Price Information
| Cat No. | Plasmid Name | Availability | Buy one, get one free! (?) |
|---|---|---|---|
| V009690 | pA2 | In stock, instant shipping |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pA2
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5143 bp
- Type:
- Phagemid cloning vector
- Replication origin:
- ori
- Source/Author:
- Fagerlund A, Myrset AH, Kulseth MA.
- Promoter:
- araBAD
- Growth Strain(s):
- DH10B
- Growth Temperature:
- 37℃
pA2 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.
pA2 vector Sequence
LOCUS 40924_2987 5143 bp DNA circular SYN 17-DEC-2018
DEFINITION Phagemid cloning vector pA2, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5143)
AUTHORS Fagerlund A, Myrset AH, Kulseth MA.
TITLE Construction and characterization of a 9-mer phage display
pVIII-library with regulated peptide density
JOURNAL Appl. Microbiol. Biotechnol. 80 (5), 925-936 (2008)
PUBMED 18716770
REFERENCE 2 (bases 1 to 5143)
AUTHORS Fagerlund A, Myrset AH, Kulseth MA.
TITLE Direct Submission
JOURNAL Submitted (12-FEB-2008) GE Healthcare Medical Diagnostics, P.O. Box
4220 Nydalen, Oslo N-0401, Norway
REFERENCE 3 (bases 1 to 5143)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 5143)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Appl.
Microbiol. Biotechnol."; date: "2008"; volume: "80"; issue: "5";
pages: "925-936"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(12-FEB-2008) GE Healthcare Medical Diagnostics, P.O. Box 4220
Nydalen, Oslo N-0401, Norway"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5143
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 1..285
/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)"
gene 318..891
/gene="gVIII"
/label=gVIII
CDS 318..401
/codon_start=1
/gene="gVIII"
/product="major coat protein pVIII"
/label=gVIII
/protein_id="ACA60871.1"
/translation="MKKSLVLKASVAVATLVPMLSFAAEGEF"
sig_peptide 318..398
/gene="gVIII"
misc_feature 396..401
/gene="gVIII"
/label=unique EcoRI cloning site
/note="unique EcoRI cloning site"
misc_feature 402..749
/gene="gVIII"
/note="spacer fragment; derived from lambda phage"
misc_feature 750..755
/gene="gVIII"
/label=unique BglII cloning site
/note="unique BglII cloning site"
CDS 751..891
/codon_start=1
/gene="gVIII"
/product="major coat protein pVIII"
/label=gVIII
/protein_id="ACA60872.1"
/translation="DLAKAAFDSLQASATEYIGYAWAMVVVIVGATIGIKLFKKFTSKA
S"
primer_bind complement(891..907)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 1348..1861
/label=M13 ori
/note="M13 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 1925..2029
/label=AmpR promoter
CDS 2030..2887
/label=AmpR
/note="beta-lactamase"
rep_origin 3061..3649
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature complement(3835..3975)
/label=bom
/note="basis of mobility region from pBR322"
CDS complement(4242..5117)
/label=araC
/note="L-arabinose regulatory protein"