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pDG364 vector (V013018) Gene synthesis in pDG364 backbone

Price Information

Cat No. Plasmid Name Availability Buy one, get one free! (?)
V013018 pDG364 In stock, instant shipping

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

Basic Vector Information

The plasmid pDG364 is a protein expression vector designed for use in Bacillus subtilis. With a size of 6257 bp, it features an ampicillin resistance gene (AmpR) for selection in prokaryotic systems and is primarily used for recombinant protein expression in its bacterial host.

Vector Name:
pDG364
Antibiotic Resistance:
Ampicillin
Length:
6277 bp
Type:
Protein expression
Replication origin:
ori
Host:
Bacillus subtilis
Selection Marker:
Chl
Growth Strain(s):
DH10B
Growth Temperature:
37℃

pDG364 vector Map

Copy Sequence View Map
pDG3646277 bp30060090012001500180021002400270030003300360039004200450048005100540057006000Chloramphenicol acetyltransferaseamyE-RoriAmpRAmpR promoteramyE-F

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.

References

  • Li J, Yang M, Chen B, Wang Z, Cao Y, Yang Y, Zhang M, Zhang D, Ni X, Zeng Y, Pan K. Evaluation of the Immunity Responses in Mice to Recombinant Bacillus subtilis Displaying Newcastle Disease Virus HN Protein Truncations. Microorganisms. 2024 Feb 21;12(3):439. doi: 10.3390/microorganisms12030439. PMID: 38543490; PMCID: PMC10972176.

pDG364 vector Sequence

LOCUS       Exported                6277 bp DNA     circular SYN 13-OCT-2025
DEFINITION  Exported.
ACCESSION   V013018
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6277)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6277)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 6277)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6277
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             226..873
                     /gene="cat"
                     /label=Chloramphenicol acetyltransferase
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     CDS             1256..2272
                     /codon_start=1
                     /label=amyE-R
                     /label=alpha-amylase
                     /translation="MSDDDIRLGWAVIASRSGSTPLFFSRPEGGGNGVRFPGKSQIGDR
                     GSALFEDQAITAVNRFHNVMAGQPEELSNPNGNNQIFMNQRGSHGVVLANAGSSSVSIN
                     TATKLPDGRYDNKAGAGSFQVNDGKLTGTINARSVAVLYPDDIAKAPHVFLENYKTGVT
                     HSFNDQLTITLRADANTTKAVYQINNGPETAFKDGDQFTIGKGDPFGKTYTIMLKGTNS
                     DGVTRTEKYSFVKRDPASAKTIGYQNPNHWSQVNAYIYKHDGSRVIELTGSWPGKPMTK
                     NADGIYTLTLPADTDTTNAKVIFNNGSAQVPGQNQPGFDYVLNGLYNDSGLSGSLPH"
     rep_origin      complement(3577..4165)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4339..5196)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5197..5301)
                     /label=AmpR promoter
     CDS             5521..6063
                     /codon_start=1
                     /label=amyE-F
                     /translation="MFAKRFKTSLLPLFAGFLLLFHLVLAGPAAASAETANKSNELTAP
                     SIKSGTILHAWNWSFNTLKHNMKDIHDAGYTAIQTSPINQVKEGNQGDKSMSNWYWLYQ
                     PTSYQIGNRYLGTEQEFKEMCAAAEEYGIKVIVDAVINHTTSDYAAISNEVKSIPNWTH
                     GNTQIKNWSDRWDVTQN"