Addgene - pRK793 Plasmid Data

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Plasmid 8827: pRK793

Gene/insert name:		TEV protease, S219V mutant
Alternative names:		tobacco etch virus protease
Insert size (bp):		750
Species of gene(s):		Other
Relevant mutations/deletions:		S219V mutation
Fusion proteins or tags:		His
Terminal:		N terminal on insert
Fusion proteins or tags:		polyarginine
Terminal:		C terminal on insert
Vector backbone:		pMal-C2 (Search Vector Database)
Backbone manufacturer:		New England Biolabs
Type of vector:		Bacterial expression
Backbone size (bp):		6700
Cloning site 5':		SacI
Site destroyed during cloning:		No
Cloning site 3':		BamHI
Site destroyed during cloning:		No
5' Sequencing primer:		N/A (List of Sequencing Primers)
3' Sequencing primer:		M13-F20
Bacteria resistance:		Ampicillin for p793; Chloramphenicol for pRIL.
High or low copy:		Low Copy
Grow in standard E. coli @ 37C:		No
Please specify bacterial strain for growth and growth condition:		Shifting the temperature from 37C to 30C during induction maximizes the yield of soluble TEV protease.
If you did not originally clone this gene, from whom and where did you receive the plasmid used to derive this plasmid:		ATCC
Sequence:		View sequence
Plasmid Provided In:		BL21(DE3)-RIL
Principal Investigator:		David S. Waugh
Terms and Licenses:		MTA

Comments: Bacterial strain: E.coli BL21(DE3)-RIL (Stratagene).

Addgene has sequenced a portion of this plasmid for verification. Click here for the sequencing result.

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Selected features

lacI

72 - 1163

Orf frame 3

204 - 1163

tac_promoter

1407 - 1435

M13_pUC_rev_primer

1447 - 1469

Orf frame 1

2793 - 1867

Orf frame 1

1528 - 3459

MBP_F_primer

2602 - 2625

6xHis

2713 - 2730

6xHis

2716 - 2733

M13_forward20_primer

3514 - 3498

M13_pUC_fwd_primer

3529 - 3507

lacZ_a

3507 - 3650

pBAD_rev_primer

3710 - 3693

pTrcHis_rev_primer

3710 - 3693

rrnB_terminator

3743 - 3900

rrnB_T1_terminator

3866 - 3909

rrnB_T2_terminator

4041 - 4068

AmpR_promoter

4110 - 4138

Orf frame 1

4180 - 5040

Ampicillin

4180 - 5040

f1_origin

5578 - 5272

pBR322_origin

5690 - 6309

pGEX_3_primer

6706 - 6728

ROP

6913 - 6722

Unique restriction sites

ApaI		642
HpaI		937
NarI		1071
BglII		1887
NcoI		2489
SacI		2638
SpeI		3115
BamHI		3463
XbaI		3469
SalI		3475
PstI		3485
MscI		7383

Article: Tobacco etch virus protease: mechanism of autolysis and rational design of stable mutants with wild-type catalytic proficiency. Kapust RB et al. (Protein Eng. 2001 Dec . 14(12):993-1000. Pubmed)

Please acknowledge the principal investigator and cite this article if you use this plasmid in a publication.

Also, please include the text "Addgene plasmid 8827" in your Materials and Methods section. This information allows Addgene to create a link from the plasmid page to your publication.