Price Information
| Cat No. | Plasmid Name | Availability | Buy one, get one free! (?) |
|---|---|---|---|
| V012891 | pVE5523 | In stock, instant shipping |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
pVE5523 is a secretory expression plasmid for Lactobacillus.
- Vector Name:
- pVE5523
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8315 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Lactic acid bacteria
- Selection Marker:
- Ery
- Promoter:
- P59
- Growth Strain(s):
- DH10B
- Growth Temperature:
- 37℃
pVE5523 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.
pVE5523 vector Sequence
LOCUS Exported 8315 bp DNA circular SYN 23-APR-2025
DEFINITION Exported.
ACCESSION V012891
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8315)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 8315)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 8315)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..8315
/mol_type="other DNA"
/organism="synthetic DNA construct"
source 5550..5553
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 831..2321
/codon_start=1
/label=AMP-binding protein
/translation="MNIPFVVETVLHDGLLKYKFKNSKIRSITTKPGKSKGAIFAYRSK
SSMIGGRGVVLTSEEAIQENQDTFTHWTPNVYRYGTYADENRSYTKGHSENNLRQINTF
FIDFDIHTAKETISASDILTTAIDLGFMPTMIIKSDKGYQAYFVLETPVYVTSKSEFKS
VKAAKIISQNIREYFGKSLPVDLTCNHFGIARIPRTDNVEFFDPNYRYSFKEWQDWSFK
QTDNKGFTRSSLTVLSGTEGKKQVDEPWFNLLLHETKFSGEKGLIGRNNVMFTLSLAYF
SSGYSIETCEYNMFEFNNRLDQPLEEKEVIKIVRSAYSENYQGANREYITILCKAWVSS
DLTSKDLFVRQGWFKFKKKRSERQRVHLSEWKEDLMAYISEKSDVYKPYLVTTKKEIRE
VLGIPERTLDKLLKVLKANQEIFFKIKPGRNGGIQLASVKSLLLSIIKVKKEEKESYIK
ALTNSFDLEHTFIQETLNKLAERPKTDTQLDLFSYDTG"
primer_bind complement(2818..2834)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 3308..3412
/label=AmpR promoter
CDS 3413..4270
/label=AmpR
/note="beta-lactamase"
rep_origin 4444..5032
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 5320..5341
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 5356..5386
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5394..5410
/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 5418..5434
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 5455..5473
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 5549..5659
/label=P59
/note="Streptococcus cremoris promoter 59"
promoter 5550..5553
/label=P59
/note="Streptococcus cremoris promoter 59"
misc_feature 5624..5629
/label=minus_35_signal
misc_feature 5647..5652
/label=minus_10_signal
CDS 5737..5817
/codon_start=1
/label=signal peptide of Usp45
/translation="MKKKIISAILMSTVILSAAAPLSGVYA"
CDS 5863..6303
/label=MNase
/note="micrococcal nuclease from Staphylococcus aureus"
terminator 6342..6428
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 6447..6474
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
CDS 7091..7825
/gene="ermBP"
/label=rRNA adenine N-6-methyltransferase
/note="rRNA adenine N-6-methyltransferase from Enterococcus
faecalis. Accession#: P0A4D5"