Addgene - pNIC-ZB Plasmid Data

Plasmid 26107: pNIC-ZB

Gene/insert name:
None
GenBank ID:
GU452710
Fusion protein or tag:
His6 - Z-basic - TEV
Terminal:
N terminal on backbone
Vector backbone:
pET-28-a
(Search Vector Database)
Backbone manufacturer:
Novagen
Vector type:
Bacterial Expression
Backbone size (bp):
5515
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 expression, use host cells that express T7 RNA pol, e.g. BL21(DE3).
High or low copy:
High Copy
Sequence: View sequences (4)
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 26107" in your Materials and Methods section.

Price: $65.00

Available to academic and non-profits only

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