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hal 390 myc tag fmc63 28 wpre3 sv40pa har  (Addgene inc)


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    Structured Review

    Addgene inc hal 390 myc tag fmc63 28 wpre3 sv40pa har
    Hal 390 Myc Tag Fmc63 28 Wpre3 Sv40pa Har, supplied by Addgene inc, used in various techniques. Bioz Stars score: 94/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/sv40pa/PBRM1+(Plasmid+%2339028)/10__1016_slash_j__omton__2026__201126-192-26-48
    Average 94 stars, based on 3 article reviews
    hal 390 myc tag fmc63 28 wpre3 sv40pa har - by Bioz Stars, 2026-10
    94/100 stars

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    Related Articles

    Bioprocessing:

    Article Title: Targeted long-read sequencing captures CRISPR editing and AAV integration outcomes in brain
    Article Snippet: For in vivo mouse studies, four different recombinant AAV (rAAV) vectors were generated by the Research Vector Core at the Raymond G. Perelman Center for Cellular and Molecular Therapeutics at The Children’s Hospital of Philadelphia. .. PX551 AAV shuttle plasmid expressed SpCas9 under the control of the neuronal-specific Mecp2 promoter and upstream of an SV40pA (PX551 was a gift from Feng Zhang; Addgene plasmid # 60957; http://n2t.net/addgene:60957; RRID:Addgene_60957).46 gRNA expression cassettes were moved into the G0619 AAV shuttle plasmid with eGFP gene under the control of the CMV promoter and upstream of an SV40pA signal (G0619 was from the University of Iowa Viral Vector Core). .. The non-targeting ctrl-gRNA sequence used was from the GeCKO v2 CRISPR screening library.47 All rAAV plasmid shuttles have AAV2 ITR sequences. rAAV vectors were produced by the standard calcium phosphate transfection method in HEK293 cells with the AdHelper plasmid, AAV1 Rep2/Cap1 packaging plasmid, and AAV shuttle plasmids with double CsCl purification.48 Vector titers were determined by ddPCR and were 1E13 vg/mL.

    Plasmid Preparation:

    Article Title: Targeted long-read sequencing captures CRISPR editing and AAV integration outcomes in brain
    Article Snippet: For in vivo mouse studies, four different recombinant AAV (rAAV) vectors were generated by the Research Vector Core at the Raymond G. Perelman Center for Cellular and Molecular Therapeutics at The Children’s Hospital of Philadelphia. .. PX551 AAV shuttle plasmid expressed SpCas9 under the control of the neuronal-specific Mecp2 promoter and upstream of an SV40pA (PX551 was a gift from Feng Zhang; Addgene plasmid # 60957; http://n2t.net/addgene:60957; RRID:Addgene_60957).46 gRNA expression cassettes were moved into the G0619 AAV shuttle plasmid with eGFP gene under the control of the CMV promoter and upstream of an SV40pA signal (G0619 was from the University of Iowa Viral Vector Core). .. The non-targeting ctrl-gRNA sequence used was from the GeCKO v2 CRISPR screening library.47 All rAAV plasmid shuttles have AAV2 ITR sequences. rAAV vectors were produced by the standard calcium phosphate transfection method in HEK293 cells with the AdHelper plasmid, AAV1 Rep2/Cap1 packaging plasmid, and AAV shuttle plasmids with double CsCl purification.48 Vector titers were determined by ddPCR and were 1E13 vg/mL.

