Plasmid_Backbone
LT

Part:BBa_K2144101

Designed by: Sigrun Stulz   Group: iGEM16_Stockholm   (2016-10-14)


Vector containing Hisx6 and Sortase tag LPETGG

This vector is constructed to simplify the usage of the transpeptidase Sortase A for future iGEM teams. Following a BamHI restriction site we constructed a synthetic sequence consisting of three main elements:

1) A linker (GSSS) to allow for limited mobility of the attached peptides and facilitate correct folding

2) A Sortase A tag (LPETGG) that is used to conjugate the proteins to poly-glycines

3) A standard Hisx6 tag commonly used for Immobilized Metal Affinity Chromatography (IMAC)


This sequence follows a BamHI restriction site which can be used by teams to clone in whichever peptide they would like to conjugate using Sortase A.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2029
    Illegal SpeI site found at 2099
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2029
    Illegal SpeI site found at 2099
    Illegal NotI site found at 2106
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2029
    Illegal BamHI site found at 2038
    Illegal XhoI site found at 1013
    Illegal XhoI site found at 1905
  • 23
    INCOMPATIBLE WITH RFC[23]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2029
    Illegal SpeI site found at 2099
  • 25
    INCOMPATIBLE WITH RFC[25]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2029
    Illegal SpeI site found at 2099
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.


[edit]
Categories
//chassis/prokaryote/ecoli
//function/conjugation
//proteindomain/affinity
Parameters
resistance