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Revision as of 15:35, 9 October 2018


pSAD 

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]

Introduction

For the silver medal criteria “Validated Part” we submit a basic part: constitutive promoter pSAD from Chlamydomonas reinhardtii. It is a high constitutive expression promoter that encodes for a ferrodoxin-binding protein of photosystem I. This pSAD promoter was submitted in 2014 by the Concordia team (Bba_K1527005). To be compatible with the PhytoBrick standard (RFC1000), an illegal BpiI restriction site was removed (A289T) 1 . We cloned P pSAD by PCR amplification in A2-A3 fusion site to use it for our retrotransposon design. Acceptor plasmid is the MoClo Universal pL0 acceptor plasmid pAGM9121 2 . We sequenced again the sequence to be sure there is no unwanted mutations.

1- Biological background

pSAD is a strong constitutive promotor in Chlamydomonas. It regulates a gene which encodes an abundant chloroplast protein located on the stromal side of the Photosystem I complex.

2- Usage in iGEM projects


Characterization