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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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