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Genetic Insert: STAgR Insert gRNAScaffold_mU6
Vector Backbone Description: Backbone Marker:Stricker Lab; Vector Backbone:PCR Template for STAgR Reactions; Vector Types:PCR Template for STAgR Inserts; Bacterial Resistance:Kanamycin
Defining Citation: PMID:29702666
Proper citation: RRID:Addgene_102844 Copy
Species: Mus musculus
Genetic Insert: CTCF
Vector Backbone Description: Vector Backbone:NEB DHFR control plasmid; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:29159284
Comments: This plasmid encodes the DNA binding domain of mouse CTCF protein (F1 to F9) for E. coli expression.
Proper citation: RRID:Addgene_102859 Copy
Species: Mus musculus
Genetic Insert: PSD-95
Vector Backbone Description: Backbone Size:4141; Vector Backbone:pCMV; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:25948262
Proper citation: RRID:Addgene_102949 Copy
Species: Gallus gallus
Genetic Insert: SIINFEKL
Vector Backbone Description: Backbone Size:8200; Vector Backbone:PresentER; Vector Types:Mammalian Expression, Retroviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:30499773
Comments: Please visit https://www.biorxiv.org/content/early/2018/09/22/267047 for bioRxiv preprint.
Proper citation: RRID:Addgene_102945 Copy
Species: Plasmodium falciparum
Genetic Insert: PfSSB
Vector Backbone Description: Vector Backbone:pET22B; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:29466468
Comments: The expressed protein is a Plasmodium falciparum single-stranded DNA-binding protein, PfSSB, for subsequent labeling at the single, native cysteine with one IDCC (N-[2-(iodoacetamido)ethyl]-7-diethylaminocoumarin-3-carboxamide) per subunit. This adduct is then used as a fluorescent single-stranded DNA (ssDNA) biosensor, having ~20-fold fluorescence increase on binding ssDNA with very high affinity. PfSSB was expressed without the N-terminal apicoplast localisation sequence (76 amino acids) but amino acids are numbered for the full-length protein, that is the first amino acid in the expressed protein is M77.
Protocol
See Hedgethorne and Webb (2012), which is also followed for PfSSB purification, except with the omission of the PEI precipitation step.
Hedgethorne, K.; Webb, M. R., Fluorescent SSB as a reagentless biosensor for single-stranded DNA. Methods Mol. Biol. 2012, 922, 219-233.
Proper citation: RRID:Addgene_103000 Copy
Species: Synthetic
Genetic Insert: Synthetic construct isolate AAV-PHP.B2 VP1 gene
Vector Backbone Description: Vector Backbone:pUC57-mini; Vector Types:Mammalian Expression, AAV; Bacterial Resistance:Ampicillin
Defining Citation: PMID:26829320
Comments: Please note that this is a non-standard rep-cap construct. It uses a tTA-TRE amplification loop to increase Capsid expression and AAV production. We find that this system increases AAV titers 1.5-5 fold (Ben Deverman, Bryan Simpson and Paul Patterson, unpublished data). This is a tet-off system, so no dox or tet is needed to turn it on. It can be used like any other rep-cap plasmid. *This system should only present a problem if the rAAV genome to be packaged has a tet responsive element that directs expression of a protein that affected the health of the production cells. The introduction of an XbaI restriction site for ease of cloning introduces a K449R mutation, which does not have an overt effect on vector production or transduction.
Proper citation: RRID:Addgene_103003 Copy
Species: Synthetic
Genetic Insert: Synthetic construct isolate AAV-PHP.B VP1 gene
Vector Backbone Description: Vector Backbone:pUC57-mini; Vector Types:Mammalian Expression, AAV; Bacterial Resistance:Ampicillin
Defining Citation: PMID:26829320
Comments: Please note that this is a non-standard rep-cap construct. It uses a tTA-TRE amplification loop to increase Capsid expression and AAV production. We find that this system increases AAV titers 1.5-5 fold (Ben Deverman, Bryan Simpson and Paul Patterson, unpublished data). This is a tet-off system, so no dox or tet is needed to turn it on. It can be used like any other rep-cap plasmid. *This system should only present a problem if the rAAV genome to be packaged has a tet responsive element that directs expression of a protein that affected the health of the production cells. The introduction of an XbaI restriction site for ease of cloning introduces a K449R mutation, which does not have an overt effect on vector production or transduction.
Proper citation: RRID:Addgene_103002 Copy
Species: Synthetic
Genetic Insert: Synthetic construct isolate AAV-PHP.eB VP1 gene
Vector Backbone Description: Vector Backbone:pUC57-mini; Vector Types:Mammalian Expression, AAV; Bacterial Resistance:Ampicillin
Defining Citation: PMID:28671695
Comments: Please note that this is a non-standard rep-cap construct. It uses a tTA-TRE amplification loop to increase Capsid expression and AAV production. We find that this system increases AAV titers 1.5-5 fold (Ben Deverman, Bryan Simpson and Paul Patterson, unpublished data). This is a tet-off system, so no dox or tet is needed to turn it on. It can be used like any other rep-cap plasmid. This system should only present a problem if the rAAV genome to be packaged has a tet responsive element that directs expression of a protein that affected the health of the production cells. The introduction of an XbaI restriction site for ease of cloning introduces a K449R mutation, which does not have an overt effect on vector production or transduction.
