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Species: Homo sapiens
Genetic Insert: RB shRNA
Vector Backbone Description: Backbone Size:13700; Vector Backbone:pSLIK-EGFP; Vector Types:Mammalian Expression, Lentiviral, RNAi; Bacterial Resistance:Ampicillin
Comments: The whole sequence for the shRNA including the loop should be from 3872-3931 in the vector: AGCGAAACGATTATCCATTCAAATAGTGAAGCCACAGATGTATTTGAAT
The shRNA is downstream of the EGFP
GGATAATCGTT
pSLIK vector from Ian Frasier
Proper citation: RRID:Addgene_31500 Copy
Species: fluorescent proteins
Genetic Insert: eYFP and mCherry
Vector Backbone Description: Backbone Marker:Clontech; Backbone Size:2850; Vector Backbone:pTRETightBI; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:21857679
Comments: 7 tandem miR-20 binding sites in mCherry 3'UTR
Proper citation: RRID:Addgene_31466 Copy
Species: Homo sapiens
Genetic Insert: Actin related protein 2/3 complex, subunit 3
Vector Backbone Description: Backbone Size:3460; Vector Backbone:pQStrep2; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:15998469
Comments: The vector pQTEV is derived from Qiagen pQE-2 and contains an inactive chloramphenicol resistance gene sequence.
Please note that Addgene's quality control sequence shows a small deletion in the vector region downstream of insert; the deletion does not affect any features in this construct.
Proper citation: RRID:Addgene_31580 Copy
Genetic Insert: lactose repressor protein
Vector Backbone Description: Backbone Size:4540; Vector Backbone:pEMBL9'; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:11266612
Comments: Parent plasmid was pEMBL9' with insert of extended LacI sequence from pIQ (Hare & Sadler, Gene 3 (1978) 269-278.
For expression: E. coli BLIM cells (or any E. coli WITHOUT lactose repressor in the genome or accompanying plasmid)
Proper citation: RRID:Addgene_31490 Copy
Species: T7 phage
Genetic Insert: RNA polymerase
Vector Backbone Description: Backbone Size:5400; Vector Backbone:pBAD33; Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:10610690
Comments: For expression: E. coli CV2
N823D mutation should not affect plasmid function.
Proper citation: RRID:Addgene_31491 Copy
Species: Gallus gallus
Genetic Insert: OVA
Vector Backbone Description: Backbone Marker:Reff et al., 1994; Backbone Size:9209; Vector Backbone:TCAE (modified); Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:9364701
Comments: *The backbone (TCAE) was modified to eliminate the immunoglobulin variable and constant regions, the neomycin resistance gene, and the DHFR gene. See paper for more details.
Proper citation: RRID:Addgene_31598 Copy
Species: Homo sapiens
Genetic Insert: Snail
Vector Backbone Description: Backbone Size:5598; Vector Backbone:pGL2_Basic; Vector Types:Mammalian Expression, Luciferase; Bacterial Resistance:Ampicillin
Defining Citation: PMID:12705869
Comments: Contains Snail promoter (-1045 to -56) sequence upstream of luciferase
Proper citation: RRID:Addgene_31694 Copy
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: 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: 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
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
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: 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
Species: Mus musculus
Genetic Insert: kinesin-1A
Vector Backbone Description: Backbone Marker:Clontech; Backbone Size:3940; Vector Backbone:pEGFP-C1 (EGFP removed); Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:19812246
Proper citation: RRID:Addgene_31607 Copy
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