LOCUS       sequence-308938-        6856 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pGREEN-based cloning construct for expression in planta.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6856)
  AUTHORS   Pratt AI, Knoblauch J, Kunz HH
  TITLE     An updated pGREEN-based vector suite for cost-effective cloning in 
            plant molecular biology.
  JOURNAL   MicroPubl Biol. 2020 Oct 7;2020. doi: 
            10.17912/micropub.biology.000317.
  PUBMED    33274321
REFERENCE   2  (bases 1 to 6856)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "MicroPubl 
            Biol."; date: "2020-10-7"; volume: "2020. doi"; pages: " 
            10.17912/micropub.biology.000317"
COMMENT     Sequence Label: pG20_GUS_Blp
FEATURES             Location/Qualifiers
     source          1..6856
                     /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    625..647
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     polyA_signal    655..831
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(1063..1614)
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     promoter        complement(1634..1979)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(1671..1691)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             2707..4518
                     /codon_start=1
                     /gene="uidA (or gusA) from E. coli"
                     /product="beta-glucuronidase"
                     /label=GUS
                     /translation="MLRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
                     AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
                     MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
                     FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ
                     VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGEQF
                     LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
                     ADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
                     KNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISD
                     LFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMY
                     TDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRK
                     PKSAAFLLQKRWTGMNFGEKPQQGGKQ"
     misc_feature    4939..4963
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5054..5642)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5134..5153)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5813..6628)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(5813..6628)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     6536..6555
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6620..6639
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
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 tcgagcgcct ttgcccacaa cccggcggcc ggccgcaaca gatcgtttta
      541 taaatttttt tttttgaaaa agaaaaagcc cgaaaggcgg caacctctcg ggcttctgga
      601 tttccgatcc ccggaattag atcttggcag gatatattgt ggtgtaacgt tatcgatctg
      661 gattttagta ctggattttg gttttaggaa ttagaaattt tattgataga agtattttac
      721 aaatacaaat acatactaag ggtttcttat atgctcaaca catgagcgaa accctataag
      781 aaccctaatt tcccttatcg ggaaactact cacacattat ttatggagaa aatagagaga
      841 gatagatttg tagagagaga ctggtgattt cagcgtaccg aattaattcc tgaatcactg
      901 cgaccggccc tcccgcgacc cagccgagcg agcttagcga actgtggacg agaactgtgc
      961 caccaagcgt aaggccgttc tctcgcattt gccttgctag gctcgcgcga gttgctggct
     1021 gaggcgttct cgaaatcagc tcttgttcgg tcggcatcta cttcagattt cggtgacggg
     1081 caggaccgga cggggcggca ccggcaggct gaagtccagc tgccagaaac ccacgtcatg
     1141 ccagttcccg tgcttgaagc cggccgcccg cagcatgccg cggggggcat atccgagcgc
     1201 ctcgtgcatg cgcacgctcg ggtcgttggg cagcccgatg acagcgacca cgctcttgaa
     1261 gccctgtgcc tccagggact tcagcaggtg ggtgtagagc gtggagccca gtcccgtccg
     1321 ctggtggcgg ggggagacgt acacggtgga ctcggccgtc cagtcgtagg cgttgcgtgc
     1381 cttccaggga cccgcgtagg cgatgccggc gacctcgccg tccacctcgg cgacgagcca
     1441 gggatagcgc tcccgcagac ggacgaggtc gtccgtccac tcctgcggtt cctgcggctc
     1501 ggtacggaag ttgaccgtgc ttgtctcgat gtagtggttg acgatggtgc agaccgccgg
     1561 catgtccgcc tcggtggcac ggcggatgtc ggccgggcgt cgttctgggc tcataggggg
     1621 gatcagcttg ggctgtcctc tccaaatgaa atgaacttcc ttatatagag gaagggtctt
     1681 gcgaaggata gtgggattgt gcgtcatccc ttacgtcagt ggagatgtca catcaatcca
     1741 cttgctttga agtcgtggtt ggaacgactt ctttttccac gatgctcctc gtgggtgggg
     1801 gtccatcttt gggaccactg tcggcagagg catcttgaat gatagccttt cctttatcgc
     1861 aatgatggca tttgtaggag ccaccttcct tttctactgt cctttcgatg aagtgacaga
     1921 tagctgggca atggaatccg aggaggtttc ccgaaattac cctttgttga aaagtctcaa
     1981 taaagcttcg acgagtcagt aataaacggc gtcaaagtgg ttgcagccgg cacacacgag
     2041 tcgtgtttat caactcaaag cacaaatact tttcctcaac ctaaaaataa ggcaattagc
     2101 caaaaacaac tttgcgtgta aacaacgctc aatacacgtg tcattttatt attagctatt
     2161 gcttcaccgc cttagctttc tcgtgaccta gtcgtcctcg tcttttcttc ttcttcttct
     2221 ataaaacaat acccaaagac ctcttcttct tcacaattca gatttcaatt tctcaaaatc
     2281 ttaaaaactt tctctcaatt ctctctaccg tgatcaaggt aaatttctgt gttccttatt
     2341 ctctcaaaat cttcgatttt gttttcgttc gatcccaatt tcgtatatgt tctttggttt
     2401 agattctgtt aatcttagat cgaagtcgat tttctgggtt tgatcgttag atatcatctt
