YEp353 vector (V010187)

Price Information

Cat No. Plasmid Name Availability Add to cart
V010187 YEp353 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:
YEp353
Antibiotic Resistance:
Ampicillin
Length:
7944 bp
Type:
Yeast Plasmids
Replication origin:
ori
Host:
Yeast
Source/Author:
Myers AM, Tzagoloff A, Kinney DM, Lusty CJ.
Copy Number:
High copy number
Promoter:
URA3
Growth Strain(s):
Top10
Growth Temperature:
37℃

YEp353 vector Map

YEp3537944 bp30060090012001500180021002400270030003300360039004200450048005100540057006000630066006900720075007800MCSlacZURA3URA3 promoter2u oriAmpR promoterAmpRori

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.

YEp353 vector Sequence

LOCUS       YEp353.        7944 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Yeast episomal vector with a URA3 marker for construction of lacZ 
            fusions. For other reading frames, use YEp354 or YEp355.
ACCESSION   .
VERSION     .
KEYWORDS    YEp353
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7944)
  AUTHORS   Myers AM, Tzagoloff A, Kinney DM, Lusty CJ.
  TITLE     Yeast shuttle and integrative vectors with multiple cloning sites 
            suitable for construction of lacZ fusions.
  JOURNAL   Gene 1986;45:299-310.
  PUBMED    3026915
REFERENCE   2  (bases 1 to 7944)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 7944)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Gene"; 
            date: "1986"; volume: "45"; pages: "299-310"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     The GenBank sequence was edited at base 3044 to eliminate a second 
            EcoRI site.
FEATURES             Location/Qualifiers
     source          1..7944
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..36
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             45..3092
                     /label=lacZ
                     /note="beta-galactosidase"
     CDS             complement(3293..4093)
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     promoter        complement(4094..4304)
                     /label=URA3 promoter
     rep_origin      complement(4541..5705)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        5987..6091
                     /label=AmpR promoter
     CDS             6092..6949
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      7123..7711
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"