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phage vector  (New England Biolabs)


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    Structured Review

    New England Biolabs phage vector
    Phage Vector, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 96/100, based on 395 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/phage+vector/BsrGI-HF/pm40739340-458-22-28
    Average 96 stars, based on 395 article reviews
    phage vector - by Bioz Stars, 2026-09
    96/100 stars

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    Related Articles

    Plasmid Preparation:

    Article Title: Retinal pigmented epithelium derived neurotrophic factor
    Article Snippet: Positive clones are isolated, and the DNA purified, with reagents supplied under the trade name “Qiagen Maxi” by Qiagen Inc. of Studio City, Calif. .. The inserts within the phage vector are excised with an appropriate restriction enzyme, circularized with T4 DNA ligase supplied by New England Biolabs of Beverly, Mass. and transformed into competent E. coli Sure cells supplied by Stratagene. ..

    Article Title: PEX39 facilitates the peroxisomal import of PTS2 proteins
    Article Snippet: Materials were obtained from the indicated sources: puromycin (InvivoGen ant-pr-1); doxycycline hyclate (Thermo Scientific Chemicals 446060050); antibodies to ACAA1 (HPA006764), AGPS (HPA030211), C6ORF226 (HPA045350), PEX5 (HPA039260), Hs PEX13 (HPA032142) were from Sigma-Aldrich; the antibody to ACTB (sc-69879) was from Santa Cruz Biotechnology; antibodies to PEX7 (20614-1-AP), PHYH (12858-1-AP), SCP2 (23006-1-AP) were from Proteintech; antibodies to CANX (2433), CS (14309), GAPDH (2118), HA (3724), Histone H3 (3638), and RPS6KB1 (2708), as well as HRP-conjugated anti-rabbit secondary antibody (7074) were from Cell Signaling Technology; HRP-conjugated anti-mouse secondary antibody for IP (ab131368) was from Abcam. .. pHAGE vectors encoding the following were obtained as indicated: C-terminally FLAG-HA-tagged human ACAA1, PHYH, PEX7, and wild-type Hs PEX39 were obtained from the human orfeome collection (version 8); C-terminally FLAG-HA-tagged human GAPDH and KPWE mutants of Hs PEX39 were generated by cloning gblocks (IDT) containing the corresponding cDNA into a pHAGE vector containing a C-terminal FLAG-HA tag [linearized with BsrGI-HF (NEB R3575)] via NEBuilder HiFi DNA assembly (NEB E2621); N-terminally FLAG-HA-tagged EGFP was a gift from Alban Ordureau; human GAPDH, wild-type and KPWE-mutated Hs PEX39 were generated by cloning gblocks containing the corresponding cDNA into a pHAGE vector (linearized with BsrGI-HF) via NEBuilder HiFi DNA assembly. .. An sgRNA against AAVS1 (sg AAVS1 ) and 3 independent sgRNAs against HsPEX39 (sg HsPEX39 _1-3) were cloned into a lentiCRISPR-v2 vector (TLCV2, doxycycline-inducible Cas9, Addgene 87360 ) linearized with BsmBI-v2 (NEB R0739) by ligation of annealed oligonucleotides (IDT) with Quick Ligase (NEB M2200).

    Article Title: Mathematical model to reduce loop mediated isothermal amplification (LAMP) false‐positive diagnosis
    Article Snippet: .. This is a double stranded phage vector that originated from bacteriophage M13 and was purchased from New England Biolabs (Ipswich, MA). ..

    Article Title: PEX39 facilitates the peroxisomal import of PTS2-containing proteins
    Article Snippet: .. Human GAPDH, wild-type Hs PEX39 and Hs PEX39 with a substituted KPWE motif were generated by cloning gblocks containing the corresponding cDNA into a pHAGE vector (linearized with BsrGI-HF) via NEBuilder HiFi DNA assembly. .. Three independent sgRNAs against HsPEX39 (sg HsPEX39 _1–3) and sgRNAs against PEX7 (sg PEX7 ) and AAVS1 (sg AAVS1 ) were cloned into a lentiCRISPR v2 vector (TLCV2; doxycycline-inducible Cas9; 87360; Addgene; ref. ) linearized with BsmBI-v2 (R0739; NEB) by ligation of annealed oligonucleotides (Integrated DNA Technologies) with Quick Ligase (M2200; NEB).

