Price Information
| Cat No. | Plasmid Name | Availability | Buy one, get one free! (?) |
|---|---|---|---|
| V009282 | pBAMD1-4 | In stock, instant shipping |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pBAMD1-4
- Antibiotic Resistance:
- Ampicillin
- Length:
- 4730 bp
- Type:
- Cloning vector
- Replication origin:
- R6K γ ori
- Source/Author:
- Martinez-Garcia E, Aparicio T, de Lorenzo V, Nikel PI.
- Growth Strain(s):
- GT115
pBAMD1-4 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.
pBAMD1-4 vector Sequence
LOCUS 40924_5899 4730 bp DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector pBAMD1-4, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4730)
AUTHORS Martinez-Garcia E, Aparicio T, de Lorenzo V, Nikel PI.
TITLE New transposon tools tailored for metabolic engineering of
gram-negative microbial cell factories
JOURNAL Front Bioeng Biotechnol 2, 46 (2014)
PUBMED 25389526
REFERENCE 2 (bases 1 to 4730)
AUTHORS Martinez-Garcia E, Aparicio T, de Lorenzo V, Nikel PI.
TITLE Direct Submission
JOURNAL Submitted (26-AUG-2014) Systems Biology Program, Centro Nacional de
Biotecnologia (CNB-CSIC), C/Darwin 3, Madrid, Madrid 28049, Spain
REFERENCE 3 (bases 1 to 4730)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 4730)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Front
Bioeng Biotechnol 2, 46 (2014)"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(26-AUG-2014) Systems Biology Program, Centro Nacional de
Biotecnologia (CNB-CSIC), C/Darwin 3, Madrid, Madrid 28049, Spain"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..4730
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 10..28
/label=Tn5 ME
/note="hyperactive mosaic end for Tn5 transposase
recognition (Reznikoff et al., 2004)"
misc_feature 62..118
/label=MCS
/note="pUC18/19 multiple cloning site"
regulatory 127..156
/label=terminator T500
/note="terminator T500"
/regulatory_class="terminator"
CDS 362..1150
/label=SmR
/note="aminoglycoside adenylyltransferase (Murphy, 1985)"
regulatory 1164..1235
/label=terminator of gene 32 (T32)
/note="terminator of gene 32 (T32)"
/regulatory_class="terminator"
misc_feature complement(1246..1264)
/label=Tn5 ME
/note="hyperactive mosaic end for Tn5 transposase
recognition (Reznikoff et al., 2004)"
terminator 1275..1369
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS 1474..2901
/label=Tn5 transposase
/note="transposase from the bacterial Tn5 transposon
(Reznikoff, 1993)"
promoter 2958..3049
/label=AmpR promoter
CDS 3050..3907
/label=AmpR
/note="beta-lactamase"
terminator 3914..3953
/label=fd terminator
/note="central terminator from bacteriophage fd (Otsuka and
Kunisawa, 1982)"
oriT 4101..4209
/label=oriT
/note="incP origin of transfer"
rep_origin 4220..4608
/label=R6K gamma ori
/note="gamma replication origin from E. coli plasmid R6K;
requires the R6K initiator protein pi for replication"
terminator complement(4639..4725)
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"