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Species: Synthetic
Genetic Insert: dCas9-p65HSF1
Vector Backbone Description: Vector Backbone:pAC90-pmax-DEST; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:26768771
Comments: Please visit http://casil.io for more information, updates, and protocols. For more information on Cheng Lab CRISPR Plasmids please refer to: http://www.addgene.org/crispr/albert-cheng/
Proper citation: RRID:Addgene_71907 Copy
Species: Synthetic
Genetic Insert: PUFc-p65HSF1
Vector Backbone Description: Vector Backbone:pAC90-pmax-DEST; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:26768771
Comments: Please visit http://casil.io for more information, updates, and protocols. For more information on Cheng Lab CRISPR Plasmids please refer to: http://www.addgene.org/crispr/albert-cheng/
Proper citation: RRID:Addgene_71908 Copy
Species: Synthetic
Genetic Insert: NLSPUFa_VP64
Vector Backbone Description: Vector Backbone:pAC90-pmax-DEST; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:26768771
Comments: Please visit http://casil.io for more information, updates, and protocols. For more information on Cheng Lab CRISPR Plasmids please refer to: http://www.addgene.org/crispr/albert-cheng/
Proper citation: RRID:Addgene_71881 Copy
Species: Synthetic
Genetic Insert: dCas9-mCBPHAT
Vector Backbone Description: Vector Backbone:pAC90-pmax-DEST; Vector Types:Mammalian Expression, CRISPR; Bacterial Resistance:Kanamycin
Defining Citation: PMID:26768771
Comments: Please visit http://casil.io for more information, updates, and protocols. For more information on Cheng Lab CRISPR Plasmids please refer to: http://www.addgene.org/crispr/albert-cheng/
Proper citation: RRID:Addgene_71887 Copy
Species: Synthetic
Genetic Insert: dCas9-VP64
Vector Backbone Description: Vector Backbone:pAC90-pmax-DEST; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:26768771
Comments: Please visit http://casil.io for more information, updates, and protocols. For more information on Cheng Lab CRISPR Plasmids please refer to: http://www.addgene.org/crispr/albert-cheng/
Proper citation: RRID:Addgene_71879 Copy
Species: Synthetic
Genetic Insert: dCas9
Vector Backbone Description: Vector Backbone:pAC90-pmax-DEST; Vector Types:Mammalian Expression, CRISPR; Bacterial Resistance:Kanamycin
Defining Citation: PMID:26768771
Comments: Please visit http://casil.io for more information, updates and protocols
Proper citation: RRID:Addgene_73169 Copy
Species: Synthetic
Genetic Insert: Clover_PUFc
Vector Backbone Description: Vector Backbone:pAC90-pmax-DEST; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:26768771
Comments: Please visit http://casil.io for more information, updates and protocols
Proper citation: RRID:Addgene_73689 Copy
Species: Synthetic
Genetic Insert: Clover_PUFa
Vector Backbone Description: Vector Backbone:pAC90-pmax-DEST; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:26768771
Comments: Please visit http://casil.io for more information, updates and protocols
Proper citation: RRID:Addgene_73688 Copy
Species: Synthetic
Genetic Insert: mRuby2_PUFa
Vector Backbone Description: Vector Backbone:pAC90-pmax-DEST; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:26768771
Comments: Please visit http://casil.io for more information, updates and protocols
Proper citation: RRID:Addgene_73690 Copy
Species: Synthetic
Genetic Insert: PUFb_p65HSF1
Vector Backbone Description: Vector Backbone:pAC90-pmax-DEST; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:26768771
Comments: Please visit http://casil.io for more information, updates and protocols
Proper citation: RRID:Addgene_73697 Copy
Species: Synthetic
Genetic Insert: None
Vector Backbone Description: Vector Backbone:DVK; Vector Types:Synthetic Biology, Other, Destination Vector; Bacterial Resistance:Kanamycin
Defining Citation: PMID:26479688
Comments: parts.igem.org/Part:pSB1K3
Proper citation: RRID:Addgene_66069 Copy
Species: Synthetic
Genetic Insert: None
Vector Backbone Description: Vector Backbone:DVK; Vector Types:Synthetic Biology, Other, Destination Vector; Bacterial Resistance:Kanamycin
Defining Citation: PMID:26479688
Comments: parts.igem.org/Part:pSB1K3
Proper citation: RRID:Addgene_66067 Copy
Species: Synthetic
Genetic Insert: None
Vector Backbone Description: Vector Backbone:DVK; Vector Types:Gateway Cloning, Synthetic Biology; Bacterial Resistance:Kanamycin
Defining Citation: PMID:26479688
Comments: parts.igem.org/Part:pSB1K3
Proper citation: RRID:Addgene_66071 Copy
Genetic Insert: 2xtRNA-SupD
Vector Backbone Description: Vector Backbone:pKD; Vector Types:Bacterial Expression, Synthetic Biology; Bacterial Resistance:Kanamycin
Defining Citation: PMID:26350500
Comments: See supplementary information accompanying article for additional cloning information. We recommend introducing this plasmid into a recoded strain of E. coli (no genomic TAG codons) for high-efficiency phoshoprotein expression.
