Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V010932 | pETcoco-1 | In stock, 1 week for quality controls |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pETcoco-1
- Antibiotic Resistance:
- Chloramphenicol
- Length:
- 12474 bp
- Type:
- pET & Duet Vectors (Novagen)
- Replication origin:
- ori2
- Source/Author:
- Novagen (EMD Millipore)
- Copy Number:
- Low copy number
- Promoter:
- araBAD
pETcoco-1 vector Map
Plasmid 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 ℃.
6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it
General Plasmid Transform Protocol
1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.
2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.
3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.
4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.
5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.
pETcoco-1 vector Sequence
LOCUS pETcoco-1. 12474 bp DNA circular SYN 01-JAN-1980
DEFINITION Bacterial vector with a chloramphenicol resistance marker that
allows single-copy replication, on-demand amplification, and tightly
regulated expression.
ACCESSION .
VERSION .
KEYWORDS pETcoco-1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12474)
AUTHORS Novagen (EMD Millipore)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 12474)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
COMMENT The copy number can be increased to 20-50 copies per cell by adding
arabinose to the growth medium.
FEATURES Location/Qualifiers
source 1..12474
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 3..27
/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 42..64
/label=RBS
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 72..74
/codon_start=1
/product="start codon"
/label=start codon
/note="ATG"
/translation="M"
CDS 75..92
/label=6xHis
/note="6xHis affinity tag"
CDS 102..146
/label=S-Tag
/note="affinity and epitope tag derived from pancreatic
ribonuclease A"
CDS 162..176
/label=enterokinase site
/note="enterokinase recognition and cleavage site"
CDS 249..281
/label=HSV tag
/note="HSV (herpes simplex virus) epitope tag"
CDS 326..343
/label=6xHis
/note="6xHis affinity tag"
terminator 410..457
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(834..1709)
/label=araC
/note="L-arabinose regulatory protein"
promoter 1736..2020
/label=araBAD promoter
/note="promoter of the L-arabinose operon of E. coli; the
araC regulatory gene is transcribed in the opposite
direction (Guzman et al., 1995)"
CDS 2080..3225
/label=trfA
/note="trans-acting replication protein that binds to and
activates oriV"
CDS complement(3399..4055)
/label=CmR
/note="chloramphenicol acetyltransferase"
promoter complement(4056..4158)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
rep_origin complement(5022..5734)
/direction=LEFT
/label=oriV
/note="incP origin of replication"
rep_origin 5813..6032
/label=ori2
/note="secondary origin of replication for the bacterial F
plasmid; also known as oriS"
CDS 6123..6875
/label=repE
/note="replication initiation protein for the bacterial F
plasmid"
misc_feature 6881..7131
/label=incC
/note="incompatibility region of the bacterial F plasmid"
CDS 7457..8629
/label=sopA
/note="partitioning protein for the bacterial F plasmid"
CDS 8632..9600
/label=sopB
/note="partitioning protein for the bacterial F plasmid"
misc_feature 9676..10149
/gene="sopC"
/label=sopC
/note="sopC"
/note="centromere-like partitioning region of the bacterial
F plasmid"
misc_feature complement(10409..10807)
/label=cos
/note="lambda cos site; allows packaging into phage lambda
particles"
protein_bind complement(10825..10858)
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
protein_bind complement(10951..10972)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
CDS complement(10988..12067)
/label=lacI
/note="lac repressor"
promoter complement(12068..12145)
/label=lacI promoter