Part:BBa_K2623014
Cytosolic-abundant heat soluble protein "CAHS " (promoter, RBS, RFP, lacI and double terminator)
SUMMARY
This part contains the coding region of the Hypsibius dujardini (Water bear) (Macrobiotus dujardini) CAHS gene. Cytosolic abundant heat soluble proteins acting as a molecular shield in water-deficient condition. Tardigrade-specific intrinsically disordered proteins (TDPs) are essential for desiccation tolerance by forming non-crystalline amorphous solids upon desiccation, and this vitrified state mirrors their protective capabilities.
CAHS is a cytoplasmic-abundant protein which can strength the bacteria stress tolerance under the conditions such as dehydration. So taking biosafety into account, we add a constant promoter BBa_J23100 into the CAHS circuit to express Lac I BBa_S0100 in order to inhabit the initiation of the Lac O BBa_R0011 promoter which can only be activated and then expressing CAHS and mRFP BBa_J04650 after IPTG is added.
The separate fragment of CAHS is 714bp, and the whole circuit is 2934bp.The plasmid skeleton is pSB1C3.
Identification
When we are building this circuit, we are doing the nucleic acid gel electrophoresis experiment to verify. After the circuit is built, we send the plasmid to sequence, and get the correct sequencing.
After new molecular cloning experiments, we do Enzyme-Cut identification to certify the plasmid is correct. We use the EcoR I and Pst I to cut the plasmid, then we getting two fragments - one is 2932bp, and the other is 2029bp.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 375
Illegal NheI site found at 1679
Illegal NheI site found at 1702 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 2880
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1376
Illegal AgeI site found at 1488 - 1000COMPATIBLE WITH RFC[1000]
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