Price Information
| Cat No. | Plasmid Name | Availability | Buy one, get one free! (?) |
|---|---|---|---|
| V017641 | pBAD33-Tet | In stock, 1 week for quality controls |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pBAD33-Tet
- Antibiotic Resistance:
- Chloramphenicol, Tetracycline
- Length:
- 6774 bp
- Type:
- Protein expression, Tetracycline inducible
- Replication origin:
- p15A ori
- Host:
- E. coli
- Promoter:
- BAD
- 5' Primer:
- PBAD30.F
- Growth Strain(s):
- DH5a
- Growth Temperature:
- 37℃
pBAD33-Tet vector Map
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.
pBAD33-Tet vector Sequence
LOCUS . 6774 bp DNA circular UNK 01-JAN-1980
DEFINITION synthetic circular DNA.
ACCESSION
VERSION
KEYWORDS .
SOURCE .
ORGANISM .
.
FEATURES Location/Qualifiers
CDS complement(99..974)
/label="araC"
/note="L-arabinose regulatory protein"
promoter 1001..1285
/label="araBAD promoter"
/note="promoter of the L-arabinose operon of E. coli; the
araC regulatory gene is transcribed in the opposite
direction (Guzman et al., 1995)"
misc_feature 1306..1362
/label="MCS"
/note="pUC18/19 multiple cloning site"
terminator 1565..1651
/label="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 1743..1770
/label="rrnB T2 terminator"
/note="transcription terminator T2 from the E. coli rrnB
gene"
promoter 1789..1880
/label="AmpR promoter"
promoter 2130..2234
/label="AmpR promoter"
CDS 2309..3496
/label="TcR"
/note="tetracycline efflux protein"
rep_origin 3756..4211
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
CDS complement(4769..5425)
/label="CmR"
/note="chloramphenicol acetyltransferase"
promoter complement(5426..5528)
/label="cat promoter"
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
rep_origin complement(6054..6599)
/label="p15A ori"
/note="Plasmids containing the medium-copy-number p15A
origin of replication can be propagated in E. coli cells
that contain a second plasmid with the ColE1 origin."