BBa_K300003

BBa_K300003 Version 1

Component

Source: http://parts.igem.org/Part:BBa_K300003

Generated By: https://synbiohub.org/public/igem/igem2sbol/1

Created by: Giacomo Zambianchi, Alessandro Ranieri, Manuel Lupotto, Paolo Magni

Date created: 2010-10-02 11:00:00

Date modified: 2015-05-08 01:11:51

Phasin (PhaP) - internal domain

Details

• Types: DnaRegion
• Roles: Protein_Domain; polypeptide_domain
• Sequences: BBa_K300003_sequence (Version 1)

Description

-

Notes

Ralstonia eutropha genomic DNA.

Source

It is identical to BBa_K208001, but it lacks the stop codon in order to support protein fusions.

It has been designed as a head domain:

The Prefix sequence is 5'-GAATTCGCGGCCGCTTCTAG-3' (RFC10 Prefix)

The Suffix sequence is 5'-ACTAGTAGCGGCCGCTGCAG-3' (RFC23 Suffix)

For these reasons, a tail domain or an internal domain (compatible with RFC23) are ready to be assembled downstream to create protein fusions.


CONSTRUCTION METHOD:

BBa_K208001 (provided by iGEM HQ in pSB3K3 in RFC23 standard) was PCR-amplified/mutagenized with primers:

phaPSF: 5'GCTTCTAGAATGATCCTCACCCCGGAACA-3'
phaPSR: 5'-GCTACTAGTGGCAGCCGTCGTCTTCTTTG-3'

in order to delete the stop codon.

The PCR product was ran on a 1% agarose gel, gel-extracted, cut with XbaI-SpeI and ligated with pSB1A3 (previously cut with XbaI-SpeI and dephosphorylated).
Positive clones were found through digestion screening/sequencing.

Other Properties

• igem#experience: None
• igem#sampleStatus: It's complicated
• igem#status: Planning
• synbiohub#ownedBy: user/james
• synbiohub#ownedBy: user/myers
• synbiohub#topLevel: BBa_K300003/1

Member of these Collections

• iGEM Parts Registry