Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V009284 | pBAM1 | 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:
- pBAM1
- Antibiotic Resistance:
- Ampicillin
- Length:
- 4384 bp
- Type:
- Cloning vector
- Replication origin:
- R6K γ ori
- Source/Author:
- Martinez-Garcia E, Calles B, Arevalo-Rodriguez M, de Lorenzo V.
pBAM1 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.
pBAM1 vector Sequence
LOCUS 40924_5889 4384 bp DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector pBAM1, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4384)
AUTHORS Martinez-Garcia E, Calles B, Arevalo-Rodriguez M, de Lorenzo V.
TITLE pBAM1: an all-synthetic genetic tool for analysis and construction
of complex bacterial phenotypes
JOURNAL BMC Microbiol. 11 (1), 38 (2011)
PUBMED 21342504
REFERENCE 2 (bases 1 to 4384)
AUTHORS Martinez-Garcia E, Calles B, Arevalo-Rodriguez M, de Lorenzo V.
TITLE Direct Submission
JOURNAL Submitted (20-JAN-2011) Systems Biology Program, Centro Nacional de
Biotecnologia, C/ Darwin 3, Madrid, Madrid 28049, Spain
REFERENCE 3 (bases 1 to 4384)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 4384)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "BMC
Microbiol."; date: "2011"; volume: "11"; issue: "1"; pages: "38"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(20-JAN-2011) Systems Biology Program, Centro Nacional de
Biotecnologia, C/ Darwin 3, Madrid, Madrid 28049, Spain"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..4384
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin complement(4..392)
/direction=LEFT
/label=R6K gamma ori
/note="gamma replication origin from E. coli plasmid R6K;
requires the R6K initiator protein pi for replication"
oriT complement(411..519)
/direction=LEFT
/label=oriT
/note="incP origin of transfer"
terminator complement(666..705)
/label=fd terminator
/note="central terminator from bacteriophage fd (Otsuka and
Kunisawa, 1982)"
CDS complement(712..1569)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(1570..1661)
/label=AmpR promoter
CDS complement(1718..3145)
/codon_start=1
/label=Tn5 transposase
/note="transposase from the bacterial Tn5 transposon
(Reznikoff, 1993)"
/translation="MITSALHRAADWAKSVFSSAALGDPRRTARLVNVAAQLAKYSGKS
ITISSEGSKAMQEGAYRFIRNPNVSAEAIRKAGAMQTVKLAQEFPELLAIEDTTSLSYR
HQVAEELGKLGSIQDKSRGWWVHSVLLLEATTFRTVGLLHQEWWMRPDDPADADEKESG
KWLAAAATSRLRMGSMMSNVIAVCDREADIHAYLQDKLAHNERFVVRSKHPRKDVESGL
YLYDHLKNQPELGGYQISIPQKGVVDKRGKRKNRPARKASLSLRSGRITLKQGNITLNA
VLAEEINPPKGETPLKWLLLTSEPVESLAQALRVIDIYTHRWRIEEFHKAWKTGAGAER
QRMEEPDNLERMVSILSFVAVRLLQLRESFTPPQALRAQGLLKEAEHVESQSAETVLTP
DECQLLGYLDKGKRKRKEKAGSLQWAYMAIARLGGFMDSKRTGIASWGALWEGWEALQS
KLDGFLAAKDLMAQGIKI"
misc_feature 3239..3257
/label=Tn5 ME
/note="hyperactive mosaic end for Tn5 transposase
recognition (Reznikoff et al., 2004)"
CDS 3365..4177
/codon_start=1
/label=KanR
/note="aminoglycoside phosphotransferase"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
misc_feature complement(4267..4323)
/label=MCS
/note="pUC18/19 multiple cloning site"
misc_feature complement(4355..4373)
/label=Tn5 ME
/note="hyperactive mosaic end for Tn5 transposase
recognition (Reznikoff et al., 2004)"