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Species: Homo sapiens
Genetic Insert: MCU
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:5485; Vector Backbone:pDEST40-pcDNA; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:21685886
Proper citation: RRID:Addgene_31731 Copy
Species: Homo sapiens
Genetic Insert: Snail
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:5428; Vector Backbone:pcDNA3.1+; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:15314165
Proper citation: RRID:Addgene_31697 Copy
Species: Mus musculus
Genetic Insert: Smo
Vector Backbone Description: Backbone Marker:Hawley et al., 1994; Backbone Size:7000; Vector Backbone:pMSCV-pac; Vector Types:Mammalian Expression, Retroviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:19948480
Proper citation: RRID:Addgene_31619 Copy
Species: Drosophila melanogaster
Genetic Insert: Rab5
Vector Backbone Description: Backbone Marker:Drosophila Gateway Collection; Vector Backbone:pAGW; Vector Types:Insect Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:21169635
Proper citation: RRID:Addgene_31733 Copy
Species: Homo sapiens
Genetic Insert: ATR
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:5400; Vector Backbone:pcDNA3.1; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:16354678
Comments: Plasmid contains an R1631H polymorphism. This is not thought to affect plasmid function.
Proper citation: RRID:Addgene_31611 Copy
Species: Homo sapiens
Genetic Insert: MCU
Vector Backbone Description: Backbone Marker:Invitrogen (Cat#V355-20); Backbone Size:5784; Vector Backbone:pcDNA6.2/C-EmGFP-DEST; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:21685886
Comments: *A2T aa residue substitution in MCU, which is not known to affect protein function
Proper citation: RRID:Addgene_31732 Copy
Species: Drosophila melanogaster
Genetic Insert: Rab8
Vector Backbone Description: Backbone Marker:Drosophila Gateway Collection; Vector Backbone:pAGW; Vector Types:Insect Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:21169635
Proper citation: RRID:Addgene_31735 Copy
Species: Drosophila melanogaster
Genetic Insert: Rab6
Vector Backbone Description: Backbone Marker:Drosophila Gateway Collection; Vector Backbone:pAGW; Vector Types:Insect Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:21169635
Proper citation: RRID:Addgene_31734 Copy
Species: Drosophila melanogaster
Genetic Insert: Rab10
Vector Backbone Description: Backbone Marker:Drosophila Gateway Collection; Vector Backbone:pAGW; Vector Types:Insect Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:21169635
Proper citation: RRID:Addgene_31737 Copy
Species: Drosophila melanogaster
Genetic Insert: Rab9
Vector Backbone Description: Backbone Marker:Drosophila Gateway Collection; Vector Backbone:pAGW; Vector Types:Insect Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:21169635
Proper citation: RRID:Addgene_31736 Copy
Species: Mus musculus
Genetic Insert: Smo
Vector Backbone Description: Backbone Marker:RZPD; Backbone Size:4095; Vector Backbone:pCS2+; Vector Types:Mammalian Expression, Worm Expression, Zebrafish, Avian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:19948480
Comments: Wild-type mouse Smo was tagged at the N terminus (with the insertion C terminal to the signal sequence) with YFP.
Proper citation: RRID:Addgene_31618 Copy
Species: Mus musculus
Genetic Insert: Smo
Vector Backbone Description: Backbone Marker:RZPD; Backbone Size:4095; Vector Backbone:pCS2+; Vector Types:Mammalian Expression, Worm Expression, Zebrafish, Avian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:19948480
Comments: Wild-type mouse Smo was tagged at the N terminus (with the insertion C terminal to the signal sequence) with SNAP.
Proper citation: RRID:Addgene_31617 Copy
Species: Homo sapiens
Genetic Insert: p21-activated protein kinase 2 S20A
Vector Backbone Description: Backbone Size:2900; Vector Backbone:pECE, Addgene plasmid 26453; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:22137581
Proper citation: RRID:Addgene_31681 Copy
Species: Homo sapiens
Genetic Insert: ALK5
Vector Backbone Description: Backbone Size:4716; Vector Backbone:pRK5; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Comments: This construct is beginning from a Signal Peptide of human TbRI/ALK5, followed by a HA tag, the extracellular cDNA sequence of human TbRI/ALK, and ended by a Flag tag. This vector to produces a secreted TbRI/ALK5 extracellular form, with two individual tags on each side.
