Source:
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 decarboxylaseSource
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 |