Price Information
Cat No. | Plasmid Name | Availability | Add to cart |
---|---|---|---|
V011387 | pLSLZ | 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:
- pLSLZ
- Antibiotic Resistance:
- Kanamycin
- Length:
- 15655 bp
- Type:
- Plant Expression Vectors
- Replication origin:
- ori
- Host:
- Plants
- Promoter:
- CaMV 35S (enhanced)
pLSLZ 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.
pLSLZ vector Sequence
LOCUS V011387 15655 bp DNA circular SYN 13-JAN-2022 DEFINITION Exported. ACCESSION V011387 VERSION V011387 KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct . REFERENCE 1 (bases 1 to 15655) TITLE Direct Submission REFERENCE 2 (bases 1 to 15655) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article" FEATURES Location/Qualifiers source 1..15655 /mol_type="other DNA" /organism="synthetic DNA construct" protein_bind 529..553 /label="attB1" /note="recombination site for the Gateway(R) BP reaction" CDS 587..607 /label="SV40 NLS" /note="nuclear localization signal of SV40 (simian virus 40) large T antigen" CDS 641..817 /gene="EGR1" /label="Early growth response protein 1" /note="Early growth response protein 1 from Serinus canaria. Accession#: O73694" CDS 875..1462 /label="FokI cleavage domain" /note="nonspecific DNA cleavage domain of the FokI endonuclease (Li et al., 1992)" terminator 1476..1728 /label="NOS terminator" /note="nopaline synthase terminator and poly(A) signal" protein_bind 1773..1793 /label="attB5" /note="core recombination site for the Gateway(R) BP reaction" CDS 2824..3612 /label="NeoR/KanR" /note="aminoglycoside phosphotransferase" terminator 3642..3894 /note="NOS terminator" /note="nopaline synthase terminator and poly(A) signal" protein_bind complement(4907..4931) /label="attB2" /note="recombination site for the Gateway(R) BP reaction" promoter 5697..6041 /label="CaMV 35S promoter" /note="strong constitutive promoter from cauliflower mosaic virus" primer_bind complement(6752..6768) /label="M13 rev" /note="M13 rev" /note="common sequencing primer, one of multiple similar variants" protein_bind 6776..6792 /label="lac repressor encoded by lacI binding site" /bound_moiety="lac repressor encoded by lacI" /note="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)." promoter complement(6800..6830) /label="lac promoter" /note="promoter for the E. coli lac operon" protein_bind complement(6845..6866) /label="CAP binding site" /note="CAP binding activates transcription in the presence of cAMP." promoter 7057..7734 /label="CaMV 35S promoter (enhanced)" /note="cauliflower mosaic virus 35S promoter with a duplicated enhancer region" CDS 7802..8824 /label="HygR" /note="aminoglycoside phosphotransferase from E. coli" polyA_signal 8868..9042 /label="CaMV poly(A) signal" /note="cauliflower mosaic virus polyadenylation signal" misc_feature complement(9120..9144) /label="LB T-DNA repeat" /note="left border repeat from nopaline C58 T-DNA" CDS 9569..10360 /label="KanR" /note="aminoglycoside phosphotransferase" rep_origin 10450..11038 /label="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" misc_feature complement(11224..11364) /label="bom" /note="basis of mobility region from pBR322" rep_origin complement(11708..11902) /direction=LEFT /label="pVS1 oriV" /note="origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" CDS complement(11971..13035) /label="pVS1 RepA" /note="replication protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" CDS complement(13472..14098) /label="pVS1 StaA" /note="stability protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" misc_feature complement(15398..15422) /label="RB T-DNA repeat" /note="right border repeat from nopaline C58 T-DNA" primer_bind 15625..15641 /label="M13 fwd" /note="common sequencing primer, one of multiple similar variants"