Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V000186 | pTA7001-DEST | 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:
- pTA7001-DEST
- Antibiotic Resistance:
- Kanamycin, Chloramphenicol
- Length:
- 16023 bp
- Type:
- Plant Expression
- Replication origin:
- oriV
- Host:
- Plants
- Selection Marker:
- Hygromycin
- Copy Number:
- Low Copy
- Promoter:
- NOS
- Cloning Method:
- Restriction Enzyme
pTA7001-DEST vector 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.
pTA7001-DEST vector Sequence
LOCUS 40924_42414 16023 bp DNA circular SYN 13-MAY-2021
DEFINITION GATEWAY compatible derivative of plant binary vector pTA7001 from
Nam Chua's laboratory at The Rockefeller University, Dexamethasone
inducible.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 16023)
AUTHORS Gu Y, Innes RW
TITLE The KEEP ON GOING protein of Arabidopsis recruits the ENHANCED
DISEASE RESISTANCE1 protein to trans-Golgi network/early endosome
vesicles.
JOURNAL Plant Physiol. 2011 Apr;155(4):1827-38. doi: 10.1104/pp.110.171785.
Epub 2011 Feb 22.
PUBMED 21343429
REFERENCE 2 (bases 1 to 16023)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 16023)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1104/pp.110.171785"; journalName: "Plant Physiol"; date:
"2011-04"; volume: "155"; issue: "4"; pages: "1827-38"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..16023
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(181..828)
/codon_start=1
/label=TetR
/note="tetracycline resistance regulatory protein"
/translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
misc_feature complement(1293..1317)
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
promoter 1832..2177
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
CDS 2439..2672
/codon_start=1
/label=VP16 AD
/note="transcriptional activation domain of herpes simplex
virus protein VP16 (Triezenberg et al., 1988; Cousens et
al., 1989)"
/translation="APPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGF
TPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
promoter 3996..4179
/label=NOS promoter
/note="nopaline synthase promoter"
CDS 4231..5241
/codon_start=1
/label=HygR
/note="aminoglycoside phosphotransferase from E. coli"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRP"
terminator 5294..5546
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
promoter 5766..5812
/label=minimal CaMV 35S promoter
/note="minimal 35S promoter from cauliflower mosaic virus"
protein_bind 5846..5970
/label=attR1
/note="recombination site for the Gateway(R) LR reaction"
promoter 5995..6025
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
CDS 6079..6735
/codon_start=1
/label=CmR
/note="chloramphenicol acetyltransferase"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
CDS 7080..7382
/codon_start=1
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
protein_bind complement(7426..7550)
/label=attR2
/note="recombination site for the Gateway(R) LR reaction"
primer_bind complement(8092..8109)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(8092..8108)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(8101..8123)
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind complement(8248..8267)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 8458..8479
/label=F1ori-F
/note="F1 origin, forward primer"
misc_feature complement(8726..8750)
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
CDS complement(10349..11494)
/codon_start=1
/label=trfA
/note="trans-acting replication protein that binds to and
activates oriV"
/translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
CDS complement(11796..12587)
/codon_start=1
/label=KanR
/note="aminoglycoside phosphotransferase"
/translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
mobile_element 12779..13546
/label=IS1
/note="prokaryotic transposable element"
rep_origin complement(14169..14880)
/direction=LEFT
/label=oriV
/note="incP origin of replication"
CDS complement(15010..15378)
/codon_start=1
/label=traJ
/note="oriT-recognizing protein"
/translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
KQDELGKVMMGVVRPRAEP"
oriT complement(15411..15520)
/direction=LEFT
/label=oriT
/note="incP origin of transfer"
primer_bind 15743..15762
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 15996..16013
/label=L4440
/note="L4440 vector, forward primer"