The cDNAs are expressed under the control of a CMV promoter. An additional PGK promoter controls the expression of a Blasticidin resistance selection marker.
All the cDNAs are V5-tagged.
The cDNAs were incorporated into the lenti cassette via the gateway system. Thus, the relevant cDNAs can be mobilized via the gateway system to gateway-compatible cassettes.
More library related info:
The Lenti-cDNA clones are now available to UNC investigators and their collaborators outside UNC. To verify the content of the entry clones, pools of full entry clone plasmids were Illumina sequenced and those that were matched to the original MGC cDNA were chosen. The library is premised on the CCSB - Broad lentiviral ORF collection and was tested via V5 epitope tag expression.
~90% of ORF lentivirus induced expression greater than the control mean. Further details can be found in Figure 2 of: A public genome-scale lentiviral expression library of human ORFs.
Nat Methods. 2011 Jun 26;8(8):659-61
There are a total of 15,801 viable clones in this collection, representing 13,016 unique gene IDs. The collection consists of fully-sequenced clones, as well as known mutants, partially sequenced, and unsequenced clones.
Summary of the clone count for each of these categories:
Fully-sequenced, no mutations: 12,467 clones
Mutant Subcollection: 1742 clones
Partially Sequenced Subcollection: 798 clones
Unsequenced Subcollection: 794 clones