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Pentr1a

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PENTR1A is a plasmid vector used in molecular biology for the Gateway cloning system. It serves as a donor vector for the recombination-based transfer of DNA sequences into compatible destination vectors. The core function of PENTR1A is to provide a standardized entry clone format for further subcloning and expression experiments, but no further details on its intended use are provided.

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Lab products found in correlation

Lentiguide puro, by Addgene (1 mentions) Pentr1a, by Thermo Fisher Scientific (1 mentions) Pinducer20, by Addgene (1 mentions) Gateway lr clonase enzyme, by Thermo Fisher Scientific (1 mentions) Plenti cmv puro dest, by Addgene (1 mentions) Genejet plasmid maxiprep kit, by Thermo Fisher Scientific (1 mentions) Migr1, by Addgene (1 mentions)

Spelling variants of this product

PENTR1A vector (3 citations) PENTR1A no CCDB (w48-1) vector (2 citations) PENTR1A no ccDB (2 citations) PENTR1A-GFP-N2 (2 citations)

3 protocols using pentr1a

1

Plasmid Constructs for Protein Interactions

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For YFPN–COUP-TFII and SMAD4-YFPC, COUP-TFII and SMAD4 were first cloned into pENTR1A (Invitrogen) and then cloned into pBabeCMV-YFPn-DEST and pCL-CMV-DEST-YFPc using Gateway (Invitrogen), respectively. For HA-tagged FANCD2 and its mutants, Flag-tagged COUP-TFII and its mutants, and Flag-TR4 were cloned into pcDNA3.1 by polymerase chain reaction (PCR). For inducible HA-FANCD2, CRISPR-Cas9–resistant FANCD2 and their mutants were cloned into pENTR1A first and then cloned into pInducer20 (Addgene #44012) by Gateway. For sgRNA expression, DNA oligos were synthesized in Sigma and cloned into lentiGuide-puro (Addgene #52963).
Xu M., Qin J., Wang L., Lee H.J., Kao C.Y., Liu D., Songyang Z., Chen J., Tsai M.J, & Tsai S.Y. (2019). Nuclear receptors regulate alternative lengthening of telomeres through a novel noncanonical FANCD2 pathway. Science Advances, 5(10), eaax6366.
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2

Generating Lentiviral Vectors for DNAJB6 Variants

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GFP-DNAJB6b-WT and GFP-DNAJB6b-M3 DNA sequences were amplified from plasmids containing pcDNA5/FRT/TO GFP-DNAJB6b-WT and pcDNA5/FRT/TO GFP-DNAJB6b-M326 (a kind gift from Prof. Harrie H. Kampinga), adding BsmBI site at both the ends of the sequence. The amplified segments were inserted into an entry vector pENTR1A (a gift from Eric Campeau and Paul Kaufman; Addgene, plasmid# 17398; PRID:Addgene_17398) by golden gate assembly. The resultant plasmids were recombined with pLenti-CMV-Puro-DEST (a gift from Eric Campeau and Paul Kaufman; Addgene, plasmid# 17452; PRID:Addgene_17452) using a gateway LR clonase enzyme (ThermoFisher Scientific, 11791100) to achieve the expression vector pLenti-CMV-GFP-DNAJB6-WT-Puro-DEST and pLenti-CMV-GFP-DNAJB6-M3-Puro-DEST. pEGFP-Q74 was a gift from David Rubinsztein (Addgene plasmid #40262; PRID:Addgene_40262). Full sequences of used proteins are provided in the Supplementary Materials. Amino acid residues changed in M3 with respect to WT DNAJB6 sequence are marked yellow.
Joshi B.S., Garcia Romeu H., Aliyandi A., de Vries M.P, & Zuhorn I.S. (2022). DNAJB6-Containing Extracellular Vesicles as Chaperone Delivery Systems: A Proteomic Analysis. Pharmaceutics, 14(11), 2485.
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Generation of Plasmid Constructs for FAK Expression

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The plasmids pENTR1A (#17398) and MIGR1 (#27490) were obtained from Addgene. pENTR1A-IRES-GFP was generated by cloning IRES-GFP (digested with EcoR1 and Sal1) into the pENTR1A vector. The pBabe FAK and pBabe FAK (K454R) plasmids were kind gifts from Dr. Fillippo Giancotti17 (link). The pENTR1A-FAK-IRES-GFP and pENTR1A- FAK(K454R)-IRES-GFP plasmids were generated by cloning FAK, FAK(K454R) and FAK (FERM) coding region into the pENTR1A-IRES-GFP vector. The pT3EF1α plasmid was obtained from Dr. Xin Chen (UCSF)14 (link). The pT3- IRES-GFP, pT3- FAK-IRES-GFP, and pT3- FAK(K454R)-IRES-GFP were then generated by gateway cloning using pENTR1A-IRES-GFP, pENTR1A-FAK-IRES-GFP, and pENTR1A- FAK(K454R)-IRES-GFP as entry vectors and pT3EF1α as destination vector, respectively. The plasmids were purified using GeneJET Plasmid Maxiprep Kit (Thermo Fisher Scientific) for hydrodynamic tail vein injection.
Shang N., Arteaga M., Zaidi A., Cotler S.J., Breslin P., Ding X., Kuo P., Nishimura M., Zhang J, & Qiu W. (2016). FAK Kinase Activity Is Required for the Progression of c-MET/β-Catenin-Driven Hepataocellular Carcinoma. Gene Expression, 17(1), 79-88.
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