BBa_K300002

BBa_K300002 Version 1

Component

Source:
http://parts.igem.org/Part:BBa_K300002
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) - head domain



    • Details

    Types
    DnaRegion

    Roles
    Protein_Domain

    polypeptide_domain

    Sequences BBa_K300002_sequence (Version 1)

    Description

    -

    Notes

    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:
    phaP10F 5'-GCTTCTAGATGATCCTCACCCCGGAACA-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 pSB1AK3 (previously cut with XbaI-SpeI and dephosphorylated).
    Positive clones were found through digestion screening/sequencing.

    Source

    Ralstonia eutropha genomic DNA.

    Sequence Annotation Location Component / Role(s)
    Start
    		1,3
    		start_codon feature/start

    igem#experience
    None
     
    igem#sampleStatus
    It's complicated
    igem#status
    Planning
     
    synbiohub#ownedBy
    user/james
     
    synbiohub#ownedBy
    user/myers
     
    synbiohub#topLevel
    BBa_K300002/1
     

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