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Species: Synthetic
Genetic Insert: GCaMP6s
Vector Backbone Description: Backbone Marker:custom; Backbone Size:5095; Vector Backbone:pAAV-CaMKIIa-WPRE-BGHpA; Vector Types:Mammalian Expression, AAV; Bacterial Resistance:Ampicillin
Comments: The addition of the P2A-nls-dTomato allows for easy identification of GCaMP6 expressing cells exhibiting nuclear red fluorescence that does not significantly overlap with the cytosolic GCaMP signal. The GCaMP and fluorophore are physically uncoupled.
Permissions were obtained from Douglas Kim and the Clontech licensing office for depositing this new plasmid.
Proper citation: RRID:Addgene_51086 Copy
Genetic Insert: LifeAct-mRFPruby
Vector Backbone Description: Backbone Marker:L. Naldini; Vector Backbone:pLenti.PGK:EGFP; Vector Types:Mammalian Expression, Lentiviral; Bacterial Resistance:Ampicillin
Proper citation: RRID:Addgene_51009 Copy
Species: Synthetic
Genetic Insert: H2B-Dendra2
Vector Backbone Description: Vector Backbone:pLenti.CAG; Vector Types:Mammalian Expression, Lentiviral; Bacterial Resistance:Ampicillin
Comments: Please note that the H2B sequence in this plasmid was derived from H2B-GFP (Addgene plasmid# 11680) and contains the same differences (D26G and V119I) compared to GenBank reference sequences, such as NP_066402.2.
Proper citation: RRID:Addgene_51005 Copy
Species: Homo sapiens
Genetic Insert: H2B-EGFP
Vector Backbone Description: Backbone Size:8344; Vector Backbone:pLenti.Syn(0.5).eGFP; Vector Types:Mammalian Expression, Lentiviral; Bacterial Resistance:Ampicillin
Comments: Please note that the H2B sequence in this plasmid was derived from H2B-GFP (Addgene plasmid# 11680) and contains the same differences (D26G and V119I) compared to GenBank reference sequences, such as NP_066402.2.
Proper citation: RRID:Addgene_51004 Copy
Genetic Insert: H2B-ChFP
Vector Backbone Description: Backbone Size:6686; Vector Backbone:pRRLsin.cPPT.PGKp.eGFP.wpre; Vector Types:Mammalian Expression, Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:26209648
Comments: Please note that the H2B sequence in this plasmid was derived from H2B-GFP (Addgene plasmid# 11680) and contains the same differences (D26G and V119I) compared to GenBank reference sequences, such as NP_066402.2.
Proper citation: RRID:Addgene_51007 Copy
Vector Backbone Description: Backbone Marker:Clontech; Backbone Size:8089; Vector Backbone:pSIR; Vector Types:Mammalian Expression, Retroviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:25051498
Proper citation: RRID:Addgene_51128 Copy
Species: Homo sapiens
Genetic Insert: CRAF
Vector Backbone Description: Backbone Size:5200; Vector Backbone:pBABEpuro; Vector Types:Mammalian Expression, Retroviral; Bacterial Resistance:Ampicillin
Proper citation: RRID:Addgene_51127 Copy
Vector Backbone Description: Backbone Marker:Bert Vogelstein lab (Addgene Plasmid # 16405); Backbone Size:8935; Vector Backbone:pAdTrack-CMV; Vector Types:Adenoviral; Bacterial Resistance:Kanamycin
Proper citation: RRID:Addgene_50957 Copy
Species: Mus musculus
Genetic Insert: Gli2
Vector Backbone Description: Backbone Marker:Invitrogen; Vector Backbone:pEF5-FRT-DEST; Vector Types:Mammalian Expression, Flpin system; Bacterial Resistance:Ampicillin
Defining Citation: PMID:24373970
Proper citation: RRID:Addgene_51250 Copy
Genetic Insert: Co-InCreN
Vector Backbone Description: Backbone Marker:CLONTECH; Backbone Size:4814; Vector Backbone:pDIRECT; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:24413561
Comments: Please note that the complementary plasmid pCAG-Co-InCreC (www.addgene.org/51268) must be ordered with pCAG-Co-InCreN in order to successfully use the Co-Driver system as described in the associated publication.
Proper citation: RRID:Addgene_51267 Copy
Genetic Insert: b3STOPb3-ZsGreen
Vector Backbone Description: Backbone Marker:CLONTECH; Backbone Size:4814; Vector Backbone:pDIRECT; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:24413561
Proper citation: RRID:Addgene_51266 Copy
Species: Synthetic
Genetic Insert: 3xFLAG-dCas9
Vector Backbone Description: Backbone Marker:Hodaka Fujii; Backbone Size:6238; Vector Backbone:pMXs-neo; Vector Types:Mammalian Expression, Retroviral, CRISPR; Bacterial Resistance:Ampicillin
Defining Citation: PMID:25051498
Comments: The coding sequence of 3xFLAG-dCas9 can be cleaved with Pac I and Not I.
