Basic Vector Information
Unavailable, please do not order
- Vector Name:
- pHELLSGATE
- Antibiotic Resistance:
- Chloramphenicol
- Length:
- 18691 bp
- Type:
- Gateway Cloning Vectors
- Replication origin:
- oriV
- Host:
- Plants
- Source/Author:
- Wesley SV, Helliwell CA, Smith NA, Wang MB, Rouse DT, Liu Q, Gooding
- Copy Number:
- High copy number
- Promoter:
- CaMV 35S
pHELLSGATE vector Map
pHELLSGATE vector Sequence
LOCUS pHELLSGATE. 18691 bp DNA circular SYN 01-JAN-1980
DEFINITION High-throughput binary Gateway(R) donor vector for silencing genes
in plants using intron-containing hairpin RNA.
ACCESSION .
VERSION .
KEYWORDS pHELLSGATE
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 18691)
AUTHORS Wesley SV, Helliwell CA, Smith NA, Wang MB, Rouse DT, Liu Q, Gooding
PS, Singh SP, Abbott D, Stoutjesdijk PA, Robinson SP, Gleave AP,
Green AG, Waterhouse PM.
TITLE Construct design for efficient, effective and high-throughput gene
silencing in plants.
JOURNAL Plant J. 2001;27:581-90.
PUBMED 11576441
REFERENCE 2 (bases 1 to 18691)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 18691)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Plant J.";
date: "2001"; volume: "27"; pages: "581-90"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..18691
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter complement(65..83)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(90..106)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 264..447
/label=NOS promoter
/note="nopaline synthase promoter"
CDS 448..1266
/label=KanR
/note="fusion between an N-terminal peptide of nopaline
synthase and Tn5 aminoglycoside phosphotransferase"
terminator 1894..2146
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
misc_feature complement(2268..2292)
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
oriT 2846..2955
/label=oriT
/note="incP origin of transfer"
mobile_element 3015..3782
/label=IS1
/note="prokaryotic transposable element"
CDS 3782..4132
/label=traJ
/note="oriT-recognizing protein"
CDS 4407..5552
/label=trfA
/note="trans-acting replication protein that binds to and
activates oriV"
rep_origin complement(6915..7503)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS 8600..9388
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase"
/label=aadA
/note="SmR"
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEDRLASRADQLEEFVHYVKGEITKVVGK"
rep_origin complement(9993..10705)
/direction=LEFT
/label=oriV
/note="incP origin of replication"
misc_feature complement(11182..11206)
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
protein_bind 11462..11483
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 11498..11528
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 11536..11552
/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 11560..11576
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 11594..11612
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
promoter 12672..13017
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
protein_bind 13048..13279
/label=attP1
/note="recombination site for the Gateway(R) BP reaction"
CDS complement(13678..13980)
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
protein_bind complement(14387..14618)
/label=attP2
/note="recombination site for the Gateway(R) BP reaction
(pDONR(TM)221 version)"
intron 14662..16258
/label=PDK intron
/note="modified pyruvate orthophosphate dikinase intron
from Flaveria trinervia"
protein_bind 16320..16551
/label=attP2
/note="recombination site for the Gateway(R) BP reaction
(pDONR(TM)221 version)"
CDS complement(16858..17160)
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
protein_bind complement(17659..17890)
/label=attP1
/note="recombination site for the Gateway(R) BP reaction"
terminator 17922..18628
/label=OCS terminator
/note="octopine synthase terminator"
promoter complement(18669..18687)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
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