Part:BBa_K1795023
GalK Integrator Cassette
We created a modified backbone that will allow users to quickly and easily create parts that can be integrated onto the E. coli genome. This backbone is designed as follows (where H1 refers to the first region homologous to the E. coli genome, and H2 refers to the second homologous region)
In order to insert a part onto the genome, the user simply needs to put a part onto this backbone (between the prefix and suffix), and PCR up the region between (and including) H1 and H2.
Users can put their parts onto this backbone with their preferred method of assembly. This part exhibits resistance to chloramphenicol and kanamycin. The parts integrated with this cassette will contain kanamycin resistance, and will be integrated in the galK region of the E. coli genome.
Homologous regions modeled after those in Elowitz et al 2002.
This part has been successfully used to integrate fluorescent proteins onto the E. coli genome. Visual confirmation of integration of CFP at the galK locus is shown via confocal microscopy: [1]
![[1]](https://static.igem.org/mediawiki/2015/8/89/WM150914_r0011_no_saturation_replicate_1_picture_4.png){kind=link}
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal prefix found at 2078
Illegal suffix found at 2100 - 12INCOMPATIBLE WITH RFC[12]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 2078
Illegal SpeI site found at 2101
Illegal PstI site found at 2115
Illegal NotI site found at 2084
Illegal NotI site found at 2108 - 21INCOMPATIBLE WITH RFC[21]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 2078
Illegal XhoI site found at 1012
Illegal XhoI site found at 1904 - 23INCOMPATIBLE WITH RFC[23]Illegal prefix found at 2078
Illegal suffix found at 2101 - 25INCOMPATIBLE WITH RFC[25]Illegal prefix found at 2078
Plasmid lacks a suffix.
Illegal XbaI site found at 2093
Illegal SpeI site found at 2101
Illegal PstI site found at 2115 - 1000INCOMPATIBLE WITH RFC[1000]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal SapI site found at 2069
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