Generator

Part:BBa_K4325026

Designed by: Laiyi Teng   Group: iGEM22_SZPT-CHINA   (2022-10-02)
Revision as of 06:11, 10 October 2022 by Taaa (Talk | contribs)


pDawn-RBS070-LKD-T1

Description

This composite part is a generator containing pDawn (CI-LVA) and LKD.

Biology

This composite part is a generator containing pDawn (CI-LVA) (BBa_K1075044) and LKD (BBa_K4325004).


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

Biology

This composite part is a generator containing pDawn (CI-LVA) (BBa_K1075044) and LKD (BBa_K4325004).

Characterization

1.Characterization in E. coli TOP10

As shown in Figure1, 14 colonies meet our expectation and one of them was picked and culture in the dark and under blue light. Growth curves shows the bacteria continue growing after 8 hours, indicating this system is not robust.

Figure 1:

(a) Colony growth of E. coli TOP10 containing pSEVA331-pDawn(cI-LVA)-RBS070-LKD-T1 in the dark

(a2) Colony growth of E. coli TOP10 containing pSEVA331-pDawn(cI-LVA)-RBS070-LKD-T1 under blue light

(a3) Colony growth of E. coli TOP10-pSEVA331-pDawn(cI-LVA)-RBS070-LKD-T1;

2.Characterization in G. hansenii ATCC53582

The picked plasmids from E. coli TOP10 were transformed into G. hansenii ATCC53582. It shows that these two plasmids still maintain their excellent lysis effect in G. hansenii ATCC53582.

Figure 2:

(a1) Colony growth of G. hansenii ATCC53582 containing pSEVA331-pDawn(cI-LVA)-RBS070-LKD-T1 in the dark

(a2) Colony growth of G. hansenii ATCC53582 containing pSEVA331-pDawn(cI-LVA)-RBS070-LKD-T1 under blue light
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Categories
Parameters
None