    Article Title: Decoding RNA Metabolism by RNA-linked CRISPR Screening in Human Cells
    Article Snippet: The attP landing pad vector (pHPHS232) was created by using the plasmid backbone, AAVS1 homology arms, and cHS4 insulator from pASHS11 (pAAVS1P-iCAG.copGFP /Addgene 66577); the Tet-responsive promoter, attP , mTagBFP2, P2A, iCasp9, T2A, blasticidin S deaminase, and pCMV-rTTA from pHPHS111 (Addgene 200630); NeoR from pHPHS27 (mtk8b_LA_AAVS1_SA_neoR /Addgene 123742); and SV40pA from pHPHS5 (mtk4b_002_tSV40 / Addgene 123843). .. The attP* landing pad vector with Cas9 (pHPHS800) was created using the plasmid backbone from pYTK089 (Addgene 65196); the cHS4 insulator from pASHS11 (Addgene 66577); the EF1α promoter from pHPHS3 (MTK2_007_pEF1α /Addgene 123702); attP* encoded on oAS1848; attB encoded on oAS1482/oAS1540; SpCas9-NLS-FLAG from lentiCas9-Blast (Addgene 52962); T2A from pPBHS126 (pRRL U6-empty-gRNA-MND-Cas9-t2A-Blast ); Hygromycin phosphotransferase (HPH) from pHPHS7 (MTK6_009 CMV-Hygro-bgPA /Addgene 123863); and SV40pA from pHPHS5 (Addgene 123843). .. A base vector for reporter cloning (pHPHS806) was created using the plasmid backbone from pYTK089 (Addgene 65196); the cHS4 insulator, TRE3GV promoter, and T2A-PuroR from pASHS11 (pAAVS1P-iCAG.copGFP/Addgene 66577); EYFP-bGHpA from pPBHS285 ; attB* encoded on oAS1853/oAS1854; mCherry from pHPHS109 (Addgene 171598); and SV40pA from pHPHS5 (Addgene 123843).

    Article Title: Decoding RNA Metabolism by RNA-linked CRISPR Screening in Human Cells
    Article Snippet: The attP* landing pad vector with Cas9 (pHPHS800) was created using the plasmid backbone from pYTK089 (Addgene 65196); the cHS4 insulator from pASHS11 (Addgene 66577); the EF1α promoter from pHPHS3 (MTK2_007_pEF1α /Addgene 123702); attP* encoded on oAS1848; attB encoded on oAS1482/oAS1540; SpCas9-NLS-FLAG from lentiCas9-Blast (Addgene 52962); T2A from pPBHS126 (pRRL U6-empty-gRNA-MND-Cas9-t2A-Blast ); Hygromycin phosphotransferase (HPH) from pHPHS7 (MTK6_009 CMV-Hygro-bgPA /Addgene 123863); and SV40pA from pHPHS5 (Addgene 123843). .. A base vector for reporter cloning (pHPHS806) was created using the plasmid backbone from pYTK089 (Addgene 65196); the cHS4 insulator, TRE3GV promoter, and T2A-PuroR from pASHS11 (pAAVS1P-iCAG.copGFP/Addgene 66577); EYFP-bGHpA from pPBHS285 ; attB* encoded on oAS1853/oAS1854; mCherry from pHPHS109 (Addgene 171598); and SV40pA from pHPHS5 (Addgene 123843). ..

    Control:

    Article Title: Targeted long-read sequencing captures CRISPR editing and AAV integration outcomes in brain
    Article Snippet: For in vivo mouse studies, four different recombinant AAV (rAAV) vectors were generated by the Research Vector Core at the Raymond G. Perelman Center for Cellular and Molecular Therapeutics at The Children’s Hospital of Philadelphia. .. PX551 AAV shuttle plasmid expressed SpCas9 under the control of the neuronal-specific Mecp2 promoter and upstream of an SV40pA (PX551 was a gift from Feng Zhang; Addgene plasmid # 60957; http://n2t.net/addgene:60957; RRID:Addgene_60957).46 gRNA expression cassettes were moved into the G0619 AAV shuttle plasmid with eGFP gene under the control of the CMV promoter and upstream of an SV40pA signal (G0619 was from the University of Iowa Viral Vector Core). .. The non-targeting ctrl-gRNA sequence used was from the GeCKO v2 CRISPR screening library.47 All rAAV plasmid shuttles have AAV2 ITR sequences. rAAV vectors were produced by the standard calcium phosphate transfection method in HEK293 cells with the AdHelper plasmid, AAV1 Rep2/Cap1 packaging plasmid, and AAV shuttle plasmids with double CsCl purification.48 Vector titers were determined by ddPCR and were 1E13 vg/mL.

