pET30Xa-SBL1(160S,NoCys)
(Plasmid
#66877)
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PurposeExpresses a cysteine-free version of WTsoybean lipoxygenase-1 160S (4Cys to Ser) with N-term Histag
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 66877 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepETE30Xa-LIC
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Backbone manufacturerNovagen
- Backbone size w/o insert (bp) 5444
- Total vector size (bp) 7978
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Modifications to backboneC-term extension: TAAACTAGTCATGGGGC
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Vector typeBacterial Expression
Growth in Bacteria
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Bacterial Resistance(s)Kanamycin, 50 μg/mL
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Growth Temperature37°C
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Growth Strain(s)DH5alpha
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Growth instructions37C before induction, 30 deg for 14-16+ hrs after induction
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Copy numberUnknown
Gene/Insert
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Gene/Insert namesoybean lipoxygenase-1
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SpeciesGlycine max
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Insert Size (bp)2517
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Mutationfour (all) cysteines mutated to serine
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GenBank IDAAA33986.1
- Promoter T7lac
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Tag
/ Fusion Protein
- Histag (N terminal on backbone)
Cloning Information
- Cloning method Ligation Independent Cloning
- 5′ sequencing primer tca tca ttc ttc tgg tct gg
- 3′ sequencing primer ttg ccg act ctc att agc c (Common Sequencing Primers)
Resource Information
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Supplemental Documents
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
Depositor Comments
prepared by four sequential mutations of Cys to Ser, from pET30Xa-SBL1(160S,NoCys)
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
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For your Materials & Methods section:
pET30Xa-SBL1(160S,NoCys) was a gift from Betty Gaffney (Addgene plasmid # 66877 ; http://n2t.net/addgene:66877 ; RRID:Addgene_66877) -
For your References section:
On the relationships of substrate orientation, hydrogen abstraction, and product stereochemistry in single and double dioxygenations by soybean lipoxygenase-1 and its Ala542Gly mutant. Coffa G, Imber AN, Maguire BC, Laxmikanthan G, Schneider C, Gaffney BJ, Brash AR. J Biol Chem. 2005 Nov 18;280(46):38756-66. Epub 2005 Sep 12. 10.1074/jbc.M504870200 PubMed 16157595