Part:BBa_K4335024

Rbcs2 Promotor+StayGold+Rbcs2 Term

BBa_K4335024 is a composite part consisting of a promoter, a fluorescent protein gene and a terminator. StayGold, a green fluorescent protein (GFP) , fluoresces in expression. StayGold has an order of magnitude more photostability than any currently available fluorescent protein. Commonly used to label and trace cellular components. BBa_K4335002 is our StayGold gene optimized for the codon of Chlamydomonas reinhardtii.

Usage

We introduced the StayGold gene into the pTX2040 plasmid. PCrRBCS2 is used as the promoter and TCrRBCS2 is used as the terminator to achieve the expression of the StayGold gene.

Result

Plasmid construction

We designed two primers: Staygold-F and Staygold-R, and confirmed the successful assembly by PCR amplification.

The primers

Primers targeted location

The electrophoretogram of StayGold after amplification

Functional Identification

Fluorescence excitation of positive clones transferred into pTX2040 vector compared with wild-type mCherry, together with DIC field and chlorophyll excitation as fluorescence controls.

Reference

[1] Hirano, M. et al. A highly photostable and bright green fluorescent protein. Nat. Biotechnol. 40, 1132–1142 (2022).

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
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 895
  
• 1000
  COMPATIBLE WITH RFC[1000]