Plasmid
Part:BBa_K1312004:Design
Designed by: Yutaka Murata Group: iGEM14_Kyoto (2014-10-04)
pSB1C3 plus 6x His tag
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal prefix found in sequence at 2070
Illegal suffix found in sequence at 1 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 2070
Illegal SpeI site found at 2
Illegal PstI site found at 16
Illegal NotI site found at 9
Illegal NotI site found at 2076 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 2070
Illegal XhoI site found at 1054
Illegal XhoI site found at 1946 - 23INCOMPATIBLE WITH RFC[23]Illegal prefix found in sequence at 2070
Illegal suffix found in sequence at 2 - 25INCOMPATIBLE WITH RFC[25]Illegal prefix found in sequence at 2070
Illegal XbaI site found at 2085
Illegal SpeI site found at 2
Illegal PstI site found at 16 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
We added 6xHistag and stop codon UAA to after Biobrick Suffix. When you use this part, you need to take care of cloning method. When you clone coding protein region, you have to clone this before stop codon. This is because if coding protein region has stop codon, ribosome stop transcripting at stop codon, so 6xHistag which is after Biobrick suffix isn't transcribed. If you clone protein coding gene just before stop codon, 6xHistag and stop codon which is after 6xHistag is transcribed in adequate reading frame.
Source
pSB1C3