pHELLSGATE 4 vector (V011790)

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LOCUS       pHELLSGATE_4.       17861 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Improved high-throughput binary Gateway(R) donor vector for 
            silencing genes in plants using intron-containing hairpin RNA.
ACCESSION   .
VERSION     .
KEYWORDS    pHELLSGATE 4
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 17861)
  AUTHORS   Helliwell CA, Wesley SV, Wielopolska AJ, Waterhouse PM.
  TITLE     High-throughput vectors for efficient gene silencing in plants.
  JOURNAL   Funct. Plant Biol. 2002;29:1217-25.
REFERENCE   2  (bases 1 to 17861)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 17861)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Funct. 
            Plant Biol."; date: "2002"; volume: "29"; pages: "1217-25"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..17861
                     /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)"
     misc_feature    14644..14654
                     /label=MCS 1
                     /note="MCS 1"
                     /note="multiple cloning site 1"
     intron          14662..15428
                     /label=PDK intron
                     /note="pyruvate orthophosphate dikinase intron from
                     Flaveria trinervia"
     misc_feature    15448..15454
                     /label=MCS 2
                     /note="MCS 2"
                     /note="multiple cloning site 2"
     protein_bind    15490..15721
                     /label=attP2
                     /note="recombination site for the Gateway(R) BP reaction 
                     (pDONR(TM)221 version)"
     CDS             complement(16028..16330)
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(16829..17060)
                     /label=attP1
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      17092..17798
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(17839..17857)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"