Plasmid_Backbone
K592200
Part:BBa_K592200:Design
Designed by: Erik Gullberg, Antonio Ascue Avalos, Erik Lundin Group: iGEM11_Uppsala-Sweden (2011-09-21)
Low to medium copy BioBrick standard vector
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 3116
Illegal NheI site found at 1403
Illegal SpeI site found at 2
Illegal PstI site found at 16
Illegal NotI site found at 9
Illegal NotI site found at 3122 - 21INCOMPATIBLE WITH RFC[21]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 3116 - 23INCOMPATIBLE WITH RFC[23]Illegal prefix found at 3116
Illegal suffix found at 2 - 25INCOMPATIBLE WITH RFC[25]Illegal prefix found at 3116
Plasmid lacks a suffix.
Illegal XbaI site found at 3131
Illegal SpeI site found at 2
Illegal PstI site found at 16
Illegal NgoMIV site found at 1969
Illegal AgeI site found at 994
Illegal AgeI site found at 1317 - 1000INCOMPATIBLE WITH RFC[1000]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal BsaI site found at 289
Design Notes
The plasmid sequence is based on pSB3T5, but to make the plasmid usable for Lambda Red recombineering (Datsenko and Wanner, 2000), all sequences with homologies to the E coli chromosome had to be removed. To accomplish this, the E coli His operon terminator BBa_B0053 has been replaced with the late terminator of the Salmonella phage P22.
Source
The P22 late terminator come from the genome of the Salmonella phage P22 (GenBank: AF217253.1, 19662..19702).
References
Datsenko, K. A. and B. L. Wanner (2000). "One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products." Proc Natl Acad Sci U S A 97(12): 6640-5.