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Size (kb): 4.320
Vector: pFN476 (plasmid)
Promoters: Promoter lac
Construction: pGEM3Z, pKA101-6
Construct size (kb): 4.320
Features: insert detection: lacZ'
promoter for expression: T7
promoter for in vitro transcription: SP6
replicon: rep (pSC101)
MCS: pGEM-3Z; HindIII...EcoRI
restriction site: BamHI
coding sequence: tnpR
expression vector T7 promoter
vector permitting RNA synthesis in vitro
vector permitting visual detection of recombinants beta-galactosidase beta-D-galactosidase
vector with low copy number
Restriction digests of the clone give the following sizes (kb): EcoRI--4.2; HindIII--4.2; PstI--3.1, 1.1.
Low copy number vector to express transcripts in vivo from the T7 promoter and to minimize toxic gene effects.
Plasmid contains the pSC101 replicon and is maintained at 1-5 copies per cell.
Low copy number minimizes the effect of uninduced expression of T7 RNA polymerase, reducing transcription of potentially toxic inserts.
T7 RNA polymerase can also be provided by infection of F' hosts with M13mGP1-2 (ATCC 40303)
Sankar P, et al. Expression analysis of cloned chromosomal segments of Escherichia coli. J. Bacteriol. 175: 5145-5152, 1993. PubMed: 8349554