LOCUS       sequence-322858-        6409 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 6409)
  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 6409)
  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_FLAG_Blp
FEATURES             Location/Qualifiers
     source          1..6409
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..346)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(38..58)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             1074..1097
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    1521..1545
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(1636..2224)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1716..1735)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3733..4548)
                     /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(3733..4548)
                     /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     4456..4475
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4540..4559
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4839..5274
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    5401..5423
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     polyA_signal    5431..5607
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(5839..6390)
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
ORIGIN
        1 tgtcctctcc aaatgaaatg aacttcctta tatagaggaa gggtcttgcg aaggatagtg
       61 ggattgtgcg tcatccctta cgtcagtgga gatgtcacat caatccactt gctttgaagt
      121 cgtggttgga acgacttctt tttccacgat gctcctcgtg ggtgggggtc catctttggg
      181 accactgtcg gcagaggcat cttgaatgat agcctttcct ttatcgcaat gatggcattt
      241 gtaggagcca ccttcctttt ctactgtcct ttcgatgaag tgacagatag ctgggcaatg
      301 gaatccgagg aggtttcccg aaattaccct ttgttgaaaa gtctcaataa agcttcgacg
      361 agtcagtaat aaacggcgtc aaagtggttg cagccggcac acacgagtcg tgtttatcaa
      421 ctcaaagcac aaatactttt cctcaaccta aaaataaggc aattagccaa aaacaacttt
      481 gcgtgtaaac aacgctcaat acacgtgtca ttttattatt agctattgct tcaccgcctt
      541 agctttctcg tgacctagtc gtcctcgtct tttcttcttc ttcttctata aaacaatacc
      601 caaagacctc ttcttcttca caattcagat ttcaatttct caaaatctta aaaactttct
      661 ctcaattctc tctaccgtga tcaaggtaaa tttctgtgtt ccttattctc tcaaaatctt
      721 cgattttgtt ttcgttcgat cccaatttcg tatatgttct ttggtttaga ttctgttaat
      781 cttagatcga agacgatttt ctgggtttga tcgttagata tcatcttaat tctcgattag
      841 ggtttcatag atatcatccg atttgttcaa ataatttgag ttttgtcgaa taattactct
      901 tcgatttgtg atttctatct agatctggtg ttagtttcta gtttgtgcga tcgaatttgt
      961 cgattaatct gagtttttct gattaacaga ctagagttaa ccgggctcag gcctggcgcg
     1021 ccactagtgg atccatcgat agtactgtcg acctcgaggg taccgctccc ggggactaca
     1081 aggatgacga tgacaagtaa gagctcatat gaagatgaag atgaaatatt tggtgtgtca
     1141 aataaatagc ttgtgtgctt aagtttgtgt ttttttcttg gcttgttgtg ttatgaattt
     1201 gtggcttttt ctaatattaa atgaatgtaa gatctcatta taatgaataa acaaatgttt
     1261 ctataatcca ttgtgaatgt tttgttggat ctcttctgca gcatataact actgtatgtg
     1321 ctatggtatg gactatggaa tatgattaaa gataaggaat tcagatcggc tgagtggctc
     1381 cttcaacgtt gcggttctgt cagttccaaa cgtaaaacgg cttgtcccgc gtcatcggcg
     1441 ggggtcataa cgtgactccc ttaattctcc gctcatgatc agattgtcgt ttcccgcctt
     1501 cagtttaaac tatcagtgtt tgacaggata tattggcggg taaactaagt cgctgtatgt
     1561 gtttgtttga gatctcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     1621 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     1681 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     1741 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     1801 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     1861 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     1921 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     1981 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     2041 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     2101 gctgaagcca gttaccttcg gaagaagagt tggtagctct tgatccggca aacaaaccac
     2161 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     2221 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     2281 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     2341 aaaatgaagt tttaaatcaa tctaaagtac tgatgaatcc cctaatgatt tttatcaaaa
     2401 tcattaagtt aaggtagata cacatcttgt catatgatca aatggtttcg ccaaaaatca
     2461 ataatcagac aacaaaatgt gcgaactcga tattttacac gactctcttt accaattctg
     2521 ccccgaatta cacttaaaac gactcaacag cttaacgttg gcttgccacg ccttacttga
     2581 ctgtaaaact ctcactctta ccgaacttgg ccgtaacctg ccaaccaaag cgagaacaaa
     2641 acataacatc aaacgaatcg accgattgtt aggtaatcgt cacctccaca aagagcgact
