LOCUS       sequence-314408-        5065 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 5065)
  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 5065)
  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_6xHis_Blp
FEATURES             Location/Qualifiers
     source          1..5065
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     133..152
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    323..347
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(771..788)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        1516..1861
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     1804..1824
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             1881..2432
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     polyA_signal    2664..2840
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    2848..2870
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     rep_origin      2997..3432
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     primer_bind     complement(3712..3731)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3723..4538
                     /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             3723..4538
                     /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     complement(3796..3815)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(4709..5065,1..232)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
       61 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
      121 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
      181 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
      241 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgagatctca
      301 aacaaacaca tacagcgact tagtttaccc gccaatatat cctgtcaaac actgatagtt
      361 taaactgaag gcgggaaacg acaatctgat catgagcgga gaattaaggg agtcacgtta
      421 tgacccccgc cgatgacgcg ggacaagccg ttttacgttt ggaactgaca gaaccgcaac
      481 gttgaaggag ccactcagcc gatctgaatt ccttatcttt aatcatattc catagtccat
      541 accatagcac atacagtagt tatatgctgc agaagagatc caacaaaaca ttcacaatgg
      601 attatagaaa catttgttta ttcattataa tgagatctta cattcattta atattagaaa
      661 aagccacaaa ttcataacac aacaagccaa gaaaaaaaca caaacttaag cacacaagct
      721 atttatttga cacaccaaat atttcatctt catcttcata tgagctctta gtgatggtga
      781 tggtgatgcc cgggagcggt accctcgagg tcgacagtac tatcgatgga tccactagtg
      841 gcgcgccagg cctgagcccg gttaactcta gtctgttaat cagaaaaact cagattaatc
      901 gacaaattcg atcgcacaaa ctagaaacta acaccagatc tagatagaaa tcacaaatcg
      961 aagagtaatt attcgacaaa actcaaatta tttgaacaaa tcggatgata tctatgaaac
     1021 cctaatcgag aattaagatg atatctaacg atcaaaccca gaaaatcgtc ttcgatctaa
     1081 gattaacaga atctaaacca aagaacatat acgaaattgg gatcgaacga aaacaaaatc
     1141 gaagattttg agagaataag gaacacagaa atttaccttg atcacggtag agagaattga
     1201 gagaaagttt ttaagatttt gagaaattga aatctgaatt gtgaagaaga agaggtcttt
     1261 gggtattgtt ttatagaaga agaagaagaa aagacgagga cgactaggtc acgagaaagc
     1321 taaggcggtg aagcaatagc taataataaa atgacacgtg tattgagcgt tgtttacacg
     1381 caaagttgtt tttggctaat tgccttattt ttaggttgag gaaaagtatt tgtgctttga
     1441 gttgataaac acgactcgtg tgtgccggct gcaaccactt tgacgccgtt tattactgac
     1501 tcgtcgaagc tttattgaga cttttcaaca aagggtaatt tcgggaaacc tcctcggatt
     1561 ccattgccca gctatctgtc acttcatcga aaggacagta gaaaaggaag gtggctccta
     1621 caaatgccat cattgcgata aaggaaaggc tatcattcaa gatgcctctg ccgacagtgg
     1681 tcccaaagat ggacccccac ccacgaggag catcgtggaa aaagaagtcg ttccaaccac
     1741 gacttcaaag caagtggatt gatgtgacat ctccactgac gtaagggatg acgcacaatc
     1801 ccactatcct tcgcaagacc cttcctctat ataaggaagt tcatttcatt tggagaggac
     1861 agcccaagct gatcccccct atgagcccag aacgacgccc ggccgacatc cgccgtgcca
     1921 ccgaggcgga catgccggcg gtctgcacca tcgtcaacca ctacatcgag acaagcacgg
     1981 tcaacttccg taccgagccg caggaaccgc aggagtggac ggacgacctc gtccgtctgc
     2041 gggagcgcta tccctggctc gtcgccgagg tggacggcga ggtcgccggc atcgcctacg
     2101 cgggtccctg gaaggcacgc aacgcctacg actggacggc cgagtccacc gtgtacgtct
     2161 ccccccgcca ccagcggacg ggactgggct ccacgctcta cacccacctg ctgaagtccc
     2221 tggaggcaca gggcttcaag agcgtggtcg ctgtcatcgg gctgcccaac gacccgagcg
     2281 tgcgcatgca cgaggcgctc ggatatgccc cccgcggcat gctgcgggcg gccggcttca
     2341 agcacgggaa ctggcatgac gtgggtttct ggcagctgga cttcagcctg ccggtgccgc
     2401 cccgtccggt cctgcccgtc accgaaatct gaagtagatg ccgaccgaac aagagctgat
     2461 ttcgagaacg cctcagccag caactcgcgc gagcctagca aggcaaatgc gagagaacgg
     2521 ccttacgctt ggtggcacag ttctcgtcca cagttcgcta agctcgctcg gctgggtcgc
     2581 gggagggccg gtcgcagtga ttcaggaatt aattcggtac gctgaaatca ccagtctctc
     2641 tctacaaatc tatctctctc tattttctcc ataaataatg tgtgagtagt ttcccgataa
     2701 gggaaattag ggttcttata gggtttcgct catgtgttga gcatataaga aacccttagt
     2761 atgtatttgt atttgtaaaa tacttctatc aataaaattt ctaattccta aaaccaaaat
     2821 ccagtactaa aatccagatc gataacgtta caccacaata tatcctgcca agatctaatt
     2881 ccggggatcg gaaatccaga agcccgagag gttgccgcct ttcgggcttt ttctttttca
     2941 aaaaaaaaaa tttataaaac gatctgttgc ggccggccgc cgggttgtgg gcaaaggcgc
     3001 tcgacggtgg gcaaccgctt gcggttgtcc acgggcggag ccggtgcgcg tagcgcattg
     3061 tccacaagcc aagggcgacc aataattgat atatatattc ataattgaaa agctaattga
     3121 acatactact tgctgtaact acttgccgga gcgaggggtg tttgcaagct gttgatctga
     3181 aagggctatt agcgttctca cgtgcctttt tgattagcga tttcacgtga ccttattagc
     3241 gatttcacgt actccgatta gcgatttcac gtaccctgat tagcgatttc acgtggatag
     3301 tttttggagc gggccggaaa gccccgtgaa tcaaggcttt gcggggcatt agcggtttca
     3361 cgtggataac taccctctat ccacaggctt ccggggataa aaaagcccgc tcgacggcgg
     3421 gctgttggat ggggatcgcc tgaatcgccc catcatccag ccagaaagtg agggagccac
     3481 ggttgatgag agctttgttg taggtggacc agttggtgat tttgaacttt tgctttgcca
     3541 cggaacggtc tgcgttgtcg ggaagatgcg tgatctgatc cttcaactca gcaaaagttc
     3601 gatttattca acaaagccac gttgtgtctc aaaatctctg atgttacatt gcacaagata
     3661 aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg
     3721 ttatgagcca tattcaacgg gaaacgtctt gctcaaggcc gcgattaaat tccaacatgg
     3781 atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa
     3841 tctaccgatt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta
     3901 gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc
     3961 ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg
     4021 cgatcccagg gaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata
     4081 ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc
     4141 cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt
     4201 tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga
     4261 aagaaatgca taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct
     4321 cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag
     4381 tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt
     4441 ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata
     4501 aattgcagtt tcatttgatg ctcgatgagt ttttctaatc actagaccaa tgttacacat
     4561 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4621 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4681 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4741 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4801 caactcttct tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     4861 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4921 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4981 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5041 gcacacagcc cagcttggag cgaac
//
