Basic Vector Information
pUC19RP12 vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
pUC19RP12 vector Sequence
LOCUS 40924_45143 3474 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pUC19RP12, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3474)
AUTHORS Posfai G, Kolisnychenko V, Bereczki Z, Blattner FR.
TITLE Markerless gene replacement in Escherichia coli stimulated by a
double-strand break in the chromosome
JOURNAL Nucleic Acids Res. 27 (22), 4409-4415 (1999)
PUBMED 10536150
REFERENCE 2 (bases 1 to 3474)
AUTHORS Posfai G, Kolisnychenko V, Bereczki Z, Blattner FR.
TITLE Direct Submission
JOURNAL Submitted (21-JUL-1999) Institute of Biochemistry, Biol. Res. Center
Hung. Acad.Sci., Temesvari krt.,62, Szeged H-6726, Hungary
REFERENCE 3 (bases 1 to 3474)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 3474)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic
Acids Res."; date: "1999"; volume: "27"; issue: "22"; pages:
"4409-4415"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(21-JUL-1999) Institute of Biochemistry, Biol. Res. Center Hung.
Acad.Sci., Temesvari krt.,62, Szeged H-6726, Hungary"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..3474
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 379..395
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 450..505
/label=tetR/tetA promoters
/note="overlapping promoters for bacterial tetR and tetA"
CDS 519..1211
/gene="SCEI"
/label=SCEI
/note="Intron-encoded endonuclease I-SceI from
Saccharomyces cerevisiae (strain ATCC 204508 / S288c).
Accession#: P03882"
primer_bind complement(1253..1269)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(1277..1293)
/label=lac operator
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(1301..1331)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(1346..1367)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin 1652
/label=ColE1
/note="ColE1"
rep_origin complement(1655..2243)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2417..3274)
/label=AmpR
/note="beta-lactamase"
promoter complement(3275..3379)
/label=AmpR promoter
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