    Expressing:

    Article Title: Targeted long-read sequencing captures CRISPR editing and AAV integration outcomes in brain
    Article Snippet: For in vivo mouse studies, four different recombinant AAV (rAAV) vectors were generated by the Research Vector Core at the Raymond G. Perelman Center for Cellular and Molecular Therapeutics at The Children’s Hospital of Philadelphia. .. PX551 AAV shuttle plasmid expressed SpCas9 under the control of the neuronal-specific Mecp2 promoter and upstream of an SV40pA (PX551 was a gift from Feng Zhang; Addgene plasmid # 60957; http://n2t.net/addgene:60957; RRID:Addgene_60957).46 gRNA expression cassettes were moved into the G0619 AAV shuttle plasmid with eGFP gene under the control of the CMV promoter and upstream of an SV40pA signal (G0619 was from the University of Iowa Viral Vector Core). .. The non-targeting ctrl-gRNA sequence used was from the GeCKO v2 CRISPR screening library.47 All rAAV plasmid shuttles have AAV2 ITR sequences. rAAV vectors were produced by the standard calcium phosphate transfection method in HEK293 cells with the AdHelper plasmid, AAV1 Rep2/Cap1 packaging plasmid, and AAV shuttle plasmids with double CsCl purification.48 Vector titers were determined by ddPCR and were 1E13 vg/mL.

    Amplification:

    Article Title: Generation of primordial germ cell-like cells by two germ plasm components, dnd1 and nanos3 , in medaka ( Oryzias latipes )
    Article Snippet: .. SV40pA was amplified from the cyto-YFP-FKBPx5 vector (#103777; Addgene) and cloned into the BamHI and KpnI sites of pGGEV_2′_XcmI-LacZ (#49303; Addgene). .. Similarly, nanos3 and dnd1 3′ UTR were amplified from the cDNA of medaka ovaries and inserted into BamHI and KpnI sites of pGGEV_2′_XcmI-LacZ. nanos3 and dnd1 were inserted upstream of SV40pA or their 3′ UTR into the pGGDestSC-ATG vector (#49322; Addgene) via Golden GATE cloning to generate pGGDestSC- nanos3 - SV40pA , pGGDestSC- dnd1 - SV40pA, pGGDestSC- nanos3 - nanos3 3′ UTR , pGGDestSC- dnd1 - dnd1 3′ UTR ( nanos3 and dnd1 were derived from medaka) , pGGDestSC-Z nanos3 - SV40pA and pGGDestSC-Z dnd1 - SV40pA (nanos3 and dnd1 were derived from zebrafish) as previously described.

    Article Title: Generation of primordial germ cell-like cells by two germ plasm components, dnd1 and nanos3 , in medaka ( Oryzias latipes ).
    Article Snippet: They were cloned into the BamHI and KpnI sites of pGGEV1_XcmI-LacZ (gifted by Dr. JoachimWittbrodt, #49296; Addgene).62 Similarly, the protein-coding regions of zebrafish nanos3 and dnd1were amplified from the cDNA of zebrafish ovaries and cloned into the BamHI and KpnI of the pGGEV_–1_XcmI-LacZ vector. .. SV40pA was amplified from the cyto-YFP-FKBPx5 vector (#103777; Addgene)63 and cloned into the BamHI and KpnI sites of pGGEV_20_XcmI-LacZ (#49303; Addgene). .. Similarly, nanos3 and dnd1 30 UTR were amplified from the cDNA of medaka ovaries and inserted into BamHI and KpnI sites of pGGEV_20_XcmI-LacZ. nanos3 and dnd1 were inserted upstream of SV40pA or their 30 UTR into the pGGDestSC-ATG vector (#49322; Addgene) via Golden GATE cloning to generate pGGDestSC-nanos3-SV40pA, pGGDestSC-dnd1-SV40pA, pGGDestSC-nanos3-nanos3 30 UTR, pGGDestSCdnd1-dnd1 30 UTR (nanos3 and dnd1were derived frommedaka), pGGDestSC-Znanos3-SV40pA and pGGDestSC-Zdnd1-SV40pA (nanos3 and dnd1 were derived from zebrafish) as previously described.62 To protect CRISPR-Cas13d mediated knockdown, medaka dnd1 and nanos3 gRNA-targeted sites were mutated by mutagenesis PCR according to the protocol of the PrimeSTAR Mutagenesis Basal Kit (Figures S1A and S1B, Takara Bio, Japan).