Proper citation: RRID:Addgene_31720 Copy
Species: Homo sapiens
Genetic Insert: MCU
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:2787; Vector Backbone:pDONR223; Vector Types:pDONR Gateway; Bacterial Resistance:Spectinomycin
Defining Citation: PMID:21685886
Proper citation: RRID:Addgene_31729 Copy
Vector Backbone Description: Backbone Marker:Khavari Lab; Backbone Size:11100; Vector Backbone:LZRS-RfA; Vector Types:Retroviral; Bacterial Resistance:Chloramphenicol and Ampicillin
Defining Citation: PMID:15342384
Proper citation: RRID:Addgene_31601 Copy
Species: Mus musculus
Genetic Insert: ΔN331 kinesin-1B
Vector Backbone Description: Vector Backbone:pCGN; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:19812246
Proper citation: RRID:Addgene_31603 Copy
Genetic Insert: BoNT/A LC
Vector Backbone Description: Backbone Size:0; Vector Backbone:pBN3; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:18940613
Comments: The author states that although the construct is from a toxic protein, it only contains one third of the biologically active protein,i.e., it is inactive and harmless per se, and it does not require BL4 level for its expression/manipulation.
Toxic BoNT/A is made of two 'chains' (i.e. polypeptides) linked to each other by one disulfide bond. The light chain (LC) is 50KDa long, whereas the heavy chain (HC) is 100 KDa. The HC is necessary for the toxin to target and enter motorneurons. This construct is the LC of BoNTA, and it completely lacks the HC, so it is unable to intoxicate neurons.
Protein sequence of BoNT/A fragment present in this plasmid (corresponds to YP_001386738.1)
MSGLEVSFEELRTFGGHDAKFIDSLQENEFRLYYYNKFKDIASTLNKAKSIVGTTASLQYMKNVFKEKYLLSEDTSGKFSVDKLKFDKLYKMLTEIYTEDNFVKFFKVLNRKTYLNFDKAVFKINIVPKVNYTIYDGFNLRNTNLAANFNGQNTEINNMNFTKLKNFT
Nucleotide sequence of BoNT/A fragment present in this plasmid based on Addgene's Sanger sequencing results:
ATGAGTGGGTTAGAAGTAAGCTTTGAGGAACTTAGAACATTTGGGGGACATGATGCAAAGTTTATAGATAGTTTACAGGAAAACGAATTTCGTCTATATTATTATAATAAGTTTAAAGATATAGCAAGTACACTTAATAAAGCTAAATCAATAGTAGGTACTACTGCTTCATTACAGTATATGAAAAATGTTTTTAAAGAGAAATATCTCCTATCTGAAGATACATCTGGAAAATTTTCGGTAGATAAATTAAAATTTGATAAGTTATACAAAATGTTAACAGAGATTTACACAGAGGATAATTTTGTTAAGTTTTTTAAAGTACTTAACAGAAAAACATATTTGAATTTTGATAAAGCCGTATTTAAGATAAATATAGTACCTAAGGTAAATTACACAATATATGATGGATTTAATTTAAGAAATACAAATTTAGCAGCAAACTTTAATGGTCAAAATACAGAAATTAATAATATGAATTTTACTAAACTAAAAAATTTTACT
Proper citation: RRID:Addgene_31602 Copy
Species: Mus musculus
Genetic Insert: kinesin-1C
Vector Backbone Description: Backbone Marker:Clontech; Backbone Size:4001; Vector Backbone:pEGFP-C1 (EGFP removed); Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:19812246
Proper citation: RRID:Addgene_31605 Copy
Species: Homo sapiens
Genetic Insert: MCU
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:2787; Vector Backbone:pDONR223; Vector Types:pDONR Gateway; Bacterial Resistance:Spectinomycin
Defining Citation: PMID:21685886
Comments:
*A3T aa residue substitution in MCU, which is not known to affect protein function
Proper citation: RRID:Addgene_31726 Copy
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