Construction strategy of gRNA retroviral vectors
1. Cleave gBlock from a gRNA vector constructed using gRNA cloning vector (Addgene #41824) with appropriate restriction enzymes (eg. [Xho I + Hind III], EcoR I).
2. Insert the cleaved gBlock into pSIR-based self-inactivating retroviral vectors.
Vectors & Sites of insertion
pSIR-neo (Addgene #51128): eg. [Xho I + Hind III]
pSIR-GFP (Addgene #51134): eg. [Xho I + Hind III], EcoR I
pSIR-DsRed-Express2 (Addgene #51135): eg. [Xho I + Hind III], EcoR I
pSIR-hCD2 (Addgene #51143): eg. EcoR I
Proper citation: RRID:Addgene_51260 Copy
Species: Synthetic
Genetic Insert: Pyronic
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:5419; Vector Backbone:pcDNA3.1(-); Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:24465702
Proper citation: RRID:Addgene_51308 Copy
Genetic Insert: Roxed-Cre
Vector Backbone Description: Vector Backbone:pTS(2); Vector Types:; Bacterial Resistance:Ampicillin
Defining Citation: PMID:24413561
Proper citation: RRID:Addgene_51276 Copy
Genetic Insert: Dre
Vector Backbone Description: Backbone Marker:CLONTECH; Backbone Size:4814; Vector Backbone:pDIRECT; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:24413561
Comments: Please note that the complementary plasmid pCAG-Roxed-Cre (www.addgene.org/51273) must be ordered with pCAG-NLS-HA-Dre in order to successfully use the Co-Driver recombinase system as described in the associated publication.
Proper citation: RRID:Addgene_51272 Copy
Species: Homo sapiens
Genetic Insert: cofilin 1
Vector Backbone Description: Backbone Marker:R.Y. Tsien, PNAS 2002 v99 pp7877–7882; Vector Backbone:pmRFP-N1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:24391794
Comments: This cofilin plasmid contains some silent mutations to make it resistant to an siRNA used in the Bamburg lab. The mutations change the wildtype sequence (AAG TCT TCA ACG CCA GAG GAG) into AAG AGC TCA ACC CCA GAG GAG.
Proper citation: RRID:Addgene_51279 Copy
Species: Homo sapiens
Genetic Insert: U6
Vector Backbone Description: Backbone Marker:Addgene plasmid 51283; Vector Backbone:pAluYa5-neo-TET; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:19390602
Comments: U6 is tagged with the self splicing intron neo cassette such that G418 resistance will be obtained only if retrotransposition occurs.
Please note that the cassette is in the reverse orientation within the MCS relative to the full sequence--This has no functional consequence.
Proper citation: RRID:Addgene_51286 Copy
Species: Homo sapiens
Genetic Insert: L1 ORF2
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:4595; Vector Backbone:pBudCE4.1; Vector Types:Mammalian Expression; Bacterial Resistance:Bleocin (Zeocin)
Defining Citation: PMID:21572950
Comments: This plasmid was created using the codon optimized L1RP as a source for the ORF2 coding sequence.
Proper citation: RRID:Addgene_51289 Copy
Species: Homo sapiens
Genetic Insert: L1
Vector Backbone Description: Backbone Marker:Stratagene; Vector Backbone:pBluescript II; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:21572950
Comments: L1 is tagged with neoI indicator cassette containing an inverted neomycin resistance gene. The SV40 promoter (SV40p) drives transcription of the neomycin gene which is disrupted by an intron in the opposite orientation that renders it non-functional. The intron is only spliced out from tagged L1 transcripts generated by the CMV promoter. When retrotransposition of the spliced L1 RNA occurs, the new insert will contain a functional neo gene that is expressed from the SV40 promoter.
The L1 open reading frames were codon optimized using Primo Optimum 3.4 (http://www.changbioscience.com/primo/primoo.html), which makes synonymous changes to optimal codons within the nucleotide sequence. A 63 bp inter-ORF with 92% sequence identity to L1RP was utilized for all constructs. Sequence changes to this region were generated to introduce restriction enzyme sites used for plasmid construction. The optimized nucleotide sequence is 74.9% identical to the sequence of L1RP.
Proper citation: RRID:Addgene_51288 Copy
Species: Micromonospora echinospora
Genetic Insert: CalR1
Vector Backbone Description: Vector Backbone:pET21a; Vector Types:Bacterial Expression, Synthetic Biology; Bacterial Resistance:Ampicillin
Defining Citation: PMID:24316737
Comments: UCSF Case No. SF11-016-2; U.S. Patent Application 13/489,205. GeneArt inserted the synthesized fragment into a pET21a vector.
Proper citation: RRID:Addgene_51161 Copy
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