pKM999

Description:	A Gateway™ destination vector for the production of recombinant proteins as fusions to the C-terminus of Thermococcus litoralis maltose-binding protein.
Origin:	A derivative of pET11c (Novagen)
Replicon:	ColE1
Antibotic Resistance:	Ampicillin
Promotor:	bacteriophage T7
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:	Fox, J. D., Routzahn, K. M., Bucher, M. H., and Waugh, D. S. (2003). Maltodextrin-binding proteins from diverse bacteria and archaea are potent solubility enhancers. FEBS Lett. 537: 53-57. Horlacher, R., Xavier, K. B., Santos, H., DiRuggiero, J., Kossmann, M., and Boos, W. (1998). Archaeal binding protein-dependent ABC transporter: Molecular and biochemical analysis of the trehalose/maltose transport system of the hyperthermophilic archaeon Thermococcus litoralis. J. Bacteriol. 180: 680-689.
Nucleotide Sequence:

Amino Acid Sequence: *

MKIEEGKIVF AVGGAPNEIE YWKGVIAEFE KKYPGVTVEL KRQATDTEQR RLDLVNALRG KSSDPDVFLM DVAWLGQFIA SGWLEPLDDY VQKDNYDLSV FFQSVINLAD KQGGKLYALP VYIDAGLLYY RKDLLEKYGY SKPPETWQEL VEMAQKIQSG ERETNPNFWG FVWQGKQYEG LVCDFVEYVY SNGGSLGEFK DGKWVPTLNK PENVEALQFM VDLIHKYKIS PPNTYTEMTE EPVRLMFQQG NAAFERNWPY AWGLHNADDS PVKGKVGVAP LPHFPGHKSA ATLGGWHIGI SKYSDNKALA WEFVKFVESY SVQKGFAMNL GWNPGRVDVY DDPAVVSKSP HLKELRAVFE NAVPRPIVPY YPQLSEIIQK YVNSALAGKI SPQEALDKAQ KEAEELVKQY SKDQTSLYKK AG...passenger protein. * Amino acid residues encoded by the attR1 site in the Gateway™ cloning cassette are underlined.