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Species: Rattus norvegicus
Genetic Insert: LAMP1
Vector Backbone Description: Vector Backbone:pCAG; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:23791179
Proper citation: RRID:Addgene_168513 Copy
Species: Rattus norvegicus
Genetic Insert: rat LAMP1-EGFP
Vector Backbone Description: Backbone Size:4125; Vector Backbone:pCS2; Vector Types:Unspecified; Bacterial Resistance:Ampicillin
Defining Citation: PMID:33296658
Proper citation: RRID:Addgene_169003 Copy
Species: Rattus norvegicus
Genetic Insert: GBAi
Vector Backbone Description: Backbone Size:6000; Vector Backbone:pLIC-His; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:29133411
Proper citation: RRID:Addgene_171755 Copy
Species: Rattus norvegicus
Genetic Insert: GBAi
Vector Backbone Description: Backbone Size:6000; Vector Backbone:pLIC-myc; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:29133411
Proper citation: RRID:Addgene_171753 Copy
Species: Rattus norvegicus
Genetic Insert: PV-DsRed-NLS-CD
Vector Backbone Description: Backbone Size:33400; Vector Backbone:Adenoviral Recombinant (pAdEasy); Vector Types:Mammalian Expression, Adenoviral; Bacterial Resistance:Kanamycin
Defining Citation: PMID:17420246
Comments: AdEasy is a registered trademark of the Johns Hopkins University.
Proper citation: RRID:Addgene_17287 Copy
Species: Rattus norvegicus
Genetic Insert: PV-GFP-NLS
Vector Backbone Description: Backbone Size:5900; Vector Backbone:pCMV-Myc-Cyto-GFP; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:11971908
Proper citation: RRID:Addgene_17300 Copy
Species: Rattus norvegicus
Genetic Insert: DHFR-GFP-th-GluA1
Vector Backbone Description: Backbone Size:8420; Vector Backbone:pLenti; Vector Types:Mammalian Expression, Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:34241635
Comments: Please visit https://www.biorxiv.org/content/10.1101/2020.09.02.278770v1 for bioRxiv preprint.
Proper citation: RRID:Addgene_173008 Copy
Species: Rattus norvegicus
Genetic Insert: jGCaMP7c variant 1513-WPRE
Vector Backbone Description: Backbone Size:4764; Vector Backbone:pAAV-GFAP-EGFP; Vector Types:AAV; Bacterial Resistance:Ampicillin
Proper citation: RRID:Addgene_173159 Copy
Species: Rattus norvegicus
Genetic Insert: GLUT1
Vector Backbone Description: Backbone Marker:Stratagene; Backbone Size:3200; Vector Backbone:pBS+; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:3016720
Proper citation: RRID:Addgene_15993 Copy
Species: Rattus norvegicus
Genetic Insert: AMPK alpha2
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:5400; Vector Backbone:pcDNA3; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:11389854
Proper citation: RRID:Addgene_15991 Copy
Species: Rattus norvegicus
Genetic Insert: ARHGEF9
Vector Backbone Description: Vector Backbone:mu6pro; Vector Types:RNAi; Bacterial Resistance:Ampicillin
Defining Citation: PMID:33316079
Comments: Targets all collybistin mRNAs (target is located in the protein coding region. Nucleotides 1316-1335 of AJ302676 [CB2SH3-] or 1496-1515 of AJ250425 [CB1SH3+]. Targets all collybistin isoforms).
Proper citation: RRID:Addgene_160065 Copy
Species: Rattus norvegicus
Genetic Insert: ARHGEF9
Vector Backbone Description: Vector Backbone:mu6pro; Vector Types:RNAi; Bacterial Resistance:Ampicillin
Defining Citation: PMID:33316079
Comments: 3-point mutation negative control for CBSH3+ shRNA-1 targeting collybistin isoforms with SH3+ domain
Proper citation: RRID:Addgene_160067 Copy
Species: Rattus norvegicus
Genetic Insert: ARHGEF9
Vector Backbone Description: Vector Backbone:mu6pro; Vector Types:RNAi; Bacterial Resistance:Ampicillin
Defining Citation: PMID:33316079
Comments: Targets collybistin SH3+ mRNAs (target is located in the protein coding region of SH3+. Nucleotides 498-518 of AJ250425 [CB1SH3+]. Targets collybistin isoforms with SH3 domain).
