pDonor plasmids were generated by cloning the Donor fragments into either an EcoRV-digested pBluescript KS+ vector (pDonor_hU6) or into the Topo Blunt II plasmid (Invitrogene). The sU6 promoter was generated by replacing the regulatory elements of mU6 for those of hU6 (that is, the Octamer motif, the Proximal sequence element and the TATA-box). See Supplementary Fig. 2a for a schematic representation of these plasmids. The sequences contained in each of the pDonors are provided in Supplementary Fig. 2 , along with the sequence of the DNA oligos used for cloning gRNA pairs under the various promoters.
Pbluescript ks
Manufactured by Thermo Fisher Scientific
PBluescript KS is a plasmid vector commonly used in molecular biology experiments. It serves as a cloning and expression platform for genetic sequences. The vector contains various functional elements, including a multiple cloning site, lac promoter, and antibiotic resistance markers, which facilitate the insertion, maintenance, and selection of DNA fragments of interest.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using pbluescript ks
1
Plasmid Construction for gRNA Expression
2
Generation of pDonor Plasmids and sU6 Promoter
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