Price Information
| Cat No. | Plasmid Name | Availability | Buy one, get one free! (?) |
|---|---|---|---|
| V017343 | ePop | In stock, instant shipping |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
Expresses E lysis protein that causes in subpopulation death when threshold cell density is reached, resulting in oscillations. The plasmid tends to be unstable, with the region containing Lysis protein E and mutated LuxR being prone to deletion. Please screen at least 5 small colonies by restriction digest and culture the clones at 30 ℃ if possible
- Vector Name:
- ePop
- Antibiotic Resistance:
- Chloramphenicol
- Length:
- 4198 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- E. coli
- Growth Strain(s):
- DH10a
- Growth Temperature:
- 37℃
ePop 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.
ePop vector Sequence
LOCUS Exported 4198 bp DNA circular SYN 05-JUN-2025
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS ePop
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4198)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..4198
/mol_type="other DNA"
/organism="synthetic DNA construct"
source 152..424
/mol_type="other DNA"
/organism="synthetic DNA construct"
source 1744..2322
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 152..424
/gene="E"
/label=Lysis protein E
/note="Lysis protein E from Enterobacteria phage phiX174.
Accession#: P03639"
protein_bind 443..459
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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 661..690
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
protein_bind 867..886
/label=lac operator (symmetric)
/bound_moiety="lac repressor encoded by lacI"
/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). The
symmetric lac operator was optimized for tight binding of
lac repressor."
protein_bind 900..916
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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 935..946
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 954..1265
/codon_start=1
/product="Early stop codon introduced in mutated luxR
results in loss of luxR functionality; the regulation of
this gene is not required for oscillating behavior observed
in the study."
/label=mutated LuxR
/translation="MKNINADDTYRIINKIKACRSNNDINQCLSDMTKMVHCEYYLLAI
IYPHSMVKSDISILDNYPKKWRQYYDDANLIKYDPIVDYSNSNHSPINWNIFENKCCK"
CDS 1744..2322
/gene="luxI"
/label=Acyl-homoserine-lactone synthase
/note="Acyl-homoserine-lactone synthase from Aliivibrio
fischeri. Accession#: P12747"
terminator 2371..2457
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
rep_origin complement(2622..3210)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
terminator complement(3298..3392)
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS complement(3413..4072)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
promoter complement(4073..4175)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"