Proper citation: RRID:Addgene_160068 Copy
Species: Rattus norvegicus
Genetic Insert: ARHGEF9
Vector Backbone Description: Backbone Marker:Modified from Bryan Roth (Addgene plasmid # 65417); Vector Backbone:pAAV2; Vector Types:AAV; Bacterial Resistance:Ampicillin
Defining Citation: PMID:34083383
Proper citation: RRID:Addgene_160069 Copy
Species: Rattus norvegicus
Genetic Insert: ARHGEF9
Vector Backbone Description: Vector Backbone:mu6pro; Vector Types:RNAi; Bacterial Resistance:Ampicillin
Defining Citation: PMID:33316079
Comments: 3-point mutation negative control for CBSH3- shRNA3 targeting collybistin MYD-CBSH3- isoforms (Addgene Plasmid #160212).
Proper citation: RRID:Addgene_160213 Copy
Species: Rattus norvegicus
Genetic Insert: soluble adenylyl cyclase
Vector Backbone Description: Backbone Marker:Stratagene; Backbone Size:4500; Vector Backbone:pBK-CMV; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:9874775
Comments: Original library clone from Lambda-Zap rat testis library. CTCT repeats inserted between EcoRI sites in 5' UTR. Clone starts at nucleotide 241 of sAC sequence. EcoRI/XhoI digest: 0.7, 1.1, 1.5, 2.0, 4.5 kb.
pSAC5.02 is derived from an original library clone obtained in the cloning of rat sAC.
The isolation of this cDNA clone seems outdated now, but it was still a current method back in 1999, when we cloned sAC.
We obtained peptide sequences from purified sAC protein, and designed PCR primers to those peptides. These resulted in isolation of a 1 kb fragment encoding a portion of the sAC cDNA, but there was open reading frame on each end of this PCRd clone. We then screened a rat testis library we generated in the lamdba zap cloning system. We screened the library by hybridization using the 1 kb PCR fragment as probe and we identified three, independent cDNA clones.
THe lambda zap cloning system that we used enabled isolation of plasmids from the lambda phage library using helper phage. THe resultant plasmids contained library inserts (in our case, a sAC cDNA) in the pBK-CMV cloning vector. In this cloning vector, the cDNAs were inserted directionally, meaning that the vectors were already suitable for bacterial expression using the T7 promoter. We were unable to get sufficient protein via bacterial expression, so we deleted the T7 promoter region by digesting with NheI-SpeI and reclosing the vector; this brought the CMV promoter closer to the inserted sAC cDNA and facilitated high level expression in mammalian cells (i.e., HEK293 cells). This is how plasmid psAC5.02 was generated.
THe CTCT repeats are within the 5' UTR of the sAC cDNA - they may have been a cloning artifact during generation of the library, or they may represent some 5' control element. The cDNA clone can not be moved (in its entirety) as an EcoR1 fragment (the pBK-CMV vector has EcoRI sites flanking the inserted cDNA) because there is an internal EcoR1 site within the sAC cDNA. EcoR1 can be used as a diagnostic digest to confirm the validity of the plasmid.
Proper citation: RRID:Addgene_16077 Copy
Species: Rattus norvegicus
Genetic Insert: PKC beta 1
Vector Backbone Description: Backbone Size:5400; Vector Backbone:pHACE; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:12794082
Comments: From user-provided sequence (see 'Reviews'), there are five amino acid differences from one of two NCBI reference sequences (AAA41868.1 and NP_001165776.1): 25A, 62F, 64K, 69C, and 294G. These "mutations" are also present in human, bovine, and xenopus sequences and are therefore not believed to affect function.
Proper citation: RRID:Addgene_16381 Copy
Species: Rattus norvegicus
Genetic Insert: LAMP1
Vector Backbone Description: Vector Backbone:N/A; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:31539493
Proper citation: RRID:Addgene_164209 Copy
Species: Rattus norvegicus
Genetic Insert: rat FL Annexin A11
Vector Backbone Description: Backbone Marker:Gunter Stier; Vector Backbone:pET; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:32344647
Proper citation: RRID:Addgene_164496 Copy
Species: Rattus norvegicus
Genetic Insert: GCaMP6s
Vector Backbone Description: Backbone Marker:Clontech; Backbone Size:8513; Vector Backbone:pLVX-IRES-Hygro; Vector Types:Mammalian Expression, Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:31346185
Proper citation: RRID:Addgene_164591 Copy
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