LOCUS       Exported                4446 bp DNA     circular SYN 21-OCT-2025
DEFINITION  Agrobacterium binary vector with kanamycin-and 
            bialophos/phosphinothricin-resistance genes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4446)
  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 4446)
  AUTHORS   pGreen website
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 4446)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4446)
  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     SGRef: number: 3; 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..4446
                     /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)"
     terminator      complement(679..930)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(953..1501)
                     /codon_start=1
                     /label=BlpR
                     /note="phosphinothricin acetyltransferase"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIQTSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     promoter        complement(1544..1721)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     1968..1984
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1991..2009
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    complement(2018..2124)
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     promoter        complement(2137..2155)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2176..2192)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2200..2216)
                     /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(2224..2254)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2269..2290)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     misc_feature    2529..2553
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(2644..3232)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3406..4218)
                     /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 tcagcttgca tgccggtcga tctagtaaca tagatgacac cgcgcgcgat aatttatcct
      721 agtttgcgcg ctatattttg ttttctatcg cgtattaaat gtataattgc gggactctaa
      781 tcaaaaaacc catctcataa ataacgtcat gcattacatg ttaattatta catgcttaac
      841 gtaattcaac agaaattata tgataatcat cgcaagaccg gcaacaggat tcaatcttaa
      901 gaaactttat tgccaaatgt ttgaacgatc tgcttgactc taggggtcat cagatttcgg
      961 tgacgggcag gaccggacgg ggcggcaccg gcaggctgaa gtccagctgc cagaaaccca
     1021 cgtcatgcca gttcccgtgc ttgaagccgg ccgcccgcag catgccacgg ggggcatatc
     1081 cgagcgcctc gtgcatgcgc acgctcgggt cgttgggcag cccgatgaca gcgaccacgc
     1141 tcttgaagcc ctgtgcctcc agggacttca gcaggtgggt gtagagcgtg gagcccagtc
     1201 ccgtccgctg gtggcggggg gagacgtaca cggttgactc ggccgtccag tcgtaggcgt
     1261 tgcgtgcctt ccagggaccc gcgtaggcga tgccggcgac ctcgccgtcc acctcggcga
     1321 cgagccaggg atagcgctcc cgcagacgga cgaggtcgtc cgtccactcc tgcggttcct
     1381 gcggctcggt acggaagttg accgtgcttg tctggatgta gtggttgacg atggtgcaga
     1441 ccgccggcat gtccgcctcg gtggcacggc ggatgtcggc cgggcgtcgt tctgggctca
     1501 tggtagatcc ccctcgatcg agttgagagt gaatatgaga ctctaattgg ataccgaggg
     1561 gaatttatgg aacgtcagtg gagcattttt gacaagaaat atttgctagc tgatagtgac
     1621 cttaggcgac ttttgaacgc gcaataatgg tttctgacgt atgtgcttag ctcattaaac
     1681 tccagaaacc cggctgagtg gctccttcaa cgttgcggtt ctgtcagttc caaacgtaaa
     1741 acggcttgtc ccgcgtcatc ggcgggggtc ataacgtgac tcccttaatt ctcatgtatc
     1801 gataacatta acgtttacaa tttcgcgcca ttcgccattc aggctgcgca actgttggga
     1861 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     1921 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
     1981 cagtgaattg taatacgact cactataggg cgaattgggt accgggcccc ccctcgaggt
     2041 cgacggtatc gataagcttg atatcgattc ctgcagcccg ggggatccac tagttctaga
     2101 gcggccgcca ccgcggtgga gctccagctt ttgttccctt tagtgagggt taattccgag
     2161 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     2221 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     2281 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     2341 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     2401 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     2461 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     2521 gaaggccttg acaggatata ttggcgggta aactaagtcg ctgtatgtgt ttgtttgaga
     2581 tctcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     2641 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     2701 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     2761 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2821 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2881 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2941 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3001 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3061 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     3121 taccttcgga agaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3181 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3241 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3301 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     3361 taaatcaatc taaagtatat atgtgtaaca ttggtctagt gattagaaaa actcatcgag
     3421 catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag
     3481 ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg
     3541 gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc
     3601 aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg
     3661 caaaagttta tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc
     3721 aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa
     3781 tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa
     3841 cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa
     3901 tgctgttttc cctgggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa
     3961 atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc
     4021 tgtaacaaca ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg
     4081 cttcccatac aatcggtaga ttgtcgcacc tgattgcccg acattatcgc gagcccattt
     4141 atacccatat aaatcagcat ccatgttgga atttaatcgc ggccttgagc aagacgtttc
     4201 ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag acagttttat
     4261 tgttcatgat gatatatttt tatcttgtgc aatgtaacat cagagatttt gagacacaac
     4321 gtggctttgt tgaataaatc gaacttttgc tgagttgaag gatcagatca cgcatcttcc
     4381 cgacaacgca gaccgttccg tggcaaagca aaagttcaaa atcaccaact ggtccaccta
     4441 caacaa
//
