pDawn-B0034-S105-rrnB T1

Description

This composite part is a generator consisting of the pDawn promoter and S105.

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

2021 SZPT-China

Biology

This composite part is a generator consisting of the pDawn (BBa_K1075044) promoter and S105 (BBa_K3740024).

Usage

The target gene of pDawn blue light response system (BBa_K1075044) and Lambda Lysis Cassette S105 (BBa_K3740024) were inserted into the pSEVA331 expression vector. Then, random primer guided mutagenesis method to modulate the strength of the RBS (BBa_B0034) located upstream of S105. After introducing into E. coli DH5α, a drop plate assay was performed to screen the bacterial isolate that can grow normally in the dark but not under the blue light irradiation.

Figure 1, Gene circuit of S105(BBa_K3740024)

Characterization

Although S105 grew in the dark and did not grow under blue light, the growth state of plaque in S105 was abnormal in the dark. We speculate that it should be caused by the background expression of the pDawn promoter in the dark. (The blue box is invalid data.)

Figure 2. The growth status of pDawn-RBSNNN-S105-rrnB T1-pSEVA331-DH5α under blue light (a) and dark condition (b).

Due to limited time, we have not been able to verify the lytic function of S105 in G. hansenii ATCC 53582.

References

[1] Ceyssens P J, Lavigne R, Mattheus W, et al. Genomic Analysis of Pseudomonas aeruginosa Phages LKD16 and LKA1: Establishment of the KMV Subgroup within the T7 Supergroup[J]. Journal of Bacteriology, 2006.