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pET28a-DDX43-FL-G84D

Plasmid

#134577

Purpose

Express human DDX43 mutant (full-length with G84D mutation) in E. coli

Depositor

Yuliang Wu

Article

Talwar et al J Biol Chem. 2017 Jun 23;292(25):10429-1

Insert

DDX43-G84D (DDX43 Human)

Use

Tags

His

Expression

Bacterial

Mutation

full-length (G84D)

Promoter

T7

Available since

Jan. 6, 2020

Availability

Academic Institutions and Nonprofits only

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pET28a-clbH-C-A2-PCP

Plasmid

#114157

Purpose

Expresses ClbH-C-A2-PCP

Depositor

Emily Balskus

Article

Zha et al Nat Chem Biol. 2017 Oct;13(10):1063-1065

Insert

clbH

Use

Tags

6xHis-thrombin-T7 tag

Expression

Bacterial

Mutation

Only contains the C-A2-PCP domains of ClbH

Promoter

Available since

Aug. 31, 2018

Availability

Academic Institutions and Nonprofits only

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pET28a_6HT-TBP6.7(P46A)

Plasmid

#112721

Purpose

This mutation (P46A) marks the first residue within the evolved region of U1A.

Depositor

Joseph Wedekind

Article

Belashov et al Nucleic Acids Res. 2018 Jul 27;46(13):64

Insert

6His-TEV-TBP6.7(P46A)

Use

Tags

6His-TEV

Expression

Bacterial

Mutation

P46A

Promoter

T7

Available since

Aug. 8, 2018

Availability

Academic Institutions and Nonprofits only

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pET28a_6HT-TBP6.7(R47A)

Plasmid

#112722

Purpose

Arg47 within TBP6.7 is the most critical residue in terms of forming interactions with WT TAR. Mutating this amino acid to Ala results in ~600-fold reduction in Kd.

Depositor

Joseph Wedekind

Article

Belashov et al Nucleic Acids Res. 2018 Jul 27;46(13):64

Insert

6His-TEV-TBP6.7(R47A)

Use

Tags

6His-TEV

Expression

Bacterial

Mutation

R47A

Promoter

T7

Available since

Aug. 8, 2018

Availability

Academic Institutions and Nonprofits only

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pET28a_6HT-TBP6.7(R47K)

Plasmid

#112723

Purpose

Arg47 of TBP6.7 interacts with Hoogsteen edge of TAR's Gua26. Mutating this amino acid to Lys reduces binding to the WT TAR RNA ~330-fold.

Depositor

Joseph Wedekind

Article

Belashov et al Nucleic Acids Res. 2018 Jul 27;46(13):64

Insert

6His-TEV-TBP6.7(R47K)

Use

Tags

6His-TEV

Expression

Bacterial

Mutation

R47K

Promoter

T7

Available since

Aug. 8, 2018

Availability

Academic Institutions and Nonprofits only

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pET28a_6HT-TBP6.7(Q48A)

Plasmid

#112724

Purpose

Gln48 contacts the backbone of TAR RNA as well as mediates intramoecular interaction to stabilize the conformation of beta2-beta3 loop within TBP6.7.

Depositor

Joseph Wedekind

Article

Belashov et al Nucleic Acids Res. 2018 Jul 27;46(13):64

Insert

6His-TEV-TBP6.7(Q48A)

Use

Tags

6His-TEV

Expression

Bacterial

Mutation

Q48A

Promoter

T7

Available since

Aug. 8, 2018

Availability

Academic Institutions and Nonprofits only

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pET28a_6HT-TBP6.7(Q48T)

Plasmid

#112725

Purpose

This mutant of TBP6.7 binds to TAR very similarly to TBP6.7(WT), with only 1.4-fold reduced Kd.

Depositor

Joseph Wedekind

Article

Belashov et al Nucleic Acids Res. 2018 Jul 27;46(13):64

Insert

6His-TEV-TBP6.7(Q48T)

Use

Tags

6His-TEV

Expression

Bacterial

Mutation

Q48T

Promoter

T7

Available since

Aug. 8, 2018

Availability

Academic Institutions and Nonprofits only

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pET28a_6HT-TBP6.7(T50A)

Plasmid

#112727

Purpose

Lav-evolved amino acid T50 within TBPs is important in engaging intramolecularly to steer also lab-evolved R49 into conformation compatible binding to RNA.

Depositor

Joseph Wedekind

Article

Belashov et al Nucleic Acids Res. 2018 Jul 27;46(13):64

Insert

6His-TEV-TBP6.7(T50A)

Use

Tags

6His-TEV

Expression

Bacterial

Mutation

T50A

Promoter

T7

Available since

Aug. 8, 2018

Availability

Academic Institutions and Nonprofits only

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pET28a_6HT-TBP6.7(P51A)

Plasmid

#112728

Purpose

Similar to P46, P51 was selected and is present in all evolved TBPs. Mutating this position into Ala has a modest effect on binding.

Depositor

Joseph Wedekind

Article

Belashov et al Nucleic Acids Res. 2018 Jul 27;46(13):64

Insert

6His-TEV-TBP6.7(P51A)

Use

Tags

6His-TEV

Expression

Bacterial

Mutation

P51A

Promoter

T7

Available since

Aug. 8, 2018

Availability

Academic Institutions and Nonprofits only

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pET28a_6HT-TBP6.7(R52A)

Plasmid

#112729

Purpose

52nd position is Arg in both wild-type U1A and TBPs, but the RNA recognition by this residue in both types of proteins is fundamentally different.

