Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V002520 | pTRKH2 | 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:
- pTRKH2
- Length:
- 6721 bp
- Type:
- Cloning vector
- Replication origin:
- p15A ori
- Source/Author:
- Welker DL, Hughes JE, Steele JL, Broadbent JR.
pTRKH2 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.
pTRKH2 vector Sequence
LOCUS V002520 6721 bp DNA circular SYN 18-DEC-2018
DEFINITION Exported.
ACCESSION V002520
VERSION V002520
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 6721)
AUTHORS Welker DL, Hughes JE, Steele JL, Broadbent JR.
TITLE High efficiency electrotransformation of Lactobacillus casei
JOURNAL FEMS Microbiol. Lett. 362 (2), 1-6 (2015)
PUBMED 25670703
REFERENCE 2 (bases 1 to 6721)
AUTHORS Welker DL, Hughes JE, Steele JL, Broadbent JR.
TITLE Direct Submission
JOURNAL Submitted (03-NOV-2017) Biology, Utah State University, 5305 Old
Main Hill, Logan, UT 84322-5305, USA
REFERENCE 3 (bases 1 to 6721)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6721)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
SGRef: number: 1; type: "Journal Article"; journalName: "FEMS
Microbiol. Lett."; date: "2015"; volume: "362"; issue: "2"; pages:
"1-6"
SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(03-NOV-2017) Biology, Utah State University, 5305 Old Main Hill,
Logan, UT 84322-5305, USA"
SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6721
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 102..123
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 138..168
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 176..192
/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 200..216
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(295..311)
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
CDS 992..1726
/gene="ermBP"
/label="rRNA adenine N-6-methyltransferase"
/note="rRNA adenine N-6-methyltransferase from Enterococcus
faecalis. Accession#: P0A4D5"
regulatory 2384..5298
/label="Enterococcus origin region from pAMBeta1"
/note="Enterococcus origin region from pAMBeta1"
/regulatory_class="replication_regulatory_region"
CDS 3312..4802
/codon_start=1
/gene="repR"
/product="RepR"
/label="repR"
/note="DNA replication protein"
/protein_id="AXU41049.1"
/translation="MNIPFVVETVLHDGLLKYKFKNSKIRSITTKPGKSKGAIFAYRSK
SSMIGGRGVVLTSEEAIQENQDTFTHWTPNVYRYGTYADENRSYTKGHSENNLRQINTF
FIDFDIHTAKETISASDILTTAIDLGFMPTMIIKSDKGYQAYFVLETPVYVTSKSEFKS
VKAAKIISQNIREYFGKSLPVDLTCNHFGIARIPRTDNVEFFDPNYRYSFKEWQDWSFK
QTDNKGFTRSSLTVLSGTEGKKQVDEPWFNLLLHETKFSGEKGLIGRNNVMFTLSLAYF
SSGYSIETCEYNMFEFNNRLDQPLEEKEVIKIVRSAYSENYQGANREYITILCKAWVSS
DLTSKDLFVRQGWFKFKKKRSERQRVHLSEWKEDLMAYISEKSDVYKPYLVTTKKEIRE
VLGIPERTLDKLLKVLKANQEIFFKIKPGRNGGIQLASVKSLLLSIIKVKKEEKESYIK
ALTNSFDLEHTFIQETLNKLAERPKTDTQLDLFSYDTG"
gene 3312..4802
/gene="repR"
/label="repR"
promoter complement(5512..5614)
/label="cat promoter"
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
rep_origin complement(6140..6685)
/direction=LEFT
/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."