p18sheLL vector (V011769)

Price Information

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V011769 p18sheLL In stock, 1 week for quality controls

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Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.

Basic Vector Information

Vector Name:
p18sheLL
Antibiotic Resistance:
Ampicillin
Length:
10723 bp
Type:
Mammalian Expression
Replication origin:
ori
Promoter:
CMV
5' Primer:
NA
3' Primer:
NA

p18sheLL vector Map

p18sheLL10723 bp5001000150020002500300035004000450050005500600065007000750080008500900095001000010500Kozak sequenceMinor capsid protein L2WPREbGH poly(A) signalf1 oripBABE 3'BleoRSV40 poly(A) signalIn lacZ genelac promoterCAP binding siteL4440oriAmpRAmpR promoterpRS-markerCMV enhancerCMV promotertet operatortet operatorLNCX

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.

p18sheLL vector Sequence

LOCUS       V011769                10723 bp    DNA     circular SYN 13-MAY-2021
DEFINITION  Exported.
ACCESSION   V011769
VERSION     V011769
KEYWORDS    p18sheLL
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 10723)
  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 10723)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 10723)
  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..10723
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      7..16
                     /label="Kozak sequence"
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2229..3614
                     /gene="L2"
                     /label="Minor capsid protein L2"
                     /note="Minor capsid protein L2 from Human papillomavirus
                     type 18. Accession#: P06793"
     misc_feature    3659..4247
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     polyA_signal    4290..4514
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4560..4988
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4997..5017)
                     /label="pBABE 3'"
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             6946..7311
                     /label="BleoR"
                     /note="antibiotic-binding protein"
     polyA_signal    7444..7577
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7614..7630)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     primer_bind     complement(7614..7630)
                     /label="M13 Reverse"
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7627..7649)
                     /label="M13/pUC Reverse"
                     /note="In lacZ gene"
     protein_bind    7638..7654
                     /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(7662..7692)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(7707..7728)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     primer_bind     complement(7845..7862)
                     /label="L4440"
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8016..8601)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(8775..9632)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(9633..9737)
                     /label="AmpR promoter"
     primer_bind     complement(9812..9831)
                     /label="pRS-marker"
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
     enhancer        10003..10382
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        10383..10586
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     protein_bind    10588..10606
                     /label="tet operator"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    10609..10627
                     /gene="tetO"
                     /label="tet operator"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     10637..10661
                     /label="LNCX"
                     /note="Human CMV promoter, forward primer"