Depositor

Joseph Wedekind

Article

Belashov et al Nucleic Acids Res. 2018 Jul 27;46(13):64

Insert

6His-TEV-TBP6.7(R52A)

Use

Tags

6His-TEV

Expression

Bacterial

Mutation

R52A

Promoter

T7

Available since

Aug. 8, 2018

Availability

Academic Institutions and Nonprofits only

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pET28a_6HT-TBP6.7(Q54A)

Plasmid

#112730

Purpose

Although not evolved, this residue within TBPs participates in stabilizing the polypeptide loop responsible for RNA recognition.

Depositor

Joseph Wedekind

Article

Belashov et al Nucleic Acids Res. 2018 Jul 27;46(13):64

Insert

6His-TEV-TBP6.7(Q54A)

Use

Tags

6His-TEV

Expression

Bacterial

Mutation

Q54A

Promoter

T7

Available since

Aug. 8, 2018

Availability

Academic Institutions and Nonprofits only

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pET28a-RupA_T1-NHis

Plasmid

#109259

Purpose

NRPS T domain with N-terminal 6His tag

Depositor

Emily Balskus

Article

Schneider et al Tetrahedron

Insert

RupA_T1

Use

Tags

His tag

Expression

Bacterial

Mutation

contains aa 917..1025 of WP_015523502.1

Promoter

T7

Available since

July 6, 2018

Availability

Academic Institutions and Nonprofits only

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pET28a-RupA_R-NHis

Plasmid

#109257

Purpose

NRPS R domain with N-terminal 6His tag

Depositor

Emily Balskus

Article

Schneider et al Tetrahedron

Insert

RupA_R

Use

Tags

His tag

Expression

Bacterial

Mutation

contains aa 2038..2443 of WP_015523502.1

Promoter

T7

Available since

July 6, 2018

Availability

Academic Institutions and Nonprofits only

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pET28A-p53 (1-73) (PPP)

Plasmid

#62067

Purpose

Expression of human p53 (1-73) (P12A, P13A, P27A) in e. coli

Depositor

Gary Daughdrill

Article

Borcherds et al Nat Chem Biol. 2014 Dec;10(12):1000-2. d

Insert

p53 (TP53 Human)

Use

Tags

His

Expression

Bacterial

Mutation

Contains TAD residues 1-73; P12A, P13A, P27A

Promoter

T7

Available since

Jan. 29, 2015

Availability

Academic Institutions and Nonprofits only
WT C-His Ana GvpC in pET28a

Plasmid

#85732

Purpose

Expresses His-tagged GvpC from Anabaena flos-aquae in E.coli

Depositor

Mikhail Shapiro

Article

Lakshmanan et al ACS Nano. 2016 Aug 23;10(8):7314-22. doi

Insert

Gas vesicle protein C (his-tagged)

Use

Tags

His-tag

Expression

Bacterial

Mutation

Added an extra glycine after the methionine at th…

Promoter

T7

Available since

March 29, 2017

Availability

Academic Institutions and Nonprofits only

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pET28a-TS2126 RnlA-Strep

Plasmid

#159350

Purpose

Express TS2126 RNA ligase with His6 tag and Strep tag at N and C terminal, respectively

Depositor

Takashi Ito

Article

Hisano et al BMC Biol. 2021 Oct 14;19(1):225. doi: 10

Insert

TS2126 RNA ligase

Use

Tags

His6 and Strep

Expression

Bacterial

Mutation

Promoter

T7

Available since

Sept. 27, 2022

Availability

Academic Institutions and Nonprofits only

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pET28a T7pCONS TIR-2 sfGFP

Plasmid

#154464

Purpose

pET28a with a design flaw in the T7 promoter fixed, and a synthetically evolved TIR. Contains the coding sequence for sfGFP

Depositor

Daniel Daley

Article

Shilling et al Commun Biol. 2020 May 7;3(1):214. doi: 1

Insert

super folder GFP

Use

Tags

His-thrombin

Expression

Bacterial

Mutation

Promoter

T7

Available since

July 20, 2020

Availability

Academic Institutions and Nonprofits only

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pET28a-MBP-CBX1-eReader

Plasmid

#135089

Purpose

Bacterial expression of engineered CBX1 with enhanced binding to H3K9me

Depositor

Marcey Waters

Article

Albanese et al ACS Chem Biol. 2020 Jan 17;15(1):103-111

Insert

CBX1 chromobox protein 1, engineered

Use

Tags

6xhis-tag and MBP

Expression

Bacterial

Mutation

lysine 43 mutated to alanine; aspartic acid 59 mu…

Promoter

T7

Available since

March 15, 2022

Availability

Academic Institutions and Nonprofits only

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pET28a SnoopTag-mEGFP-SpyTag

Plasmid

#72325

Purpose

Bacterial expression of monomeric enhanced green fluorescent protein (mEGFP) containing SnoopTag at the N-terminus and SpyTag at the C-terminus, for programmed polyprotein synthesis.

Depositor

Mark Howarth

Article

Veggiani et al Proc Natl Acad Sci U S A. 2016 Jan 19. p

Insert

pET28a SnoopTag-mEGFP-SpyTag

Use

Tags

N-terminal His6 tag

Expression

Bacterial

Mutation

Promoter

Available since

Feb. 12, 2016

Availability

Academic Institutions and Nonprofits only

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pET28a-His6-Ran(Q69L)

Plasmid

#171563

Purpose

Encodes human Ran protein

Depositor

Yan Zeng

Insert

Ran (RAN Human)

Use

Tags

Expression

Bacterial

Mutation

Promoter

Available since

July 19, 2021

Availability

Academic Institutions and Nonprofits only

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