Basic Vector Information
- Vector Name:
- pET-21-VRE_thyA-cat
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8428 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Chen V, Griffin ME, Maguin P, Varble A
pET-21-VRE_thyA-cat 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 ℃.
pET-21-VRE_thyA-cat vector Sequence
LOCUS V015997 8428 bp DNA circular SYN 20-JUL-2021
DEFINITION Exported.
ACCESSION V015997
VERSION V015997
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 8428)
AUTHORS Chen V, Griffin ME, Maguin P, Varble A, Hang HC.
TITLE RecT recombinase expression enables efficient gene editing in
Enterococcus
JOURNAL Appl Environ Microbiol, AEM0084421 (2021) In press
PUBMED 34232061
REFERENCE 2 (bases 1 to 8428)
AUTHORS Chen V, Griffin ME, Hang HC.
TITLE Direct Submission
JOURNAL Submitted (04-MAR-2021) Laboratory of Chemical Biology and Microbial
Pathogenesis, The Rockefeller University, 1230 York Ave., New York,
NY 10065, USA
REFERENCE 3 (bases 1 to 8428)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
SGRef: number: 1; type: "Journal Article"; journalName: "Appl
Environ Microbiol, AEM0084421 (2021) In press"
SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(04-MAR-2021) Laboratory of Chemical Biology and Microbial
Pathogenesis, The Rockefeller University, 1230 York Ave., New York,
NY 10065, USA"
FEATURES Location/Qualifiers
source 1..8428
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin 12..467
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 494..598
/label="AmpR promoter"
CDS 599..1456
/label="AmpR"
/note="beta-lactamase"
rep_origin 1630..2218
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature complement(2404..2546)
/label="bom"
/note="basis of mobility region from pBR322"
CDS complement(2651..2839)
/label="rop"
/note="Rop protein, which maintains plasmids at low copy
number"
protein_bind complement(3614..3635)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
CDS complement(3651..4730)
/label="lacI"
/note="lac repressor"
promoter complement(4731..4808)
/label="lacI promoter"
promoter 5117..5135
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 5136..5160
/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)."
RBS 5175..5197
/label="RBS"
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
misc_feature 5204..6303
/label="homology arm 1"
/note="homology arm 1"
gene 5204..6255
/gene="AX771_05170"
/label="AX771_05170"
gene 6267..6303
/gene="thyA"
/label="thyA"
regulatory 6334..6386
/regulatory_class="promoter"
regulatory 6344..6349
/regulatory_class="minus_35_signal"
regulatory 6368..6373
/regulatory_class="minus_10_signal"
regulatory 6404..6409
/regulatory_class="ribosome_binding_site"
CDS 6416..7063
/gene="cat"
/label="Chloramphenicol acetyltransferase"
/note="Chloramphenicol acetyltransferase from
Staphylococcus aureus. Accession#: P00485"
gene 7091..8001
/gene="thyA"
/label="thyA"
gene 8021..8190
/gene="AX771_05180"
/label="AX771_05180"
CDS 8190..8222
/label="T7 tag (gene 10 leader)"
/note="leader peptide from bacteriophage T7 gene 10"
CDS 8272..8289
/label="6xHis"
/note="6xHis affinity tag"
terminator 8356..8403
/label="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
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