Price Information
| Cat No. | Plasmid Name | Availability | Buy one, get one free! (?) |
|---|---|---|---|
| V009847 | Mini-Tn4001PSpuro | In stock, instant shipping |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- Mini-Tn4001PSpuro
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5321 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Algire MA, Lartigue C, Thomas DW, Assad-Garcia N, Glass JI, Merryman C.
- Promoter:
- T7
- Growth Strain(s):
- JM109
Mini-Tn4001PSpuro 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.
Mini-Tn4001PSpuro vector Sequence
LOCUS 40924_1984 5321 bp DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector Mini-Tn4001PSpuro, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5321)
AUTHORS Algire MA, Lartigue C, Thomas DW, Assad-Garcia N, Glass JI, Merryman
C.
TITLE New selectable marker for manipulating the simple genomes of
Mycoplasma species
JOURNAL Antimicrob. Agents Chemother. 53 (10), 4429-4432 (2009)
PUBMED 19687239
REFERENCE 2 (bases 1 to 5321)
AUTHORS Merryman C.
TITLE Direct Submission
JOURNAL Submitted (30-MAR-2009) Synthetic Biology, J. Craig Venter
Institute, 9704 Medical Center Dr, Rockville, MD 20850, USA
REFERENCE 3 (bases 1 to 5321)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 5321)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Antimicrob.
Agents Chemother."; date: "2009"; volume: "53"; issue: "10"; pages:
"4429-4432"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(30-MAR-2009) Synthetic Biology, J. Craig Venter Institute, 9704
Medical Center Dr, Rockville, MD 20850, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5321
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(4..600)
/codon_start=1
/label=PuroR
/note="puromycin N-acetyltransferase"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
CDS 1105..2277
/codon_start=1
/gene="tnp"
/product="transposase"
/label=tnp
/protein_id="ACR82773.1"
/translation="MTQVHFTLKSEEIQSIIEYSVKDDVSKNILTTVFNQLMENQRTEY
IQAKEYERTENRQSQRNGYYERSFTTRVGTLELKVPRTRDGHFSPTVFERYQRNEKALM
ASMLEMYVSGVSTRKVSKIVEELCGKSVSKSFVSSLTEQLEPMVNEWQNRLLSEKNYPY
LMTDVLYIKVREENRVLSKSCHIAIGITKDGDREIIGFMIQSGESEETWTTFFEYLKER
GLQGTELVISDAHKGLVSAIRKSFTNVSWQRCQVHFLRNIFTTIPKKNSKSFREAVKGI
FKFTDINLAREAKNRLIHDYIDQPKYSKACASLDDGFEDAFQYTVQGNSHNRLKSTNLI
ERLNQEVRRREKIIRIFPNQTSANRLIGAVLMDLHDEWIYSSRKYINFDK"
gene 1105..2277
/gene="tnp"
/label=tnp
promoter complement(2298..2316)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(2323..2339)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 2481..2936
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 2962..3066
/label=AmpR promoter
CDS 3067..3924
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 4098..4686
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 4974..4995
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 5010..5040
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5048..5064
/label=lac operator
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
primer_bind 5072..5088
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 5109..5127
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"