| Vector Information |
Size (kb): 9.0000000000000000 Vector: p519gfp (plasmid) Promoters: Promoter for expression lac Marker(s):kanR Features: marker(s): kanR, neoR
other: mob
promoter for expression: lac
MCS: HindIII/.../XbaI
restriction site: EcoRI
restriction site: NdeI
ribosome-binding site: GFP
coding sequence: GFPmut2 |
| Comments |
Restriction digests of the clone give the following sizes (kb): EcoRI/XbaI--9.0; EcoRI--9.2; EcoRI/HindIII--6.7, 1.7, 0.7; PstI--5.6, 1.8, 1.2. The promoterless reporter GFPmut2 cassette, with its ribosome binding site, can be recovered as a ndIII/EcoRI (5'/3') fragment (approximately 740 bp). A broad host range mob+ plasmid with GFPmut2 expression controlled by a lac promoter. Cloning into the SphI, PstI, AccI, HincII, SalI, XbaI, or NdeI sites will probably affect GFPmut2* expression as it would separate the CDS or RBS from the lac promoter. This has not yet been tested. |
| References |
Matthysse AG, et al. Construction of GFP vectors for use in gram negative bacteria other than E. coli. FEMS Microbiol. Lett. 145: 87-94, 1996. PubMed: 8931331
Keen NT, et al. Improved broad-host-range plasmids for DNA cloning in gram-negative bacteria. Gene 70: 191-197, 1988. PubMed: 2853689
Cormack BP, et al. FACS-optimized mutants of the green fluorescent protein (GFP). Gene 173: 33-38, 1996. PubMed: 8707053
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