    Article Title: PEX39 facilitates the peroxisomal import of PTS2-containing proteins.
    Article Snippet: .. Human GAPDH, wild-type HsPEX39 and HsPEX39 with a substituted KPWE motif were generated by cloning gblocks containing the corresponding cDNA into a pHAGE vector (linearized with BsrGI-HF) via NEBuilder HiFi DNA assembly. .. Three independent sgRNAs against HsPEX39 (sgHsPEX39_1–3) and sgRNAs against PEX7 (sgPEX7) and AAVS1 (sgAAVS1) were cloned into a lentiCRISPR v2 vector (TLCV2; doxycycline-inducible Cas9; 87360; Addgene; ref. 93) linearized with BsmBI-v2 (R0739; NEB) by ligation of annealed oligonucleotides (Integrated DNA Technologies) with Quick Ligase (M2200; NEB).

    Article Title: Comprehensive assessment of anaplastic lymphoma kinase in localized and metastatic prostate cancer reveals targetable alterations
    Article Snippet: A.M. DeMarzo reports grants from Janssen R&D; personal fees from Merck, and personal fees from Cepheid outside the submitted work. .. The coding sequence representative of exons 16 to 29 of ALK (ALK16–29) was amplified by PCR using the full-length vector as a template and was cloned into the pHAGE vector after XhoI digestion using the HiFi DNA assembly kit (New England Biolabs) following manufacturer’s instructions. .. Nelson reports grants from Janssen; personal fees from Janssen, Bristol Myers Squibb, Veneble Fitzpatrick, and personal fees from Pfizer outside the submitted work.

    Transformation Assay:

    Article Title: Retinal pigmented epithelium derived neurotrophic factor
    Article Snippet: Positive clones are isolated, and the DNA purified, with reagents supplied under the trade name “Qiagen Maxi” by Qiagen Inc. of Studio City, Calif. .. The inserts within the phage vector are excised with an appropriate restriction enzyme, circularized with T4 DNA ligase supplied by New England Biolabs of Beverly, Mass. and transformed into competent E. coli Sure cells supplied by Stratagene. ..

    Generated:

    Article Title: PEX39 facilitates the peroxisomal import of PTS2 proteins
    Article Snippet: Materials were obtained from the indicated sources: puromycin (InvivoGen ant-pr-1); doxycycline hyclate (Thermo Scientific Chemicals 446060050); antibodies to ACAA1 (HPA006764), AGPS (HPA030211), C6ORF226 (HPA045350), PEX5 (HPA039260), Hs PEX13 (HPA032142) were from Sigma-Aldrich; the antibody to ACTB (sc-69879) was from Santa Cruz Biotechnology; antibodies to PEX7 (20614-1-AP), PHYH (12858-1-AP), SCP2 (23006-1-AP) were from Proteintech; antibodies to CANX (2433), CS (14309), GAPDH (2118), HA (3724), Histone H3 (3638), and RPS6KB1 (2708), as well as HRP-conjugated anti-rabbit secondary antibody (7074) were from Cell Signaling Technology; HRP-conjugated anti-mouse secondary antibody for IP (ab131368) was from Abcam. .. pHAGE vectors encoding the following were obtained as indicated: C-terminally FLAG-HA-tagged human ACAA1, PHYH, PEX7, and wild-type Hs PEX39 were obtained from the human orfeome collection (version 8); C-terminally FLAG-HA-tagged human GAPDH and KPWE mutants of Hs PEX39 were generated by cloning gblocks (IDT) containing the corresponding cDNA into a pHAGE vector containing a C-terminal FLAG-HA tag [linearized with BsrGI-HF (NEB R3575)] via NEBuilder HiFi DNA assembly (NEB E2621); N-terminally FLAG-HA-tagged EGFP was a gift from Alban Ordureau; human GAPDH, wild-type and KPWE-mutated Hs PEX39 were generated by cloning gblocks containing the corresponding cDNA into a pHAGE vector (linearized with BsrGI-HF) via NEBuilder HiFi DNA assembly. .. An sgRNA against AAVS1 (sg AAVS1 ) and 3 independent sgRNAs against HsPEX39 (sg HsPEX39 _1-3) were cloned into a lentiCRISPR-v2 vector (TLCV2, doxycycline-inducible Cas9, Addgene 87360 ) linearized with BsmBI-v2 (NEB R0739) by ligation of annealed oligonucleotides (IDT) with Quick Ligase (NEB M2200).

