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http://www.addgene.org/24368

Species: Neurospora crassa
Genetic Insert: QF
Vector Backbone Description: Backbone Size:7700; Vector Backbone:pattB-hsp70; Vector Types:Insect Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:20434990
Comments: A newer version of this plasmid is now available - pattB-hsp70P-QF#7-hsp70T www.addgene.org/46134 (Addgene plasmid # 46134)

Proper citation: RRID:Addgene_24368 Copy   

•   Source: Addgene
  

http://www.addgene.org/24361

Genetic Insert: mCD8-GFP
Vector Backbone Description: Backbone Size:9000; Vector Backbone:pQUAST; Vector Types:Insect Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:20434990
Comments: The FRT-Stop-FRT cassette from pUAS>Stop>mCD8-GFP was excised by BglII/NotI digestion. The mCD8-GFP cassette was excised from pQUAST-mCD8GFP by NotI digestion. The two inserts were ligated into the BglII/NotI sites of the pQUAST vector.

Proper citation: RRID:Addgene_24361 Copy   

•   Source: Addgene
  

http://www.addgene.org/24362

Species: Drosophila melanogaster
Genetic Insert: Shibire (ts1)
Vector Backbone Description: Backbone Size:9000; Vector Backbone:pQUAST; Vector Types:Insect Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:20434990
Comments: Shibire was PCR amplified from genomic DNA of UAS-shibire transgenic flies (Kitamoto, 2001) using genUASFOR and genUASREVsv40 oligos, and ligated into the NotI/KpnI sites of pQUAST.

Proper citation: RRID:Addgene_24362 Copy   

•   Source: Addgene
  

http://www.addgene.org/24363

Species: Drosophila melanogaster
Genetic Insert: Shibire (ts1)
Vector Backbone Description: Backbone Size:9000; Vector Backbone:pQUAS>stop>mCD8-GFP; Vector Types:Insect Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:20434990
Comments: The NotI fragment containing the mCD8-GFP cassette was excised from pQUAS>stop>mCD8- GFP, the vector was blunted, and ligated to a blunted NotI/KpnI shibirets1 isolated from pQUAST-shibirets1. ts1

Proper citation: RRID:Addgene_24363 Copy   

•   Source: Addgene
  

http://www.addgene.org/24364

Species: Neurospora crassa
Genetic Insert: QF
Vector Backbone Description: Backbone Size:8000; Vector Backbone:pDonor-M2; Vector Types:Insect Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:20434990
Comments: An enhancer trap was constructed that allows recombination mediated cassette exchange (Oberstein et al., 2005). A loxM2 site was inserted immediately following the 5P promoter, and a LoxP site was placed after the selectable white marker to generate the vector pDonor-M2, which contains elements in the following order: 5P- Ppromoter-LoxM2-MCS-SV40 terminator-white-LoxP-3P. QF was PCR amplified from pAC-QF adding 5’ EcoRI and 3’ AatII restriction sites, and insertedinto the EcoRI/AatII cloning sites of pDonor-M2 to yield pQF-M2ET.

Proper citation: RRID:Addgene_24364 Copy   

•   Source: Addgene
  

http://www.addgene.org/24418

Vector Backbone Description: Backbone Marker:Soriano Lab (Addgene Plasmid# 21714); Backbone Size:9846; Vector Backbone:pRosa26-1; Vector Types:Mouse Targeting; Bacterial Resistance:Chloramphenicol and Ampicillin
Defining Citation: PMID:21375737
Comments: Murine Rosa26 targeting vector with R1R2 Gateway sites. Includes a ccdB cassette, Rex-Neo selection marker cassette, and DTA selection cassette. Plasmid was constructed as follows: The pRosa26-1 plasmid (Addgene plasmid number 21714) was digested at the XbaI cloning site and a PCR product containing the attR1-ccdb-attR2 sequences from pDest27 and an additional PacI site was introduced. The Rex-Neo resistance cassette from the αMHC-eGFP-Rex-Neo lentivirus plasmid was PCR-cloned into the PacI site to allow for antibiotic selection in eukaryotic cells with G418

