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Other IDs
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Nucleotide (GenBank) :
U25270
Expression cloning vector pACYC177/ET3d/yNMT, complete sequence.
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Host
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Distribution host: Escherichia coli BL21(DE3)
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Vector Information
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Size (kb): 4.9429998397827150 DESCRIPTION OF VECTOR COMPONENT:
Name of vector: pACYC177/ET3d
Intact vector size:
Type of vector: plasmid
Vector end: NcoI
Vector end: BamHI
Cloning sites: NcoI, NcoI+BamHI
Polylinker sites:
Construction: pACYC177,pET3d
Host range: Escherichia coli
Features (with orientation and position when available):
replicon: p15A
marker(s): kanR, ->
ribosome-binding site: gene 10
promoter: T7 (phi10), ->
Cross references: Vector: pACYC177/ET3d/yNMT (plasmid) Promoters: Promoter for expression T7 (phi10) Construction: pACYC177,pET3d Marker(s):kanR Construct size (kb): 4.900000095367432 Features: marker(s): kanR
promoter for expression: T7 (phi10)
replicon: p15A
coding sequence: NMT1
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Applications
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produces protein peptide N-myristoyl transferase glycylpeptide N-tetradecanoyltransferase
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Comments
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Restriction digests of the clone give the following sizes (kb): NdeI--5.5; NcoI--5.5; StuI--3.9, 1.5. Expression system compatible with ColE1 plasmids, for cotransformation and production of myristoylated proteins. Introduction of the NcoI site during amplification introduced a single base substitution T->G at nt 4. Expression of this gene in the presence of 100 uM myristic acid allows myristoylation of recombinant proteins produced by a second ColE1 plasmid.
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References
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Haun RS, et al. Effect of myristoylation on GTP-dependent binding of ADP-ribosylation factor to Golgi. J. Biol. Chem. 268: 7064-7068, 1993. PubMed: 8463239
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Shipping Information
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Distributed: freeze-dried
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