pAR1219 BioVector NTCC质粒载体菌种细胞基因保藏中心

pAR1219

pAR1219


Organism
Escherichia coli (Migula) Castellani and Chalmers
Classification
Enterobacteriaceae, Escherichia
Clone type
Clone
Applications
Molecular biology
Specific applications
Produces protein RNA polymerase
Genotype
pAR1219 T7gene1 lacI F- hsdR19 recA1 rpoB331 IN(rrnD-rrnE)1 lambda-
Comments
Restriction digests of the clone give the following sizes (kb): BamHI--4.4 (doublet); BglII-uncut; EcoRI--8.8; PstI--8.8; HindIII--8.8.
The T7 RNA polymerase expression construct, encoding gene 1 and lac regulatory sequences, can easily be excised as a 4.4 kb BamHI fragment for insertion into another construct.
The insert size given is for that of the sum of the lac and gene 1 sequences.
Gene1 was sequenced from T7 wild-type DNA. The junctions between gene 1 and lac were sequenced. Lac sequence is taken from GenBank J01637.
pAR1219 produces large amounts of T7 RNA polymerase in a suitable host upon induction with IPTG. Large amounts of active T7 RNA polymerase can be purified from the strain BL21/pAR1219.
The lac fragment contains the lacI promoter, lacI gene, lacUV5 promoter, lac operator, and the translation start site and first 147 amino acids of B-galactosidase.
pAR1219 was made by inserting a 1724 bp HincII fragment from pMC1, containing lac control elements, into the BglII site of pAR1173.
The construction consists of a fragment containing the T7 RNA polymerase gene (gene 1, including nucleotides 3145 to 5841 of T7 DNA) under the transcriptional regulation of lacI and the lacUV5 promoter.
Construct size (kb)
8.8
Intact vector size
4.363
Vector name
pBR322
Type of vector
plasmid
Construction
pBR313
Host range
Escherichia coli
Vector end
BamHI
Vector information
Cross references: DNA Seq. Acc.: J01749
Cloning sites
EcoRI; ClaI; HindIII; EcoRV; BamHI; SphI; SalI; XmaIII; NruI; BspMI; BsmI; StyI; AvaI; BalI; BspMII; PvuII; Tth111I; NdeI; AflIII; PpaI; PstI; PvuI; ScaI; SspI; AatII
Markers
ampR; tetR
Replicon
pMB1
Insert information
Insert size (kb)
4.4400000000000004
Type of DNA
genomic
Insert information
DESCRIPTION OF INSERT COMPONENT:
Genomic copy number: unique
Cross references: DNA Seq. Acc.: J02518
Genome
bacteriophage T7
Target gene
RNA polymerase
Gene name
RNA polymerase
Gene product
RNA polymerase [gene 1]
Gene symbol
gene 1
Contains complete coding sequence
Unknown
Insert end
Modification: BamHI linkers

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