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Vector Backbone Description: Backbone Marker:Novagen; Backbone Size:7266; Vector Backbone:pET, pCoofy3; 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_55186 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
Vector Backbone Description: Backbone Marker:Mullins, E.D., Kang, S., 2001. Transformation: a tool for studying fungal pathogens of plants. Cell. Mol. Life Sci. 58, 2043–205; Backbone Size:7483; Vector Backbone:pDHt; Vector Types:fungal expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:15893252
Proper citation: RRID:Addgene_63615 Copy
Species: T7 bacteriophage
Genetic Insert: CTGA-R13-AKSRV RNAP
Vector Backbone Description: Vector Backbone:pQE; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:25279711
Proper citation: RRID:Addgene_63622 Copy
Species: T7 bacteriophage
Genetic Insert: T3 RRVH RNAP
Vector Backbone Description: Vector Backbone:pQE; Vector Types:; Bacterial Resistance:Kanamycin
Defining Citation: PMID:25279711
Proper citation: RRID:Addgene_63623 Copy
Species: T7 bacteriophage
Genetic Insert: CGG-R12-KIRV RNAP
Vector Backbone Description: Vector Backbone:pQE; Vector Types:; Bacterial Resistance:Kanamycin
Defining Citation: PMID:25279711
Proper citation: RRID:Addgene_63668 Copy
Species: Homo sapiens
Genetic Insert: XBP1
Vector Backbone Description: Vector Backbone:pShuttle2-Flag; Vector Types:Mammalian Expression, Adenoviral; Bacterial Resistance:Kanamycin
Defining Citation: PMID:19416856
Proper citation: RRID:Addgene_63679 Copy
Vector Backbone Description: Backbone Marker:Novagen; Backbone Size:5731; Vector Backbone:pET28a; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Proper citation: RRID:Addgene_63723 Copy
Vector Backbone Description: Vector Backbone:pShuttle-CMV; Vector Types:Mammalian Expression, Adenoviral; Bacterial Resistance:Kanamycin
Proper citation: RRID:Addgene_63681 Copy
Vector Backbone Description: Backbone Size:9231; Vector Backbone:pSN40; Vector Types:C. albicans bait plasmid; Bacterial Resistance:Kanamycin
Defining Citation: PMID:20719741
Comments: Note--the ccdB gene contained is inactive as it is missing the start codon so no ccd survival strain is necessary.
Proper citation: RRID:Addgene_63734 Copy
Species: T7 bacteriophage
Genetic Insert: K1F IKR RNAP
Vector Backbone Description: Vector Backbone:pQE; Vector Types:; Bacterial Resistance:Kanamycin
Defining Citation: PMID:25279711
Proper citation: RRID:Addgene_63626 Copy
Species: T7 bacteriophage
Genetic Insert: CTGA-R13-AKSIRV RNAP
Vector Backbone Description: Vector Backbone:pQE; Vector Types:; Bacterial Resistance:Kanamycin
Defining Citation: PMID:25279711
Proper citation: RRID:Addgene_63627 Copy
Species: Saccharomyces cerevisiae
Genetic Insert: Dbp9
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:4762; Vector Backbone:pDONR221; Vector Types:Gateway Cloning; Bacterial Resistance:Kanamycin
Defining Citation: PMID:25877921
Proper citation: RRID:Addgene_63743 Copy
Species: Saccharomyces cerevisiae
Genetic Insert: Drs1
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:4762; Vector Backbone:pDONR221; Vector Types:Gateway Cloning; Bacterial Resistance:Kanamycin
Defining Citation: PMID:25877921
Proper citation: RRID:Addgene_63744 Copy
Species: Saccharomyces cerevisiae
Genetic Insert: Nsr1
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:4762; Vector Backbone:pDONR221; Vector Types:Gateway Cloning; Bacterial Resistance:Kanamycin
Defining Citation: PMID:25877921
Proper citation: RRID:Addgene_63769 Copy
Species: Saccharomyces cerevisiae
Genetic Insert: Nog2
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:4762; Vector Backbone:pDONR221; Vector Types:Gateway Cloning; Bacterial Resistance:Kanamycin
Defining Citation: PMID:25877921
Proper citation: RRID:Addgene_63764 Copy
Species: Saccharomyces cerevisiae
Genetic Insert: Nop2
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:4762; Vector Backbone:pDONR221; Vector Types:Gateway Cloning; Bacterial Resistance:Kanamycin
Defining Citation: PMID:25877921
Proper citation: RRID:Addgene_63766 Copy
Species: Saccharomyces cerevisiae
Genetic Insert: Mak16
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:4762; Vector Backbone:pDONR221; Vector Types:Gateway Cloning; Bacterial Resistance:Kanamycin
Defining Citation: PMID:25877921
Proper citation: RRID:Addgene_63762 Copy
Species: Saccharomyces cerevisiae
Genetic Insert: Tma16
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:4762; Vector Backbone:pDONR221; Vector Types:Gateway Cloning; Bacterial Resistance:Kanamycin
Defining Citation: PMID:25877921
Proper citation: RRID:Addgene_63779 Copy
Species: Saccharomyces cerevisiae
Genetic Insert: Rrp1
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:4762; Vector Backbone:pDONR221; Vector Types:Gateway Cloning; Bacterial Resistance:Kanamycin
Defining Citation: PMID:25877921
Proper citation: RRID:Addgene_63773 Copy
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