Part:BBa_K1424003
Kanamycin resistance for Synechocystis
The kanamycin resistance gene tn903 aph from E. coli for use with Synechocystis sp. PCC 6803.
The kanamycin gene to allow transformation in Synechocystis sp. PCC 6803 (Poteete et al, 2006).
Usage and Biology
BBa_K1424000 [[1]] & BBa_K1424001 [[2]] were used (flanking a kanamycin resistance cassette designed for Synechocystis sp. PCC 6803; BBa_K1424003) as double homologous recombination sequences for insertion of exogenous DNA into the genome of Synechocystis sp. PCC 6803. Colonies were picked and segregated on kanamycin containing BG11 growth media. The inclusion of exogenous DNA into the host genome was verified by performing a band-shift colony PCR. Using left homologous region forward primer and right homologous region reverse primer, PCR amplification of this genomic region can be performed. The increase in band size (band shift) of the mutant strain respective of the wild type strain confirms integration of exogenous kanamycin resistance cassette into the host genome.
ADVANTAGES IN GENOME ENGINEERING APPLICATIONS: This part can be used to select positive integrants in the Synechocystis sp. PCC 6803 genome.
Improved Function
This part is an improvement to the documented part BBa_K300989. It has been optimized for use in the organism Synechocystis sp. PCC 6803, and also optimized to be compatible with iGEM assembly.
Future Improvements
This part was used as the basis for the design of the part BBa_K4047019 by the MiamiU_OH 2021 project. The part was codon optimized for use in a similar but differing cyanobacteria, Synechococcus elongatus PCC 7942. Resulting transformants successfully grew on BG11 media containing kanamycin.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 135
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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