Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V017316 | pN565 | 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:
- pN565
- Antibiotic Resistance:
- Streptomycin
- Length:
- 9856 bp
- Type:
- Protein expression
- Replication origin:
- pSa ori
- Host:
- E. coli
- Copy Number:
- Low
- Promoter:
- T7
- Growth Temperature:
- 37℃
pN565 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.
pN565 vector Sequence
LOCUS V017316 9856 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V017316
VERSION V017316
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 9856)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..9856
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 127..2772
/note="T7 RNA polymerase from Escherichia phage T7.
Accession#: P00573"
/label="T7 RNA polymerase"
misc_feature 2776..2796
/label="BioBrick suffix"
/note="universal suffix for all parts"
terminator 2832..2863
/label="tonB terminator"
/note="bidirectional E. coli tonB-P14 transcription
terminator"
CDS 3725..4693
/label="RepA"
/note="replication protein of plasmid pSa"
rep_origin 4733..5168
/label="pSa ori"
/note="origin of replication from bacterial plasmid pSa"
CDS 6843..7631
/label="SmR"
/note="aminoglycoside adenylyltransferase (Murphy, 1985)"
terminator 7792..7851
/label="his operon terminator"
/note="This putative transcriptin terminator from the E.
coli his operon has a 2-bp deletion introduced during
synthesis. Its efficiency has not been determined."
terminator complement(7911..7940)
/label="T3Te terminator"
/note="phage T3 early transcription terminator"
CDS complement(8046..9125)
/label="lacI"
/note="lac repressor"
promoter complement(9126..9203)
/label="lacIq promoter"
/note="In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about 10-fold."
promoter 9722..9750
/label="tac promoter"
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind complement(9756..9775)
/label="lac operator (symmetric)"
/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). The
symmetric lac operator was optimized for tight binding of
lac repressor."