Part:BBa_K2984025

AR Promoter A1-B1

This part is a part of the Chlamy-HUB-Collection. Another strong constitutive promoter is the AR promoter that is composed of HSP70A and RBCS2 building a tandem promoter. Therefore the HSP70A (A) promoter is fused upstream of another promoter resulting in a stronger transgene expression (Schroda et al. 2000). The combination of the HSP70A (A) promoter and the RBCS2 (R) promoter building _PAR has the positive effect that transcriptional gene silencing (TGS) is strongly reduced making this promoter popular for transgene expression (Schroda et al. 2002). This part was designed for C. reinhardtii.

Usage and Biology

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 282
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Characterization

Fig.1 - Successful transformation of L0-AR_A1-B1 into E. coli.

The AR Promotor was used for two different paromomycin constructs. The first carries the registry part BBa_K2703008 which we wanted to compare to our improved paromomycin construct which also feautres this promoter. The improvement was done as part of the 2019 gold criteria. The colony growth proves the functionality of the promoter. This part was also used for characterization purposes of the ARS secretion signal in the level 1 construct L1c-AR-ARS-YFP-Rbcs2

Fig.1 - TAP-agar plates containing paromomycin. The C.Reinhardtii colonies growing on the plate were tranformed with our improved paromomycin construct which is flanked by this AR promoter and the Rbcs2 terminator

References

1. Schroda, M., Blöcker, D., & Beck, C. F. (2000). The HSP70A promoter as a tool for the improved expression of transgenes in Chlamydomonas. The plant journal, 21(2), 121-131.