URL: http://www.addgene.org/80624
Proper Citation: RRID:Addgene_80624
Insert Name: empty control
Bacterial Resistance: Ampicillin
Defining Citation: PMID:25027842
Vector Backbone Description: Backbone Marker:Kristoffer Ricken, Boris Fehse; Backbone Size:8300; Vector Backbone:LeGO-Cer; Vector Types:Lentiviral; Bacterial Resistance:Ampicillin
Comments: The LeGO-Cer/BSD backbone was cut with XbaI and XhoI to remove the mouse U6 promoter. The linearized plasmid was blunted by a proof-read Polymerase and religated. The resulting LeGO-Cer-noU6 was cut with BamHI and the mouse phosphoglycerate kinase 1 promoter (PGK) -amplified from the MSCV-puro vector (Clonetech/Takara Bio) - was inserted in sense orientation to drive expression of the Cerulean-BSD cassette. An SV40 consensus polyadenylation signal (pA) with a PacI and a PstI site at the 3' end was gene synthesized and inserted into the NsiI site before the PGK promoter. The SFFV promoter from the LeGO-Cer backbone was removed by excision with NotI and PacI and a cloning oligo with an MCS for lncRNA insertion (5'-NotI->AanI->XmaJI->XhoI->XbaI->HpaI->PacI-3') was inserted with NotI and PacI. The SFFV fragment -amplified from LeGO-Cer- was reinserted into the NotI site to drive expression of the lnRNA-pA cassette.
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Source: Addgene
