PpcleAmiR1-3 vector (V004169)

Basic Vector Information

      • Vector Name:
      • PpcleAmiR1-3
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 7676 bp
      • Type:
      • UNVERIFIED: Expression vector
      • 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.

PpcleAmiR1-3 vector Vector Map

PpcleAmiR1-37676 bp3006009001200150018002100240027003000330036003900420045004800510054005700600063006600690072007500M13 fwdf1 oriAmpR promoterAmpRoriCAP binding sitelac promoterlac operatorM13 revT7 promoterCaMV 35S promoterNeoR/KanRCaMV poly(A) signalattB2OCS terminatorSP6 promoter

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 ℃.

PpcleAmiR1-3 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                7676 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  UNVERIFIED: Expression vector PpcleAmiR1-3, complete sequence.
ACCESSION   MH310734
VERSION     .
KEYWORDS    UNVERIFIED.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7676)
  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 7676)
  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 7676)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     GenBank staff is unable to verify sequence and/or annotation 
            provided by the submitter.
            ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7676
                     /organism="Expression vector PpcleAmiR1-3"
                     /mol_type="other DNA"
                     /db_xref="taxon:2293867"
     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
                     /gene="bla"
                     /label=AmpR promoter
     CDS             773..1633
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1804..2392
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2680..2701
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /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
                     /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     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             3297..4091
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERGRTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTQGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4193..4369
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     protein_bind    complement(4431..4455)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      6907..7613
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(7646..7664)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"

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