Basic Vector Information
- Vector Name:
- PpcleAmiR4-7
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8026 bp
- Type:
- UNVERIFIED: Expression vector
- Replication origin:
- ori
- Source/Author:
- Whitewoods CD, Cammarata J, Nemec Venza Z, Sang S, Crook AD, Aoyama T, Wang XY, Waller M, Kamisugi Y, Cuming AC, Szovenyi P, Nimchuk ZL, Roeder AH, Scanlon MJ, Harrison CJ.
- Promoter:
- Ubi
PpcleAmiR4-7 vector Vector Map
Plasmid Resuspension 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 ℃.
PpcleAmiR4-7 vector Sequence
LOCUS 40924_34146 8026 bp DNA circular SYN 18-DEC-2018
DEFINITION UNVERIFIED: Expression vector PpcleAmiR4-7, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8026)
AUTHORS Whitewoods CD, Cammarata J, Nemec Venza Z, Sang S, Crook AD, Aoyama
T, Wang XY, Waller M, Kamisugi Y, Cuming AC, Szovenyi P, Nimchuk ZL,
Roeder AH, Scanlon MJ, Harrison CJ.
TITLE CLAVATA Was a Genetic Novelty for the Morphological Innovation of 3D
Growth in Land Plants
JOURNAL Curr. Biol. 28 (15), 2365-2376 (2018)
PUBMED 30033333
REFERENCE 2 (bases 1 to 8026)
AUTHORS Whitewoods CD, Cammarata J, Nemec Venza Z, Sang S, Crook AD, Aoyama
T, Waller M, Kamisugi Y, Cuming AC, Szovenyi P, Nimchuk ZL, Roeder
AHK., Scanlon MJ, Harrison CJ.
TITLE Direct Submission
JOURNAL Submitted (30-APR-2018) Biological Sciences, University of Bristol,
24 Tyndall Avenue, Bristol BS81TQ, UK
REFERENCE 3 (bases 1 to 8026)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 8026)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Curr.
Biol."; date: "2018"; volume: "28"; issue: "15"; pages: "2365-2376"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(30-APR-2018) Biological Sciences, University of Bristol, 24 Tyndall
Avenue, Bristol BS81TQ, UK"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
GenBank staff is unable to verify sequence and/or annotation
provided by the submitter.
FEATURES Location/Qualifiers
source 1..8026
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(17..33)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin complement(213..641)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 668..772
/label=AmpR promoter
CDS 773..1630
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 1804..2392
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 2680..2701
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2716..2746
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2754..2770
/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 2778..2794
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 2825..2843
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 2926..3271
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
CDS 3288..4310
/codon_start=1
/label=HygR
/note="aminoglycoside phosphotransferase from E. coli"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
E"
polyA_signal 4523..4699
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
protein_bind complement(4765..4785)
/label=attB2
/note="core recombination site for the Gateway(R) BP
reaction"
promoter 4808..6790
/label=Ubi promoter
/note="maize polyubiquitin gene promoter"
terminator 7257..7963
/label=OCS terminator
/note="octopine synthase terminator"
promoter complement(7996..8014)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
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