pSico vector (V001038)

Price Information

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V001038 pSico In stock, instant shipping

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

Basic Vector Information

pSico is a vector for conditional (Cre-Lox), stable expression of shRNAs for RNA interference in cells and transgenic mice. Addition of Cre TURNS ON shRNA expression.

Vector Name:
pSico
Antibiotic Resistance:
Ampicillin
Length:
7563 bp
Type:
Mammalian Expression, Lentiviral, RNAi, Cre/Lox
Replication origin:
ori
Copy Number:
High Copy
Cloning Method:
Restriction Enzyme
5' Primer:
EGFP-C
Growth Strain(s):
Top10
Growth Temperature:
37℃

pSico vector Map

pSico7563 bp3006009001200150018002100240027003000330036003900420045004800510054005700600063006600690072007500gp41 peptideProtein TatcPPT/CTSmU6-FloxP511CMV enhancerCMV promoterEGFPloxP511KS primerWPREKS primer5' LTR (truncated)oriAmpRAmpR promoterpRS-markerCMV enhancerCMV promoter5' LTR (truncated)HIV-1 PsiRRE

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.

References

  • Ventura A, Meissner A, Dillon CP, McManus M, Sharp PA, Van Parijs L, Jaenisch R, Jacks T. Cre-lox-regulated conditional RNA interference from transgenes. Proc Natl Acad Sci U S A. 2004 Jul 13;101(28):10380-5.

pSico vector Sequence

LOCUS       V001038                 7563 bp    DNA     circular SYN 13-MAY-2021
DEFINITION  Exported.
ACCESSION   V001038
VERSION     V001038
KEYWORDS    pSico
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7563)
  AUTHORS   Ventura A, Meissner A, Dillon CP, McManus M, Sharp PA, Van Parijs L,
            Jaenisch R, Jacks T
  TITLE     Cre-lox-regulated conditional RNA interference from transgenes.
  JOURNAL   Proc Natl Acad Sci U S A. 2004 Jul 13. 101(28):10380-5.
   PUBMED   15240889
REFERENCE   2  (bases 1 to 7563)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 7563)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl
            Acad Sci U S A. 2004 Jul 13. 101(28):10380-5."
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7563
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             337..381
                     /label="gp41 peptide"
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
                     al., 2013)"
     CDS             530..571
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    679..796
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     primer_bind     1086..1106
                     /label="mU6-F"
                     /note="Mouse U6 promoter, forward primer"
     protein_bind    1124..1157
                     /label="loxP511"
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (GTATACAT). loxP511 sites are compatible with each
                     other, but incompatible with wild-type loxP sites."
     enhancer        1257..1560
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1561..1764
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             1809..2525
                     /label="EGFP"
                     /note="enhanced GFP"
     protein_bind    complement(2536..2569)
                     /label="loxP511"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATAC) (Shaw et al., 2021). loxP511 sites are
                     compatible with each other, but incompatible with wild-type
                     loxP sites."
     primer_bind     2596..2612
                     /label="KS primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     misc_feature    2687..3275
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     primer_bind     complement(3278..3294)
                     /label="KS primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     LTR             3804..3984
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(4046..4634)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(4808..5665)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(5666..5770)
                     /label="AmpR promoter"
     primer_bind     complement(5845..5864)
                     /label="pRS-marker"
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
     enhancer        6036..6415
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6417..6615
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     LTR             6633..6813
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6860..6985
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    7482..7563
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."