pGreen 000

LOCUS       Exported File           3228 bp ds-DNA    circular SYN 13-七月-2016
DEFINITION  Compact Agrobacterium binary vector with a kanamycin-resistance 
            gene. Also known as pGreen 0000.
ACCESSION   AJ007829
VERSION     .
KEYWORDS    pGreen
SOURCE      http://www.biofeng.com/
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3228)
  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 3228)
  AUTHORS   pGreen website
  TITLE     Direct Submission
  JOURNAL   Exported 2016-07-13 from SnapGene Viewer 1.5.3
            http://www.snapgene.com
COMMENT     This plasmid must be transformed into an Agrobacterium strain that 
            also carries a vector such as pSoup.
FEATURES             Location/Qualifiers
     source          1..3228
                     /organism="synthetic DNA construct"
                     /lab_host="Plant Cells"
                     /mol_type="other DNA"
     misc_feature    540..562
                     /note="LB"
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)
                     The T-DNA region is flanked by?BglII sites, these sites are
                     useful in post-transformation analysis.?The polylinker is 
                     based on?pBluescript, and all associated plasmids and 
                     cassettes have been modified, to remove restriction sites 
                     included in this polylinker.Further unique sites?HpaI 
                     and?StuI are located internal to the Left and Right border 
                     sequence respectively. These blunt sites are designed to 
                     facilitate the cloning of selection and marker cassettes, 
                     which are flanked by blunt?EcoRV sites.The various 
                     combinations of marker genes, selection genes, promoters 
                     and the sequences removed by site directed mutagenesis are 
                     catalogued in the cassette directory."
                     /note="color: #ffe4c4"
     CDS             complement(578..940)
                     /codon_start=1
                     /gene="lacZ (truncated)"
                     /product="lacZ-alpha fragment of?beta-galactosidase"
                     /function="
                     "
                     /note="lacZ-alpha"
                     /note="color: #993366"
                     /protein_id="
                     "
                     /translation="MTMITPSSELTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWKL"
     primer_bind     727..743
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: RIGHT"
     promoter        750..768
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     misc_feature    777..884
                     /note="MCS"
                     /note="pBluescript multiple cloning site"
                     /note="color: #99ccff"
     promoter        complement(897..915)
                     /note="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
                     /note="color: #ffffff; direction: LEFT"
     primer_bind     complement(936..952)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    960..976
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="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)."
                     /note="color: #31849b"
     promoter        complement(984..1015)
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                     ???1:??984?..??990?/?#ffffff?/?-10
                     ???2:??991?..?1009?/?#ffffff
                     ???3:?1010?..?1015?/?#ffffff?/?-35"
     misc_feature    1290..1314
                     /note="RB"
                     /note="?RB T-DNA repeat?
                     right border repeat from nopaline C58 T-DNA
                     The T-DNA region is flanked by?BglII sites, these sites are
                     useful in post-transformation analysis.?The polylinker is 
                     based on?pBluescript, and all associated plasmids and 
                     cassettes have been modified, to remove restriction sites 
                     included in this polylinker.Further unique sites?HpaI 
                     and?StuI are located internal to the Left and Right border 
                     sequence respectively. These blunt sites are designed to 
                     facilitate the cloning of selection and marker cassettes, 
                     which are flanked by blunt?EcoRV sites.The various 
                     combinations of marker genes, selection genes, promoters 
                     and the sequences removed by site directed mutagenesis are 
                     catalogued in the cassette directory."
                     /note="color: #ffe4c4"
     rep_origin      complement(1405..1993)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(2094..2909)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /note="KanR"
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /note="color: #ccffcc"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYGLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      3200..407
                     /gene="
                     "
                     /note="pSa ori"
                     /note="origin of replication from bacterial plasmid pSa
                     The RepA gene acts?in trans?upon the pSa Ori sequence. The 
                     RepA is therefore resident on a seperate plasmid pSoup. 
                     This plasmid can be co-electroporated with pGreen vector, 
                     or?Agrobacterium?cells with the plasmid can be made 
                     competent, for independent electroporation."
                     /note="color: #ffff00"
ORIGIN
        1 tttttatccc cggaagcctg tggatagagg gtagttatcc acgtgaaacc gctaatgccc
       61 cgcaaagcct tgattcacgg ggctttccgg cccgctccaa aaactatcca cgtgaaatcg
      121 ctaatcaggg tacgtgaaat cgctaatcgg agtacgtgaa atcgctaata aggtcacgtg
      181 aaatcgctaa tcaaaaaggc acgtgagaac gctaatagcc ctttcagatc aacagcttgc
      241 aaacacccct cgctccggca agtagttaca gcaagtagta tgttcaatta gcttttcaat
      301 tatgaatata tatatcaatt attggtcgcc cttggcttgt ggacaatgcg ctacgcgcac
      361 cggctccgcc cgtggacaac cgcaagcggt tgcccaccgt cgagcgccag cgcctttgcc
      421 cacaacccgg cggccggccg caacagatcg ttttataaat tttttttttt gaaaaagaaa
      481 aagcccgaaa ggcggcaacc tctcgggctt ctggatttcc gatccccgga attagatctt
      541 ggcaggatat attgtggtgt aacgttaaca ttaacgttta caatttccat tcgccattca
      601 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
      661 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
      721 gacgttgtaa aacgacggcc agtgaattgt aatacgactc actatagggc gaattgggta
      781 ccgggccccc cctcgaggtc gacggtatcg ataagcttga tatcgaattc ctgcagcccg
      841 ggggatccac tagttctaga gcggccgcca ccgcggtgga gctccagctt ttgttccctt
      901 tagtgagggt taattccgag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
      961 tgttatccgc tcacaattcc acacaacata cgagccggaa ghcataaagt gtaaagcctg
     1021 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     1081 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     1141 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     1201 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     1261 ggataacgca ggaaagaaca tgaaggcctt gacaggatat attggcgggt aaactaagtc
     1321 gctgtatgtg tttgtttgag atctcatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     1381 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     1441 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     1501 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     1561 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     1621 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     1681 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     1741 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     1801 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     1861 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     1921 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     1981 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     2041 aaactcacgt taagggattt tggtcatggt tacaaccaat taaccaattc tgattagaaa
     2101 aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat
     2161 ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg
     2221 gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat
     2281 ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc
     2341 ggtgagaatg gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta
     2401 cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga
     2461 gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac
     2521 cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct
     2581 aatacctgga atgctgtttt ccctgggatc gcagtggtga gtaaccatgc atcatcagga
     2641 gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg
     2701 accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct
     2761 ggcgcatcgg gcttcccata caatccatag attgtcgcac ctgattgccc gacattatcg
     2821 cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg cggcctggag
     2881 caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca
     2941 gacagtttta ttgttcatga tgatatattt ttatcttgtg caatgtaaca tcagagattt
     3001 tgagacacaa cgtggctttg ttgaataaat cgaacttttg ctgagttgaa ggatcagatc
     3061 acgcatcttc ccgacaacgc agaccgttcc gtggcaaagc aaaagttcaa aatcaccaac
     3121 tggtccacct acaacaaagc tctcatcaac cgtggctccc tcactttctg gctggatgat
     3181 ggggcgattc aggcgatccc catccaacag cccgccgtcg agcgggct
//