Part:BBa_K3407017:Design
Cry7Ca1 toxin with inducible T7 promoter, RBS and T7 terminator
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 688
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1388
Illegal XhoI site found at 367 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
The Cry protein, Cry7Ca1 (GenBank accession no. EF486523) from the Bt strain BTH-13 is a three-domain Cry protein with a molecular weight of 129,196.50 kDa, whereas the activated toxin has a molecular weight of 68 kDa. This part contains the activated toxin. The gene was codon-optimized for expression in Escherichia coli using the GenSmartTM Codon Optimization Tool. The forbidden restriction enzymes sites are the following: BioBrick forbidden restriction sites (EcoRI, XbaI, SpeI, PstI) and Type IIS forbidden restriction sites (BsaI, SapI and Bpil).
The expression cassette of this part was synthesized in the backbone vector pTWIST_Amp_HighCopy from Twist bioscience, containing the viral promoter and terminator from the T7 bacteriophage. This synthetically synthetised plasmid is referred as pTWIST_Cry7Ca1.
This part was designed as if it was cloned using Modular Cloning Toolkit [1][2], so it contians the scars left after assembly.
Source
Synthetically synthesised.
References