BBa_K260003

BBa_K260003 Version 1

Component

Source:
http://parts.igem.org/Part:BBa_K260003
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Kaj Bernhardt
Date created: 2009-10-16 11:00:00
Date modified: 2015-05-08 01:11:42

pRhaFlp. [L-rhamnose] -> FLP(F70L)



Types
DnaRegion

Roles
Plasmid_Backbone

plasmid_vector

Sequences BBa_K260003_sequence (Version 1)

Description

This plasmid is based on pSC101 and contains the rhamnose operon (rhaR and rhaS genes) as well as the F70L variant of FLP recombinase under rhamnose-promoter control. Therefore L-rhamnose can be used to induce FLP(F70L) expression.

The pSC101 origin of replication is temperature sensitive due to a mutation of repA. Permisive temperature is 30 degC. 37 degC should be avoided as the plasmid can not replicate.

Notes

This plasmid comes from Sarov M, Schneider S, Pozniakovski A, Roguev A, Ernst S, Zhang Y, Hyman AA, Stewart AF. A recombineering pipeline for functional genomics applied to Caenorhabditis elegans. Nat Methods. 2006 Oct;3(10):839-44. However, it was modified by deletion of dhfr gene and RedABGRecA operon through insertion of a kanamycin resistance cassette.

Source

dhfr had to be deleted as the same gene for trimpethoprim resistance was going to be used elsewhere in the same cell.

Sequence Annotation Location Component / Role(s)
suffix
repA
pSC101 origin
KanR
T4_32 terminator
FLP(F70L)
rhaS
rhaR
T4_32 terminator
rhamnose-promoter
prefix
1,21
108,1058
1442,1803
1854,2648
2902,2933
2954,4225
4513,5349
5333,6271
6272,6303
4226,4512
6324,6345
engineered_region feature/BioBrick
feature/cds CDS
feature/misc sequence_feature
CDS feature/cds
stem_loop feature/stem_loop
feature/cds CDS
CDS feature/cds
feature/cds CDS
stem_loop feature/stem_loop
feature/promoter promoter
feature/BioBrick engineered_region
igem#experience
None
 
igem#sampleStatus
Not in stock
igem#status
Unavailable
 
synbiohub#ownedBy
user/james
 
synbiohub#ownedBy
user/myers
 
synbiohub#topLevel
BBa_K260003/1