pxylA-BmSQS-ScFPP-6×His vector (V014159)

Price Information

Cat No. Plasmid Name Availability Add to cart
V014159 pxylA-BmSQS-ScFPP-6×His In stock, 1 week for quality controls

Buy one, get one free!

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

Basic Vector Information

Vector Name:
pxylA-BmSQS-ScFPP-6×His
Antibiotic Resistance:
Ampicillin
Length:
9748 bp
Type:
Protein expression
Replication origin:
ori
Host:
Bacillus subtilis
Selection Marker:
Erm
Promoter:
xylA
Growth Strain(s):
DH5a
Growth Temperature:
37℃

pxylA-BmSQS-ScFPP-6×His vector Map

pxylA-BmSQS-ScFPP-6×His9748 bp4008001200160020002400280032003600400044004800520056006000640068007200760080008400880092009600ropbomoriAmpRlambda t0 terminatorFarnesyl pyrophosphate synthase6xHisAmpR promoterrRNA adenine N-6-methyltransferaserrnB T1 terminatorrrnB T2 terminator

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.

pxylA-BmSQS-ScFPP-6×His vector Sequence

LOCUS       V014159                 9748 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014159
VERSION     V014159
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9748)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9748
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1087..1275
                     /label="rop"
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     misc_feature    1380..1520
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(1706..2294)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(2468..3325)
                     /label="AmpR"
                     /note="beta-lactamase"
     terminator      4074..4108
                     /label="lambda t0 terminator"
                     /note="minimal transcription terminator from phage lambda
                     (Scholtissek and Grosse, 1987)"
     CDS             6445..7500
                     /gene="ERG20"
                     /label="Farnesyl pyrophosphate synthase"
                     /note="Farnesyl pyrophosphate synthase from Saccharomyces
                     cerevisiae (strain ATCC 204508 / S288c). Accession#:
                     P08524"
     CDS             7501..7518
                     /label="6xHis"
                     /note="6xHis affinity tag"
     promoter        complement(7567..7671)
                     /label="AmpR promoter"
     CDS             8050..8784
                     /gene="ermBP"
                     /label="rRNA adenine N-6-methyltransferase"
                     /note="rRNA adenine N-6-methyltransferase from Enterococcus
                     faecalis. Accession#: P0A4D5"
     terminator      8957..9043
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      9062..9089
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"