Difference between revisions of "Part:BBa K3866000"
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===Verification of Cloning=== | ===Verification of Cloning=== | ||
− | [[File:T--Thessaly-- | + | [[File:T--Thessaly--arac.png|700px|thumb|none|<i><b>Fig.3:</b>(U=Uncut C=Cut) Restriction Digestion of araC:ParaBAD with EcoRI+ BamHI. Positive clones 1, 2 Expected bands 2160, 1184</i>]] |
===Source=== | ===Source=== |
Revision as of 12:07, 24 September 2021
araC-ParaBAD:RBS GB compatible with A1-B2
Usage and Biology
The PBAD promoter, derived from the arabinose operon andis an inducible promoter for arabinose. The use of an inducible promoter provides a most reversible and flexible gene circuit and at the same time exhibits a higher efficiency and lower side effects as cell death.(Kallunki, Barisic, Jäättelä and Liu, 2019). The araC gene, is a regulatory gene and is located upstream of the L-arabinose operon, encodes a positive regulatory protein AraC, required for L-arabinose utilization in Escherichia coli. The araC gene has a constitutive promoter.
Design Notes
The coding sequence was domesticated . We removed BsmBI ,BsaI , BtgZI, BpiI sites in order to be compatible with GoldenBraid and MoClo. The sequence is present in pUPD2 BBa_K3505007 and has overhangs compatible for GoldenBraid cloning. The CDS has position A1-B2
Verification of Cloning
Source
From the iGEM Distribution Kit 2019.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1148
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 983
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 965
References
- Kallunki, Barisic, Jäättelä and Liu, 2019. How to Choose the Right Inducible Gene ExpressionSystem for Mammalian Studies?Cells, 8(8), p.796.