This part can be used to construct cloning vectors that are compatible with Golden Gate assembly. Two diverging BsaI cloning sites are situated inbetween a strong constitutive promoter+RBS and double terminator. One or more linear fragments can be cloned into this vector in an efficient one-pot restriction and ligation reaction. The outer sticky ends are GATG and TACT and the resulting constructs are indistinguishable from those created by standard Registry assembly methods, since the same scars are generated.
Sequence and Features
- 10COMPATIBLE WITH RFC
- 12Illegal NheI site found at 7
Illegal NheI site found at 30
- 21COMPATIBLE WITH RFC
- 23COMPATIBLE WITH RFC
- 25COMPATIBLE WITH RFC
- 1000Illegal BsaI site found at 77
Illegal BsaI.rc site found at 66
- Engler, C., Gruetzner, R., Kandzia, R., & Marillonnet, S. (2009). Golden gate shuffling: a one-pot DNA shuffling method based on type IIs restriction enzymes. PloS one, 4(5), e5553.