LOCUS       40924_22593        5138 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Agrobacterium binary vector with kanamycin- and 
            hygromycin-resistance genes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5138)
  AUTHORS   Hellens RP, Edwards EA, Leyland NR, Bean S, Mullineaux PM.
  TITLE     pGreen: a versatile and flexible binary Ti vector for 
            Agrobacterium-mediated plant transformation.
  JOURNAL   Plant Mol. Biol. 2000;42:819-32.
  PUBMED    10890530
REFERENCE   2  (bases 1 to 5138)
  AUTHORS   pGreen website
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 5138)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plant Mol. 
            Biol."; date: "2000"; volume: "42"; pages: "819-32"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     This plasmid must be transformed into an Agrobacterium strain that 
            also carries a vector such as pSoup.
FEATURES             Location/Qualifiers
     source          1..5138
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      63..498
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    633..655
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA
                     (truncated)"
     polyA_signal    complement(663..839)
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(1052..2074)
                     /codon_start=1
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     promoter        complement(2091..2436)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     2658..2674
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2681..2699
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    complement(2708..2815)
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     promoter        complement(2828..2846)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2867..2883)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2891..2907)
                     /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)."
     promoter        complement(2915..2946)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2961..2982)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     misc_feature    3221..3245
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(3336..3924)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4098..4910)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANVVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
ORIGIN
        1 agctctcatc aaccgtggct ccctcacttt ctggctggat gatggggcga ttcaggcgat
       61 ccccatccaa cagcccgccg tcgagcgggc ttttttatcc ccggaagcct gtggatagag
      121 ggtagttatc cacgtgaaac cgctaatgcc ccgcaaagcc ttgattcacg gggctttccg
      181 gcccgctcca aaaactatcc acgtgaaatc gctaatcagg gtacgtgaaa tcgctaatcg
      241 gagtacgtga aatcgctaat aaggtcacgt gaaatcgcta atcaaaaagg cacgtgagaa
      301 cgctaatagc cctttcagat caacagcttg caaacacccc tcgctccggc aagtagttac
      361 agcaagtagt atgttcaatt agcttttcaa ttatgaatat atatatcaat tattggtcgc
      421 ccttggcttg tggacaatgc gctacgcgca ccggctccgc ccgtggacaa ccgcaagcgg
      481 ttgcccaccg tcgagcgcca gcgcctttgc ccacaacccg gcggccggcc gcaacagatc
      541 gttttataaa tttttttttt tgaaaaagaa aaagcccgaa aggcggcaac ctctcgggct
      601 tctggatttc cgatccccgg aattagagat cttggcagga tatattgtgg tgtaacgtta
      661 tcgatctgga ttttagtact ggattttggt tttaggaatt agaaatttta ttgatagaag
      721 tattttacaa atacaaatac atactaaggg tttcttatat gctcaacaca tgagcgaaac
      781 cctataagaa ccctaatttc ccttatcggg aaactactca cacattattt atggagaaaa
      841 tagagagaga tagatttgta gagagagact ggtgatttca gcgtaccgaa ttaattctcc
      901 cgcgaccagc cgagcgagct tagcgaactg tggacgagaa ctgtgccacc aagcgtaagg
      961 ccgttctctc gcatttgcct tgctaggctc gcgcgagttg ctggctgagg cgttctcgaa
     1021 atcagctctt gttcggtcgg catctactct attcctttgc cctcggacga gtgctggggc
     1081 gtcggtttcc actatcggcg agtacttcta cacagccatc ggtccagacg gccgcgcttc
     1141 tgcgggcgat ttgtgtacgc ccgacagtcc cggctccgga tcggacgatt gcgtcgcatc
     1201 gaccctgcgc ccaagctgca tcatcgaaat tgccgtcaac caagctctga tagagttggt
     1261 caagaccaat gcggagcata tacgcccgga ggcgtggcga tcctgcaagc tccggatgcc
     1321 tccgctcgaa gtagcgcgtc tgctgctcca tacaagccaa ccacggcctc cagaagaaga
     1381 tgttggcgac ctcgtattgg gaatccccga acatcgcctc gctccagtca atgaccgctg
     1441 ttatgcggcc attgtccgtc aggacattgt tggagccgaa atccgcgtgc acgaggtgcc
