BBa_K2013020

BBa_K2013020 Version 1

Component

Source:
http://parts.igem.org/Part:BBa_K2013020
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Mei Deng
Date created: 2016-10-11 11:00:00
Date modified: 2016-10-15 08:18:39

Sc Alcohol dehydrogenase 2



Types
DnaRegion

Roles
Translational_Unit

engineered_region

Sequences BBa_K2013020_sequence (Version 1)

Description

This part contains the sequence of coding Sc Alcohol dehydrogenase 2 that catalyzes Ketoisovalerate into isobutanol.In addition,We carried out codon optimization on this part


Experimental Validation


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


Amplification


Enzyme:KOD


Primer-F:5GAATTCGCGGCCGCTTCTAGAGTACTAGAGAAAGAGGAGAAATACTAGATGAGCATCCCG-3′


Primer-R:5′- GCGGGTCGTTATGTTGTGGATACCAGCAAATAATAATACTAGTAGCG-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 EcoRV 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 Sc Alcohol dehydrogenase 2

Source

This sequence is from Saccharomyces cerevisiae YJM1386

Sequence Annotation Location Component / Role(s)
RBS
Sc Alcohol dehydrogenase 2
1,13
20,1068
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_K2013020/1