Price Information
| Cat No. | Plasmid Name | Availability | Buy one, get one free! (?) |
|---|---|---|---|
| V003849 | pQLinkN | In stock, instant shipping |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pQLinkN
- Antibiotic Resistance:
- Ampicillin
- Length:
- 4823 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Scheich C, Kummel D, Soumailakakis D, Heinemann U, Bussow K.
- Growth Strain(s):
- Top10
pQLinkN 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.
pQLinkN vector Sequence
LOCUS 40924_36233 4823 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pQLinkN, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4823)
AUTHORS Scheich C, Kummel D, Soumailakakis D, Heinemann U, Bussow K.
TITLE Vectors for co-expression of an unrestricted number of proteins
JOURNAL Nucleic Acids Res. 35 (6), E43 (2007)
PUBMED 17311810
REFERENCE 2 (bases 1 to 4823)
AUTHORS Kuemmel D, Scheich C, Buessow K.
TITLE Direct Submission
JOURNAL Submitted (22-DEC-2006) Vertebrate Genomics, Max Planck Institute
for Molecular Genetics, Ihnestr. 63-73, Berlin 14195, Germany
REFERENCE 3 (bases 1 to 4823)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 4823)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic
Acids Res."; date: "2007"; volume: "35"; issue: "6"; pages: "E43"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(22-DEC-2006) Vertebrate Genomics, Max Planck Institute for
Molecular Genetics, Ihnestr. 63-73, Berlin 14195, Germany"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..4823
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 7..27
/label=LIC cassette with PacI restriction site
/note="LIC cassette with PacI restriction site"
promoter 36..80
/label=T5 promoter
/note="bacteriophage T5 promoter for E. coli RNA
polymerase, with embedded lac operator"
protein_bind 88..104
/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 123..134
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
regulatory 127..133
/regulatory_class="ribosome_binding_site"
misc_feature 141
/note="translation start site for inserts cloned into the
multiple cloning site"
protein_bind complement(184..208)
/label=attB2
/note="recombination site for the Gateway(R) BP reaction"
terminator 230..324
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
misc_feature 329..378
/label=LIC cassette with SwaI, PacI restriction sites
/note="LIC cassette with SwaI, PacI restriction sites"
CDS 418..1074
/label=CmR
/note="chloramphenicol acetyltransferase"
terminator 1142..1228
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
protein_bind complement(1287..1308)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
CDS complement(1324..2403)
/label=lacI
/note="lac repressor"
promoter complement(2404..2481)
/label=lacIq promoter
/note="In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about 10-fold."
misc_feature 2673..2813
/label=bom
/note="basis of mobility region from pBR322"
rep_origin 2997
/label=ColE1 ori
/note="ColE1 ori"
rep_origin complement(2999..3587)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(3761..4618)
/label=AmpR
/note="beta-lactamase"
promoter complement(4619..4723)
/label=AmpR promoter