pFS102-SV40-Galpha15 vector (V006187)

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Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V006187	pFS102-SV40-Galpha15	In stock, instant shipping	Buy one, get one free!

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

Basic Vector Information

Vector Name:: pFS102-SV40-Galpha15

Antibiotic Resistance:: Ampicillin

Length:: 4714 bp

Type:: Cloning vector

Replication origin:: ori

Source/Author:: Sedlmayer F, Hell D, Mueller M, Auslaender D, Fussenegger M.

pFS102-SV40-Galpha15 vector Map

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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.

pFS102-SV40-Galpha15 vector Sequence

LOCUS       V006187                 4714 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V006187
VERSION     V006187
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 4714)
  AUTHORS   Sedlmayer F, Hell D, Mueller M, Auslaender D, Fussenegger M.
  TITLE     Designer cells programming quorum-sensing interference with human
            pathogens
  JOURNAL   Nat Commun (2018) In press
REFERENCE   2  (bases 1 to 4714)
  AUTHORS   Sedlmayer F, Hell D, Mueller M, Auslaender D, Fussenegger M.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-AUG-2017) Department of Biosystems Science and
            Engineering (D-BSSE), ETH Zurich, Mattenstrasse 26, Baseel 4058,
            Switzerland
REFERENCE   3  (bases 1 to 4714)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4714)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "Nat Commun
            (2018) In press"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (21-AUG-2017) Department of Biosystems Science and Engineering
            (D-BSSE), ETH Zurich, Mattenstrasse 26, Baseel 4058, Switzerland"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4714
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             15..1136
                     /gene="GNA15"
                     /label="Guanine nucleotide-binding protein subunit
                     alpha-15"
                     /note="Guanine nucleotide-binding protein subunit alpha-15
                     from Homo sapiens. Accession#: P30679"
     polyA_signal    complement(1186..1307)
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(1972..2560)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(2734..3591)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(3592..3696)
                     /label="AmpR promoter"
     rep_origin      3723..4178
                     /direction=RIGHT
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     polyA_signal    4309..4357
                     /label="poly(A) signal"
                     /note="synthetic polyadenylation signal"
     misc_feature    4371..4462
                     /label="pause site"
                     /note="RNA polymerase II transcriptional pause signal from
                     the human alpha-2 globin gene"
     promoter        4517..4713
                     /label="SV40 promoter"
                     /note="SV40 early promoter"