Part:BBa_K812051
Golden brick version of pSB1K3 plasmid for Golden Gate and Golden Brick cloning
This is a modified version of pSB1K3 from the registry with the J4450 negative control. The biobrick sites are modified to incorporate a BsaI restriction site that enables the use of golden gate cloning and golden brick cloning to assemble full contructs in a single shot of digestion/ligation.
The complete protocol of the goldengate can be find in the original paper: Engler C, Gruetzner R, Kandzia R, Marillonnet S (2009) Golden Gate Shuffling: A One-Pot DNA Shuffling Method Based on Type IIs Restriction Enzymes. PLoS ONE 4(5): e5553. doi:10.1371/journal.pone.0005553
The Golden Brick assembly is an invention of the iGEM Evry 2012 Team. A complete description of the procedure can be find on our website: http://2012.igem.org/Team:Evry/GB
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 2190
Illegal XbaI site found at 2212
Illegal SpeI site found at 1
Illegal PstI site found at 23 - 12INCOMPATIBLE WITH RFC[12]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 2190
Illegal SpeI site found at 1
Illegal PstI site found at 23 - 21INCOMPATIBLE WITH RFC[21]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 2190
Illegal XhoI site found at 1040
Illegal XhoI site found at 2066 - 23INCOMPATIBLE WITH RFC[23]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 2190
Illegal XbaI site found at 2212
Illegal SpeI site found at 1
Illegal PstI site found at 23 - 25INCOMPATIBLE WITH RFC[25]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 2190
Illegal XbaI site found at 2212
Illegal SpeI site found at 1
Illegal PstI site found at 23 - 1000INCOMPATIBLE WITH RFC[1000]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal BsaI site found at 7
Illegal BsaI.rc site found at 2206
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