

Source

This sequence is from Escherichia coli strain K-12 NEB 5-alpha

Sequence Annotation Location Component / Role(s)
RBS
protocatechuate decarboxylase
1,13
20,1462
feature/rbs ribosome_entry_site
feature/protein CDS
igem#experience
None
 
igem#status
Planning
 
synbiohub#ownedBy
user/james
 
synbiohub#ownedBy
user/myers
 
synbiohub#topLevel
BBa_K2013015/1