Difference between revisions of "Part:BBa K1648004"
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Expression of RFP in Insertion is shown(Fig 2.).
 
Expression of RFP in Insertion is shown(Fig 2.).
  
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[[File:20150919_021936.jpg]]
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[[File:20150919_021936.jpg350px]]
  
 
Fig 2. Insertion kit with RFP(BBa_K1648004) is expressed in E. coli (DH5 alpha)
 
Fig 2. Insertion kit with RFP(BBa_K1648004) is expressed in E. coli (DH5 alpha)

Revision as of 12:42, 19 September 2015

Insertion Kit for Fusing Protein of Interest to Magnetosome Membrane

Construct Map for K1648004

Consist of a gene (mamC) from magnetosome island of Magnetospirillum gryphiswaldense genome, a promotor & RBS(J13002) in front of multiple restriction sites and a double terminator(B0015) at the back of multiple restriction sites.

Different genes can be added into this part at multiple restriction sites, so that different proteins will be expressed on the magnetosome after transformation of the part into A. vinelandii genome via random integration.

This Part were verified by double digestion (Fig. 1).

K1648004gel.jpg

Fig 1. The photo of 1% agarose gel electrophoresis showing PCR products of operons in Magnetosome Island (MAI). L: DNA ladder. Lane 1: mamHIEJKLMN. Lane 2: mamOPQRBSTU. Lane 3: mamPQRBSTU. Lane 4: mamYXZftsZ-like. Lane 5: mamGFDC. Lane 6: mamGFD. Lane 7: mms6.

RFP is inserted in Insertion Kit for biobrick standard. The Insertion kit with RFP is transformed in E. coli (DH5 alpha). Expression of RFP in Insertion is shown(Fig 2.).

File:20150919 021936.jpg350px

Fig 2. Insertion kit with RFP(BBa_K1648004) is expressed in E. coli (DH5 alpha)

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2759
    Illegal XbaI site found at 2765
    Illegal SpeI site found at 1
    Illegal PstI site found at 7
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2759
    Illegal SpeI site found at 1
    Illegal PstI site found at 7
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2759
    Illegal XhoI site found at 1162
    Illegal XhoI site found at 2188
  • 23
    INCOMPATIBLE WITH RFC[23]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2759
    Illegal XbaI site found at 2765
    Illegal SpeI site found at 1
    Illegal PstI site found at 7
  • 25
    INCOMPATIBLE WITH RFC[25]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2759
    Illegal XbaI site found at 2765
    Illegal SpeI site found at 1
    Illegal PstI site found at 7
    Illegal NgoMIV site found at 2546
    Illegal NgoMIV site found at 2636
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal BsaI site found at 2611