Generator

Part:BBa_K4325025

Designed by: Laiyi Teng   Group: iGEM22_SZPT-CHINA   (2022-10-02)
Revision as of 12:11, 10 October 2022 by Kuaikuai (Talk | contribs)

pDawn(cl)-RBS070-LKD-T1

Description

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

Usage

We inserted the pDawn (without LVA tag)(pDawn without LVA tag)blue light response system and the lysis gene LKD (LKD) into the pSEVA331 expression vector, which was inserted into E. coli TOP10 screened out the colonies 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:

Figure1 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.

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