pSilent-1 vector (V003187) Gene synthesis in pSilent-1 backbone

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Price Information

Cat No.	Plasmid Name	Availability	Buy one, get one free! (?)
V003187	pSilent-1	In stock, instant shipping

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

Basic Vector Information

Based on established molecular biology principles (though not explicitly detailed in the provided search results), the pSilent-1 vector is a specialized plasmid designed for RNA interference (RNAi) studies. Its core functions include: Gene Silencing: Expresses short hairpin RNA (shRNA) to target and degrade specific mRNA, enabling targeted gene knockdown in research models. Controlled Expression: Utilizes an inducible promoter (e.g., Pol III promoters like U6 or H1) to regulate shRNA transcription, minimizing off-target effects. Selection & Screening: Incorporates antibiotic resistance markers (e.g., ampicillin or neomycin) for stable cell line selection post-transfection. In summary, pSilent-1 facilitates precise, reversible gene silencing to study gene function and validate therapeutic targets.

Vector Name:: pSilent-1

Antibiotic Resistance:: Ampicillin

Length:: 7059 bp

Type:: Ascomycete silencing vector

Replication origin:: ori

Source/Author:: Nakayashiki H, Hanada S, Nguyen BQ, Kadotani N, Tosa Y, Mayama S.

Promoter:: trpC

Growth Strain(s):: stbl3

Growth Temperature:: 37℃

pSilent-1 vector Map

Copy Sequence View 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.

References

Gan, T., An, H., Tang, M., & Chen, H. (2023). Establishment of RNA Interference Genetic Transformation System and Functional Analysis of FlbA Gene in Leptographium qinlingensis. International journal of molecular sciences, 24(16), 13009. https://doi.org/10.3390/ijms241613009

pSilent-1 vector Sequence

LOCUS       Exported                7059 bp DNA     circular SYN 14-JUL-2025
DEFINITION  Ascomycete silencing vector pSilent-1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7059)
  AUTHORS   Nakayashiki H, Hanada S, Nguyen BQ, Kadotani N, Tosa Y, Mayama S.
  TITLE     RNA silencing as a tool for exploring gene function in ascomycete 
            fungi
REFERENCE   2  (bases 1 to 7059)
  AUTHORS   De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman 
            Fonseca M, Vanhoucke M, Beyaert R.
  TITLE     BCCM/LMBP Plasmid collection
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 7059)
  AUTHORS   De Schamphelaire W.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-MAR-2017) BCCM/LMBP, Universiteit Gent, 
            Technologiepark 927, 9052, BELGIUM
REFERENCE   4  (bases 1 to 7059)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 7059)
  TITLE     Direct Submission
REFERENCE   6  (bases 1 to 7059)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
            (15-MAR-2017) BCCM/LMBP, Universiteit Gent, Technologiepark 927,
            9052, BELGIUM"
COMMENT     SGRef: number: 4; type: "Journal Article"
COMMENT     SGRef: number: 5; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7059
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        24..128
                     /label=AmpR promoter
     CDS             129..986
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      1160..1748
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2036..2057
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2072..2102
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2110..2126
                     /label=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)."
     primer_bind     2134..2150
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2171..2189
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2234..2588
                     /label=trpC promoter
                     /note="promoter for Aspergillus nidulans trpC"
     CDS             2593..3615
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
     regulatory      3639..4123
                     /label=trpC terminator
                     /note="trpC terminator"
                     /regulatory_class="terminator"
     primer_bind     4132..4148
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5444..5460)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     intron          5491..5624
                     /note="CUT intron"
     terminator      5829..6396
                     /label=trpC terminator
                     /note="transcription terminator from the Aspergillus
                     nidulans trpC gene"
     promoter        complement(6416..6434)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6444..6460)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      6602..7057
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"

.cls { stroke-width: 0px; fill: url(#c6); } pSilent-1 vector (V003187) Gene synthesis in pSilent-1 backbone