pFCcGi vector (V000516)

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Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V000516	pFCcGi	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:: pFCcGi

Antibiotic Resistance:: Ampicillin

Length:: 7471 bp

Type:: Bacterial Expression

Replication origin:: ori

Promoter:: araBAD

5' Primer:: PBAD-F

3' Primer:: PBAD-R

pFCcGi 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.

pFCcGi vector Sequence

LOCUS       V000516                 7471 bp    DNA     circular SYN 22-DEC-2021
DEFINITION  Exported.
ACCESSION   V000516
VERSION     V000516
KEYWORDS    pFCcGi
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7471)
  AUTHORS   Figueira R, Watson KG, Holden DW, Helaine S
  TITLE     Identification of salmonella pathogenicity island-2 type III
            secretion system effectors involved in intramacrophage replication
            of S. enterica serovar typhimurium: implications for rational
            vaccine design.
  JOURNAL   MBio. 2013 Apr 16;4(2):e00065. doi: 10.1128/mBio.00065-13.
   PUBMED   23592259
REFERENCE   2  (bases 1 to 7471)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 7471)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "MBio.";
            date: "2013-04-16"; pages: "
            10.1128/mBio.00065-13"
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7471
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             41..748
                     /label="mCherry"
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     CDS             complement(1016..1891)
                     /label="araC"
                     /note="L-arabinose regulatory protein"
     promoter        1918..2202
                     /label="araBAD promoter"
                     /note="promoter of the L-arabinose operon of E. coli; the
                     araC regulatory gene is transcribed in the opposite
                     direction (Guzman et al., 1995)"
     CDS             2280..2993
                     /label="yeGFP"
                     /note="yeast-enhanced green fluorescent protein"
     oriT            3472..3559
                     /label="RSF1010 oriT"
                     /note="origin of transfer of the broad-host-range plasmid
                     RSF1010 (Scholz et al., 1989)"
     primer_bind     complement(4872..4889)
                     /label="L4440"
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5043..5631)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(5805..6662)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(6663..6767)
                     /label="AmpR promoter"
     primer_bind     6835..6853
                     /label="pBRforEco"
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"
     CDS             7096..7209
                     /gene="rpmJ"
                     /label="Large ribosomal subunit protein bL36A"
                     /note="Large ribosomal subunit protein bL36A from
                     Escherichia coli (strain K12). Accession#: P0A7Q6"