Plasmid_Backbone
Part:BBa_K864014
Designed by: Erik Lundin Group: iGEM12_Uppsala_University (2012-09-24)
Revision as of 21:23, 26 September 2012 by Fredrikbostrom (Talk | contribs)
Low copy BioBrick standard vector with LacI
pSB4K15Iq(FRT) is a BioBrick standard vector with low copy pSC101 replication origin BBa_K864050 (~5 copies per cell) and kanamycin antibiotic resistance marker, especially usable for Lambda Red recombineering in E coli. The backbone sequence is based on pSB3T5, but the E coli His operon terminator BBa_B0053 has been replaced with the late terminator of the Salmonella phage P22.
This vector carries a LacIq cassette on the backbone for tight repression of any lac promoter cloned into the cloning site.
The resistance cassette is flanked by FRT sites (BBa_J61020) which allows for subsequent removal of the resistance cassette from the chromosome by the FLP recombinase.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 4891
Illegal NheI site found at 2425
Illegal SpeI site found at 2
Illegal PstI site found at 16
Illegal NotI site found at 9
Illegal NotI site found at 4897 - 21INCOMPATIBLE WITH RFC[21]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 4891 - 23INCOMPATIBLE WITH RFC[23]Illegal prefix found at 4891
Illegal suffix found at 2 - 25INCOMPATIBLE WITH RFC[25]Illegal prefix found at 4891
Plasmid lacks a suffix.
Illegal XbaI site found at 4906
Illegal SpeI site found at 2
Illegal PstI site found at 16
Illegal NgoMIV site found at 2319 - 1000INCOMPATIBLE WITH RFC[1000]Plasmid lacks a prefix.
Plasmid lacks a suffix.
[edit]
Categories
Parameters
//plasmidbackbone/assembly
//plasmidbackbone/copynumber/low
//plasmidbackbone/operation
//plasmidbackbone/version/5
//plasmidbackbone/copynumber/low
//plasmidbackbone/operation
//plasmidbackbone/version/5
chassis | |
copies | ~5 |
mcs | BioBrick |
origin | pSC101 |
resistance | K |