pAX01 vector (V000803)

Price Information

Cat No. Plasmid Name Availability Add to cart
V000803 pAX01 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:
pAX01
Antibiotic Resistance:
Ampicillin, Erythromycin
Length:
7781 bp
Type:
Bacillus subtilis expression vectors
Replication origin:
ori

pAX01 vector Vector Map

pAX017781 bp3006009001200150018002100240027003000330036003900420045004800510054005700600063006600690072007500lambda t0 terminatorrrnB T2 terminatorrrnB T1 terminatorrRNA adenine N-6-methyltransferaseAmpR promoterlambda t0 terminatorbla promoterAmpRoribomrop

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.

pAX01 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       V000803                 7781 bp    DNA     circular SYN 13-JAN-2022
DEFINITION  Exported.
ACCESSION   V000803
VERSION     V000803
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7781)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 7781)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7781
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      546..580
                     /note="lambda t0 terminator"
                     /note="minimal transcription terminator from phage lambda
                     (Scholtissek and Grosse, 1987)"
     terminator      complement(660..687)
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(706..792)
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(965..1699)
                     /gene="ermBP"
                     /label="rRNA adenine N-6-methyltransferase"
                     /note="rRNA adenine N-6-methyltransferase from Enterococcus
                     faecalis. Accession#: P0A4D5"
     promoter        2078..2182
                     /label="AmpR promoter"
     terminator      complement(3674..3708)
                     /label="lambda t0 terminator"
                     /note="minimal transcription terminator from phage lambda
                     (Scholtissek and Grosse, 1987)"
     promoter        4352..4456
                     /gene="bla"
                     /label="bla promoter"
                     /note="AmpR promoter"
     CDS             4457..5314
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      5488..6076
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     misc_feature    complement(6262..6402)
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     CDS             complement(6507..6695)
                     /label="rop"
                     /note="Rop protein, which maintains plasmids at low copy
                     number"