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pET51b(+)-LUCIDRS-MTX

Plasmid

#113925

Purpose

Plasmid encoding the bacterial expression of the red-shifted LUCID semi-synthetic biosensor for methotrexate

Depositor

Kai Johnsson

Article

Hiblot et al Angew Chem Int Ed Engl. 2017 Nov 13;56(4

Insert

LUCID_RS-MTX

Use

Luciferase

Tags

Expression

Bacterial

Mutation

Promoter

T7

Available since

Sept. 17, 2018

Availability

Academic Institutions and Nonprofits only

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pET29b-clbI S178A

Plasmid

#114159

Purpose

Expresses ClbI S178A

Depositor

Emily Balskus

Article

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

Insert

clbI

Use

Tags

6xHis and 6xHis-Thrombin site

Expression

Bacterial

Mutation

Mutated S178 to Ala

Promoter

Available since

Sept. 5, 2018

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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