Part:BBa_K4628043

pGreenⅡ -TMV RNA dependent RNA polymerase-MP-subPromoter CP-EGFP-TLS(Ara.Met.full)-UTR

On the basis of pGreen Ⅱ0800-TMV RNA dependent RNA polymerase-movement protein-subPromoter CP-EGFP, We inserted a tRNA-like sequence (TLS (Ara.Met.full)) before TMV 3 'UTR.

Usage and Biology

To enable specific Rnas to be both long-lived and mobile, we utilize the plasmid to bind TMV RNA-dependent RNA polymerase and TLS.

Further, we try to use RNA fold pairs to change the secondary structure of 3’UTR and TLS after insertion into TLS. The results showed that the secondary structure of TLS and 3’UTR was not affected after insertion, and the most important stem-ring structure was not destroyed（Fig.1）. The above provides evidence that the modification does not affect biological function.

Figure 1 Prediction of RNA secondary structure

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 258
  Illegal EcoRI site found at 2663
  Illegal EcoRI site found at 4242
  Illegal XbaI site found at 987
  Illegal XbaI site found at 3347
  Illegal SpeI site found at 1206
  Illegal SpeI site found at 3209
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 258
  Illegal EcoRI site found at 2663
  Illegal EcoRI site found at 4242
  Illegal NheI site found at 1910
  Illegal SpeI site found at 1206
  Illegal SpeI site found at 3209
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 258
  Illegal EcoRI site found at 2663
  Illegal EcoRI site found at 4242
  Illegal BglII site found at 2609
  Illegal BglII site found at 3344
  Illegal BglII site found at 4172
  Illegal BglII site found at 4267
  Illegal BglII site found at 4978
  Illegal BamHI site found at 3320
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 258
  Illegal EcoRI site found at 2663
  Illegal EcoRI site found at 4242
  Illegal XbaI site found at 987
  Illegal XbaI site found at 3347
  Illegal SpeI site found at 1206
  Illegal SpeI site found at 3209
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 258
  Illegal EcoRI site found at 2663
  Illegal EcoRI site found at 4242
  Illegal XbaI site found at 987
  Illegal XbaI site found at 3347
  Illegal SpeI site found at 1206
  Illegal SpeI site found at 3209
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 4726