Device

Part:BBa_K4628043

Designed by: Linghao Xia   Group: iGEM23_ZJU-China   (2023-10-11)


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.

Enhancer

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


[edit]
Categories
Parameters
biologyArtificial synthesis
chassisNicotiana benthamiana