Price Information
Cat No. | Plasmid Name | Availability | Add to cart |
---|---|---|---|
V005891 | p58sheLL | 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:
- p58sheLL
- Antibiotic Resistance:
- Ampicillin
- Length:
- 10730 bp
- Type:
- Mammalian Expression
- Replication origin:
- ori
- Promoter:
- CMVd2
- 5' Primer:
- NA
- 3' Primer:
- NA
p58sheLL 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.
p58sheLL vector Sequence
LOCUS V005891 10730 bp DNA circular SYN 13-MAY-2021 DEFINITION Exported. ACCESSION V005891 VERSION V005891 KEYWORDS p58sheLL SOURCE synthetic DNA construct ORGANISM synthetic DNA construct . REFERENCE 1 (bases 1 to 10730) AUTHORS Kondo K, Ishii Y, Ochi H, Matsumoto T, Yoshikawa H, Kanda T TITLE Neutralization of HPV16, 18, 31, and 58 pseudovirions with antisera induced by immunizing rabbits with synthetic peptides representing segments of the HPV16 minor capsid protein L2 surface region. JOURNAL Virology. 2007 Feb 20;358(2):266-72. Epub 2006 Sep 28. PUBMED 17010405 REFERENCE 2 (bases 1 to 10730) TITLE Direct Submission REFERENCE 3 (bases 1 to 10730) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Virology."; date: "2007-02-20"; volume: "358(2)"; pages: "266-72. Epub 2006 Sep 28" SGRef: number: 2; type: "Journal Article" FEATURES Location/Qualifiers source 1..10730 /mol_type="other DNA" /organism="synthetic DNA construct" CDS 20..1513 /gene="L1" /label="Major capsid protein L1" /note="Major capsid protein L1 from Human papillomavirus 58. Accession#: P26535" CDS 2200..3615 /gene="L2" /label="Minor capsid protein L2" /note="Minor capsid protein L2 from Human papillomavirus 58. Accession#: P26538" misc_feature 3667..4255 /label="WPRE" /note="woodchuck hepatitis virus posttranscriptional regulatory element" polyA_signal 4298..4522 /label="bGH poly(A) signal" /note="bovine growth hormone polyadenylation signal" rep_origin 4568..4996 /label="f1 ori" /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5005..5025) /label="pBABE 3'" /note="SV40 enhancer, reverse primer for pBABE vectors" CDS 6954..7319 /label="BleoR" /note="antibiotic-binding protein" polyA_signal 7452..7585 /label="SV40 poly(A) signal" /note="SV40 polyadenylation signal" primer_bind complement(7622..7638) /label="M13 rev" /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7622..7638) /label="M13 Reverse" /note="In lacZ gene. Also called M13-rev" primer_bind complement(7635..7657) /label="M13/pUC Reverse" /note="In lacZ gene" protein_bind 7646..7662 /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)." promoter complement(7670..7700) /label="lac promoter" /note="promoter for the E. coli lac operon" protein_bind complement(7715..7736) /label="CAP binding site" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(7853..7870) /label="L4440" /note="L4440 vector, forward primer" rep_origin complement(8024..8609) /direction=LEFT /label="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(8783..9640) /label="AmpR" /note="beta-lactamase" promoter complement(9641..9745) /label="AmpR promoter" primer_bind complement(9820..9839) /label="pRS-marker" /note="pRS vectors, use to sequence yeast selectable marker" enhancer 10011..10390 /label="CMV enhancer" /note="human cytomegalovirus immediate early enhancer" promoter 10391..10594 /label="CMV promoter" /note="human cytomegalovirus (CMV) immediate early promoter" protein_bind 10596..10614 /label="tet operator" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 10617..10635 /gene="tetO" /label="tet operator" /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind 10645..10669 /label="LNCX" /note="Human CMV promoter, forward primer"