     1501 ggacttcggg gcagtcctcg gcccaaagca tcagctcatc gagagcctgc gcgacggacg
     1561 cactgacggt gtcgtccatc acagtttgcc agtgatacac atggggatca gcaatcgcgc
     1621 atatgaaatc acgccatgta gtgtattgac cgattccttg cggtccgaat gggccgaacc
     1681 cgctcgtctg gctaagatcg gccgcagcga tcgcatccat agcctccgcg accggctgaa
     1741 gaacagcggg cagttcggtt tcaggcaggt cttgcaacgt gacaccctgt gcacggcggg
     1801 agatgcaata ggtcaggctc tcgctgaact ccccaatgtc aagcacttcc ggaatcggga
     1861 gcgcggccga tgcaaagtgc cgataaacat aacgatcttt gtagaaacca tcggcgcagc
     1921 tatttacccg caggacatat ccacgccctc ctacatcgaa gctgaaagca cgagattctt
     1981 cgccctccga gagctgcatc aggtcggaga cgctgtcgaa cttttcgatc agaaacttct
     2041 cgacagacgt cgcggtgagt tcaggctttt tcatagggat cagcttgggc tgtcctctcc
     2101 aaatgaaatg aacttcctta tatagaggaa gggtcttgcg aaggatagtg ggattgtgcg
     2161 tcatccctta cgtcagtgga gatgtcacat caatccactt gctttgaaga cgtggttgga
     2221 acgtcttctt tttccacgat gctcctcgtg ggtgggggtc catctttggg accactgtcg
     2281 gcagaggcat cttgaatgat agcctttcct ttatcgcaat gatggcattt gtaggagcca
     2341 ccttcctttt ctactgtcct ttcgatgaag tgacagatag ctgggcaatg gaatccgagg
     2401 aggtttcccg aaattaccct ttgttgaaaa gtctcaatag ccctttggtc ttctgagact
     2461 gtatctttga catttttgga gtaggggtac gataacatta acgtttacaa tttcgcgcca
     2521 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     2581 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     2641 ttcccagtca cgacgttgta aaacgacggc cagtgaattg taatacgact cactataggg
     2701 cgaattgggt accgggcccc ccctcgaggt cgacggtatc gataagcttg atatcgaatt
     2761 cctgcagccc gggggatcca ctagttctag agcggccgcc accgcggtgg agctccagct
     2821 tttgttccct ttagtgaggg ttaattccga gcttggcgta atcatggtca tagctgtttc
     2881 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga aghcataaag
     2941 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     3001 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     3061 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     3121 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     3181 acagaatcag gggataacgc aggaaagaac atgaaggcct tgacaggata tattggcggg
     3241 taaactaagt cgctgtatgt gtttgtttga gatctcatgt gagcaaaagg ccagcaaaag
     3301 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3361 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3421 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3481 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3541 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3601 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3661 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3721 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     3781 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaagaagagt tggtagctct
     3841 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3901 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3961 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     4021 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgtgtaa
     4081 cattggtcta gtgattagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata
     4141 tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca
     4201 ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca
     4261 acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca
     4321 ccatgagtga cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc tttccagact
     4381 tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta
     4441 ttcattcgtg attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta
     4501 caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca
     4561 cctgaatcag gatattcttc taatacctgg aatgctgttt tccctgggat cgcagtggtg
     4621 agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat
     4681 tccgtcagcc agtttagtct gaccatctca tctgtaacaa cattggcaac gctacctttg
     4741 ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcggta gattgtcgca
     4801 cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg
     4861 gaatttaatc gcggccttga gcaagacgtt tcccgttgaa tatggctcat aacacccctt
     4921 gtattactgt ttatgtaagc agacagtttt attgttcatg atgatatatt tttatcttgt
     4981 gcaatgtaac atcagagatt ttgagacaca acgtggcttt gttgaataaa tcgaactttt
     5041 gctgagttga aggatcagat cacgcatctt cccgacaacg cagaccgttc cgtggcaaag
     5101 caaaagttca aaatcaccaa ctggtccacc tacaacaa
//