     2701 cgctgtatac cgttggcatg ctagctttat ctgttcgggc aatacgatgc ccattgtact
     2761 tgttgactgg tctgatatcc gtgagcaaaa acggcttatg gtattgcgag cttcagtcgc
     2821 actacacggt cgttctgtta ctctttatga gaaagcgttc ccgctttcag agcaatgttc
     2881 aaagaaagct catgaccaat ttctagccga ccttgcgagc attctaccga gtaacaccac
     2941 accgctcatt gtcagtgatg ctggctttaa agtgccatgg tataaatccg ttgagaagct
     3001 gggttggtac tggttaagtc gagtaagagg aaaagtacaa tatgcagacc taggagcgga
     3061 aaactggaaa cctatcagca acttacatga tatgtcatct agtcactcaa agactttagg
     3121 ctataagagg ctgactaaaa gcaatccaat ctcatgccaa attctattgt ataaatctcg
     3181 ctctaaaggc cgaaaaaatc agcgctcgac acggactcat tgtcaccacc cgtcacctaa
     3241 aatctactca gcgtcggcaa aggagccatg gattctagca actaacttac ctgttgaaat
     3301 tcgaacaccc aaacaacttg ttaatatcta ttcgaagcga atgcagattg aagaaacctt
     3361 ccgagacttg aaaagtcctg cctacggact aggcctacgc catagccgaa cgagcagctc
     3421 agagcgtttt gatatcatgc tgctaatcgc cctgatgctt caactaacat gttggcttgc
     3481 gggcgttcat gctcagaaac aaggttggga caagcacttc caggctaaca cagtcagaaa
     3541 tcgaaacgta ctctcaacag ttcgcttagg catggaagtt ttgcggcatt ctggctacac
     3601 aataacaagg gaagacttac tcgtggctgc aaccctacta gctcaaaatt tattcacaca
     3661 tggttacgct ttggggaaat tatgagggga tctctcagtc taaagtatat atgtgtaaca
     3721 ttggtctagt gattagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc
     3781 aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc
     3841 gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac
     3901 atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc
     3961 atgagtgacg actgaatccg gtgagaatgg caaaagttta tgcatttctt tccagacttg
     4021 ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt
     4081 cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca
     4141 aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc
     4201 tgaatcagga tattcttcta atacctggaa tgctgttttc cctgggatcg cagtggtgag
     4261 taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc
     4321 cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc
     4381 atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcggtaga ttgtcgcacc
     4441 tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga
     4501 atttaatcgc ggccttgagc aagacgtttc ccgttgaata tggctcataa caccccttgt
     4561 attactgttt atgtaagcag acagttttat tgttcatgat gatatatttt tatcttgtgc
     4621 aatgtaacat cagagatttt gagacacaac gtggctttgt tgaataaatc gaacttttgc
     4681 tgagttgaag gatcagatca cgcatcttcc cgacaacgca gaccgttccg tggcaaagca
     4741 aaagttcaaa atcaccaact ggtccaccta caacaaagct ctcatcaacc gtggctccct
     4801 cactttctgg ctggatgatg gggcgattca ggcgatcccc atccaacagc ccgccgtcga
     4861 gcgggctttt ttatccccgg aagcctgtgg atagagggta gttatccacg tgaaaccgct
     4921 aatgccccgc aaagccttga ttcacggggc tttccggccc gctccaaaaa ctatccacgt
     4981 gaaatcgcta atcagggtac gtgaaatcgc taatcggagt acgtgaaatc gctaataagg
     5041 tcacgtgaaa tcgctaatca aaaaggcacg tgagaacgct aatagccctt tcagatcaac
     5101 agcttgcaaa cacccctcgc tccggcaagt agttacagca agtagtatgt tcaattagct
     5161 tttcaattat gaatatatat atcaattatt ggtcgccctt ggcttgtgga caatgcgcta
     5221 cgcgcaccgg ctccgcccgt ggacaaccgc aagcggttgc ccaccgtcga gcgcctttgc
     5281 ccacaacccg gcggccggcc gcaacagatc gttttataaa tttttttttt tgaaaaagaa
     5341 aaagcccgaa aggcggcaac ctctcgggct tctggatttc cgatccccgg aattagatct
     5401 tggcaggata tattgtggtg taacgttatc gatctggatt ttagtactgg attttggttt
     5461 taggaattag aaattttatt gatagaagta ttttacaaat acaaatacat actaagggtt
     5521 tcttatatgc tcaacacatg agcgaaaccc tataagaacc ctaatttccc ttatcgggaa
     5581 actactcaca cattatttat ggagaaaata gagagagata gatttgtaga gagagactgg
     5641 tgatttcagc gtaccgaatt aattcctgaa tcactgcgac cggccctccc gcgacccagc
     5701 cgagcgagct tagcgaactg tggacgagaa ctgtgccacc aagcgtaagg ccgttctctc
     5761 gcatttgcct tgctaggctc gcgcgagttg ctggctgagg cgttctcgaa atcagctctt
     5821 gttcggtcgg catctacttc agatttcggt gacgggcagg accggacggg gcggcaccgg
     5881 caggctgaag tccagctgcc agaaacccac gtcatgccag ttcccgtgct tgaagccggc
     5941 cgcccgcagc atgccgcggg gggcatatcc gagcgcctcg tgcatgcgca cgctcgggtc
     6001 gttgggcagc ccgatgacag cgaccacgct cttgaagccc tgtgcctcca gggacttcag
     6061 caggtgggtg tagagcgtgg agcccagtcc cgtccgctgg tggcgggggg agacgtacac
     6121 ggtggactcg gccgtccagt cgtaggcgtt gcgtgccttc cagggacccg cgtaggcgat
     6181 gccggcgacc tcgccgtcca cctcggcgac gagccaggga tagcgctccc gcagacggac
     6241 gaggtcgtcc gtccactcct gcggttcctg cggctcggta cggaagttga ccgtgcttgt
     6301 ctcgatgtag tggttgacga tggtgcagac cgccggcatg tccgcctcgg tggcacggcg
     6361 gatgtcggcc gggcgtcgtt ctgggctcat aggggggatc agcttgggc
//