    Clone Assay:

    Article Title: Generation of primordial germ cell-like cells by two germ plasm components, dnd1 and nanos3 , in medaka ( Oryzias latipes )
    Article Snippet: .. SV40pA was amplified from the cyto-YFP-FKBPx5 vector (#103777; Addgene) and cloned into the BamHI and KpnI sites of pGGEV_2′_XcmI-LacZ (#49303; Addgene). .. Similarly, nanos3 and dnd1 3′ UTR were amplified from the cDNA of medaka ovaries and inserted into BamHI and KpnI sites of pGGEV_2′_XcmI-LacZ. nanos3 and dnd1 were inserted upstream of SV40pA or their 3′ UTR into the pGGDestSC-ATG vector (#49322; Addgene) via Golden GATE cloning to generate pGGDestSC- nanos3 - SV40pA , pGGDestSC- dnd1 - SV40pA, pGGDestSC- nanos3 - nanos3 3′ UTR , pGGDestSC- dnd1 - dnd1 3′ UTR ( nanos3 and dnd1 were derived from medaka) , pGGDestSC-Z nanos3 - SV40pA and pGGDestSC-Z dnd1 - SV40pA (nanos3 and dnd1 were derived from zebrafish) as previously described.

    Article Title: Generation of primordial germ cell-like cells by two germ plasm components, dnd1 and nanos3 , in medaka ( Oryzias latipes ).
    Article Snippet: They were cloned into the BamHI and KpnI sites of pGGEV1_XcmI-LacZ (gifted by Dr. JoachimWittbrodt, #49296; Addgene).62 Similarly, the protein-coding regions of zebrafish nanos3 and dnd1were amplified from the cDNA of zebrafish ovaries and cloned into the BamHI and KpnI of the pGGEV_–1_XcmI-LacZ vector. .. SV40pA was amplified from the cyto-YFP-FKBPx5 vector (#103777; Addgene)63 and cloned into the BamHI and KpnI sites of pGGEV_20_XcmI-LacZ (#49303; Addgene). .. Similarly, nanos3 and dnd1 30 UTR were amplified from the cDNA of medaka ovaries and inserted into BamHI and KpnI sites of pGGEV_20_XcmI-LacZ. nanos3 and dnd1 were inserted upstream of SV40pA or their 30 UTR into the pGGDestSC-ATG vector (#49322; Addgene) via Golden GATE cloning to generate pGGDestSC-nanos3-SV40pA, pGGDestSC-dnd1-SV40pA, pGGDestSC-nanos3-nanos3 30 UTR, pGGDestSCdnd1-dnd1 30 UTR (nanos3 and dnd1were derived frommedaka), pGGDestSC-Znanos3-SV40pA and pGGDestSC-Zdnd1-SV40pA (nanos3 and dnd1 were derived from zebrafish) as previously described.62 To protect CRISPR-Cas13d mediated knockdown, medaka dnd1 and nanos3 gRNA-targeted sites were mutated by mutagenesis PCR according to the protocol of the PrimeSTAR Mutagenesis Basal Kit (Figures S1A and S1B, Takara Bio, Japan).

    Cloning:

    Article Title: Decoding RNA Metabolism by RNA-linked CRISPR Screening in Human Cells
    Article Snippet: The attP* landing pad vector with Cas9 (pHPHS800) was created using the plasmid backbone from pYTK089 (Addgene 65196); the cHS4 insulator from pASHS11 (Addgene 66577); the EF1α promoter from pHPHS3 (MTK2_007_pEF1α /Addgene 123702); attP* encoded on oAS1848; attB encoded on oAS1482/oAS1540; SpCas9-NLS-FLAG from lentiCas9-Blast (Addgene 52962); T2A from pPBHS126 (pRRL U6-empty-gRNA-MND-Cas9-t2A-Blast ); Hygromycin phosphotransferase (HPH) from pHPHS7 (MTK6_009 CMV-Hygro-bgPA /Addgene 123863); and SV40pA from pHPHS5 (Addgene 123843). .. A base vector for reporter cloning (pHPHS806) was created using the plasmid backbone from pYTK089 (Addgene 65196); the cHS4 insulator, TRE3GV promoter, and T2A-PuroR from pASHS11 (pAAVS1P-iCAG.copGFP/Addgene 66577); EYFP-bGHpA from pPBHS285 ; attB* encoded on oAS1853/oAS1854; mCherry from pHPHS109 (Addgene 171598); and SV40pA from pHPHS5 (Addgene 123843). ..



