pDawn(cl)-RBS070-LKD-T1

Description

The composite part is a generator consisting of pDawn (without LVA tag) (BBa_K4325010) and LKD (BBa_K4325004).

Usage

We inserted the pDawn (without LVA tag) (BBa_K4325010)blue light response system and the lysis gene LKD (BBa_K4325004) into the pSEVA331 expression vector, which was inserted into E. coli TOP10 and screened out the colonies that grew in the dark but did not grow in blue light to verify the responsiveness of pDawn (without LVA tag) 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 pDawn(without LVA tag)-RBS070-LKD-T1-pSEVA331 in response to blue light lysis in E. coli.

Figure 1 was the growth condition of pSEVA331-pDawn (without LVA tag) -RBS070-LKD-T1-TOP10 both in the dark and under the light.

As shown in Figure 1, four bacterial colony of pSEVA331-pDawn (without LVA tag) -RBS070-LKD-T1-TOP10 grew in the dark, but it did not grow under the light.

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.