Generator

Part:BBa_K4325025

Designed by: Laiyi Teng   Group: iGEM22_SZPT-CHINA   (2022-10-02)
Revision as of 17:54, 11 October 2022 by Taaa (Talk | contribs)

pDawn(cl)-RBS070-LKD-T1

Description

The composite part is a generator consisting of pDawn(cI)(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(cI)-RBS070-LKD-T1 in response to blue light lysis in E. coli.

Figure 1 was the growth condition of pDawn(cI)-RBS070-LKD-T1-TOP10 both in the dark and under the light.

As shown in Figure 1, ten bacterial colonies of pDawn(cI)-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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