Part:BBa_K5218016

pS1300-RcC3H-GFP

Source：plasmid derived from pCAMBIA1300 backbone with Gibson Cloning system.

Lengths：12296 bp

Usage：Functional characterization of RcC3H CDS in transgenic tobacco.

The RcC3H CDS is described in basic part BBa_K52180015. In the design of the construct, RcC3H CDS, driven by a super promoter (CaMV 35S), is fused with eGFP reporter gene (3' end) followed by NOS terminator.

Figure 1. Designed map of pS1300-RcC3H-GFP construct.

Team BGI-MammothEdu-South 2024 tried to extract total RNA from Rosa chinensis of different tissues and stages, unfortunately couldn't get proper RNA samples for reverse transcription. We noticed that the Rosa chinensis precipitated RNA was brown color, which was caused by secondary metabolites. The absorbance curves of both total RNA and cDNA were abnormal, indicating the failure of the extraction and RT. Due to limited time, Team BGI-MammothEdu-South did not work on this gene.

Figure 2: Abnormality in Rosa chinensis total RNA and reverse transcription.

A. Nanodrop-2000 measurement of total RNA extracted from Rosa chinensis.

B. Nanodrop-2000 measurement of cDNA from reverse transcription.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 652
  Illegal PstI site found at 938
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 652
  Illegal PstI site found at 938
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 652
  Illegal BamHI site found at 241
  Illegal BamHI site found at 1257
  Illegal XhoI site found at 1077
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 652
  Illegal PstI site found at 938
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 652
  Illegal PstI site found at 938
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 779
  Illegal SapI site found at 34
  Illegal SapI.rc site found at 805