Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V012109 | p45sheLL | 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:
- p45sheLL
- Antibiotic Resistance:
- Ampicillin
- Length:
- 10814 bp
- Type:
- Mammalian Expression
- Replication origin:
- ori
- Promoter:
- CMV
- 5' Primer:
- NA
- 3' Primer:
- NA
p45sheLL 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.
p45sheLL vector Sequence
LOCUS V012109 10814 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V012109
VERSION V012109
KEYWORDS p45sheLL
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 10814)
AUTHORS Roberts JN, Buck CB, Thompson CD, Kines R, Bernardo M, Choyke PL,
Lowy DR, Schiller JT
TITLE Genital transmission of HPV in a mouse model is potentiated by
nonoxynol-9 and inhibited by carrageenan.
JOURNAL Nat Med. 2007 Jul;13(7):857-61. Epub 2007 Jul 1.
PUBMED 17603495
REFERENCE 2 (bases 1 to 10814)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 10814)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat Med.";
date: "2007-07"; volume: "13(7)"; pages: "857-61. Epub 2007 Jul 1"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..10814
/mol_type="other DNA"
/organism="synthetic DNA construct"
regulatory 19..28
/label="Kozak sequence"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 2266..3654
/gene="L2"
/label="Minor capsid protein L2"
/note="Minor capsid protein L2 from Human papillomavirus
45. Accession#: P36761"
protein_bind complement(3665..3689)
/label="attB2"
/note="recombination site for the Gateway(R) BP reaction"
misc_feature 3751..4339
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
polyA_signal 4382..4606
/label="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
rep_origin 4652..5080
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5089..5109)
/label="pBABE 3'"
/note="SV40 enhancer, reverse primer for pBABE vectors"
CDS 7038..7403
/label="BleoR"
/note="antibiotic-binding protein"
polyA_signal 7536..7669
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
primer_bind complement(7706..7722)
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(7706..7722)
/label="M13 Reverse"
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(7719..7741)
/label="M13/pUC Reverse"
/note="In lacZ gene"
protein_bind 7730..7746
/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(7754..7784)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(7799..7820)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(7937..7954)
/label="L4440"
/note="L4440 vector, forward primer"
rep_origin complement(8108..8693)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(8867..9724)
/label="AmpR"
/note="beta-lactamase"
promoter complement(9725..9829)
/label="AmpR promoter"
primer_bind complement(9904..9923)
/label="pRS-marker"
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 10095..10474
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 10475..10678
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
protein_bind 10680..10698
/label="tet operator"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 10701..10719
/gene="tetO"
/label="tet operator"
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
primer_bind 10729..10753
/label="LNCX"
/note="Human CMV promoter, forward primer"