pHELLSGATE vector (V011788)

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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"