pAX01-ECFP-GABABR1a VTF-Ypet-2 vector (V013460)

Price Information

Cat No. Plasmid Name Availability Add to cart
V013460 pAX01-ECFP-GABABR1a VTF-Ypet-2 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-ECFP-GABABR1a VTF-Ypet-2
Antibiotic Resistance:
Ampicillin
Length:
10967 bp
Type:
Protein expression
Replication origin:
ori
Host:
Bacillus subtilis
Selection Marker:
Ery
Promoter:
xylA
Growth Strain(s):
DH5a
Growth Temperature:
37℃

pAX01-ECFP-GABABR1a VTF-Ypet-2 vector Vector Map

pAX01-ECFP-GABABR1a VTF-Ypet-210967 bp5001000150020002500300035004000450050005500600065007000750080008500900095001000010500lambda t0 terminatorrrnB T2 terminatorrrnB T1 terminatorrRNA adenine N-6-methyltransferaseAmpR promoterECFPlambda t0 terminatorAmpRoribomrop

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-ECFP-GABABR1a VTF-Ypet-2 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       V013460                10967 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V013460
VERSION     V013460
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 10967)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10967
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(546..580)
                     /label="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"
     CDS             complement(4672..5388)
                     /label="ECFP"
                     /note="enhanced CFP"
     terminator      complement(6860..6894)
                     /label="lambda t0 terminator"
                     /note="minimal transcription terminator from phage lambda
                     (Scholtissek and Grosse, 1987)"
     CDS             7643..8500
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      8674..9262
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     misc_feature    complement(9448..9588)
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     CDS             complement(9693..9881)
                     /label="rop"
                     /note="Rop protein, which maintains plasmids at low copy
                     number"