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    Image Search Results


    (a) Schematic of the bicistronic GPCR-RUSH integrated via the PiggyBac transposon system. (b, e) Flow cytometry histograms showing total receptor expression based on GFP fluorescence in DOR-RUSH (b) or MOR-RUSH (e) cells, compared with HeLa wild-type (wt) cells. (c, f) Flow cytometry histograms of surface DOR (c) or MOR (f) levels at indicated time points after biotin addition. Surface receptors were stained with AF647-conjugated GFP-Nb. HeLa wt cells stained in parallel depicted as control. (d) Kinetics of DOR and MOR PM arrival, expressed as fold change in surface receptor levels relative to the signal at 0 h. Data are mean ± SD of 3 independent experiments. (g) Confocal time-series of MOR-RUSH HeLa cells imaged at the indicated time points after biotin addition (80 μM). At 22h, different cells are shown. Images are single confocal slices. Scale bar, 10 μm. (h, i) Kinetics of DOR (h) and MOR (i) PM arrival of cells with different receptor expression levels. The AF647 signal at each time point was normalized to the signal at 22 h (set to 100%). Data are related to but include gating for the top 20% (high) or bottom 20% (low) expressing cells based on GFP. Half-times of export are indicated by dashed lines. Data are mean ± SD of 3 independent experiments. (j) Confocal time-series of MOR-DOR-RUSH HeLa cells at indicated time points after biotin addition (80 μM). At 22h, different cells are shown. Images are single confocal slices. Scale bar, 10 μm. (k) Kinetics of MOR-DOR PM arrival, compared to MOR and DOR transport. The AF647 signal at each time point was normalized to the signal at 22 h (set to 100%) for each receptor. Data are mean with ± SD of 3 independent experiments.

    Journal: bioRxiv

    Article Title: CRISPR screen identifies CNIH1 as a selective driver of GPCR export

    doi: 10.1101/2025.10.27.684930

    Figure Lengend Snippet: (a) Schematic of the bicistronic GPCR-RUSH integrated via the PiggyBac transposon system. (b, e) Flow cytometry histograms showing total receptor expression based on GFP fluorescence in DOR-RUSH (b) or MOR-RUSH (e) cells, compared with HeLa wild-type (wt) cells. (c, f) Flow cytometry histograms of surface DOR (c) or MOR (f) levels at indicated time points after biotin addition. Surface receptors were stained with AF647-conjugated GFP-Nb. HeLa wt cells stained in parallel depicted as control. (d) Kinetics of DOR and MOR PM arrival, expressed as fold change in surface receptor levels relative to the signal at 0 h. Data are mean ± SD of 3 independent experiments. (g) Confocal time-series of MOR-RUSH HeLa cells imaged at the indicated time points after biotin addition (80 μM). At 22h, different cells are shown. Images are single confocal slices. Scale bar, 10 μm. (h, i) Kinetics of DOR (h) and MOR (i) PM arrival of cells with different receptor expression levels. The AF647 signal at each time point was normalized to the signal at 22 h (set to 100%). Data are related to but include gating for the top 20% (high) or bottom 20% (low) expressing cells based on GFP. Half-times of export are indicated by dashed lines. Data are mean ± SD of 3 independent experiments. (j) Confocal time-series of MOR-DOR-RUSH HeLa cells at indicated time points after biotin addition (80 μM). At 22h, different cells are shown. Images are single confocal slices. Scale bar, 10 μm. (k) Kinetics of MOR-DOR PM arrival, compared to MOR and DOR transport. The AF647 signal at each time point was normalized to the signal at 22 h (set to 100%) for each receptor. Data are mean with ± SD of 3 independent experiments.

    Article Snippet: Transgenic HeLa cell lines stably co-expressing Str-KDEL and N-terminal IL-2 signal sequence (ss) SBP-EGFP-tagged GPCRs (human), including the MOR-DOR chimera (human), were generated by cloning a “chicken β-actin promoter, Str-KDEL_SBP-EGFP-GPCR, PGK promoter, WPRE, antibiotic resistance” cassette into a PiggyBac transposon plasmid (Addgene #84239) and cotransfecting 2.5 μg of the plasmid with 1 μg of PiggyBac transposase using 3 μL Lipofectamine 2000 in a 6-well plate.

    Techniques: Flow Cytometry, Expressing, Fluorescence, Staining, Control