Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Kaj Bernhardt
Date created: 2009-10-17 11:00:00
Date modified: 2015-05-08 01:11:42
P_FRT_dhfr. (strong promoter, translated FRT site, trimethoprim resistance)
| Types | DnaRegion |
| Roles | engineered_region Intermediate |
| Sequences | BBa_K260015_sequence (Version 1) |
Description
This is the first of two parts of a FLP recombinase reporter. It has a strong promoter, a translated FRT site, and a codon-optimised gene for trimethoprim resistance (TmpR) called dhfr.The second part of this FLP recombinase reporter is the TT_FRT_GFP BioBrick ([[Part:BBa_K260016|]]). They can both be transferred to the pCC2FosM ([[Part:BBa_K260000|]]) and pCC2FosMB ([[Part:BBa_K260001|]]) backbones, at defined loci to vary the distance between both FRT sites, of which each part has exactly one.
The position of the TT_FRT_GFP BioBrick ([[Part:BBa_K260016|]]) is always the same, whereas the position of P_FRT_dhfr ([[Part:BBa_K260015|]]) can be varied to give inter-FRT distances of 500 bp, 1 kb, 2 kb, 5 kb, 10 kb. This is achieved by starting with different backbones to contain the present BioBrick part ([[Part:BBa_K260015|]]), that have specific homology arms for recombineering-mediated transfer of P_FRT_dhfr to defined positions:
[[Part:BBa_K260004|]]: pMA-@CC2FosMB-00500bp
[[Part:BBa_K260005|]]: pMA-@CC2FosMB-01000bp
[[Part:BBa_K260006|]]: pMA-@CC2FosMB-02000bp
[[Part:BBa_K260007|]]: pMA-@CC2FosMB-05000bp
[[Part:BBa_K260008|]]: pMA-RQ-@CC2FosMB-10000bp
See [[Part:BBa_K260016|]] for how this reporter works.
Notes
A strong promoter was necessary and we use [[Part:BBa_J23100|]]. An appropriate reading frame for the FRT site had to be found to prevent premature Stop-codons and hydrophobic amino acid residues. Further, the gene encoding TmpR was codon-optimised.Source
This part was synthesised by "Mr Gene" (Geneart) and is available in the following plasmid backbones:[[Part:BBa_K260004|]]: pMA-@CC2FosMB-00500bp
[[Part:BBa_K260005|]]: pMA-@CC2FosMB-01000bp
[[Part:BBa_K260006|]]: pMA-@CC2FosMB-02000bp
[[Part:BBa_K260007|]]: pMA-@CC2FosMB-05000bp
[[Part:BBa_K260008|]]: pMA-RQ-@CC2FosMB-10000bp
| Sequence Annotation | Location | Component / Role(s) |
| BBa_J23100 (strong promoter) BBa_B0034 (Elowitz RBS) ATG_Start translated FRT site optimised TmpR | 1,35 44,55 62,64 66,99 101,334 | feature/promoter promoter ribosome_entry_site feature/rbs start_codon feature/start feature/misc sequence_feature CDS feature/cds |