pFGC5941 vector (V010852)

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Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V010852	pFGC5941	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:: pFGC5941

Antibiotic Resistance:: Kanamycin

Length:: 11406 bp

Type:: Plant Vectors

Replication origin:: ori

Source/Author:: Kerschen A, Napoli CA, Jorgensen RA, Muller AE.

Copy Number:: High copy number

Promoter:: MAS

Growth Strain(s):: DH10B

Growth Temperature:: 37℃

pFGC5941 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.

pFGC5941 vector Sequence

LOCUS       pFGC5941.       11406 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Agrobacterium binary vector for expressing dsRNA, with two multiple 
            cloning sites separated by an intron.
ACCESSION   .
VERSION     .
KEYWORDS    pFGC5941
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11406)
  AUTHORS   Kerschen A, Napoli CA, Jorgensen RA, Muller AE.
  TITLE     Effectiveness of RNA interference in transgenic plants.
  JOURNAL   FEBS Lett. 2004;566:223-8.
  PUBMED    15147899
REFERENCE   2  (bases 1 to 11406)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 11406)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "FEBS 
            Lett."; date: "2004"; volume: "566"; pages: "223-8"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     The GenBank sequence was corrected by inserting a G at position 
            7514.
FEATURES             Location/Qualifiers
     source          1..11406
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..25
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(111..363)
                     /label=MAS terminator
                     /note="mannopine synthase terminator"
     CDS             complement(376..924)
                     /label=BlpR
                     /note="phosphinothricin acetyltransferase"
     promoter        complement(930..1310)
                     /label=MAS promoter
                     /note="mannopine synthase promoter (Velten et al., 1984)"
     promoter        2305..2650
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     misc_feature    2680..2734
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     misc_feature    2734..2764
                     /label=MCS 1
                     /note="MCS 1"
                     /note="multiple cloning site 1"
     intron          2767..4115
                     /label=chsA intron
                     /note="chalcone synthase A intron from Petunia hybrida"
     misc_feature    4122..4160
                     /label=MCS 2
                     /note="MCS 2"
                     /note="multiple cloning site"
     terminator      4177..4884
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     misc_feature    5128..5152
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             6452..7078
                     /label=pVS1 StaA
                     /note="stability protein from the Pseudomonas plasmid pVS1
                     (Heeb et al., 2000)"
     CDS             7510..8580
                     /label=pVS1 RepA
                     /note="replication protein from the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     rep_origin      8649..8843
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     misc_feature    9187..9327
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(9513..10101)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(10191..10982)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"