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Species: Homo sapiens
Genetic Insert: RPA1
Vector Backbone Description: Vector Backbone:pET11d; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
References:
Comments: Note: The RPA2 gene in this plasmid is a variant that also contains a T88I mutation.
Proper citation: RRID:Addgene_102617 Copy
Species:
Genetic Insert: cAMPr
Vector Backbone Description: Backbone Size:3300; Vector Backbone:P2lox; Vector Types:Mammalian Expression, Cre/Lox; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_102618 Copy
Species: Mus musculus
Genetic Insert: PPAR gamma coactivator 1 alpha
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:5500; Vector Backbone:pcDNA3.1 myc-His A; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_1026 Copy
Species: Homo sapiens
Genetic Insert: RPA1
Vector Backbone Description: Vector Backbone:pET11d; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_102613 Copy
Species: Saccharomyces cerevisiae
Genetic Insert: RPA1
Vector Backbone Description: Vector Backbone:pET11d; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
References:
Comments: RPA1 5317 RPA2 7292 RPA3 8284 (start of coding region listed)
Proper citation: RRID:Addgene_102614 Copy
Species:
Genetic Insert: J23106:B0034:spisPink:rrnBT1-T7TE
Vector Backbone Description: Backbone Marker:Self-made; Backbone Size:2195; Vector Backbone:derived from pSB1K3; Vector Types:Synthetic Biology; Bacterial Resistance:Kanamycin
References:
Comments:
Proper citation: RRID:Addgene_102701 Copy
Species:
Genetic Insert: J23106:B0034:spisPink:rrnBT1-T7TE
Vector Backbone Description: Backbone Marker:Self-made; Backbone Size:2195; Vector Backbone:derived from pSB1K3; Vector Types:Synthetic Biology; Bacterial Resistance:Kanamycin
References:
Comments:
Proper citation: RRID:Addgene_102703 Copy
Species: Homo sapiens
Genetic Insert: APPL1
Vector Backbone Description: Backbone Size:4722; Vector Backbone:mCherry-C3; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
References:
Comments: *Note: Addgene's quality control sequencing finds additional residues KPNSAVDGTAGPGSTGSR appended to the C-terminus of the APPL1 protein. These additional residues are not thought to affect protein function.
Proper citation: RRID:Addgene_102784 Copy
Species: Homo sapiens
Genetic Insert: Rab5a
Vector Backbone Description: Backbone Size:4731; Vector Backbone:EGFP-C1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
References:
Comments: mCherry-C2-Rab5a encodes wild-type human Rab5a,(NM_004162) as a fusion protein with mCherry. Rab5a was inserted between the Eco R1 and Sal 1 restriction sites. mCherry-C2 encodes a monomeric variant of DsRed. The backbone plasmid is pEGFP-C1 (Clontech). The C2 version of mCherry was created by insertion of a short piece of DNA between Xho 1 and EcoR1 restriction sites. In the resulting multiple cloning site, the Sac 1 restriction site has been eliminated and the Eco R1 restriction site is in frame.
Proper citation: RRID:Addgene_102785 Copy
Species: Other
Genetic Insert: reverse transcription xenopolymerase with 3'-5' deficient exonuclease
Vector Backbone Description: Vector Backbone:modified pET21; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_102786 Copy
Species: Homo sapiens
Genetic Insert: Zinc finger protein 57
Vector Backbone Description: Backbone Marker:NEB; Vector Backbone:NEB DHFR control plasmid; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_102716 Copy
Species:
Genetic Insert: Linker sequence
Vector Backbone Description: Backbone Marker:Self-made; Backbone Size:2267; Vector Backbone:derived from pSB1K3; Vector Types:Synthetic Biology; Bacterial Resistance:Kanamycin
References:
Comments:
Proper citation: RRID:Addgene_102714 Copy
Species:
Genetic Insert: Linker sequence
Vector Backbone Description: Backbone Marker:Self-made; Backbone Size:2267; Vector Backbone:derived from pSB1K3; Vector Types:Synthetic Biology; Bacterial Resistance:Kanamycin
References:
Comments:
Proper citation: RRID:Addgene_102710 Copy
Species:
Genetic Insert: HP1cs-Frb1x
Vector Backbone Description: Vector Backbone:pLenti; Vector Types:Mammalian Expression, Lentiviral, CRISPR; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_102808 Copy
Species:
Genetic Insert: HP1cs-Frb2x
Vector Backbone Description: Vector Backbone:pLenti; Vector Types:Mammalian Expression, Lentiviral, CRISPR; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_102809 Copy
Species:
Genetic Insert:
Vector Backbone Description: Backbone Marker:Markus Ralser (Addgene plasmid # 19407), Clontech = pTRE-T2; Vector Backbone:pTRE-Tight2; Vector Types:Mammalian Expression, RNAi; Bacterial Resistance:Ampicillin
References:
Comments: In fusion cloning was used to insert a synthetic DNA template containing the mIR cassette from the pCDNA 6.2-GW/miR into the EcoRI and XbaI site of pTRE-T2-Tight. I engineered two BsaI sites into the miR cassette so users can digest the vector with BsaI for directional cloning of Blockit microRNAs using the Block-IT RNAi designer engine from Invitrogen. To make the BsaI sites unique I mutated out Bsa I sites in the original pTRE-T2 Tight vector and also removed XhoI and XbaI sites by site-directed mutagenesis from the pTRE-T2 vector and the corresponding puromycin/hygromycin resistance genes. I also dropped in a unique NheI and EagI restriction site between the ampicillin gene and the pMB101 origin of replication sequence so that the puromycin and hygromin genes could be placed into the pTRE-T2-miR vector with the infusion cloning method. Investigators can remove the miR cassette from the pTRE-T2 puromycin/hygromycin vector and using cloning methods to insert any cDNA of interest into the site for doxycline-inducible expression of the target cDNA. A single vector can be utilized from the production of tetracycline inducible miRNAs or cDNAs.
For puromycin use .25 micrograms/ml to .50 micrograms/ml to get clones after a couple of weeks. Growth response curve tested in 293HEK cells. Suggest methylation deficient cells from NEB to alleviate methylation sensitive restriction sites like Xba I. However not recommended for long-term propagation and storage.
Proper citation: RRID:Addgene_102646 Copy
Species:
Genetic Insert:
Vector Backbone Description: Backbone Marker:Invitrogen; Vector Backbone:pcDNA3_neo; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
References:
Comments: No insert, but contains 3 HA epitopes for c-terminal tagged proteins
Proper citation: RRID:Addgene_102643 Copy
Species:
Genetic Insert:
Vector Backbone Description: Backbone Marker:Invitrogen; Vector Backbone:pcDNA3_neo; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
References:
Comments: No insert, but contains a single N-terminus Strepavidin binding peptide and Flag epitopes
Proper citation: RRID:Addgene_102644 Copy
Species: Other
Genetic Insert: C1V1::FusionRed::Kv2.1
Vector Backbone Description: Backbone Size:5366; Vector Backbone:pAAV; Vector Types:AAV; Bacterial Resistance:Ampicillin
References:
Comments:
Proper citation: RRID:Addgene_102771 Copy
Species: Homo sapiens
Genetic Insert: EPHA3
Vector Backbone Description: Backbone Marker:Invitrogen; Vector Backbone:pcDNA3.1(+)/myc-His B; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
References:
Comments: Please note last 10 amino acids in C-terminal are omitted in these constructs. These series of EPHA3 constructs carry missense mutations found in lung and colon cancer.
Proper citation: RRID:Addgene_102748 Copy
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