LOCUS       pHANNIBAL.        5824 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Vector for silencing genes in plants using intron-containing hairpin
            RNA (hpRNA).
ACCESSION   .
VERSION     .
KEYWORDS    pHANNIBAL
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5824)
  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 5824)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 5824)
  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..5824
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(19..35)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(215..643)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        670..774
                     /label=AmpR promoter
     CDS             775..1632
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      1806..2394
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2682..2703
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2718..2748
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2756..2772
                     /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     2780..2796
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2827..2845
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        3863..4208
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     misc_feature    4210..4228
                     /label=MCS 1
                     /note="MCS 1"
                     /note="multiple cloning site 1"
     intron          4236..5002
                     /label=PDK intron
                     /note="pyruvate orthophosphate dikinase intron from
                     Flaveria trinervia"
     misc_feature    5022..5045
                     /label=MCS 2
                     /note="MCS 2"
                     /note="multiple cloning site 2"
     terminator      5049..5755
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(5796..5814)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
ORIGIN
        1 tgaccaagtc agcttggcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
       61 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
      121 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggaaat
      181 tgtaaacgtt aatattttgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
      241 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
      301 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
      361 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
      421 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
      481 gggatgcccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
      541 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
      601 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg
      661 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
      721 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
      781 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
      841 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
      901 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
      961 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     1021 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     1081 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     1141 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     1201 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     1261 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     1321 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     1381 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     1441 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     1501 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     1561 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     1621 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     1681 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     1741 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     1801 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     1861 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     1921 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     1981 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     2041 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2101 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2161 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2221 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2281 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2341 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2401 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     2461 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     2521 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     2581 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     2641 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     2701 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     2761 agcggataac aatttcacac aggaaacagc tatgaccatg attacgaatt tggccaagtc
     2821 ggcctctaat acgactcact atagggagct cgtcgagcgg ccgctcgacg aattaattcc
     2881 aatcccacaa aaatctgagc ttaacagcac agttgctcct ctcagagcag aatcgggtat
     2941 tcaacaccct catatcaact actacgttgt gtataacggt ccacatgccg gtatatacga
     3001 tgactggggt tgtacaaagg cggcaacaaa cggcgttccc ggagttgcac acaagaaatt
     3061 tgccactatt acagaggcaa gagcagcagc tgacgcgtac acaacaagtc agcaaacaga
     3121 caggttgaac ttcatcccca aaggagaagc tcaactcaag cccaagagct ttgctaaggc
     3181 cctaacaagc ccaccaaagc aaaaagccca ctggctcacg ctaggaacca aaaggcccag
     3241 cagtgatcca gccccaaaag agatctcctt tgccccggag attacaatgg acgatttcct
     3301 ctatctttac gatctaggaa ggaagttcga aggtgaaggt gacgacacta tgttcaccac
     3361 tgataatgag aaggttagcc tcttcaattt cagaaagaat gctgacccac agatggttag
     3421 agaggcctac gcagcaggtc tcatcaagac gatctacccg agtaacaatc tccaggagat
     3481 caaatacctt cccaagaagg ttaaagatgc agtcaaaaga ttcaggacta attgcatcaa
     3541 gaacacagag aaagacatat ttctcaagat cagaagtact attccagtat ggacgattca
     3601 aggcttgctt cataaaccaa ggcaagtaat agagattgga gtctctaaaa aggtagttcc
     3661 tactgaatct aaggccatgc atggagtcta agattcaaat cgaggatcta acagaactcg
     3721 ccgtgaagac tggcgaacag ttcatacaga gtcttttacg actcaatgac aagaagaaaa
     3781 tcttcgtcaa catggtggag cacgacactc tggtctactc caaaaatgtc aaagatacag
     3841 tctcagaaga ccaaagggct attgagactt ttcaacaaag gataatttcg ggaaacctcc
     3901 tcggattcca ttgcccagct atctgtcact tcatcgaaag gacagtagaa aaggaaggtg
     3961 gctcctacaa atgccatcat tgcgataaag gaaaggctat cattcaagat ctctctgccg
     4021 acagtggtcc caaagatgga cccccaccca cgaggagcat cgtggaaaaa gaagacgttc
     4081 caaccacgtc ttcaaagcaa gtggattgat gtgacatctc cactgacgta agggatgacg
     4141 cacaatccca ctatccttcg caagaccctt cctctatata aggaagttca tttcatttgg
     4201 agaggacacg ctcgaggaat tcggtacccc aattggtaag gaaataatta ttttcttttt
     4261 tccttttagt ataaaatagt taagtgatgt taattagtat gattataata atatagttgt
     4321 tataattgtg aaaaaataat ttataaatat attgtttaca taaacaacat agtaatgtaa
     4381 aaaaatatga caagtgatgt gtaagacgaa gaagataaaa gttgagagta agtatattat
     4441 ttttaatgaa tttgatcgaa catgtaagat gatatactag cattaatatt tgttttaatc
     4501 ataatagtaa ttctagctgg tttgatgaat taaatatcaa tgataaaata ctatagtaaa
     4561 aataagaata aataaattaa aataatattt ttttatgatt aatagtttat tatataatta
     4621 aatatctata ccattactaa atattttagt ttaaaagtta ataaatattt tgttagaaat
     4681 tccaatctgc ttgtaattta tcaataaaca aaatattaaa taacaagcta aagtaacaaa
     4741 taatatcaaa ctaatagaaa cagtaatcta atgtaacaaa acataatcta atgctaatat
     4801 aacaaagcgc aagatctatc attttatata gtattatttt caatcaacat tcttattaat
     4861 ttctaaataa tacttgtagt tttattaact tctaaatgga ttgactatta attaaatgaa
     4921 ttagtcgaac atgaataaac aaggtaacat gatagatcat gtcattgtgt tatcattgat
     4981 cttacatttg gattgattac agttgggaaa ttgggttcga aatcgataag cttggatcct
     5041 ctagagtcct gctttaatga gatatgcgag acgcctatga tcgcatgata tttgctttca
     5101 attctgttgt gcacgttgta aaaaacctga gcatgtgtag ctcagatcct taccgccggt
     5161 ttcggttcat tctaatgaat atatcacccg ttactatcgt atttttatga ataatattct
     5221 ccgttcaatt tactgattgt accctactac ttatatgtac aatattaaaa tgaaaacaat
     5281 atattgtgct gaataggttt atagcgacat ctatgataga gcgccacaat aacaaacaat
     5341 tgcgttttat tattacaaat ccaattttaa aaaaagcggc agaaccggtc aaacctaaaa
     5401 gactgattac ataaatctta ttcaaatttc aaaaggcccc aggggctagt atctacgaca
     5461 caccgagcgg cgaactaata acgttcactg aagggaactc cggttccccg ccggcgcgca
     5521 tgggtgagat tccttgaagt tgagtattgg ccgtccgctc taccgaaagt tacgggcacc
     5581 attcaacccg gtccagcacg gcggccgggt aaccgacttg ctgccccgag aattatgcag
     5641 catttttttg gtgtatgtgg gccccaaatg aagtgcaggt caaaccttga cagtgacgac
     5701 aaatcgttgg gcgggtccag ggcgaatttt gcgacaacat gtcgaggctc agcaggacct
     5761 gcaggcatgc aagctagctt actagtgatg catattctat agtgtcacct aaatctgcgg
     5821 ccgc
//