Proper citation: RRID:Addgene_24418 Copy   

•   Source: Addgene
  

http://www.addgene.org/24414

Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:2700; Vector Backbone:pENTR2B; Vector Types:Gateway Cloning; Bacterial Resistance:Kanamycin
Defining Citation: PMID:21375737
Comments: Contains Gateway L1L3 sites. PacI and Sbf1 can be used to insert promoter of interest to drive tTA expression. Contains insulator sequence. Plasmid was constructed as follows: The pEntr2B entry vector was digested with PstI and XhoI to remove the attL2 site. Oligonucleotides containing SalI-XhoI-Bsu36I-NdeI-PacI-EcoRI-NotI flanking the attL3 sequence on the 5' side and PstI on the 3' side were ligated in to create an intermediate plasmid carrying attL1 and attL3 sites (from pDest R4-R3. A LoxP sequence was introduced at the XhoI site, and the 500-bp chicken β-globin (CBG) HS4 insulator sequence was introduced at the Bsu36I/NdeI sites. The tTA and pA sequences were subsequently cloned into the EcoRI/NotI sites by PCR cloning from pUHG15-1 to generate pEntL1L3 tTA- 2

Proper citation: RRID:Addgene_24414 Copy   

•   Source: Addgene
  

http://www.addgene.org/24416

Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:2700; Vector Backbone:pENTR2B; Vector Types:Gateway Cloning; Bacterial Resistance:Kanamycin
Defining Citation: PMID:21375737
Comments: Contains Gateway R3L2 sites. Full length (7 repeat) TetO. SbfI, FseI, AvrII, and SalI sites can be used to insert gene of interest, driven by TetO. Contains insulator sequence Plasmid was constructed as follows: The pEntr2B entry vector was digested with AflII and XhoI to remove the attL1 and ccdb genes. Oligos containing the attR3 site flanked by AflII on the 5' end and a polylinker (NotI-PacI-Bsu36I-NdeI-XhoI) on the 3' end were ligated to generate the empty pEntR3L2-MCS intermediate (Addgene plasmid # 26799). Using PCR cloning, the TetO-beta globin intron sequence from pTetO (pUHD10.3) was cloned into the NotI/PacI sites in the reverse orientation, and a 3' SbfI site was introduced. The pA sequence from pDest27 was then cloned into the NotI/SbfI sites, again in reverse orientation. A LoxP site was introduced into the NdeI/XhoI sites, allowing full excision of the targeted construct when used together with the LoxP site in the pEntl1L3 vector. The CBG core insulator was then cloned into the Bsu36I/NdeI sites to generate the starting entry vector pEntR3L2 TetO(fl)-2.

Proper citation: RRID:Addgene_24416 Copy   

•   Source: Addgene
  

http://www.addgene.org/24379

Species: Mus musculus
Genetic Insert: Foxo1
Vector Backbone Description: Backbone Size:4968; Vector Backbone:pGEX-4T3; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin

Proper citation: RRID:Addgene_24379 Copy   

•   Source: Addgene
  

http://www.addgene.org/24372

Species: Homo sapiens
Genetic Insert: CLASP2 (340-1362)
Vector Backbone Description: Backbone Marker:Agilent Technologies; Backbone Size:7469; Vector Backbone:pShuttle-CMV; Vector Types:Mammalian Expression, Adenoviral; Bacterial Resistance:Kanamycin
Defining Citation: PMID:19289791
Comments: See the attached file for the old NIH reference sequence. Numbering starts at amino acid 78 and is off by 8 amino acids because of an insertion that is not accounted for.

Proper citation: RRID:Addgene_24372 Copy   

•   Source: Addgene
  

http://www.addgene.org/24370

Species: Neurospora crassa
Genetic Insert: QS
Vector Backbone Description: Backbone Size:2950; Vector Backbone:pBluescript SK (+); Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:20434990
Comments: QS ORF was PCR amplified from pAC-QS to include EcoRI/XbaI restriction sites.

Proper citation: RRID:Addgene_24370 Copy   

•   Source: Addgene
  

http://www.addgene.org/24371

Species: Homo sapiens
Genetic Insert: GSK3beta(S9A)
Vector Backbone Description: Backbone Marker:Agilent Technologies; Backbone Size:7469; Vector Backbone:pShuttle-CMV; Vector Types:Mammalian Expression, Adenoviral; Bacterial Resistance:Kanamycin
Defining Citation: PMID:19289791
Comments: Addgene's sequence results have identified a duplicated mRFP1 and GSK S9A in this plasmid. The depositor has confirmed that this duplication should not impact plasmid functionality.

Proper citation: RRID:Addgene_24371 Copy   

•   Source: Addgene
  

http://www.addgene.org/24343

Genetic Insert: Luciferase
Vector Backbone Description: Backbone Size:4283; Vector Backbone:pGL4.23; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:20434990
Comments: Obtained by subcloning 5 copies of GAL4-binding UAS sequence and the hsp70 minimal promoter from pUAS-GG into pGL4.23 using HindIII and NcoI.

