...position do you want your gene to be in relation to the rest of the genes?
And do you want it to be in the same... simplifies vector construction by combining restriction and ligation steps together in the same tube ...the extraction and use of a coding sequence of interestCDS Level 0 part from an Addgene MoClo kit, to construct...questions to start with are:
Is your sequence of interest a promoter, a coding sequence, a terminator, or.... The overhangs are generated using type IIS restriction enzymes (BsaI, BbsI/BpiI, BsmBI), which recognize...four-base overhangs flank a coding sequence of interest (CDS1, top). Overhangs must match the Level 0 ...designing new primers to clone your sequence of interest into Level 0 parts.
Use in silico cloning tools...