    Article Title: PEX39 facilitates the peroxisomal import of PTS2-containing proteins
    Article Snippet: .. Human GAPDH, wild-type Hs PEX39 and Hs PEX39 with a substituted KPWE motif were generated by cloning gblocks containing the corresponding cDNA into a pHAGE vector (linearized with BsrGI-HF) via NEBuilder HiFi DNA assembly. .. Three independent sgRNAs against HsPEX39 (sg HsPEX39 _1–3) and sgRNAs against PEX7 (sg PEX7 ) and AAVS1 (sg AAVS1 ) were cloned into a lentiCRISPR v2 vector (TLCV2; doxycycline-inducible Cas9; 87360; Addgene; ref. ) linearized with BsmBI-v2 (R0739; NEB) by ligation of annealed oligonucleotides (Integrated DNA Technologies) with Quick Ligase (M2200; NEB).

    Article Title: PEX39 facilitates the peroxisomal import of PTS2-containing proteins.
    Article Snippet: .. Human GAPDH, wild-type HsPEX39 and HsPEX39 with a substituted KPWE motif were generated by cloning gblocks containing the corresponding cDNA into a pHAGE vector (linearized with BsrGI-HF) via NEBuilder HiFi DNA assembly. .. Three independent sgRNAs against HsPEX39 (sgHsPEX39_1–3) and sgRNAs against PEX7 (sgPEX7) and AAVS1 (sgAAVS1) were cloned into a lentiCRISPR v2 vector (TLCV2; doxycycline-inducible Cas9; 87360; Addgene; ref. 93) linearized with BsmBI-v2 (R0739; NEB) by ligation of annealed oligonucleotides (Integrated DNA Technologies) with Quick Ligase (M2200; NEB).

    Cloning:

    Article Title: PEX39 facilitates the peroxisomal import of PTS2 proteins
    Article Snippet: Materials were obtained from the indicated sources: puromycin (InvivoGen ant-pr-1); doxycycline hyclate (Thermo Scientific Chemicals 446060050); antibodies to ACAA1 (HPA006764), AGPS (HPA030211), C6ORF226 (HPA045350), PEX5 (HPA039260), Hs PEX13 (HPA032142) were from Sigma-Aldrich; the antibody to ACTB (sc-69879) was from Santa Cruz Biotechnology; antibodies to PEX7 (20614-1-AP), PHYH (12858-1-AP), SCP2 (23006-1-AP) were from Proteintech; antibodies to CANX (2433), CS (14309), GAPDH (2118), HA (3724), Histone H3 (3638), and RPS6KB1 (2708), as well as HRP-conjugated anti-rabbit secondary antibody (7074) were from Cell Signaling Technology; HRP-conjugated anti-mouse secondary antibody for IP (ab131368) was from Abcam. .. pHAGE vectors encoding the following were obtained as indicated: C-terminally FLAG-HA-tagged human ACAA1, PHYH, PEX7, and wild-type Hs PEX39 were obtained from the human orfeome collection (version 8); C-terminally FLAG-HA-tagged human GAPDH and KPWE mutants of Hs PEX39 were generated by cloning gblocks (IDT) containing the corresponding cDNA into a pHAGE vector containing a C-terminal FLAG-HA tag [linearized with BsrGI-HF (NEB R3575)] via NEBuilder HiFi DNA assembly (NEB E2621); N-terminally FLAG-HA-tagged EGFP was a gift from Alban Ordureau; human GAPDH, wild-type and KPWE-mutated Hs PEX39 were generated by cloning gblocks containing the corresponding cDNA into a pHAGE vector (linearized with BsrGI-HF) via NEBuilder HiFi DNA assembly. .. An sgRNA against AAVS1 (sg AAVS1 ) and 3 independent sgRNAs against HsPEX39 (sg HsPEX39 _1-3) were cloned into a lentiCRISPR-v2 vector (TLCV2, doxycycline-inducible Cas9, Addgene 87360 ) linearized with BsmBI-v2 (NEB R0739) by ligation of annealed oligonucleotides (IDT) with Quick Ligase (NEB M2200).

