Price Information
| Cat No. | Plasmid Name | Availability | Buy one, get one free! (?) |
|---|---|---|---|
| V018122 | pUCP24 | In stock, instant shipping |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
pUCP24 is an E. coli–Pseudomonasshuttle vector. It contains a pMB1 origin for replication in E. coliand a pRO1600 origin for maintenance in Pseudomonas, typically featuring a selectable marker like an antibiotic resistance gene for selection in both hosts.
- Vector Name:
- pUCP24
- Antibiotic Resistance:
- Gentamycin
- Length:
- 4036 bp
- Type:
- E. coli–Pseudomonas shuttle vector
- Copy Number:
- High copy number
- Growth Strain(s):
- DH10B
- Growth Temperature:
- 37℃
pUCP24 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.
References
- Findlay J, Raro OHF, Poirel L, Nordmann P; NARA Network. Molecular analysis of metallo-beta-lactamase-producing Pseudomonas aeruginosa in Switzerland 2022-2023. Eur J Clin Microbiol Infect Dis. 2024 Mar;43(3):551-557. doi: 10.1007/s10096-024-04752-8. Epub 2024 Jan 18. PMID: 38233610; PMCID: PMC10917820.
- Fournier C, Nordmann P, de la Rosa JO, Kusaksizoglu A, Poirel L. KSA-1, a naturally occurring Ambler class A extended spectrum β-lactamase from the enterobacterial species Kosakonia sacchari. J Glob Antimicrob Resist. 2024 Dec;39:6-11. doi: 10.1016/j.jgar.2024.07.008. Epub 2024 Aug 13. PMID: 39147026.
pUCP24 vector Sequence
LOCUS Exported 4036 bp DNA circular SYN 03-NOV-2025
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4036)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..4036
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin 46..397
/label=pRO1600 oriV
/note="broad-host-range origin of replication from
Pseudomonas aeruginosa plasmid pRO1600; requires the
pRO1600 Rep protein for replication (West et al., 1994)"
CDS 411..1244
/codon_start=1
/product="replication protein for the broad-host-range
plasmid pRO1600 from Pseudomonas aeruginosa "
/label=pRO1600 Rep
/translation="MASPPMVYKSNALVEAAYRLSVQEQRIVLACISQVKRSEPVTDEV
MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT
IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMDSTHAIRLYELLMQWDSIG
QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL
LFSFGPKKPAKAVGKAPAKRKAGKISDAEIAKQARPGETWEAARARLTQMPLDLA"
CDS complement(1241..1495)
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/label=lacZ-alpha
/translation="MTMITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNR
LAAHPPFASWRNSEEARTDRPSQQLRSLKGRPGRGGHGL"
primer_bind 1405..1421
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 1425..1481
/label=MCS
/note="pUC18/19 multiple cloning site"
primer_bind complement(1491..1507)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 1515..1531
/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)."
promoter complement(1539..1569)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1584..1605
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(1893..2481)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(3282..3815)
/codon_start=1
/gene="aacC1"
/product="gentamycin acetyltransferase"
/label=GmR
/note="confers resistance to gentamycin"
/translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
EEVMHFDIDPSTAT"
promoter complement(4004..4032)
/gene="intI1 (promoter lies within the coding sequence)"
/label=Pc promoter
/note="class 1 integron promoter"