pUS203

BBa_K1388000 Version 1

Component

Source:
http://parts.igem.org/Part:BBa_K1388000
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Rokiah Alford, Tom Geddes, Andreas Bachler
Date created: 2014-10-06 11:00:00
Date modified: 2015-05-08 01:10:13

IntI1 coding region under AraC-pBad control



Types
DnaRegion

Roles
Generator

engineered_region

Sequences BBa_K1388000_sequence (Version 1)

Description

This part is composed of IntI1 under control of the AraC-pBAD system from iGEM plasmid BBa_K731201.

It allows controlled expression of the IntI1 enzyme which is a vital part of the integron based recombination system we are working on. Integrase acts as a site-specific recombinase enzyme which allows insertion or excision of gene cassettes into appropriate recombination sites (AttI sites).

This part allows simple expression of Integrase with the well characterized AraC-pBAD promoter system. Integrase has a tendency to be toxic to the cell so it is recommended to move the part from the plasmid into a low copy plasmid.

Notes

The IntI1 code we found from genomic sequences had prohibited iGEM restriction sites so the sequence was modified to make it compliant with iGEM standards. We ordered the modified sequence as a gBlock (a ~1.1 Kbp DNA block) and was inserted into the AraC-pBad/plasmid backbone with Gibson Assembly. The sequence we made was further modified to include DNA to make Gibson Assembly easier.

Source

This part is built up from the BBa_K731201 iGEM plasmid (which has AraC-pBad) and the code for the IntI1 enzyme came from a genomic source.

Sequence Annotation Location Component / Role(s)
AraC
pBAD
IntI1
79,958
1104,1270
1331,2346
CDS feature/protein
promoter feature/promoter
CDS feature/protein
igem#experience
None
 
igem#sampleStatus
It's complicated
igem#status
Planning
 
synbiohub#ownedBy
user/myers
 
synbiohub#ownedBy
user/james
 
synbiohub#topLevel
BBa_K1388000/1