    Article Title: PEX39 facilitates the peroxisomal import of PTS2-containing proteins
    Article Snippet: .. Human GAPDH, wild-type Hs PEX39 and Hs PEX39 with a substituted KPWE motif were generated by cloning gblocks containing the corresponding cDNA into a pHAGE vector (linearized with BsrGI-HF) via NEBuilder HiFi DNA assembly. .. Three independent sgRNAs against HsPEX39 (sg HsPEX39 _1–3) and sgRNAs against PEX7 (sg PEX7 ) and AAVS1 (sg AAVS1 ) were cloned into a lentiCRISPR v2 vector (TLCV2; doxycycline-inducible Cas9; 87360; Addgene; ref. ) linearized with BsmBI-v2 (R0739; NEB) by ligation of annealed oligonucleotides (Integrated DNA Technologies) with Quick Ligase (M2200; NEB).

    Article Title: PEX39 facilitates the peroxisomal import of PTS2-containing proteins.
    Article Snippet: .. Human GAPDH, wild-type HsPEX39 and HsPEX39 with a substituted KPWE motif were generated by cloning gblocks containing the corresponding cDNA into a pHAGE vector (linearized with BsrGI-HF) via NEBuilder HiFi DNA assembly. .. Three independent sgRNAs against HsPEX39 (sgHsPEX39_1–3) and sgRNAs against PEX7 (sgPEX7) and AAVS1 (sgAAVS1) were cloned into a lentiCRISPR v2 vector (TLCV2; doxycycline-inducible Cas9; 87360; Addgene; ref. 93) linearized with BsmBI-v2 (R0739; NEB) by ligation of annealed oligonucleotides (Integrated DNA Technologies) with Quick Ligase (M2200; NEB).

    Sequencing:

    Article Title: Comprehensive assessment of anaplastic lymphoma kinase in localized and metastatic prostate cancer reveals targetable alterations
    Article Snippet: ALK cloning and lentivirus production Lentiviral full length ALK and the corresponding control pHAGE vectors were purchased from Addgene (Addgene IDs: 116712 and 118692). .. The coding sequence representative of exons 16 to 29 of ALK (ALK16–29) was amplified by PCR using the full-length vector as a template and was cloned into the pHAGE vector after XhoI digestion using the HiFi DNA assembly kit (New England Biolabs) following manufacturer’s instructions. ..

    Amplification:

    Article Title: Comprehensive assessment of anaplastic lymphoma kinase in localized and metastatic prostate cancer reveals targetable alterations
    Article Snippet: ALK cloning and lentivirus production Lentiviral full length ALK and the corresponding control pHAGE vectors were purchased from Addgene (Addgene IDs: 116712 and 118692). .. The coding sequence representative of exons 16 to 29 of ALK (ALK16–29) was amplified by PCR using the full-length vector as a template and was cloned into the pHAGE vector after XhoI digestion using the HiFi DNA assembly kit (New England Biolabs) following manufacturer’s instructions. ..

