pFA6a-VN-kanMX6 vector (V006307)

Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V006307	pFA6a-VN-kanMX6	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:: pFA6a-VN-kanMX6

Antibiotic Resistance:: Ampicillin

Length:: 4705 bp

Type:: Yeast tagging vector

Replication origin:: ori

Host:: Yeast

Source/Author:: Sung MK, Huh WK.

Promoter:: TEF

pFA6a-VN-kanMX6 vector Vector Map

View Map in NovoBuilder

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.

pFA6a-VN-kanMX6 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_19651        4705 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Yeast tagging vector pFA6a-VN-kanMX6, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4705)
  AUTHORS   Sung MK, Huh WK.
  TITLE     Bimolecular fluorescence complementation analysis system for in vivo
            detection of protein-protein interaction in Saccharomyces cerevisiae
  JOURNAL   Yeast 24 (9), 767-775 (2007)
  PUBMED    17534848
REFERENCE   2  (bases 1 to 4705)
  AUTHORS   Sung M-K., Huh W-K.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-JAN-2007) School of Biological Sciences, Seoul 
            National University, Seoul 151-747, Republic of Korea
REFERENCE   3  (bases 1 to 4705)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4705)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Yeast"; 
            date: "2007"; volume: "24"; issue: "9"; pages: "767-775"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (09-JAN-2007) School of Biological Sciences, Seoul National 
            University, Seoul 151-747, Republic of Korea"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4705
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             78..596
                     /codon_start=1
                     /label=VN173
                     /note="N-terminal fragment of mVenus for use in bimolecular
                     fluorescence complementation (BiFC) (Kodama and Hu, 2010)"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIE"
     terminator      620..807
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     gene            882..2238
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin
                     resistance (Wach et al., 1994)"
     promoter        complement(2343..2361)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(2619..3207)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3381..4238)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4239..4343)
                     /label=AmpR promoter
     promoter        4689..4705
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"