pDEST-HisMBP

Description:	A Gateway™ destination vector for the production of recombinant proteins as fusions to the C-terminus of a dual His6-MBP tag in the cytoplasm of E. coli.
Origin:	A derivative of pMal-C2 (New England Biolabs)
Replicon:	ColE1 (pBR322)
Antibotic Resistance:	Ampicillin
Promotor:	tac
Inducer:	IPTG (1 mM)
Notes:	Because it carries a gene (ccdB ) that encodes a lethal DNA gyrase poison, this vector must be propogated in an E. coli gyrA mutant such as DB3 or DB5 (Invitrogen), which is immune to the action of CcdB.
Literature Citations:	Nallamsetty, S., Austin, B. P., Penrose, K. J., and Waugh, D. S. (2005). Gateway vectors for the production of combinatorially-tagged His6-MBP fusion proteins in the cytoplasm and periplasm of Escherichia coli. Protein Sci. 14: 2964-2971; Tropea, J. E., Cherry, S., Nallamsetty, S., Bignon, C., and Waugh, D. S. (2006). A generic method for the production of recombinant proteins in Escherichia coli using a dual His6-MBP affinity tag. Methods Mol. Biol., in press.
Nucleotide Sequence:

Amino Acid Sequence: *

MKIHHHHHHE EGKLVIWING DKGYNGLAEV GKKFEKDTGI KVTVEHPDKL EEKFPQVAAT GDGPDIIFWA HDRFGGYAQS GLLAEITPDK AFQDKLYPFT WDAVRYNGKL IAYPIAVEAL SLIYNKDLLP NPPKTWEEIP ALDKELKAKG KSALMFNLQE PYFTWPLIAA DGGYAFKYEN GKYDIKDVGV DNAGAKAGLT FLVDLIKNKH MNADTDYSIA EAAFNKGETA MTINGPWAWS NIDTSKVNYG VTVLPTFKGQ PSKPFVGVLS AGINAASPNK ELAKEFLENY LLTDEGLEAV NKDKPLGAVA LKSYEEELAK DPRIAATMEN AQKGEIMPNI PQMSAFWYAV RTAVINAASG RQTVDEALKD AQTNSITSLY KKAG….passenger protein. * Amino acid residues encoded by the attR1 site in the Gateway™ cloning cassette are underlined.