    Polymerase Chain Reaction:

    Article Title: Comprehensive assessment of anaplastic lymphoma kinase in localized and metastatic prostate cancer reveals targetable alterations
    Article Snippet: ALK cloning and lentivirus production Lentiviral full length ALK and the corresponding control pHAGE vectors were purchased from Addgene (Addgene IDs: 116712 and 118692). .. The coding sequence representative of exons 16 to 29 of ALK (ALK16–29) was amplified by PCR using the full-length vector as a template and was cloned into the pHAGE vector after XhoI digestion using the HiFi DNA assembly kit (New England Biolabs) following manufacturer’s instructions. ..

    Clone Assay:

    Article Title: Comprehensive assessment of anaplastic lymphoma kinase in localized and metastatic prostate cancer reveals targetable alterations
    Article Snippet: ALK cloning and lentivirus production Lentiviral full length ALK and the corresponding control pHAGE vectors were purchased from Addgene (Addgene IDs: 116712 and 118692). .. The coding sequence representative of exons 16 to 29 of ALK (ALK16–29) was amplified by PCR using the full-length vector as a template and was cloned into the pHAGE vector after XhoI digestion using the HiFi DNA assembly kit (New England Biolabs) following manufacturer’s instructions. ..



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    Image Search Results


    ( A ) Chemical structure of CTX-439. ( B ) Kinome tree illustrating the selectivity profile of CTX-439. Binding affinity of CTX-439 was screened by scanMAX kinase panel and the Kd values for CDKs and the other top 6 kinases were evaluated. For CASK, DCAMKL3, DYRK2, MAP3K15, JNK3, JNK1, RSK4, ERK8, ERN1, RSK3 and CLK1, CTX-439 showed weak inhibitory effects of 50-65% at 30 nM, but it is not included in the kinome tree. ( C ) Inhibitory activities of CTX-439, THZ531 and SR4835 on transcription-related CDK kinases. ( D ) Western blot analysis of CDK12 and Pol II CTD phosphorylation in SUM149PT cells treated with the indicated concentration of CTX-439 for 6 h. ( E ) RT-qPCR analyses of BRCA1 and BRCA2 in SUM149PT cells treated with CTX-439 for 6h. ( F ) Western blot analysis of DDR proteins and apoptosis markers in SUM149PT cells treated with CTX-439 for 24h. ( G ) Drug response assay in SUM149PT cells treated with CTX-439 for 72h. ( H ) in vivo anti-tumor efficacy of CTX-439 in mice bearing SUM149PT xenografts treated with the indicated dose twice weekly (biw). ( I ) in vivo anti-tumor efficacy of CTX-439 in TNBC PDX model mice treated with the indicated dose. * p <0.05, ** p <0.01 ** p <0.001 by Dunnett’s test. Data are shown as mean ± SD (n=3, E ; n=6, H and I ).

    Journal: bioRxiv

    Article Title: CDK12/13 inhibitor, CTX-439, suppresses tumor growth and potentiates BCL-2 family blockade

