Difference between revisions of "Part:BBa K2319000"

Line 5: Line 5:
 
This part generates a fusion protein of superfolder GFP (sfGFP) and SpyCatcher (a 15.2 kDa protein) when transformed into a T7 expression system like E. coli strain BL21 (DE3).
 
This part generates a fusion protein of superfolder GFP (sfGFP) and SpyCatcher (a 15.2 kDa protein) when transformed into a T7 expression system like E. coli strain BL21 (DE3).
  
<img src="https://static.igem.org/mediawiki/2017/6/65/T--IISc-Bangalore--assembly-C1234-linear.png" width="100%">
+
[[https://static.igem.org/mediawiki/2017/6/65/T--IISc-Bangalore--assembly-C1234-linear.png]]
  
 
This part can be used as a T7 expression backbone.
 
This part can be used as a T7 expression backbone.

Revision as of 10:15, 1 November 2017


sfGFP-SpyCatcher under T7 expression system

This part generates a fusion protein of superfolder GFP (sfGFP) and SpyCatcher (a 15.2 kDa protein) when transformed into a T7 expression system like E. coli strain BL21 (DE3).

[[1]]

This part can be used as a T7 expression backbone.

Using the T7 expression backbone

Expressing any protein of interest

This T7 expression backbone can be used to express any protein if its coding sequence (with a start codon) is inserted using the HindIII and NheI sites. These sites can be added to the coding sequence using PCR with primers having 5'-overhangs.

Fusing a protein domain at the N-terminus of an existing protein

By inserting the coding sequence of a protein domain (including the start codon) using the HindIII and AgeI sites into the T7 expression backbone (which already contains a protein coding sequence), an N-terminal fusion can be performed.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1200
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 768
    Illegal XhoI site found at 1001
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 42
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 57