pShuttle vector (V003221)

Price Information

Cat No. Plasmid Name Availability Add to cart
V003221 pShuttle In stock, 1 week for quality controls

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

Basic Vector Information

Vector Name:
pShuttle
Antibiotic Resistance:
Kanamycin
Length:
6568 bp
Type:
AdEasy vector
Replication origin:
ori
Source/Author:
Grafsky AJ III.

pShuttle vector Map

pShuttle6568 bp300600900120015001800210024002700300033003600390042004500480051005400570060006300left inverted terminal repeat (LITR)Ad5 PsiMCSHexon-interlacing proteinPackaging protein 1ITRoriNeoR/KanRM13 oriITR

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.

pShuttle vector Sequence

LOCUS       V003221                 6568 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V003221
VERSION     V003221
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6568)
  AUTHORS   Grafsky AJ III.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-JAN-2001) Technical Services, Stratagene, 11011 N.
            Torrey Pines Rd, La Jolla, CA 92037, USA
REFERENCE   2  (bases 1 to 6568)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 6568)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted
            (08-JAN-2001) Technical Services, Stratagene, 11011 N. Torrey Pines
            Rd, La Jolla, CA 92037, USA"
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6568
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   4..106
                     /label="left inverted terminal repeat (LITR)"
                     /note="left inverted terminal repeat (LITR)"
     misc_signal     170..320
                     /label="Ad5 Psi"
                     /note="packaging signal for adenovirus serotype 5"
     misc_feature    359..418
                     /label="MCS"
                     /note="MCS"
     CDS             467..886
                     /gene="IX"
                     /label="Hexon-interlacing protein"
                     /note="Hexon-interlacing protein from Human adenovirus C
                     serotype 5. Accession#: P03281"
     CDS             complement(952..2286)
                     /gene="IVa2"
                     /label="Packaging protein 1"
                     /note="Packaging protein 1 from Human adenovirus C serotype
                     5. Accession#: P03271"
     repeat_region   3533..3635
                     /label="ITR"
                     /note="inverted terminal repeat of human adenovirus
                     serotype 5"
     rep_origin      complement(3888..4476)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             5289..6080
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     rep_origin      complement(6131..6510)
                     /direction=LEFT
                     /label="M13 ori"
                     /note="M13 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     repeat_region   6548..6568
                     /label="ITR"
                     /note="inverted terminal repeat of human adenovirus
                     serotype 5"