Proper citation: RRID:Addgene_24343 Copy   

•   Source: Addgene
  

http://www.addgene.org/24346

Genetic Insert: GAL80
Vector Backbone Description: Backbone Size:6400; Vector Backbone:pPac5c-PL; Vector Types:Insect Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:20434990
Comments: GAL80 cDNA was PCR amplified using primers PR511 (CCCCGGATCCCAACatggactacaacaagagatcttcgg) and PR512 (CGGTTAACGCGGCCGCttataaactataatgcgagatattgctaacg), and a lab vector, pCA-GAL80 as the template. The PCR product was cloned using BamHI and NotI into pPAC5C-PL.

Proper citation: RRID:Addgene_24346 Copy   

•   Source: Addgene
  

http://www.addgene.org/24342

Vector Backbone Description: Backbone Size:3000; Vector Backbone:pBluescript SK(+); Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:20434990
Comments: The BamHI/BglI fragment from pUAST (Brand and Perrimon, 1993), containing 5 copies of a GAL4 binding site and the Drosophila hsp70 minimal promoter was subcloned into BamHI site of pBluescript containing an optimized GFP split with an intron (GG) (Zong et al., 2005). This strategy generates a hybrid BglII/BamHI site in front of the GFP.

Proper citation: RRID:Addgene_24342 Copy   

•   Source: Addgene
  

http://www.addgene.org/24358

Genetic Insert: FLPo
Vector Backbone Description: Backbone Size:9000; Vector Backbone:pQUAST; Vector Types:Insect Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:20434990
Comments: Mammalian codon-optimized FLP recombinase (FLPo; Addgene plasmid 13792) (Raymond and Soriano, 2007) was PCR amplified to include EcoRI/XbaI restriction sites, and cloned into pQUAS-DSCP to generate pQUAS-DSCP-FLPo.

Proper citation: RRID:Addgene_24358 Copy   

•   Source: Addgene
  

http://www.addgene.org/24356

Genetic Insert: LacZ
Vector Backbone Description: Backbone Size:9000; Vector Backbone:pQUAST; Vector Types:Insect Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:20434990
Comments: The nuclear LacZ insert was PCR amplified from UAS-nucLacZ genomic flies (Bloomington Stock Center). The KpnI/XbaI digested nucLacZ PCR fragment was then ligated into pQUAST.

Proper citation: RRID:Addgene_24356 Copy   

•   Source: Addgene
  

http://www.addgene.org/24353

Species: Neuropspora crassa
Genetic Insert: QF
Vector Backbone Description: Backbone Size:18000; Vector Backbone:pBAC-GH146; Vector Types:Insect Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:20434990
Comments: pBAC-3xPDsRed-GH146-MCS with FseI-AscI-AvrII multi-cloning site has been previously described (Hong et al., 2009). QF cDNA with 5’ FseI and 3’ Avr restriction sites was  amplified from pAC-QF by PCR, and cloned into the FseI/AvrII restriction sites of pBAC-GH146 to yield pBac-GH146-QF.

Proper citation: RRID:Addgene_24353 Copy   

•   Source: Addgene
  

http://www.addgene.org/24327

Species: Homo sapiens
Genetic Insert: Tubulin
Vector Backbone Description: Backbone Marker:Agilent Technologies; Backbone Size:7469; Vector Backbone:pShuttle-CMV; Vector Types:Mammalian Expression, Adenoviral; Bacterial Resistance:Kanamycin
Defining Citation: PMID:19289791

Proper citation: RRID:Addgene_24327 Copy   

•   Source: Addgene
  

http://www.addgene.org/24336

Species: Neurospora crassa/other
Genetic Insert: 5xQUAS/GFP split with an intron (GG)
Vector Backbone Description: Backbone Size:3000; Vector Backbone:pBluescript SK(+); Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:20434990
Comments: This plasmid contains 5 copies of naturally occurring QF binding sites (each 16 bp long, shown in capital letters, with spacer sequences in small letters): GGGTAATCGCTTATCCtcGGATAAACAATTATCCtcacGGGTAATCGCTTATCCgctcGGGTAATCG CTTATCCtcGGGTAATCGCTTATCCtt This sequence was assembled from overlapping primers and cloned using XhoI and HindIII into pBluescript containing hsp70 minimal promoter (Brand and Perrimon, 1993) and an optimized GFP split with an intron (GG) (Zong et al., 2005)

Proper citation: RRID:Addgene_24336 Copy   

•   Source: Addgene