Irfp frb rab7

FKBP sequence was tagged to 3xFLAG-pDEST and mScarlet-pDEST vectors using IN-FUSION HD cloning technology (Clontech, Takara, cat #638920). LRRK2-WT, LRRK2-R1441C, LRRK2-Y1669C, LRRK2-K1906M and LRRK2-G2019S, previously cloned into pCR TM 8/GW/TOPO TM vector (ThermoFisher, cat #250020), were transferred into the 3xFLAG-FKBP-pDEST and mScarlet-FKBP-pDEST plasmids using Gateway technology (ThermoFisher, cat #11791043) . LYSO-LRRK2 was created by adding the N terminal domain of the LAMTOR1 sequence (aa 1-39) into the 3xFLAG-LRRK2 plasmid using IN-FUSION (LAMTOR1(1-39)-3xFLAG-LRRK2). CFP-FRB-LAMP1 vector (Willett et al, 2017) was a gift from Rosa Puertollano (NIH). EHD1-FRB-GFP was kindly provided by Tsukasa Okiyoneda (Kwansei Gakuin University). PM-FRB-CFP (Addgene plasmid #67517) (Varnai et al, 2006) , iRFP-FRB-RAB7 (Addgene plasmid #51613) and iRFP-FRB-RAB5 plasmids (Addgene plasmid #51612) (Hammond et al, 2014) were gifts from Tamas Balla (NIH). FRB-ECFP-Giantin was a provided by Dorus Gadella (Addgene plasmid #67903) (van Unen et al, 2015) . The LAMP1-SBP-GFP and Strep-KIFC1*-HA plasmids were a generous gift from Juan S. Bonifacino (Guardia et al, 2019) .