Proper citation: RRID:Addgene_103005 Copy
Species: Synthetic
Genetic Insert: Synthetic construct isolate AAV-PHP.B3 VP1 gene
Vector Backbone Description: Vector Backbone:pUC57-mini; Vector Types:Mammalian Expression, AAV; Bacterial Resistance:Ampicillin
Defining Citation: PMID:26829320
Comments: Please note that this is a non-standard rep-cap construct. It uses a tTA-TRE amplification loop to increase Capsid expression and AAV production. We find that this system increases AAV titers 1.5-5 fold (Ben Deverman, Bryan Simpson and Paul Patterson, unpublished data). This is a tet-off system, so no dox or tet is needed to turn it on. It can be used like any other rep-cap plasmid. *This system should only present a problem if the rAAV genome to be packaged has a tet responsive element that directs expression of a protein that affected the health of the production cells. The introduction of an XbaI restriction site for ease of cloning introduces a K449R mutation, which does not have an overt effect on vector production or transduction.
Proper citation: RRID:Addgene_103004 Copy
Species: Zaire ebolavirus, Mayinga variant
Genetic Insert: Ebolavirus minigenome
Vector Backbone Description: Vector Backbone:pCAGGs; Vector Types:Mammalian Expression, Luciferase; Bacterial Resistance:Ampicillin
Defining Citation: PMID:28807685
Proper citation: RRID:Addgene_103055 Copy
Species: Synthetic
Genetic Insert: TIMER-bac
Vector Backbone Description: Backbone Size:9262; Vector Backbone:pSC101; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:25126781
Comments: TIMER-bac was generated by exchanging TCC coding for serine 197 to ACC coding for threonine in DsRed.T3_S4T (Sörensen et al., 2003). PybaJ-Timer-bac was cloned into pUA66 with a pSC101 replicon, conferring kanamycin resistance.
Proper citation: RRID:Addgene_103057 Copy
Species: Zaire ebolavirus, Mayinga
Genetic Insert: L gene of Ebola virus
Vector Backbone Description: Vector Backbone:pCAGGs; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:28807685
Proper citation: RRID:Addgene_103052 Copy
Species: Zaire ebolavirus, Mayinga
Genetic Insert: VP30 gene of Ebola virus
Vector Backbone Description: Vector Backbone:pCAGGs; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:28807685
Proper citation: RRID:Addgene_103051 Copy
Species: Zaire ebolavirus, Mayinga
Genetic Insert: VP35
Vector Backbone Description: Vector Backbone:pCAGGs; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:27252530
Proper citation: RRID:Addgene_103053 Copy
Species: E. coli
Genetic Insert: Colicin E1 and E1 immunity protein
Vector Backbone Description: Backbone Marker:NEB; Backbone Size:2686; Vector Backbone:pUC19; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:19029376
Comments: Insert was cloned from ~base pair 5015 to ~500 of the circular ColE1 plasmid, where numbering begins within the colicin E1 gene. The ColE1 sequences include both the promoter for colicin E1 and for E1 immunity protein, which is transcribed in the opposite direction from the colicin gene. The cloned DNA does not include the intact kil (lysis protein) gene.
Proper citation: RRID:Addgene_103063 Copy
Species: Homo sapiens
Genetic Insert: Cas9 plus expressed GESTALT V1-V5 guide
Vector Backbone Description: Vector Backbone:lentiCRISPR v2; Vector Types:Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:27229144
Comments: Derived from https://www.addgene.org/52961/
Proper citation: RRID:Addgene_103062 Copy
Genetic Insert: TVA-mCherry fusion protein after CRE-mediated recombination
Vector Backbone Description: Vector Backbone:pAAV; Vector Types:AAV, Adeno Associated Viral Vector; Bacterial Resistance:Ampicillin
Defining Citation: PMID:28689641
Proper citation: RRID:Addgene_102985 Copy
Species: (Zaire ebolavirus, Mayinga)
Genetic Insert: VP35 gene of Ebola virus
Vector Backbone Description: Vector Backbone:pCAGGs; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:28807685
Proper citation: RRID:Addgene_103050 Copy
Vector Backbone Description: Backbone Marker:Stricker Lab; Vector Backbone:Human gRNA Expression Vector / PCR Template for STAgR; Vector Types:CRISPR, PCR Template for STAgR Vectors; Bacterial Resistance:Ampicillin
Defining Citation: PMID:29702666
Proper citation: RRID:Addgene_102992 Copy
Species: Homo sapiens
Genetic Insert: shNFS1_2
Vector Backbone Description: Backbone Marker:Broad Institute; Backbone Size:7466; Vector Backbone:pLKO_TRC005; Vector Types:Lentiviral, RNAi; Bacterial Resistance:Ampicillin
Defining Citation: PMID:29168506
Comments: TRCN0000229755
Proper citation: RRID:Addgene_102964 Copy
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