![](https://parts.igem.org/images/partbypart/icon_device.png)
Device
Part:BBa_K2233002:Design
Designed by: Kyosuke Kita Group: iGEM17_Kobe (2017-10-21)
lacZ for monitoring amtB gene expression in B.subtilis chromosome
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 5434
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 592
Design Notes
Restriction site for EcoRI was removed by base substitution using IDT service. This part was sequenced, and there was a synonymous point mutation in CDS of lacZ.
Source
lacZ and cat genes (with promoter+RBS) were amplified from commercialized plasmids, pMutin2 and pSweet respectively. Sequence for recombination (Recombination site B) was synthesized by IDT. Recombination site A was amplified by PCR from B.subtilis strain 168 chromosome. These DNA fragments were assembled together by Gibson assembly method.