Proper citation: RRID:Addgene_68307 Copy
Species: Homo sapiens
Genetic Insert: Zyxin
Vector Backbone Description: Backbone Size:4750; Vector Backbone:mRFP1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Comments: . Excitation = 584; Emission = 607
Proper citation: RRID:Addgene_55925 Copy
Species: Homo sapiens
Genetic Insert: H2B
Vector Backbone Description: Backbone Size:4750; Vector Backbone:rsEGFP2; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Proper citation: RRID:Addgene_56533 Copy
Species: Homo sapiens
Genetic Insert: NM_003380.3
Vector Backbone Description: Backbone Size:4750; Vector Backbone:mTagRFP; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:18454154
Comments: . Excitation = 555; Emission = 584
Proper citation: RRID:Addgene_58029 Copy
Genetic Insert: DV2 prM-E
Vector Backbone Description: Vector Backbone:pVRC8400; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:29999491
Comments: Please visit https://www.biorxiv.org/content/early/2018/03/22/286955 for bioRxiv preprint.
Proper citation: RRID:Addgene_63161 Copy
Species: Homo sapiens
Genetic Insert: Desmoglein1
Vector Backbone Description: Backbone Size:4750; Vector Backbone:mApple-C1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Comments: . Excitation = 568; Emission = 592
Proper citation: RRID:Addgene_54887 Copy
Vector Backbone Description: Backbone Marker:Novagen; Backbone Size:7701; Vector Backbone:pET, pCoofy4; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:23410102
Comments: C-terminal tags are separated by a stop codon. Depending on the LP2 primer used in SLIC, the desired C-terminal tag(s) can be fused to the target gene.
Use the one of the following linearization primers pairs for SLIC cloning. Choose one LP1 forward vector primer and one LP2 reverse vector primer:
LP1 forward vector primer:
SLIC Primer (3C site) 5' GGGCCCCTGGAACAGAACTTCCAG 3'
LP2 - select an LP2 primer below depending on which C-terminal tags you want to include fused to your gene of interest:
none SLIC Primer 5' CGCCATTAACCTGATGTTCTGGGG 3'
10His- SLIC Primer 5`GAGCATCATCATCATCACCAC 3'
StrepOne - SLIC Primer 5`AGCGCTTGGAGCCACCCGCAG 3'
S-Tag - SLIC Primer 5`AAAGAAACCGCTGCTGCTAAATTCG 3'
CBP - SLIC Primer 5`ATGAAGCGGCGGTGGAAGAAAAAC 3'
HPC4 - SLIC Primer 5`GAGGACCAGGTGGACCCCCGG 3'
Æ54CPD - SLIC Primer 5`GGCAGCGGCAAGATCCTGCAC 3'
Test each preparation of the plasmid by transforming it into non-resistant cells (ex. DH5a) to ensure negative selection by ccdB kills all the transformed cells as expected.
Proper citation: RRID:Addgene_55187 Copy
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