pDawn-B0034-RFP-rrnB T1

Description

This composite part is a generator consisting of pDawn and RFP. pDawn can induce expression in response to blue light.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 2171
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 63
Illegal NgoMIV site found at 195
Illegal NgoMIV site found at 289
Illegal NgoMIV site found at 582
Illegal NgoMIV site found at 1076
Illegal NgoMIV site found at 1094
Illegal NgoMIV site found at 1184
Illegal AgeI site found at 414
Illegal AgeI site found at 1542
Illegal AgeI site found at 3997
Illegal AgeI site found at 4109
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 1643
Illegal BsaI.rc site found at 525

2021 SZPT-China

Biology

This composite part is a generator consisting of pDawn (BBa_K1075044) and RFP(BBa_E1010). pDawn can induce expression in response to blue light.

Usage

Analyze the system by inserting the red fluorescent reporter protein RFP (BBa_E1010), assemble it on the pSEVA331 expression vector, and transfer the connection mixture to G. hansenii ATCC 53582. In this way, we can verify the blue light response function of the pDawn system in G. hansenii ATCC 53582.

Figure 1. Gene circuit of pDawn-B0034-RFP-rrnB T1

Characterization

Red fluorescence could be observed in the pDawn-B0034-RFP-rrnB T1-pSEVA331-G. hansenii ATCC 53582 under 470-nm blue light, but there was no red fluorescence under dark condition, suggesting that the pDawn promoter could respond to blue light and induce gene expression in G. hansenii ATCC 53582.

Figure 2. Red fluorescence photo of the pDawn-B0034-RFP-rrnB T1-pSEVA331-G. hansenii ATCC 53582 cultured under 470-nm blue light (a) or dark condition (b) for 24 hours.

References

[1] Robert, Ohlendorf, Roee. Corrigendum to "From Dusk Till Dawn: One-Plasmid Systems for Light-Regulated Gene Expression" [J. Mol. Biol. 416 (2012) 534–542] [J]. Journal of Molecular Biology, 2014.

Promoter(const.)-RBS34-YF1-FixJ-FixK-LambdaC-pC (pDawn)：BBa_K1075044