pDawn-X174E-LVA-T1

Description

The composite part is a generator consisting of pDawn (cI-LVA) (BBa_K1075044) and X174E-LVA (BBa_K4325007).

Usage

We inserted the pDawn (cI-LVA)(BBa_K1075044) blue light response system and the lysis gene X174E-LVA(BBa_K4325007) into the pSEVA331 expression vector, which was inserted into E.coli TOP10 and screened out the bacterial colony which grew in the dark but did not grow under the blue light to verify the responsiveness of pDawn (cI-LVA) 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
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 1643
Illegal BsaI.rc site found at 525

2022 SZPT-China

Characterization

1.Batch screening of pSEVA331-pDawn(cI-LVA)-X174E-LVA-T1 in response to blue light lysis in E. coli.

We transformed pSEVA331-pDawn (cI-LVA) -X174E-LVA-T1 into TOP10 and performed agarose gel electrophoresis, as shown in Figure 1, pSEVA331-pDawn (cI-LVA) -X174E-LVA-T1 has been successfully transformed into TOP10.

Figure 1:

pSEVA331-pDawn (cI-LVA) -RBS070-LKD-LVA-T1;and (2)pSEVA331-pDawn (cI-LVA)-X174E-LVA-T1 has been successfully identified in TOP10

References

[1]Robert Ohlendorf, Roee R. Vidavski, Avigdor Eldar et.al.From Dusk till Dawn: One-Plasmid Systems for Light-Regulated Gene Expression.Journal of Molecular Biology, 08 Jan 2012, 416(4):534-542.