     2461 aattctcgat tagggtttca tagatatcat ccgatttgtt caaataattt gagttttgtc
     2521 gaataattac tcttcgattt gtgatttcta tctagatctg gtgttagttt ctagtttgtg
     2581 cgatcgaatt tgtcgattaa tctgagtttt tctgattaac agactagagt taaccgggct
     2641 caggcctggc gcgccactag tggatccatc gatagtactg tcgacctcga gggtaccgct
     2701 cccgggatgt tacgtcctgt agaaacccca acccgtgaaa tcaaaaaact cgacggcctg
     2761 tgggcattca gtctggatcg cgaaaactgt ggaattgatc agcgttggtg ggaaagcgcg
     2821 ttacaagaaa gccgggcaat tgctgtgcca ggcagtttta acgatcagtt cgccgatgca
     2881 gatattcgta attatgcggg caacgtctgg tatcagcgcg aagtctttat accgaaaggt
     2941 tgggcaggcc agcgtatcgt gctgcgtttc gatgcggtca ctcattacgg caaagtgtgg
     3001 gtcaataatc aggaagtgat ggagcatcag ggcggctata cgccatttga agccgatgtc
     3061 acgccgtatg ttattgccgg gaaaagtgta cgtatcaccg tttgtgtgaa caacgaactg
     3121 aactggcaga ctatcccgcc gggaatggtg attaccgacg aaaacggcaa gaaaaagcag
     3181 tcttacttcc atgatttctt taactatgcc ggaatccatc gcagcgtaat gctctacacc
     3241 acgccgaaca cctgggtgga cgatatcacc gtggtgacgc atgtcgcgca agactgtaac
     3301 cacgcgtctg ttgactggca ggtggtggcc aatggtgatg tcagcgttga actgcgtgat
     3361 gcggatcaac aggtggttgc aactggacaa ggcactagcg ggactttgca agtggtgaat
     3421 ccgcacctct ggcaaccggg tgaaggttat ctctatgaac tgtgcgtcac agccaaaagc
     3481 cagacagagt gtgatatcta cccgcttcgc gtcggcatcc ggtcagtggc agtgaagggc
     3541 gaacagttcc tgattaacca caaaccgttc tactttactg gctttggtcg tcatgaagat
     3601 gcggacttgc gtggcaaagg attcgataac gtgctgatgg tgcacgacca cgcattaatg
     3661 gactggattg gggccaactc ctaccgtacc tcgcattacc cttacgctga agagatgctc
     3721 gactgggcag atgaacatgg catcgtggtg attgatgaaa ctgctgctgt cggctttaac
     3781 ctctctttag gcattggttt cgaagcgggc aacaagccga aagaactgta cagcgaagag
     3841 gcagtcaacg gggaaactca gcaagcgcac ttacaggcga ttaaagagct gatagcgcgt
     3901 gacaaaaacc acccaagcgt ggtgatgtgg agtattgcca acgaaccgga tacccgtccg
     3961 caaggtgcac gggaatattt cgcgccactg gcggaagcaa cgcgtaaact cgacccgacg
     4021 cgtccgatca cctgcgtcaa tgtaatgttc tgcgacgctc acaccgatac catcagcgat
     4081 ctctttgatg tgctgtgcct gaaccgttat tacggatggt atgtccaaag cggcgatttg
     4141 gaaacggcag agaaggtact ggaaaaagaa cttctggcct ggcaggagaa actgcatcag
     4201 ccgattatca tcaccgaata cggcgtggat acgttagccg ggctgcactc aatgtacacc
     4261 gacatgtgga gtgaagagta tcagtgtgca tggctggata tgtatcaccg cgtctttgat
     4321 cgcgtcagcg ccgtcgtcgg tgaacaggta tggaatttcg ccgattttgc gacctcgcaa
     4381 ggcatattgc gcgttggcgg taacaagaaa gggatcttca ctcgcgaccg caaaccgaag
     4441 tcggcggctt ttctgctgca aaaacgctgg actggcatga acttcggtga aaaaccgcag
     4501 cagggaggca aacaatgaga gctcatatga agatgaagat gaaatatttg gtgtgtcaaa
     4561 taaatagctt gtgtgcttaa gtttgtgttt ttttcttggc ttgttgtgtt atgaatttgt
     4621 ggctttttct aatattaaat gaatgtaaga tctcattata atgaataaac aaatgtttct
     4681 ataatccatt gtgaatgttt tgttggatct cttctgcagc atataactac tgtatgtgct
     4741 atggtatgga ctatggaata tgattaaaga taaggaattc agatcggctg agtggctcct
     4801 tcaacgttgc ggttctgtca gttccaaacg taaaacggct tgtcccgcgt catcggcggg
     4861 ggtcataacg tgactccctt aattctccgc tcatgatcag attgtcgttt cccgccttca
     4921 gtttaaacta tcagtgtttg acaggatata ttggcgggta aactaagtcg ctgtatgtgt
     4981 ttgtttgaga tctcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     5041 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     5101 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     5161 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5221 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     5281 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5341 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5401 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5461 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     5521 tgaagccagt taccttcgga agaagagttg gtagctcttg atccggcaaa caaaccaccg
     5581 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5641 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5701 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     5761 aatgaagttt taaatcaatc taaagtatat atgtgtaaca ttggtctagt gattagaaaa
     5821 actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt
     5881 tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg
     5941 caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt
     6001 tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg
     6061 gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc cagccattac
     6121 gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag
     6181 cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc
     6241 ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta
     6301 atacctggaa tgctgttttc cctgggatcg cagtggtgag taaccatgca tcatcaggag
     6361 tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga
     6421 ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg
     6481 gcgcatcggg cttcccatac aatcggtaga ttgtcgcacc tgattgcccg acattatcgc
     6541 gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc ggccttgagc
     6601 aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag
     6661 acagttttat tgttcatgat gatatatttt tatcttgtgc aatgtaacat cagagatttt
     6721 gagacacaac gtggctttgt tgaataaatc gaacttttgc tgagttgaag gatcagatca
     6781 cgcatcttcc cgacaacgca gaccgttccg tggcaaagca aaagttcaaa atcaccaact
     6841 ggtccaccta caacaa
//
