Part:BBa_K3042005

Dino III GFP Plasmid

A dinoflagellate optimized reporter plasmid. DinoIII is a dinoflagellate expression system that is constructed on the basis of genomic and transcriptomic data from many dinoflagellates. DinoIII GFP is a dinoflagellate optimized reporter plasmid which was created by the University of Connecticut. This part can be used as a reporter via fluorescence. When the transformed cells fluoresce green, this indicates the expression of the GFP (Specher, 2019). This part has been successfully transformed into O.Marina by UNAM *insert a link to paper here *. iGEM GSU wants to replicate their results with this part as well as into Symbiodinium microadriaticum, another dinoflagellate. *All sequences/info are from snapgene* Features: RNA elements Sprecher, Brittany & Zhang, Huan & Lin, Senjie. (2019). Nuclear gene transformation in a dinoflagellate. 10.1101/602821.

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 682
  Illegal NheI site found at 1781
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 1201
  Illegal XhoI site found at 1102
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 1763
  

AmpR promoter- Ampicillin resistance promoter AmpR- Product: Beta lactamase Confers resistance to ampicillin and related antibiotics

Ori- High copy number ColE1/pMB1/pBR322/pUC origin of replication Cap binding site-Cap binding activates transcription in the presence of cAMP

Lac operator- The lac repressor binds the lac operator to block transcription in E. Coli. This inhibition can be relieved by adding lactose or IPTG Lac promoter- Promoter for E. Coli lac operon Terminator- part k3042003 Cycle 3 GFP- Mammalian codon-optimized cycle 3 GFP (Crameri et al., 1996) Promoter region- part K3042002