Regulatory
lacI

Part:BBa_K4047031

Designed by: An Nguyen   Group: iGEM21_MiamiU_OH   (2021-10-06)


lac repressor

This part is the sequence for lacI, of which the product is lac repressor. This part is an improvement upon the BBa_K2936029, registered in 2019. Our team became aware the parts were twins when we registered. The sequences are identical, and our contribution is also listed on this original part page. For judging purposes, however, we have copied the content to this page and linked our own page due to spelling errors, poor documentation, and other issues with the original page registration that can not be edited.

Original part description

This protein is the key protein in lactose operon; the product of lac I is called lac repressor. Its function is to bind to the 5'-terminal manipulating gene (lac O) of the target gene, which is located between the promoter (lac P) and the target gene. When repressors bind to manipulating genes, they block the initiation of transcription on promoters.

MiamiU_OH Contribution

The function of the lac repressor is natively within E. coli, inhibiting transcription and creating a genetic switch out of the lac operon (1). The operon is diagrammatically depicted in the attached figure, the mechanism of which has been invaluable for genetic engineering by allowing user control of expression. In general, the operon is recognized by the lac repressor, the product of the LacI gene, which when bound cannot transcribe lacZ. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactoside (IPTG) to the culture.

Diagram of the operon model (1).

As explained in Messing, 1998, the lac operon is an excellent choice for genetic engineering in plant systems due minimization of plant promoter sequences, allowing control of the system even when implemented outside of the lab (2). Due to both this and the ease of use in lab, our team chose to regulate our overexpression plasmids via this operon. Our wet lab experiments also confirmed that this sequence is operational in Synechococcus elongatus PCC 7942, making the sequence a viable choice for future work in photosynthetic chassis organisms.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
//chassis/prokaryote/cyanobacterium
//function/regulation/transcriptional
Parameters
None