Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V004953 | pLP12 | 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:
- pLP12
- Antibiotic Resistance:
- Chloramphenicol
- Length:
- 3871 bp
- Type:
- Suicide vector
- Replication origin:
- R6K γ ori
- Source/Author:
- Luo P, He X, Liu Q, Hu C.
- Promoter:
- araBAD
pLP12 vector 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.
pLP12 vector Sequence
LOCUS 40924_28636 3871 bp DNA circular SYN 18-DEC-2018
DEFINITION Suicide vector pLP12, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3871)
AUTHORS Luo P, He X, Liu Q, Hu C.
TITLE Developing Universal Genetic Tools for Rapid and Efficient Deletion
Mutation in Vibrio Species Based on Suicide T-Vectors Carrying a
Novel Counterselectable Marker, vmi480
JOURNAL PLoS ONE 10 (12), E0144465 (2015)
PUBMED 26641275
REFERENCE 2 (bases 1 to 3871)
AUTHORS Luo P, He X, Hu C, Liu Q.
TITLE Direct Submission
JOURNAL Submitted (21-JUL-2015) Key Laboratory of Marine Bio-resources
Sustainable Utilization, South China Sea Institute of Oceanology,
Xingang Xi Road 164, Guangzhou, Guangdong 510301, China
REFERENCE 3 (bases 1 to 3871)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 3871)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE";
date: "2015"; volume: "10"; issue: "12"; pages: "E0144465"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(21-JUL-2015) Key Laboratory of Marine Bio-resources Sustainable
Utilization, South China Sea Institute of Oceanology, Xingang Xi
Road 164, Guangzhou, Guangdong 510301, China"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..3871
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 21..617
/codon_start=1
/product="toxin vmi480"
/label=toxin vmi480
/protein_id="ALS55902.1"
/translation="MTKKPEFYAPDLTEERLHILSENLLDVLDEAHHYSESPNATAWFK
GTANYGLPQGMLIRMHSDSAYPWLTLANQTMDYTARVGNTLVQFVVDDPHSPRKQHRLK
RNAVEKHQISLELEEDYVDTPLIWRFYLNPISNGVDYSPSISLLGFNGNGNVICSWEYD
TVVTGPVVTDKPESVEIDEPLLVRKKKVQKKVSDE"
misc_feature 667..706
/label=multiple cloning sites
/note="multiple cloning sites"
promoter complement(712..730)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
rep_origin complement(745..1133)
/direction=LEFT
/label=R6K gamma ori
/note="gamma replication origin from E. coli plasmid R6K;
requires the R6K initiator protein pi for replication"
CDS complement(1261..1917)
/codon_start=1
/label=CmR
/note="chloramphenicol acetyltransferase"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
promoter complement(1918..2020)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
promoter 2147..2165
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
oriT 2319..2428
/label=oriT
/note="incP origin of transfer"
CDS complement(2671..3546)
/codon_start=1
/label=araC
/note="L-arabinose regulatory protein"
/translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
EKVNDVAVKLS"
promoter 3573..3857
/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)"