LOCUS       Exported               10459 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pAGRIKOLA, complete sequence.
ACCESSION   AY568055
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10459)
  AUTHORS   Hilson P, Allemeersch J, Altmann T, Aubourg S, Avon A, Beynon J, 
            Bhalerao RP, Bitton F, Caboche M, Cannoot B, Chardakov V, 
            Cognet-Holliger C, Colot V, Crowe M, Darimont C, Durinck S, Eickhoff
            H, de Longevialle AF, Farmer EE, Grant M, Kuiper MT, Lehrach H, Leon
            C, Leyva A, Lundeberg J, Lurin C, Moreau Y, Nietfeld W, Paz-Ares J, 
            Reymond P, Rouze P, Sandberg G, Segura MD, Serizet C, Tabrett A, 
            Taconnat L, Thareau V, Van Hummelen P, Vercruysse S, Vuylsteke M, 
            Weingartner M, Weisbeek PJ, Wirta V, Wittink FR, Zabeau M, Small I.
  TITLE     Versatile gene-specific sequence tags for Arabidopsis functional 
            genomics: transcript profiling and reverse genetics applications
  JOURNAL   Genome Res. 14 (10B), 2176-2189 (2004)
  PUBMED    15489341
REFERENCE   2  (bases 1 to 10459)
  AUTHORS   Small ID, Hilson P.
  TITLE     pAGRIKOLA sequence
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 10459)
  AUTHORS   Small ID, Hilson P.
  TITLE     Direct Submission
  JOURNAL   Submitted (07-MAR-2004) URGV, INRA/CNRS/UEVE, 2, rue Gaston 
            Cremieux, CP5708, Evry, Essonne 91057, France
REFERENCE   4  (bases 1 to 10459)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10459
                     /organism="Cloning vector pAGRIKOLA"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
                     /label=high-throughput plant RNAi cloning vector
                     /note="high-throughput plant RNAi cloning vector"
                     /db_xref="taxon:267887"
     rep_origin      18..672
                     /label=colE1
                     /note="colE1"
     rep_origin      complement(61..649)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(820..1635)
                     /codon_start=1
                     /gene="nptI"
                     /product="APH(3)I"
                     /EC_number="2.7.1.95"
                     /label=nptI
                     /note="aminoglycoside 3-phosphotransferase; confers 
                     kanamycin resistance in bacteria"
                     /protein_id="AAS75425.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            complement(820..1635)
                     /gene="nptI"
                     /label=nptI
     rep_origin      1925..2472
                     /label=pSa
                     /note="pSa"
     rep_origin      1926..2361
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     repeat_region   2488..2512
                     /label=T-DNA left border
                     /note="T-DNA left border"
     misc_feature    2488..2510
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     regulatory      complement(2532..2791)
                     /regulatory_class="polyA_signal_sequence"
                     /note="nos terminator"
     terminator      2534..2786
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(2806..3357)
                     /codon_start=1
                     /gene="bar"
                     /product="BAR"
                     /label=bar
                     /note="phospinothricin N-acetyltransferase; confers Basta 
                     resistance in plants"
                     /protein_id="AAS75426.1"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     gene            complement(2806..3357)
                     /gene="bar"
                     /label=bar
     regulatory      complement(3378..3661)
                     /regulatory_class="promoter"
                     /note="nos promoter"
     promoter        complement(3399..3578)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     3825..3841
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3851..3869
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3895..3911
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3945..3961)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      3980..5317
                     /regulatory_class="promoter"
                     /note="CaMV 35S promoter"
     promoter        4968..5313
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     misc_feature    5322..5446
                     /label=attR1 recombination site
                     /note="attR1 recombination site"
     protein_bind    5322..5446
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(5875..6180)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CCDB"
                     /label=ccdB
                     /note="cytotoxin; counterselection of Gateway cassette"
                     /protein_id="AAS75427.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     gene            complement(5875..6180)
                     /gene="ccdB"
                     /label=ccdB
     protein_bind    complement(6619..6743)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    complement(6619..6738)
                     /label=attR2 recombination site
                     /note="attR2 recombination site"
     intron          6766..7544
                     /note="intron from Flaveria trinerva PDK gene encoding 
                     pyruvate orthophosphate dikinase"
     intron          6770..7538
                     /gene="pdk"
                     /number=3
                     /label=PDK intron
                     /note="pyruvate orthophosphate dikinase intron from 
                     Flaveria trinervia"
     intron          7573..7762
                     /label=cat1 intron
                     /note="castor bean catalase intron, modified"
     intron          complement(7581..7763)
                     /note="intron from Ricinus communis CAT1 gene encoding 
                     catalase"
     protein_bind    7786..7910
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    7791..7910
                     /label=attR2 recombination site
                     /note="attR2 recombination site"
     CDS             complement(8239..8544)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CCDB"
                     /label=ccdB
                     /note="cytotoxin; counterselection of Gateway cassette"
                     /protein_id="AAS75428.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     gene            complement(8239..8544)
                     /gene="ccdB"
                     /label=ccdB
     misc_feature    complement(9075..9199)
                     /label=attR1 recombination site
                     /note="attR1 recombination site"
     protein_bind    complement(9075..9199)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      9208..9915
                     /regulatory_class="polyA_signal_sequence"
                     /note="ocs terminator"
     terminator      9208..9915
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(9956..9974)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     promoter        complement(10013..10031)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(10052..10068)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    10076..10092
                     /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)."
     promoter        complement(10100..10130)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10145..10166
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     repeat_region   10404..10429
                     /label=T-DNA right border
                     /note="T-DNA right border"
     misc_feature    10405..10429
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"