Part:BBa_K4316010

GypsyForward in MoClo A1 position

The gypsy insulator was designed to be used with the Chlamydomonas Reinhardtii MoClo kit [1] and is in position A1. It’s 5’ scar is GGAG and it’s 3’ scar is TGAC.

The gypsy insulator from Drosophila melanogaster, when surrounding sequences with it’s reverse complementary sequence, was shown to produce insulatory effects in Arabidopsis thaliana, facilitating high and precise transgene expression [2]. Here, the sulator sequence hypothetically allows loop formation (if we are to accept the loop extrusion model) between both insulators without interfering with the transgenes.

Sequence for the insulator sequence was retreived from She et al., 2010 [2].

Sources:

[1] Crozet, Pierre & Navarro, Francisco & Willmund, Felix & Mehrshahi, Payam & Bakowski, Kamil & Lauersen, Kyle & Pérez-Pérez, María & Auroy, Pascaline & Gorchs Rovira, Aleix & Sauret-Gueto, Susana & Niemeyer, Justus & Spaniol, Benjamin & Theis, Jasmine & Trösch, Raphael & Westrich, Lisa & Vavitsas, Konstantinos & Baier, Thomas & Hübner, Wolfgang & de Carpentier, Félix & Lemaire, Stéphane. (2018). Birth of a Photosynthetic Chassis: A MoClo Toolkit Enabling Synthetic Biology in the Microalga Chlamydomonas reinhardtii. ACS Synthetic Biology. 7. 10.1021/acssynbio.8b00251.

[2] She W, Lin W, Zhu Y, Chen Y, Jin W, Yang Y, Han N, Bian H, Zhu M, Wang J. The gypsy insulator of Drosophila melanogaster, together with its binding protein suppressor of Hairy-wing, facilitate high and precise expression of transgenes in Arabidopsis thaliana. Genetics. 2010 Aug;185(4):1141-50. doi: 10.1534/genetics.110.117960. Epub 2010 Jun 1. PMID: 20516496; PMCID: PMC2922898.

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]