Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V008084 | pCXUN-FLAG | 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:
- pCXUN-FLAG
- Antibiotic Resistance:
- Kanamycin
- Length:
- 12035 bp
- Type:
- Binary vector
- Replication origin:
- ori
- Source/Author:
- Chen S, Songkumarn P, Liu J, Wang GL.
- Promoter:
- Ubi
pCXUN-FLAG 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.
pCXUN-FLAG vector Sequence
LOCUS 40924_14040 12035 bp DNA circular SYN 17-DEC-2018
DEFINITION Binary vector pCXUN-FLAG, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12035)
AUTHORS Chen S, Songkumarn P, Liu J, Wang GL.
TITLE A versatile zero background T-vector system for gene cloning and
functional genomics
JOURNAL Plant Physiol. 150 (3), 1111-1121 (2009)
PUBMED 19403729
REFERENCE 2 (bases 1 to 12035)
AUTHORS Chen S, Songkumarn P, Liu J, Wang G-L.
TITLE Direct Submission
JOURNAL Submitted (09-APR-2009) Department of Plant Pathology, The Ohio
State University, 201 Kottman Hall, 2021 Coffey Rd, Columbus, OH
43210, USA
REFERENCE 3 (bases 1 to 12035)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 12035)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Plant
Physiol."; date: "2009"; volume: "150"; issue: "3"; pages:
"1111-1121"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(09-APR-2009) Department of Plant Pathology, The Ohio State
University, 201 Kottman Hall, 2021 Coffey Rd, Columbus, OH 43210,
USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..12035
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 90..106
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
terminator complement(115..367)
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
CDS 728..1030
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
CDS complement(1089..1112)
/label=FLAG
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
regulatory complement(1128..3118)
/label=maize ubiquitin-1 promoter
/note="maize ubiquitin-1 promoter"
/regulatory_class="promoter"
promoter complement(1128..3118)
/label=Ubi promoter
/note="maize polyubiquitin gene promoter"
primer_bind complement(3140..3156)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 3359..3383
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
CDS 4682..5308
/label=pVS1 StaA
/note="stability protein from the Pseudomonas plasmid pVS1
(Heeb et al., 2000)"
CDS 5745..6809
/label=pVS1 RepA
/note="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
rep_origin 6878..7072
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
misc_feature 7416..7556
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(7742..8330)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(8420..9211)
/label=KanR
/note="aminoglycoside phosphotransferase"
misc_feature 9636..9660
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
polyA_signal complement(9738..9912)
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
CDS complement(9955..10977)
/label=HygR
/note="aminoglycoside phosphotransferase from E. coli"
promoter complement(11044..11721)
/label=CaMV 35S promoter (enhanced)
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
protein_bind 11912..11933
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 11948..11978
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 11986..12002
/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)."
primer_bind 12010..12026
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"