BBa_K2013006

BBa_K2013006 Version 1

Component

Source:
http://parts.igem.org/Part:BBa_K2013006
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 08:10:59

transcriptional regulator



Types
DnaRegion

Roles
engineered_region

Translational_Unit

Sequences BBa_K2013006_sequence (Version 1)

Description

TphRI or tphRII, aIclR-type transcriptional regulator.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:Q5


Primer-F:5′-GAATTCGCGGCCGCTTCTAGAGTACTAGAGTCACACAGGAGGGTACTAGATG-3′


Primer-R:5′- CGCTACTAGTATTATTACAGGCTACGACCCGCTG-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 NcoI and PstI 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 relates to TphRI or tphRII, aIclR-type transcriptional regulator.

Source

This sequence is from a bacterium called Ideonella sakaiensis 201-F6

Sequence Annotation Location Component / Role(s)
RBS
ISF6_0225
1,13
20,304
feature/rbs ribosome_entry_site
CDS feature/protein
igem#experience
None
 
igem#status
Planning
 
synbiohub#ownedBy
user/james
 
synbiohub#ownedBy
user/myers
 
synbiohub#topLevel
BBa_K2013006/1