Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V002574 | pTNS3 | In stock, instant shipping |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pTNS3
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9983 bp
- Type:
- Tn7 transposase expression vector
- Replication origin:
- R6K γ ori
- Source/Author:
- Choi KH, Mima T, Casart Y, Rholl D, Kumar A, Beacham IR, Schweizer HP.
- Promoter:
- Pc
- Growth Strain(s):
- DH5α
- Growth Temperature:
- 37℃
pTNS3 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.
pTNS3 vector Sequence
LOCUS Exported 9983 bp DNA circular SYN 19-JUN-2024
DEFINITION Exported.
ACCESSION V002574
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9983)
AUTHORS Choi KH, Mima T, Casart Y, Rholl D, Kumar A, Beacham IR, Schweizer
HP.
TITLE Genetic tools for select-agent-compliant manipulation of
Burkholderia pseudomallei
JOURNAL Appl. Environ. Microbiol. 74 (4), 1064-1075 (2008)
PUBMED 18156318
REFERENCE 2 (bases 1 to 9983)
AUTHORS Choi K-H., Rholl D, Schweizer HP.
TITLE A Tn7-based broad-host bacterial cloning and expression system
JOURNAL Unpublished
REFERENCE 3 (bases 1 to 9983)
AUTHORS Choi K-H., Rholl D, Schweizer HP.
TITLE Direct Submission
JOURNAL Submitted (21-SEP-2007) Microbiology, Immunology and Pathology,
Colorado State University, 1682 Campus Delivery, Ft. Collins, CO
80523-1682, USA
REFERENCE 4 (bases 1 to 9983)
TITLE Direct Submission
REFERENCE 5 (bases 1 to 9983)
TITLE Direct Submission
REFERENCE 6 (bases 1 to 9983)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Appl.
Environ. Microbiol."; date: "2008"; volume: "74"; issue: "4"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
(21-SEP-2007) Microbiology, Immunology and Pathology, Colorado State
University, 1682 Campus Delivery, Ft. Collins, CO 80523-1682, USA"
COMMENT SGRef: number: 4; type: "Journal Article"
COMMENT SGRef: number: 5; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..9983
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 96..200
/label=AmpR promoter
CDS 201..1058
/label=AmpR
/note="beta-lactamase"
rep_origin complement(1470..1854)
/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(1877..1985)
/direction=LEFT
/label=oriT
/note="incP origin of transfer"
protein_bind 2361..2382
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2397..2427
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2435..2451
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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 2459..2475
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 2512..2540
/label=Pc promoter
/note="class 1 integron promoter"
mobile_element complement(3051..3275)
/label=Tn7R
/note="mini-Tn7 element (right end of the Tn7 transposon)"
CDS 3993..6098
/gene="tnsB"
/label=Transposon Tn7 transposition protein TnsB
/note="Transposon Tn7 transposition protein TnsB from
Escherichia coli. Accession#: P13989"
CDS 6098..7762
/gene="tnsC"
/label=Transposon Tn7 transposition protein TnsC
/note="Transposon Tn7 transposition protein TnsC from
Escherichia coli. Accession#: P05846"
CDS 7768..9294
/codon_start=1
/gene="tnsD"
/product="TnsD"
/label=tnsD
/note="Tn7 transposase subunit"
/protein_id="ABW35346.1"
/translation="MRNFPVPYSNELIYSTIARAGVYQGIVSPKQLLDEVYGNRKVVAT
LGLPSHLGVIARHLHQTGRYAVQQLIYEHTLFPLYAPFVGKERRDEAIRLMEYQAQGAV
HLMLGVAASRVKSDNRFRYCPDCVALQLNRYGEAFWQRDWYLPALPYCPKHGALVFFDR
AVDDHRHQFWALGHTELLSDYPKDSLSQLTALAAYIAPLLDAPRAQELSPSLEQWTLFY
QRLAQDLGLTKSKHIRHDLVAERVRQTFSDEALEKLDLKLAENKDTCWLKSIFRKHRKA
FSYLQHSIVWQALLPKLTVIEALQQASALTEHSITTRPVSQSVQPNSEDLSVKHKDWQQ
LVHKYQGIKAARQSLEGGVLYAWLYRHDRDWLVHWNQQHQQERLAPAPRVDWNQRDRIA
VRQLLRIIKRLDSSLDHPRATSSWLLKQTPNGTSLAKNLQKLPLVALCLKRYSESVEDY
QIRRISQAFIKLKQEDVELRRWRLLRSATLSKERITEEAQRFLEMVYGEE"
gene 7768..9294
/gene="tnsD"
/label=tnsD
primer_bind complement(9589..9605)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"