    doi: 10.64898/2026.02.20.706902

    Figure Lengend Snippet: ( A ) Chemical structure of CTX-439. ( B ) Kinome tree illustrating the selectivity profile of CTX-439. Binding affinity of CTX-439 was screened by scanMAX kinase panel and the Kd values for CDKs and the other top 6 kinases were evaluated. For CASK, DCAMKL3, DYRK2, MAP3K15, JNK3, JNK1, RSK4, ERK8, ERN1, RSK3 and CLK1, CTX-439 showed weak inhibitory effects of 50-65% at 30 nM, but it is not included in the kinome tree. ( C ) Inhibitory activities of CTX-439, THZ531 and SR4835 on transcription-related CDK kinases. ( D ) Western blot analysis of CDK12 and Pol II CTD phosphorylation in SUM149PT cells treated with the indicated concentration of CTX-439 for 6 h. ( E ) RT-qPCR analyses of BRCA1 and BRCA2 in SUM149PT cells treated with CTX-439 for 6h. ( F ) Western blot analysis of DDR proteins and apoptosis markers in SUM149PT cells treated with CTX-439 for 24h. ( G ) Drug response assay in SUM149PT cells treated with CTX-439 for 72h. ( H ) in vivo anti-tumor efficacy of CTX-439 in mice bearing SUM149PT xenografts treated with the indicated dose twice weekly (biw). ( I ) in vivo anti-tumor efficacy of CTX-439 in TNBC PDX model mice treated with the indicated dose. * p <0.05, ** p <0.01 ** p <0.001 by Dunnett’s test. Data are shown as mean ± SD (n=3, E ; n=6, H and I ).

    Article Snippet: To construct HA-tagged CDK12 expression vectors, CDK12 cDNA fragment was first generated by PCR using pHAGE-CDK12 (Addgene #116723) as a template.

    Techniques: Binding Assay, Western Blot, Phospho-proteomics, Concentration Assay, Quantitative RT-PCR, In Vivo

    ( A ) Western blot analysis of cells expressing the indicated cDNA and sgRNA. mtCDK12 carries the combined mutations of TA770FL, D817N and E1041AG. ( B ) Western blot analysis showing that pS2 was maintained in mtCDK12-expressing cells under the treatment of CTX-439. ( C ) Cell survival assay. Cells with endogenous, or exogenous wt or mt CDK12 were tested. ( D ) Western blot analysis. Cells with endogenous (E), or exogenous wt or mt CDK12 were treated with the indicated drug(s) for 8h. Note that MCL1 downregulation was suppressed by mt CDK12. ( E ) RT-qPCR analysis of MCL1 expression in the nucleus. Note that nuclear MCL1 transcript upregulation was suppressed by mt CDK12. ( F-G ) RT-qPCR analyses of MCL1 ( F ) and TUBB4B readthrough ( G ) in the nucleus. Readthrough was detected in mt CDK12-expressing cells, but their level is far below than that in cells with endogenous or exogenous wt CDK12. Data are shown as mean ± SD (n=3, E-G ). * p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001 by Student’s t- test.

    Journal: bioRxiv

    Article Title: CDK12/13 inhibitor, CTX-439, suppresses tumor growth and potentiates BCL-2 family blockade

    doi: 10.64898/2026.02.20.706902

    Figure Lengend Snippet: ( A ) Western blot analysis of cells expressing the indicated cDNA and sgRNA. mtCDK12 carries the combined mutations of TA770FL, D817N and E1041AG. ( B ) Western blot analysis showing that pS2 was maintained in mtCDK12-expressing cells under the treatment of CTX-439. ( C ) Cell survival assay. Cells with endogenous, or exogenous wt or mt CDK12 were tested. ( D ) Western blot analysis. Cells with endogenous (E), or exogenous wt or mt CDK12 were treated with the indicated drug(s) for 8h. Note that MCL1 downregulation was suppressed by mt CDK12. ( E ) RT-qPCR analysis of MCL1 expression in the nucleus. Note that nuclear MCL1 transcript upregulation was suppressed by mt CDK12. ( F-G ) RT-qPCR analyses of MCL1 ( F ) and TUBB4B readthrough ( G ) in the nucleus. Readthrough was detected in mt CDK12-expressing cells, but their level is far below than that in cells with endogenous or exogenous wt CDK12. Data are shown as mean ± SD (n=3, E-G ). * p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001 by Student’s t- test.

    Article Snippet: To construct HA-tagged CDK12 expression vectors, CDK12 cDNA fragment was first generated by PCR using pHAGE-CDK12 (Addgene #116723) as a template.

    Techniques: Western Blot, Expressing, Clonogenic Cell Survival Assay, Quantitative RT-PCR