Basic Vector Information
pUC19-T7pol-lacI-lys 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 ℃.
pUC19-T7pol-lacI-lys vector Sequence
LOCUS 40924_45133 8856 bp DNA circular SYN 18-DEC-2018
DEFINITION Expression vector pUC19-T7pol-lacI-lys, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8856)
AUTHORS Kang Y, Son MS, Hoang TT.
TITLE One step engineering of T7-expression strains for protein
production: increasing the host-range of the T7-expression system
JOURNAL Protein Expr. Purif. 55 (2), 325-333 (2007)
PUBMED 17716915
REFERENCE 2 (bases 1 to 8856)
AUTHORS Kang Y, Son MS, Hoang TT.
TITLE Direct Submission
JOURNAL Submitted (30-NOV-2006) Microbiology, University of Hawaii at Manoa,
2538 The Mall - Snyder 310, Honolulu, HI 96822, USA
REFERENCE 3 (bases 1 to 8856)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 8856)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Protein
Expr. Purif."; date: "2007"; volume: "55"; issue: "2"; pages:
"325-333"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(30-NOV-2006) Microbiology, University of Hawaii at Manoa, 2538 The
Mall - Snyder 310, Honolulu, HI 96822, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..8856
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 379..395
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
CDS complement(437..889)
/label=T7 lysozyme
/note="lysozyme from bacteriophage T7"
protein_bind 1152..1199
/label=FRT
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
CDS complement(1332..1862)
/label=GmR
/note="gentamycin acetyltransferase"
promoter complement(2051..2079)
/label=Pc promoter
/note="class 1 integron promoter"
misc_feature 2120..2167
/note="FRT; Flp-recombination-target"
protein_bind 2120..2167
/label=FRT
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
CDS 2248..3327
/label=lacI
/note="lac repressor"
protein_bind 3343..3364
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3379..3409
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
protein_bind 3417..3433
/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)."
CDS 3453..3626
/label=lacZ-alpha
/note="LacZ-alpha fragment of beta-galactosidase"
CDS 3935..6583
/note="T7 RNA polymerase from Escherichia phage T7.
Accession#: P00573"
primer_bind complement(6635..6651)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(6659..6675)
/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(6683..6713)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(6728..6749)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(7037..7625)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7799..8656)
/label=AmpR
/note="beta-lactamase"
promoter complement(8657..8761)
/label=AmpR promoter
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