Addgene - pNH-TrxT Plasmid Data

Plasmid 26106: pNH-TrxT

Gene/insert name:
None
GenBank ID:
GU269914
Fusion protein or tag:
His6 - Thioredoxin - TEV
Terminal:
N terminal on backbone
Vector backbone:
pET-28-a
(Search Vector Database)
Backbone manufacturer:
Novagen
Vector type:
Bacterial Expression
Backbone size (bp):
5671
Cloning site 5':
Ligation-Independent cloning; cut with BsaI
Site destroyed during cloning:
Yes
Cloning site 3':
Ligation-Independent cloning; cut with BsaI
Site destroyed during cloning:
Yes
5' sequencing primer:
T7F List of Sequencing Primers
3' sequencing primer:
T7R
Bacterial resistance(s)
Kanamycin
Growth strain(s)
DH5alpha
Growth temperature (℃):
37
Growth instructions:
For plasmid propagation and cloning, use any E. coli strain that does not express T7 RNA polymerase. For protein expression, use a host that expresses T7 RNA polymerase, e.g. BL21(DE3).
High or low copy:
High Copy
Sequence: View sequences (3)
Principal Investigator:
Opher Gileadi
Terms and Licenses:
MTA
EMD Millipore Limited Use Label License/Brookhaven pET Assurance Letter

Comments:

Primers for LIC cloning:

Upstream: add TACTTCCAATCCATG to the 5’ end (ATG in-frame with the desired coding
sequence).

Downstream: add TATCCACCTTTACTG to 5’ end of downstream primer; add termination
codon, if necessary.

Detailed cloning method available in the paper (Savitsky et al., J Struct Biol., 2010)

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

Article: High-throughput production of human proteins for crystallization: The SGC experience. Savitsky et al (J Struct Biol. 2010 Jun 10. ():. 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 26106" in your Materials and Methods section.

Price: $65.00

Available to academic and non-profits only

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